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 .
Election/Restrictions
Applicant’s election of Group I in the reply filed on 9/2/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
Accordingly, claims 32 and 33 are withdrawn from consideration for being directed to non-elected subject matter. Claims 1-3, 7-15, 18-20, 22, 23, 26 and 31 are currently under examination.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 1/10/2024 has been considered by the examiner.
Claim Objections
Claim 31 is objected to because of the following informalities: the claim recites abbreviation “TCR” and “CAR.” It is suggested to spell out the full term when first mentioned in the claim. Appropriate correction is required.
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.
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.
Claim(s) 1, 7-9, 14 and 26 is/are rejected under 35 U.S.C. 103 as being unpatentable over Ostertag (WO 2020/051374), in view of sequences have accession number JQ048536, JQ087288 and KF420799.
Claim 1 is drawn to an HLA class Ineg cell population, wherein said cells comprise a disruption in the HLA-A, HLA-B and HLA-C genes, produced by CRISPR/Cas9 mediated gene disruption targeting a consensus sequence SEQ ID NO: 1, and do not have expressible HLA class-I protein.
Ostertag teaches a method for generating MHCI knockout cells comprising using guide RNA selective for HLA-A, HLA-B and HLA-C (paragraph ]0226]). Ostertag teaches HLA-A, HLA-B and HLA-C is reduced or eliminated in allogeneic cells (paragraph [066]). Ostertag teaches that specifically guides were designed to target a conserved region occurring in all the three MHCI protein targets, but not in HLA-E, and the MHCI knockouts were verified by surface expression of said HLA I proteins (paragraph [066]).
The only difference between the invention of claim 1 and Ostertag’s MHCI knockout cells is that the consensus sequence is not SEQ ID NO: 1 as claimed.
SEQ ID NO: 1 shares 100% sequence identity with JQ048536, JQ087288 and KF420799 (see attached alignment).
JQ048536 is from human MHC class I antigen (HLA-A) gene, JQ087288 is from HLA-B gene, and KF420799 is from HLA-C gene, so that they are consensus sequence shared between HLA-A, HLA-B and HLA-C.
It would have been obvious to an ordinary skilled in the art that SEQ ID NO:1 would be a good candidate for designing gRNA for CRISPR/Cas9 mediated disruption of all HLA class I gene, A, B and C because it is a consensus sequences shared by all three genes. Following the teaching from Ostertag, it would have been obvious to use this approach to design additional gRNA for producing HLA class I gene disruption. Substitute one consensus sequence with another to achieve a predictable result would have been within the capability of an ordinary skilled in the art. Therefore, the claimed invention of claim 1 would have been prima facie obvious to an ordinary skilled in the art at the time the application was filed.
Regarding claim 7, it would have been obvious to an ordinary skilled in the art that treating cells comprising knockouts of HLA-A, B and C with cytokines IFNγ and/or TNFα would not produce HLA-A, B and C expression because the gene is disrupted.
Regarding claim 8, since it is unclear how the cells will be “further defined,” the pan T cell (paragraph [0066]) meets the limitation of antigen presenting cells because pan T cells comprises helper T which can present antigen.
Regarding claim 9, Ostertag teaches the HLA-A, B and C knockout cells may further comprise a non-naturally occurring sequence comprising HLA-E (paragraph [0112]), which meets the limitation of an exogenous HLA class I gene.
Regarding claim 14, the method of which the expression is construct would not impart a structural difference between the prior art disclosed construct and the claimed construct. Ostertag teaches HLA-E comprises amino acids SEQ ID NO: 17066, and encoded by nucleic acid SEQ ID NO: 17067 (paragraph [[0122]). Ostertag teaches the modified cell may express HLA-E stably or transiently (paragraph [0127]). It would have been obvious to an ordinary skilled in the art that the nucleic acid sequence encoding HLA-E would have the same structure regardless whether it is assembled by Gibson assembly. Since Ostertag teaches the nucleic acid encoding HLA-E, absent evidence from the contrary, the claimed invention of claim 14 would have been obvious in view of combined teaching from Ostertag, and JQ048536, JQ087288 and KF420799.
Regarding claim 26, Ostertag teaches the T cell may be cultured in an activation supplement that comprises cytokines including IL2 and IL15 (paragraph [0311]).
Claim(s) 1-3, 7-15, 18, 19, 22-24, 26 and 31 is/are rejected under 35 U.S.C. 103 as being unpatentable over Hong (Journal of Immunotherapy, 2017, Vol.40, no.6, pages 201-210), in view of JQ048536, JQ087288 and KF420799.
Hong teaches generating HLA class I null cell line using multiplex CRISPR/Cas9 system by targeting exon 2 and 3 of HLA-A, HLA-B and HLA-C genes simultaneously. Hong teaches that artificial antigen presenting cells were generated by transfer of a single HLA class I allele and co-stimulatory molecules into HLA class I null cell line. Hong teaches artificial antigen presenting cells showed HLA-restricted antigen presentation following antigen processing and were successfully used for the efficient generation of tumor antigen specific cytotoxic T cells in vitro. Hong teaches the efficient editing of HLA genes may provide a basis for universal cellular therapies and transplantation (entire abstract).
However, Hong does not specifically teach what the consensus sequence they used to generate the HLA I null cell line.
SEQ ID NO: 1 shares 100% sequence identity with JQ048536, JQ087288 and KF420799 (see attached alignment).
JQ048536 is from human MHC class I antigen (HLA-A) gene, JQ087288 is from HLA-B gene, and KF420799 is from HLA-C gene, so that they are consensus sequence shared between HLA-A, HLA-B and HLA-C.
It would have been obvious to an ordinary skilled in the art that SEQ ID NO:1 would be a good candidate for designing gRNA for CRISPR/Cas9 mediated disruption of all HLA class I gene, A, B and C because it is a consensus sequences shared by all three genes. Substitute one consensus sequence with another to achieve a predictable result would have been within the capability of an ordinary skilled in the art. Therefore, the claimed invention of claim 1 would have been prima facie obvious to an ordinary skilled in the art at the time the application was filed.
Regarding claim 2, the cell line taught by Hong meets the limitation of immortalized cell line.
Regarding claim 3, Hong teaches the immortalized cell line is 293 T cells (page 205, 2nd col., lines 1-3).
Regarding claim 7, it would have been obvious to an ordinary skilled in the art that treating cells comprising knockouts of HLA-A, B and C with cytokines IFNγ and/or TNFα would not produce HLA-A, B and C expression because the gene is disrupted.
Regarding claim 8, the artificial antigen presenting cells may be defined as antigen presenting cells.
Regarding claims 9 and 10, Hong teaches transfer of a single HLA class I allele HLA-A and HLA-B into this HLA class null cell line (page 205, 2nd col., 1st paragraph).
Regarding claims 11 and 13, Hong teaches the modified APC showed HLA-restricted antigen presentation following antigen processing and were successfully used for the efficient generation of tumor antigen specific cytotoxic T cells in vitro. It would have been obvious to an ordinary skilled in the art for intended cancer cellular therapy, the APC would need have matched HLA class I to reduce GvH reaction to present tumor antigen. As such, the ordinary skilled in the art would be motivated to use an HLA class I gene from a cancer patient. Whether the patient has been HLA typed does not change the structure of the class I HLA gene being introduced into the APC cell.
Regarding claims 12, it would have been obvious to an ordinary skilled in the art that the nucleic acid sequence encoding the construct HLA class I antigen construct would have the same structure regardless whether it is obtained from locus specific primer pairs, which are well known method in the art. Absent evidence from the contrary, the claimed invention of claim 12 would have been obvious in view of combined teaching from Hong, and JQ048536, JQ087288 and KF420799.
Regarding claim 14, it would have been obvious to an ordinary skilled in the art that the nucleic acid sequence encoding HLA-I would have the same structure regardless whether it is assembled by Gibson assembly. Since Hong teaches the lentiviral vectors that encoding nucleic acid encoding HLA class I allele, absent evidence from the contrary, the claimed invention of claim 14 would have been obvious in view of combined teaching from Hong, and JQ048536, JQ087288 and KF420799.
Regarding claims 15, 18 and 19, Hong teaches the modified APC showed HLA-restricted antigen presentation following antigen processing and were successfully used for the efficient generation of tumor antigen specific cytotoxic T cells in vitro. Hong teaches transduction of additional genes including CD80, CD32, CD83, 4-1BBL, CD54 and CD70 and stimulated the cells with MART-126-35 peptide (page 206, 2nd col., lines 1-6). Hong teaches it generated antigen specific CTL (Figure 5 and legend). It would have been obvious to an ordinary skilled the in the art that the AAPC cells that presents tumor antigen would be loaded with MART-1 and present it at the cell surface.
Regarding claims 22 and 23, Hong teaches the stimulated cells are co-cultured with purified T cells (page 203, 2nd col., 1st paragraph).
Regarding claim 26, Hong teaches culturing medium for generating antigen specific cytotoxic T cells comprises IL2 and IL15 (page 203, 1st col., last paragraph).
Regarding claim 31, since there is no definition for “candidate TCR,” the TCR that binds MART-1 in the effector T cells meets this limitation because the purified T cells would necessarily have a TCR that specially stimulate the CTL response for antigen specific T cells.
Claim(s) 20 is/are rejected under 35 U.S.C. 103 as being unpatentable over Hong and JQ048536, JQ087288 and KF420799, as applied to claims 1 and 9 above, and further in view of Aubin (Molecular Biotechnology, 1994, Vol.1, pages 29-48).
The teaching from Hong has been discussed above. However, Hong does not teach the cells are cultured in the presence of polybrene.
Aubin teaches polybrene/DMSO assisted gene transfer is a simple and versatile transfection strategy capable of producing high numbers of stable transfects from adherent monolayer cultures with low quantities of exogeneous DNA, wherein polybrene favors uniform coating of target cells. Aubin teaches diverse cell types can be exposed to a wide range of polybrene concentrations without adverse effects (abstract). Aubin teaches optimized protocols for gene transfer in murine and human cells (abstract).
It would have been obvious to an ordinary skilled in the art to add polybrene into the culture medium for culturing engineered APC knockout taught by Hong because Aubin teaches polybrene assists gene transfer. The ordinary skilled in the art would be motivated to include polybrene because Aubin teaches polybrene is a simple and versatile transfection strategy capable of producing high numbers of stable transfectants for diverse cell types. Adding a compound into a cell culture for its predictable effect would have been prima facie obvious to an ordinary skilled in the art at the time the application was filed.
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CELINE X QIAN whose telephone number is (571)272-0777. The examiner can normally be reached M-F (8-4:00).
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/CELINE X QIAN/Primary Examiner, Art Unit 1637