DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant’s response filed 01/26/2026 has been received and considered entered. This is a response to amendments and arguments filed 01/26/2026.
Election/Restrictions
Claims 176, 178, 181, 184-191 stand withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 08/11/2025.
Claims Status
Claims 1-174, 204 is/are cancelled. Claims 175-203, 205-207 is/are currently pending with claims 176, 178, 181, 184-191 withdrawn. Claims 175, 177, 179-180, 182-183, 192-203, 205-207 is/are under examination.
Drawings
The drawings are objected to as failing to comply with 37 CFR 1.84(p)(4) because “Figures 7A-7K” has been used in paragraphs [00175] and [00177] to describe both Figs. 5A-5AE and Figs. 7A-7K. Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
Claim Rejections - 35 USC § 112
112(a):
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim 206 is rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for treatment of conditions already known to be treated by stem cell transplantation, does not reasonably provide enablement for the treatment of any disease known to be treated by any type of transplantation (including organ transplantation). The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with these claims. This rejection is amended and maintained.
The factors to be considered in determining whether a disclosure would require undue experimentation include:
A) The breadth of the claims;
(B) The nature of the invention;
(C) The state of the prior art;
(D) The level of one of ordinary skill;
(E) The level of predictability in the art;
(F) The amount of direction provided by the inventor;
(G) The existence of working examples; and
(H) The quantity of experimentation needed to make or use the invention based on the content of the disclosure.
In re Wands, 8 USPQ2d, 1400 (CAFC 1988) and MPEP 2164.01.
The breadth of the claims:
With respect to claim breadth, the standard under 35 U.S.C. §112, first paragraph, entails the determination of what the claims recite and what the claims mean as a whole. As such, the broadest reasonable interpretation of the claimed method is that it encompasses the treatment of patients in need of any transplantation, not necessarily a stem cell transplantation therapy (as “cell transplantation” encompasses transplantation of organs, wherein organs are made of cells), the claimed treatment method comprising administering an engineered cell to the patient. A skilled artisan would not know how to use the method with a reasonable expectation of success based solely on what is disclosed in the specification.
The amount of direction provided by the inventor and the level of predictability in the art:
The specification teaches a definition of “treatment” which does not describe specific diseases or conditions which could be treated using the claimed engineered cells and is sufficiently broad as to render unclear whether disease or conditions which can be “treated” include diseases not known to be treated by stem cell transplantation (see paragraphs [00204]-[00208]). The specification does teach multiple specific diseases which can be treated with cell-replacement therapies (see paragraphs [00411]-[00418]). The art at the time of filing teaches that stem cell therapies, while promising, in clinical testing, or proven for the treatment of many diseases or conditions, are not recognized as treatments for any and all diseases (see Trounson, 2015). The specification as filed does not provide guidance that overcomes this unpredictability within the art.
The existence of working examples:
What is enabled by the working examples is narrow in comparison to the breadth of the claims: The specification does not provide working examples of in vivo administration of the claimed cells.
The quantity of experimentation needed to make or use the invention:
The standard of an enabling disclosure is not the ability to make and test if the invention works but one of the ability to make and use with a reasonable expectation of success. A patent is granted for a completed invention, not the general suggestion of an idea (MPEP 2164.03 and Chiron Corp. v. Genentech Inc., 363 F.3d 1247, 1254, 70 USPQ2d 1321, 1325-26 (Fed. Cir. 2004). The instant specification is not enabling because one cannot follow the guidance presented therein, or within the art at the time of filing, and practice the claimed method without first making a substantial inventive contribution. Given that the nature of the invention is engineered stem cells, and the art provides evidence for the use of stem cells in the treatment of a limited set of diseases and conditions, a person having ordinary skill in the art would have to perform multiple further experiments, in human clinical trials and in animal models predictive of treatment in a representative number of diseases, in order to demonstrate that stem cell transplantation therapies could treat any disease or condition, and, thus, in order to demonstrate the invention could be used with a reasonable expectation of success. The amount of experimentation required for enabling guidance, commensurate in scope with what is claimed, goes beyond what is considered ‘routine' within the art, and constitutes undue further experimentation in order to use the method with a reasonable expectation of successfully treating any CNS disorder or neurodegenerative disease. Therefore, Claim 206 is rejected under 35 U.S.C. 112, first paragraph, for failing to meet the enablement requirement. This rejection can be overcome by amending “cell transplantation” in lines 1-2 of claim 206 to “stem cell transplantation”.
Response to Arguments
Applicant's arguments filed 01/26/2026 have been fully considered but they are not persuasive. Applicant has amended claim 206 to specify patients in need of “cell transplantation” because the previously-presented rejection of claim 206 argued that the applicants were enabled for stem-cell transplantation, specifically. However, the broadest reasonable interpretation of a “cell transplantation” would encompass transplantation of any cell, including the cells which make up an organ for transplantation. Amendment of “cell transplantation” to “stem cell transplantation” would be remedial.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
Section 33(a) of the America Invents Act reads as follows:
Notwithstanding any other provision of law, no patent may issue on a claim directed to or encompassing a human organism.
Claims 175, 177, 179-180, 182-183, 192-203 are rejected under 35 U.S.C. 101 and section 33(a) of the America Invents Act as being directed to or encompassing a human organism. See also Animals - Patentability, 1077 Off. Gaz. Pat. Office 24 (April 21, 1987) (indicating that human organisms are excluded from the scope of patentable subject matter under 35 U.S.C. 101). This rejection is maintained.
Claims 175, 177, 179-180, 182-183, and 192-204 are drawn to an engineered cell, the scope of the claims encompassing a human cell (claim 182 recites that the cell is a human cell). Claim 206 recites a method of treating a patient comprising administering the engineered cell to the patient; as “patient” is not defined in the specification to exclude human patients, claim 206 encompasses methods of administering a human cell to a human organism, providing evidence that the claimed cells of claims 175, 177, 179-180, 182-183, and 192-204 are intended for use in a human organism. Claim 206 is not rejected under 35 USC §101 because it claims a method, it does not claim a human organism. As such, the claims render obvious to an artisan that the claimed cells of claims 175, 177, 179-180, 182-183, and 192-204 can be present within a human organism. As such, claims 175-, 177, 179-180, 182-183, and 192-204 encompass human cells in human organisms, thus being directed to a human organism. Including the term “isolated” in the preamble of the claims (e.g. “an isolated engineered cell”) would be remedial.
Response to Arguments
Applicant's arguments filed 01/26/2026 have been fully considered but they are not persuasive.
Applicant argues that “the claimed engineered cells are not a product of nature and contain ‘markedly different characteristics from any naturally occurring counterpart’” and have “distinct modifications that distinguish them from normal human cells” (page 11). However, the claims were not rejected under 35 USC 101 because they were considered products of nature or indistinguishable from “normal” human cells. The claims were rejected under 35 USC 101 because the claimed cells can exist in a human organism, and in fact are recited in methods of transplanting said cells into a human organism. A cell in a human organism, engineered or no, is considered part of the human organism. The claimed cells, when transplanted into a human organism, would become part of the human organism, regardless of genetic differences between the engineered cells and non-engineered cells in the human organism. As noted in the rejection above, including the term “isolated” in the preamble of the claims (e.g. “an isolated engineered cell”) would be remedial.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
WO2016183041A2:
Claim(s) 175, 177, 179-180, 182-183, 192-203, 205-207 is/are rejected under 35 U.S.C. 103 as being unpatentable over Meissner (WO2016183041A2: provided by Applicant with IDS filed 10/28/2022), in view of Hawksworth (2019) and Booth (2013).
Regarding claim 175, Meissner teaches an engineered cell comprising one or more modifications fully inactivating or disrupting one or more alleles of genes encoding one or more transcriptional regulators of MHC-I or MHC-II molecules (claims 1-2, 4-6). Meissner teaches that the engineered cell further comprises increased expression of tolerogenic factors relative to a wild-type cell (claims 11-12). Meissner teaches that the one or more tolerogenic factors are selected from the following: PD-L1, HLA-E, HLA-G, and CD47 (claim 33).
Regarding claim 182, Meissner teaches that the cell is a human cell (claim 20).
Regarding claim 183, Meissner teaches that the cell is a pluripotent stem cell (claim 18), an embryonic stem cell (claim 17), or an induced pluripotent stem cell (page 27 lines 26-31).
Regarding claims 192-193, 196-197, 200-201, Meissner teaches that the one or more modifications inactivate or disrupt genes encoding transcriptional regulators of MHC-I or MHC-II molecules (claims 1-2, 4-6), specifically CIITA and B2M (claims 7, 9-10). Meissner teaches that inhibition or disruption of these transcriptional regulators of MHC-I or MHC-II molecules reduces or inhibits expression of MHC-I or MHC-II molecules, respectively (claims 3-4). Meissner further teaches that cell surface expression and trafficking of MHC-I is reduced in cells comprising inactivated transcriptional regulator B2M (page 13 lines 5-6; page 41 lines 29-32) (required by claims 192, 196-197) and cell surface expression of MHC-II is reduced in cells comprising inactivated transcriptional regulator CIITA (page 45 lines 10-14) (required by claims 193, 200). An artisan would interpret that reduced cell surface expression of an MHC-II molecule resulting from reduced expression of the MHC-II molecule would inherently be associated with reduced cell surface trafficking, as reduction in the overall expression and surface expression of the molecule would inherently result in reduced supply to a cell surface-trafficking system and reduced output from the cell surface-trafficking system (required by claim 201).
Regarding claim 194, Meissner teaches that the one or more modifications reduce expression of one or more MHC-I molecules and/or one or more MHC-II molecules (claims 3-5).
Regarding claim 195, Meissner teaches that the cell comprises one or more modifications that reduce expression of one or more of the following molecules: B2M, CIITA, NLRC5 (claims 7-10), TAP1 (page 14 lines 28-31), HLA-A, HLA-B, HLA-C (claims 66-67), RFX5, RFX-ANK, RFX-AP (page 14 lines 6-18), NFY-A, NFY-B, NFY-C (page 14 lines 19-27), and IRF1 (page 14 lines 1-5).
Regarding claim 198, Meissner teaches that the one or more modifications reduce expression of B2M (claim 21).
Regarding claim 199, the one or more modifications reduce expression of HLA-A, HLA-B, and/or HLA-C (page 2 lines 14-21).
Regarding claim 202, Meissner teaches that the one or more modifications reduce (eliminate) expression of CIITA (claim 9).
Regarding claim 203, Meissner teaches that one or more modifications reducing (eliminating) expression of CIITA eliminate expression of HLA-DR (Fig. 5C).
Regarding claim 205, Meissner teaches that the engineered cells can be used to treat patients (page 40 lines 8-26); an artisan would recognize that a pharmaceutical use of the engineered cells, such as the treatment of patients by administration of the engineered cells, would require that the engineered cells be comprised in a pharmaceutical composition.
Regarding claim 206, Meissner teaches that the engineered cells can be used in methods to treat patients in need of treatment, by administering the cells to the patients (page 40 lines 8-26).
Regarding claim 207, Meissner teaches a method of generating an engineered cell comprising: obtaining an isolated cell, wherein the cell is “isolated” in that it is removed from its normal environment (page 15 lines 2-3; page 17 lines 29-30; page77 lines 23-27; Fig. 1); introducing into the cell a CRISPR/Cas nuclease and at least one guide RNA targeting one or more alleles of one or more MHC-I molecules and/or one or more molecules which regulate expression of the one or more MHC-I molecules, or of one or more MHC-II molecules and/or one or more molecules which regulate expression of the one or more MHC-II molecules, thereby inactivating the one or more alleles (claims 105-112; page 77 line 30-page 78 line 3; Fig. 36; page 36 lines 3-29: the “pair of ribonucleic acids” of claims 105-108 and the “ribonucleic acid” of claims 109-112 are interpreted, based on the specification, to be a tracrRNA/crRNA pair or a guide RNA); thereby inactivating or disrupting the one or more alleles of the one or more MHC-I molecules, MHC-II molecules, and/or one or more molecules regulating expression of one or more MHC-I or MHC-II molecules (claims 105-112).
However, Meissner does not teach that a blood group antigen gene, including FUT1, is also inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Meissner teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is “useful for overcoming the immune rejection in cell-based transplantation therapies” (Abstract). However, Meissner does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Meissner is to create “universal donor cells” with reduced immunogenicity (Abstract), including hematopoietic stem cells (page 20 lines 3-5, Meissner creates “universal donor” hematopoietic stem cells), it would have been obvious to a person of ordinary skill in the art at the time of filing that the hematopoietic stem cells of Meissner should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US20190376045A1:
Claims 175, 177, 179-180, 182-183, 192-203, 207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US20190376045A1), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules, and comprising increased expression of one or more tolerogenic factors (including CD47), relative to a wild-type cell (claims 1, 7-8, 10).
Regarding claim 182, Deuse teaches that the cell is a human cell (claim 8).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claims 1, 7-8, 10).
Regarding claims 192-198, 200-202, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (paragraph [0045]; claim 1) and the CIITA gene (which reduces MHC-II expression) (paragraph [0045]; claim 1) and comprises increased expression of the tolerogenic factor CD47 (claim 1).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (claims 15-20).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ are knocked out or reduced (claims 26-31).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, introducing into the cell a CRISPR/Cas nuclease and a gRNA targeting an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (claims 1, 5; paragraphs [0007], [0071], [0214], [0293]).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US20220213434A1:
Claims 175, 177, 179-180, 182-183, 192-203, 206-207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US20220213434A1), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising: a fully inactivated blood group antigen gene; one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules; and increased expression of one or more tolerogenic factors (including CD47), relative to a wild-type cell (claims 1-5, 106-108).
Regarding claims 179-180, Deuse teaches that the cell is Rh-negative and type O (claim 5).
Regarding claim 182, Deuse teaches that the cell is a human cell (paragraph [0016]).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claims 1-2).
Regarding claims 192-198, 200-202, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (paragraph [0149]; claims 82-86) and the CIITA gene (which reduces MHC-II expression) (paragraph [0149]; claim 94-98) and comprises increased expression of the tolerogenic factor CD47 (claim 106-108).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (claims 19-24).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ are knocked out or reduced (claims 28-33).
Regarding claim 204, Deuse teaches that HLA-E, PD-L1, HLA-G, CD47, CD200, FASLG, CLC21, MFGE8, SERPIN B9 expression is modulated (claim 4).
Regarding claim 206, Deuse teaches that the cells can be administered to a patient in need, in a method of treatment (paragraph [0012]).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, introducing into the cell a CRISPR/Cas nuclease and a gRNA targeting an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (claims 67, 80, 82-86, 98).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US20220049227A1:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US20220049227A1), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising: a fully inactivated blood group antigen gene; one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules; and increased expression of one or more tolerogenic factors (including CD47), relative to a wild-type cell (claim 137).
Regarding claims 179-180, Deuse teaches that the cell is Rh-negative and type O (claims 147, 141, 137).
Regarding claim 182, Deuse teaches that the cell is a human cell (paragraph [0016]).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claim 137).
Regarding claims 192-198, 200-202, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (claim 146) and the CIITA gene (which reduces MHC-II expression) (claim 147) and comprises increased expression of the tolerogenic factor CD47 (claim 137).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (claim 146).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ are knocked out or reduced (claim 147).
Regarding claim 204, Deuse teaches that CD47 expression is modulated (claim 137).
Regarding claim 205, Deuse teaches that HLA-DP, HLA-DR, or HLA-DQ are knocked out (claim 147).
Regarding claim 206, Deuse teaches that the cells can be administered to a patient in need, in a method of treatment (claims 160-161).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, by reducing expression of an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (claims 158-159).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US 20250283051 A1:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US 20250283051 A1), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising: a fully inactivated blood group antigen gene; one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules; and increased expression of one or more tolerogenic factors (including CD47), relative to a wild-type cell (claim 165).
Regarding claim 182, Deuse teaches that the cell is a human cell (claim 167).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claim 167).
Regarding claims 192-198, 200-202, 204, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (paragraph [0196], claim 165) and the CIITA gene (which reduces MHC-II expression) (paragraph [0196], claim 165) and comprises increased expression of the tolerogenic factor CD47 (claim 165).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (paragraph [0142]).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ are knocked out or reduced (paragraph [0144]).
Regarding claim 205, Deuse teaches a pharmaceutical composition comprising the engineered cell (claim 175).
Regarding claim 206, Deuse teaches that the cells can be administered to a patient in need, in a method of treatment (claim 176).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, by reducing expression of an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (paragraphs [0164], [0177]).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US20210308183A1:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US20210308183A1), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising: a fully inactivated blood group antigen gene; one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules; and increased expression of one or more tolerogenic factors (including CD47), relative to a wild-type cell (claim 1).
Regarding claim 182, Deuse teaches that the cell is a human cell (claim 8).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claim 8).
Regarding claims 192-198, 200-202, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (paragraph [0229], claim 1) and the CIITA gene (which reduces MHC-II expression) (paragraph [0229], claim 1) and comprises increased expression of the tolerogenic factor CD47 (claim 1).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (paragraph [0230]).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ are knocked out or reduced (paragraph [0230]).
Regarding claim 204, Deuse teaches that CD47 expression is modulated (claim 1).
Regarding claim 205, Deuse teaches a pharmaceutical composition comprising the engineered cell (claims 47-48).
Regarding claim 206, Deuse teaches that the cells can be administered to a patient in need, in a method of treatment (claim 47).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, by reducing expression of an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (paragraph [0106]).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US11162079B2:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US11162079B2), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising: a fully inactivated blood group antigen gene; one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules; and increased expression of one or more tolerogenic factors (including CD47), relative to a wild-type cell (claims 1, 8-9).
Regarding claims 179-180, Deuse teaches that the cell is Rh-negative and type O (claims 2-3, 5-6).
Regarding claim 182, Deuse teaches that the cell is a human cell (col. 12 lines 7-14).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claim 1).
Regarding claims 192-198, 200-202, 204, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (claim 8) and the CIITA gene (which reduces MHC-II expression) (claim 9) and comprises increased expression of the tolerogenic factor CD47 (claim 1).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (claim 8).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ are knocked out or reduced (claim 9).
Regarding claim 205, Deuse teaches a composition comprising the engineered cell and a pharmaceutical carrier (col. 27 lines 23-28).
Regarding claim 206, Deuse teaches that the cells can be administered to a patient in need, in a method of treatment (col. 27 lines 23-28).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, by reducing expression of an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (col. 6 lines 36-44).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US12221622B2:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 is/are rejected under 35 U.S.C. 103 as being obvious over Deuse (US12221622B2), in view of Hawksworth (2019) and Booth (2013).
The applied reference has a common inventor (Sonja Schrepfer) with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
Regarding claim 175, Deuse teaches an engineered cell comprising: a fully inactivated blood group antigen gene; one or more modifications which inactivate or disrupt one or more alleles of one or more MHC-I and/or MHC-II molecules, or of one or more molecules which regulate expression of one or more MHC-I and/or MHC-II molecules; and increased expression of one or more tolerogenic factors, relative to a wild-type cell (claims 1, 6-19).
Regarding claims 179-180, Deuse teaches that the cell is Rh-negative and type O (claim 27).
Regarding claim 182, Deuse teaches that the cell is a human cell (claim 1).
Regarding claim 183, Deuse teaches that the cell is a pluripotent stem cell (claims 26, 28).
Regarding claims 192-198, 200-202, 204, Deuse teaches that the cell comprises inactivating modifications in the B2M gene (which reduces MHC-I expression) (claim 70; col. 24 lines 19-20) and the CIITA gene (which reduces MHC-II expression) (claim 70; col. 24 lines 19-20) and comprises increased expression of the tolerogenic factor CD47 (claim 70).
Regarding claim 199, Deuse teaches that HLA-A, HLA-B, and HLA-C are knocked out (reduced expression) (claims 7-8).
Regarding claim 203, Deuse teaches that HLA-DP, HLA-DR, HLA-DQ, HLA-DM, HLA-DO are knocked out or reduced (claims 12-13).
Regarding claim 205, Deuse teaches a composition comprising the engineered cell and a pharmaceutical carrier (col. 27 lines 51-56).
Regarding claim 206, Deuse teaches that the cells can be administered to a patient in need, in a method of treatment (col. 27 lines 51-56).
Regarding claim 207, Deuse teaches a method of generating an engineered cell comprising obtaining an isolated cell, by reducing expression of an MHC-I allele, an MHC-II allele, and/or a transcription factor allele of an MHC-I and/or MHC-II allele (claim 35).
However, Deuse does not teach that the engineered cell further comprises a modification in the FUT1 gene, rendering the FUT1 gene partially or fully inactivated.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
Deuse teaches that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creating hypoimmunogenic stem cells, is useful for avoiding the immune rejection in cell-based transplantation therapies (Abstract). However, Deuse does not disclose modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants, blood transfusion, and solid organ transplantation, as both arise from incompatibility of cell-surface antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion, hematopoietic stem cell transplantation, and solid organ transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). Booth also teaches that ABO incompatibility is a “significant barrier to solid organ transplantation” (page 1152), rendering obvious that ABO incompatibility is a major consideration in a majority of organ systems; an artisan would recognize that stem cell transplantation into solid organs would also require a consideration of ABO incompatibility. It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause loss of immature stem cells in cases of stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the stated goal of the invention of Deuse is to create “universally acceptable ‘off-the-shelf’ pluripotent stem cells” with reduced immunogenicity (Abstract), it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of Deuse should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
Response to Arguments
Applicant's arguments filed 01/26/2026 have been fully considered but they are not persuasive.
Applicant argues that an artisan would have no motivation to combine the inventions of an MHC-I- and MHC-II-deficient cell (further comprising increased CD47 expression) (as in Meissner and Deuse) and of an ABO/FUT1-deficient cell (as in Hawksworth), and argues that as Hawksworth teaches that RHAG knockout results in reduced CD47 expression, Hawskworth teaches away from the claimed invention.
As discussed above, Meissner, Deuse, and Hawksworth both teach intended uses of their respective engineered cells as universal donor cells (Meissner abstract, page 20 lines 3-5; Deuse abstract; Hawskworth page 178). Booth teaches that MHC-I and MHC-II and blood type antigens are necessary considerations in transplantation, as all can affect success of a transplant. An artisan would recognize that a “universal” donor stem cell would need to have modified MHC-I, MHC-II, and blood type antigens in order to reduce immunogenicity in a great enough proportion of the population to be considered “universal”. While Hawksworth teaches a method for reducing or eliminating FUT1 gene expression in erythroid cells, it would be obvious to an artisan that this same method can be applied to the stem cells of Meissner or Deuse, reducing or elimination expression of FUT1 and thereby providing further reduced immunogenicity.
Applicant also argues that Hawskworth teaches cells comprising 5x modifications, while the claimed cells only require modification of a single blood type antigen. While the pending claims only require modification of one blood type antigen gene (FUT1), the claims do not exclude further modification of additional blood type antigen genes. Furthermore, Hawksworth teaches that their findings can be used to create tailored or customized donor cells (pages 42, 185, 178), and teaches the individual gRNA sequences used to target each blood type antigen gene (2.1.6, pages 47-49). It would have been obvious to an artisan that the methods of Hawksworth could be used to eliminate expression of only the blood type antigen genes which a particular patient would require (e.g., eliminating expression of only FUT1 for cells for transplantation or transfusion into a Bombay phenotype patient), and such an artisan would know the precise gRNAs to knock out each individual blood type antigen gene separately.
Applicant further argues that because Hawksworth teaches that BEL-A cells exhibit a 25% reduction of CD47 expression, Hawksworth teaches away from the claimed cells exhibiting increased CD47 expression. The examiner notes that while increased CD47 expression is addressed in the rejection above, increased expression of CD47 is not required by the claim, and is an optional alternative limitation (“An engineered cell…comprises one or more modifications, wherein the one or more modifications: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility class I (MHC-I) molecules…and/or (ii) one or more MHC class II (MHC-II) molecules…and/or (b) increase expression of CD47”, claim 175). Furthermore, Hawksworth teaches that knockout of the RHAG gene resulted in the 25% reduction in CD47 expression (page 12), while Meissner teaches that increased CD47 expression is achieved by knocking-in or overexpressing CD47 (page 84 lines 11-20). As CD47 is taught by Meissner to be a tolerogenic factor (Table 2; page 63), allowing cells with CD47 knocked-in to inhibit immune rejection, it would have been obvious to an artisan that even cells engineered to be Rhnull-, such as those of Hawksworth, would benefit from knocking-in of CD47 (including under the control of a universal promoter such as CAG, as in Meissner) to increase CD47 expression and compensate for the 25% reduction of CD47 resulting from RHAG gene knockout. It therefore would have been obvious to keep the CD47 knock-in of Meissner in universal donor stem cells rendered obvious by the combined teachings of Meissner, Hawksworth, and Booth, comprising knockouts of MHC-I, MHC-II, and blood type antigen genes.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
16476794:
Claims 175, 177, 179-1180, 182-183, 192-203 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 10, 104-134, 143-147 of copending Application No. 16476794 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
17607841:
Claims 175, 177, 179-1180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 4-6, 17, 35, 51, 63, 67, 80, 83, 106, 120, 124-125 of copending Application No. 17607841 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell. The copending application additionally recites that the engineered cell comprises a modified blood group antigen gene; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of FUT1, the gene encoding the H blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
17504502:
Claims 175, 177, 179-1180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 137-141, 143-144, 146-147, 152-154, 158-167 of copending Application No. 17504502 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell. The copending application additionally recites that the engineered cell comprises a modified blood group antigen gene; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of FUT1, the gene encoding the H blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
19015552:
Claims 175, 177, 179-1180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 165-169, 174-179, 183-184 of copending Application No. 19015552 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of one or more genes which regulates expression of an MHC-I and/or MHC-II molecule (B2M, CIITA), and which increase expression of a tolerogenic factor (CD47), relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of FUT1, the gene encoding the H blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
17260222:
Claims 175, 177, 179-1180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 8, 11-12, 32-37, 53, 59-60 of copending Application No. 17260222 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of one or more genes which regulates expression of an MHC-I and/or MHC-II molecule (B2M, CIITA), and which increase expression of a tolerogenic factor (CD47), relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of FUT1, the gene encoding the H blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18021036:
Claims 175, 177, 179-1180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 312, 314-315, 317, 323-325, 333-337 of copending Application No. 18021036 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of one or more genes which regulates expression of an MHC-I and/or MHC-II molecule (B2M, CIITA), and which increase expression of a tolerogenic factor (CD47), relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of FUT1, the gene encoding the H blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
17632026:
Claims 175, 177, 179-1180, 182-183, 192-202, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-4, 8-9, 17, 26, 28, 31, 71 of copending Application No. 17632026 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of one or more genes which regulates expression of an MHC-I and/or MHC-II molecule (B2M, CIITA), and which increase expression of a tolerogenic factor (CD47), relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of FUT1, the gene encoding the H blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
17637789:
Claims 175, 177, 179-1180, 182-183, 192-202, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-3, 5, 8-10, 22-24, 27, 62-63, 160 of copending Application No. 17637789 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
17997103:
Claims 175, 177, 179-1180, 182-183, 192-203, 205-206 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 268, 280-289, 294-299 of copending Application No. 17997103 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites a method of treating diseases in a patient by administering an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18449625:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 382-384, 388-395, 400-401, 403, 408-410 of copending Application No. 18449625 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites a method of treating diseases in a patient by administering an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18561581:
Claims 175, 177, 179-180, 182-183, 192-202, 205-206 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 206-214, 217-218, 221 of copending Application No. 18561581 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18579148:
Claims 175, 177, 179-180, 182-183, 192-202, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-4, 19, 32-33, 75, 77, 98, 235-236 of copending Application No. 18579148 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18682798:
Claims 175, 177, 179-180, 182-183, 192-202, 205-206 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 9, 37-39, 47, 65, 75-76, 96, 116-117, 149, 169, 213, 312-313 of copending Application No. 18682798 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18682782:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 410, 413-416, 420-421, 423-426, 428-429 of copending Application No. 18682782 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, while the copending application does recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen, the copending application does not recite that the blood group antigen is the H antigen encoded by the FUT1 gene.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18682797:
Claims 175, 177, 179-180, 182-183, 192-202, 205-206 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 451-452, 456, 460, 464-469 of copending Application No. 18682797 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, while the copending application does recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen, the copending application does not recite that the blood group antigen is the H antigen encoded by the FUT1 gene.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18727670:
Claims 175, 177, 179-180, 182-183, 192-202, 205-206 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 518-521, 545, 548 of copending Application No. 18727670 (reference application) in view of Booth (2013) and Hawksworth (2019).
The copending application recites an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The copending application recites that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the copending application does not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the copending claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the copending application should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
This is a provisional nonstatutory double patenting rejection.
18682796:
Claims 175, 177, 179-180, 182-183, 192-202, 205-207 provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-4, 12, 14, 23, 42, 52, 86-87, 99-105 of copending Application No. 18682796 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because the copending application claims an engineered cell comprising inactivated FUT1, MHC-I and/or MHC-II alleles, and alleles encoding transcription factors of MHC-I and/or MHC-II genes, methods of using the cell to treat a patient, and methods of producing the cells using a CRISPR/Cas system.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
US11987628B2:
Claims 175, 177, 179-180, 182-183, 192-202, 205-206 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 13-17, 22, 26-29 of U.S. Patent No. 11987628B2 in view of in view of Booth (2013) and Hawksworth (2019).
The issued claims recite an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the issued claims do not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The issued claims recite that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the issued claims not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the issued claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the issued claims should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US11965022B2:
Claims 175, 177, 179-180, 182-183, 192-203, 205-206 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-2, 5-20 of U.S. Patent No. US11965022B2 in view of in view of Booth (2013) and Hawksworth (2019).
The issued claims recite an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the issued claims do not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The issued claims recite that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the issued claims not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the issued claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the issued claims should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US11802157B2:
Claims 175, 177, 179-180, 182-183, 192-203, 205-207 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 3, 14-27 of U.S. Patent No. 11802157B2 in view of in view of Booth (2013) and Hawksworth (2019).
The issued claims recite an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the issued claims do not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
Regarding claim 207, Hawksworth teaches a method of generating an engineered cell comprising: obtaining an isolated cell (BEL-A cells, see Abstract, are “isolated” because they are removed from their usual environment and immortalized); introducing into the cell a CRISPR-Cas nuclease (Cas9) and a guide RNA targeting the FUT1 gene; and selecting a cell in which the FUT1 gene is fully inactivated (page 144 Table 5-4; page 128; page 130; Fig. 5-2; Fig. 5-4 pages 134-135).
The issued claims recite that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the issued claims not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the issued claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the issued claims should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US11162079B2:
Claims 175, 177, 179-180, 182-183, 192-203 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-10, 12-20 of U.S. Patent No. 11162079B2 in view of in view of Booth (2013) and Hawksworth (2019).
The issued claims recite an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen gene FUT1.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
The issued claims recite that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the issued claims not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the issued claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the issued claims should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
US12221622B2:
Claims 175, 177, 179-180, 182-183, 192-203, 207 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-22, 27-56, 60-70 of U.S. Patent No. 12221622B2 in view of in view of Booth (2013) and Hawksworth (2019).
The issued claims recite an engineered cell comprising one or more modifications which inactivate or disrupt one or more alleles of an MHC-I molecule, an MHC-II molecule, and/or a gene which regulates expression of an MHC-I and/or MHC-II molecule, and which increase expression of a tolerogenic factor, relative to a control cell; however, the copending application does not recite that the engineered cells additionally have partially or fully inactivated expression of a blood group antigen gene FUT1.
Hawksworth teaches engineered cells (BEL-A cells, an immortalized erythroblast line, see Abstract) comprising a blood group antigen gene, FUT1, which is fully inactivated.
Regarding claim 175, Hawksworth teaches an engineered cell (BEL-A cells, see Abstract) in which expression of a blood group antigen gene (FUT1) is partially or fully inactivated (Table 5-4 page 144; pages 134-135; Fig. 5-2, pages 130-131).
Regarding claim 177, Hawksworth teaches that the blood group antigen gene is a FUT1 gene that is fully inactivated by a deletion within exon 4 (Table 5-4 page 144; Fig. 5-2).
Regarding claim 179, Hawksworth teaches that the cell is an Rh-negative cell (page 131; Fig. 5-2; Fig. 5-8; Table 5-4; page 177).
Regarding claim 180, Hawksworth teaches that the cell has a type Oh Bombay phenotype (page 177: the 5X KO BEL-A cell described on pages 130-131, 144 and in Table 5-4 is described as being Oh Bombay Rhnull phenotype on page 177).
Regarding claims 182-183, Hawksworth teaches that the cell is a human differentiated cell (BEL-A, see Abstract; page 177).
The issued claims recite that the reduced expression of MHC-I and/or MHC-II molecules, and the increased expression of tolerogenic factors, in stem cells, creates hypoimmunogenic stem cells. However, the issued claims not recite modification of blood type antigens. Booth teaches that blood type antigens must also be considered in allogeneic stem cell transplantation, in addition to the major histocompatibility complex or MHC, as blood type incompatibility between donor hematopoietic stem cells and recipient can result in complications (pages 1153-1154; Table 1). Booth draws a connection between these complications arising from hematopoietic stem cell transplants and blood transfusion, as both arise from incompatibility of blood antigens (from transfused differentiated cells or transplanted blood cell-progenitor cells, HSCs) and both can result in massive hemolysis (pages 1152-1154). Booth does not discuss the effects of H-antigen incompatibility (H-antigen is produced by the gene FUT1); however, Booth’s teachings regarding the relevance of ABO blood type antigens in both blood transfusion and hematopoietic stem cell transplantation would have rendered obvious to a person of ordinary skill in the art at the time of filing that other blood type antigens known to cause hemolysis in cases of transfusion of antigen-incompatible blood products would affect hematopoietic stem cell transplantation safety. Hawksworth teaches that the Bombay (Oh) phenotype is an extremely rare phenotype in which the H antigen—the common precursor to ABO antigens and thus required to synthesize the ABO antigens, and which forms part of the ABO(H) oligosaccharide chain (antigen) on the cell surface—is not expressed because of a loss-of-function mutation in the FUT1 gene (page 10; Fig. 1-4). Patients having the Bombay phenotype require Oh blood for transfusion because these patients generate antibodies against A, B, and H antigens, and transfusion of blood containing H antigen into a Bombay-phenotype patient results in fatal hemolysis (page 124, see page XV defining “HTR” as “haemolytic transfusion reaction”; page 10). It would therefore have been obvious to a person of ordinary skill in the art at the time of filing that the H antigen would also be necessary to consider in hematopoietic stem cell transplantation, as H antigen incompatibility was known to have the same potentially fatal hemolytic consequences as ABO antigen incompatibilities in blood transfusion because of immune system recognition of incompatible ABO/H cell-surface antigens, as described by Booth, and thus would be assumed to also cause hemolysis and loss of immature stem cells in cases of hematopoietic stem cell transplantation wherein the donor stem cells were type O (or A, B, or AB) and the recipient’s cells were Oh Bombay phenotype (see additionally Hawksworth Fig. 1-4 page 11, the H antigen forms part of the ABO antigen). As the issued claims recite cells with reduced immunogenicity, it would have been obvious to a person of ordinary skill in the art at the time of filing that the stem cells of the issued claims should be further modified by introducing a deletion into exon 4 of the FUT1 gene, inactivating the FUT1 gene and thereby preventing the synthesis of H, A, and B antigens, resulting in a stem cell with further reduced immunogenicity.
Response to Arguments
Applicant's arguments filed 01/26/2026 have been fully considered but they are not persuasive.
Applicant argues that “none of the cited reference patents teaches the inactivation of the FUT1 gene in cells” (page 19). However, as discussed in the rejections above, Hawksworth teaches FUT1-knockout cells. Applicant further argues that Hawksworth and Booth do not remedy this claimed deficiency, for the same reasons as applied to the rejection of claims under 35 USC 103.
Applicant argues that an artisan would have no motivation to combine the inventions of an MHC-I- and MHC-II-deficient cell (further comprising increased CD47 expression) (as in the copending applications and issued patents above) and of an ABO/FUT1-deficient cell (as in Hawksworth), and argues that as Hawksworth teaches that RHAG knockout results in reduced CD47 expression, Hawskworth teaches away from the claimed invention.
As discussed above, the copending and issued claims and Hawksworth both teach intended uses of their respective engineered cells as universal donor cells (Hawskworth page 178). Booth teaches that MHC-I and MHC-II and blood type antigens are necessary considerations in transplantation, as all can affect success of a transplant. An artisan would recognize that a “universal” donor stem cell would need to have modified MHC-I, MHC-II, and blood type antigens in order to reduce immunogenicity in a great enough proportion of the population to be considered “universal”. While Hawksworth teaches a method for reducing or eliminating FUT1 gene expression in erythroid cells, it would be obvious to an artisan that this same method can be applied to the stem cells of the copending and issued claims, reducing or elimination expression of FUT1 and thereby providing further reduced immunogenicity.
Applicant also argues that Hawskworth teaches cells comprising 5x modifications, while the claimed cells only require modification of a single blood type antigen. While the pending claims only require modification of one blood type antigen gene (FUT1), the claims do not exclude further modification of additional blood type antigen genes. Furthermore, Hawksworth teaches that their findings can be used to create tailored or customized donor cells (pages 42, 185, 178), and teaches the individual gRNA sequences used to target each blood type antigen gene (2.1.6, pages 47-49). It would have been obvious to an artisan that the methods of Hawksworth could be used to eliminate expression of only the blood type antigen genes which a particular patient would require (e.g., eliminating expression of only FUT1 for cells for transplantation or transfusion into a Bombay phenotype patient), and such an artisan would know the precise gRNAs to knock out each individual blood type antigen gene separately.
Applicant further argues that because Hawksworth teaches that BEL-A cells exhibit a 25% reduction of CD47 expression, Hawksworth teaches away from the claimed cells exhibiting increased CD47 expression. The examiner notes that while increased CD47 expression is addressed in the rejection above, increased expression of CD47 is not required by the claim, and is an optional alternative limitation (“An engineered cell…comprises one or more modifications, wherein the one or more modifications: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility class I (MHC-I) molecules…and/or (ii) one or more MHC class II (MHC-II) molecules…and/or (b) increase expression of CD47”, claim 175). Furthermore, Hawksworth teaches that knockout of the RHAG gene resulted in the 25% reduction in CD47 expression (page 12), while Meissner teaches that increased CD47 expression is achieved by knocking-in or overexpressing CD47 (page 84 lines 11-20). As CD47 is recited as a tolerogenic factor, it would have been obvious to an artisan that even cells engineered to be Rhnull-, such as those of Hawksworth, would benefit from knocking-in of CD47 to increase CD47 expression and compensate for the 25% reduction of CD47 resulting from RHAG gene knockout. It therefore would have been obvious to keep the CD47 knock-in of the copending and issued claims in universal donor stem cells rendered obvious by the combined teachings of the copending and issued claims, Hawksworth, and Booth, comprising knockouts of MHC-I, MHC-II, and blood type antigen genes.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/AFRICA M MCLEOD/ Examiner, Art Unit 1635
/KIMBERLY CHONG/ Primary Examiner, Art Unit 1636