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 .
Claims 1-8, 11, 17-18, 23, 29-33, 37-38, and 48-50, of record 10/22/2025, are pending and subject to prosecution. Claims 1-4, 7, 11, 23, and 29-32 are amended. Claim 21 is cancelled. Claims 48-50 are newly added.
Status of Prior Rejections/Response to Arguments
RE: Objection to the drawings:
The submission of replacement drawings is affective to obviate the objection. The objection is withdrawn.
RE: Objection to claims 4, 7, and 23:
The amendment to claims 4, 7, and 23 is effective to obviate the objection. The objection is withdrawn.
RE: Rejection of claims 2-3, 21, and 29-32 under 35 U.S.C. 112(b):
The cancellation of claim 21 renders the rejection thereto moot.
The amendment to claims 2-4 and 29-32 are effective to obviate the rejection. The rejection is withdrawn.
RE: Rejection of claims 1-3, 11, and 30 under 35 U.S.C. 103 over Cotta-Ramusino et al. (WO 2019152519 A1) in view of Kim et al. (Scientific Reports, 2017):
RE: Rejection of claims 1-8, 11, 17-18, 21, 23, 29-30, 32-33, and 37-38 under 35 U.S.C. 103 over Cotta-Ramusino et al. (WO 2019152519 A1) in view of Kim et al. (Scientific Reports, 2017), in view of Rezvani et al. (WO 2018195339 A1):
RE: Rejection of claims 1-3, 11, and 30-31 under 35 U.S.C. 103 over Cotta-Ramusino et al. (WO 2019152519 A1) in view of Kim et al. (Scientific Reports, 2017), further in view of Lange et al. (Journal of Cellular Physiology, 2007):
The cancellation of claim 21 renders the rejection thereto moot.
The amendments to claims 1 and 4 to require disruption of two or more of the enumerated genes is effective to obviate the rejections. The rejections are withdrawn. However, the applicant’s arguments are addressed to the extent that they pertain to the new or amended claims, for completeness of record.
The applicant asserts that the claimed method produces MSCs that have enhanced viability and persistence and reduced apoptosis (Applicant Remarks, page 8). The applicant also asserts that Kim et al. teach away from the use of at least five total passages (Applicant Remarks, page 8).
These arguments are not found persuasive. Evidence of nonobviousness must be fully commensurate with the scope of the claims in order to overcome a finding of obviousness. See MPEP 716.02(d). The instant specification includes working examples only for the knockout of CD47 and of PD-L1 and PD-L2 (See fig. 3-5). These examples are not sufficient to support the assertion of enhanced viability and persistence and reduced apoptosis for the full scope of the claims, nor do they provide a basis for one of ordinary skill in the art to be able to extrapolate such effects for every cell that could be produced by methods encompassed by the claims.
Regarding the teachings of Kim et al., the disclosure of alternative and nonpreferred embodiments do not constitute a teaching away. See MPEP 2123(I)-(II). Kim et al. do not expressly teach or suggest that MSCs cannot or should not be passaged more than four times. Rather, Kim et al. teach that increased genomic instability is associated with later passages and that the rate of somatic mutation accumulation increases substantially in passages 5-9, depending on cell line (See fig. 3A). The use of MSCs after passages 1-4 could therefore be considered an alternative that may be less preferred than the use of earlier-passage cells.
New Rejections
Claim Interpretation
Claim 49 recites the limitation “large-scale expansion”. This limitation is interpreted as requiring a total of at least 109 MSCs, consistent with ¶0026 of the instant application’s PG Pub.
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.
Claims 1-3, 11, 30, 48, and 50 are rejected under 35 U.S.C. 103 as being unpatentable over Cotta-Ramusino et al. (WO 2019152519 A1), of record, in view of Kim et al. (Scientific Reports, 2017), of record, Davies et al. (Stem Cells, 2016), and Ju et al. (Biomaterials, 2019).
Regarding claims 1-3, 11, and 48: Cotta-Ramusino et al. teach methods for modulating chromosomal rearrangements using editing systems (See Abstract). A cell can be edited at two or more target nucleic acids (which reads on “two or more endogenous genes”) by RNA-guided nucleases and gRNAs (which reads on “one or more guide RNAs for each of the two or more genes”) in RNP complexes (See ¶7-8, 10-12, and 42). The RNA-guided nuclease can be Cas9 or Cpf1 (See ¶172-173 and 176-177 and table 2). The cell can be an MSC (See ¶436-437). Nucleic acids encoding the genome editing system components and/or RNPs can be delivered to cells by electroporation (See ¶452 and 458-460). The RNP complexes can be delivered sequentially in any order (which reads on “two electroporation steps” and “a first delivering step comprises delivering guide RNAs that target one or more genes and a second delivering step comprises delivering guide RNAs that target one or more genes that are different from the one or more genes in the first delivering step”) (See ¶8, 10-12, 14, and 42). Oligonucleotide donor templates for repairing nuclease-mediated DNA breaks can comprise one or more stop codons (which reads on “disrupt expression”) (See ¶42 and 81). Cotta-Ramusino et al. do not expressly teach editing of the cells within a first, second, third, or fourth passage or the endogenous genes as being PD-L1 and PD-L2.
Kim et al. teach the accumulation of somatic mutations in MSCs during in vitro culture (which reads on “expansion”) (See Abstract). Kim et al. demonstrate that the abundance of mutations increases after passage 4 in two MSC lines during ex vivo expansion (See fig. 3A).
Davies et al. teach that expression and secretion of PD-L1 and PD-L2 by MSCs suppress T cell activation, downregulate IL-2 secretion, and induce hyporesponsiveness and cell death (See Abstract). Antibody blockade of PD-1 interaction with forms of PD-L1 and PD-L2 prevented suppression of proliferation, anergy, and increased apoptosis (See page 774, col. 1, full ¶1, and fig. 2 and 4).
Ju et al. teach methods and reagents for the CRISPR/Cas9-mediated simultaneous disruption of PD-L1 and PD-L2 (See Abstract and fig. 2 and 4-7). Ju et al. teach their approach as an alternative to blocking the interaction of PD-1 and PD-L1/PD-L2 with antibodies, which can have low target specificity, very long half-life, and a risk of autoimmune response in systemic administration (See page 2, col. 1, full ¶1).
It would have been obvious to one having ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method of Cotta-Ramusino et al. to comprise modification of the cells at an early passage, i.e., passage 1-4. One would be motivated to make this modification because Kim et al. teach that mutation numbers increase in at least one MSC line starting with passage 5 and that genomic instability may be responsible for malignant transformation in future in vivo use (See page 1, full ¶1 and fig. 3A). There would be a reasonable expectation of success in making this modification because the MSCs in the method of Cotta-Ramusino et al. could be readily modified during an early passage of expansion.
It also would have been obvious to modify the method of Cotta-Ramusino et al. to comprise CRISPR/Cas9-mediated editing of genes for knocking out expression of PD-L1 and PD-L2. One would have been motivated to make this modification because Davies et al. teach that PD-L1 and PD-L2 suppress T cell activation, downregulate IL-2 secretion, and induce hyporesponsiveness and cell death, which can be reduced by blocking the interaction of PD-1 with PD-L1 and PD-L2 (See Abstract; page 774, col. 1, full ¶1; and fig. 2 and 4-7). Additionally, Ju et al. teach that the use of antibodies for blocking this interaction can have low target specificity, very long half-life, and a risk of autoimmune response in systemic administration (See page 2, col. 1, full ¶1). There would be a reasonable expectation of success in making this modification because Ju et al. teach that expression of PD-L1 and PD-L2 can be successfully suppressed using CRISPR (See Abstract and fig. 2 and 4).
Regarding claim 30: Following the discission of claims 1-3, 11, and 48, Cotta-Ramusino et al. teach that the cells can be stored for late use following ex vivo modification (which reads on “the cells are expanded prior to… storage”) (See ¶439).
Regarding claim 50: Following the discussion of claims 1-3, 13, and 48, Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., do not expressly teach or suggest passaging the MSCs at least five times.
However, Kim et al. demonstrate that MSCs can be passaged about 6-8 times before population doubling time greatly increases and somatic mutations accumulate at a greater rate (See fig. 2-4). The results of Kim et al. suggest that, while MSCs have start exhibiting deleterious effects after passage 4, they could still be readily passaged one or several additional times before the effects of passaging take a significant toll on cell genotype and phenotype.
Claims 1-8, 11, 17-18, 23, 29-30, 32-33, 37-38, 48, and 50 are rejected under 35 U.S.C. 103 as being unpatentable over Cotta-Ramusino et al. (WO 2019152519 A1), of record, in view of Kim et al. (Scientific Reports, 2017), of record, Davies et al. (Stem Cells, 2016), and Ju et al. (Biomaterials, 2019), further in view of Rezvani et al. (WO 2018195339 A1), of record.
The teachings of Cotta-Ramusino et al., Kim et al., Davies et al., and Ju et al. are set forth in the rejection above and are incorporated herein in their entirety.
Regarding claims 4-8, 18, 23, 29, 32-33, and 37-38: Following the discussion of claims 1-3, 11, 30, 38, and 50, Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., render obvious the delivery of PD-L1- and PD-L2-targeting gRNAs and a gene editing nuclease to early-passage MSCs. Cotta-Ramusino et al. teach that one or more nucleic acid molecules other than the RNA-guided nuclease and gRNAs of the gene editing system can be delivered by a vector before or after one or more components of the editing system (which reads on “the steps of (b1) and (c1)” and “the steps of (b2) and (c2)” (See ¶458). Cotta-Ramusino et al. teach that the nucleic acid molecules can encode therapeutic proteins but do not teach the delivery of a vector encoding a heterologous antigen receptor or cytokine (See ¶458).
Rezvani et al. teach cells, which can be MSCs, engineered to express hIL-15 (which reads on “one or more heterologous cytokines”) and at least two antigen receptors that are CARs and/or TCRs (which read on “one or more heterologous antigen receptors”) (See ¶0006-0007, 0079, and 0097). The modified cells can be administered in an effective amount to a subject for treating an immune disorder (which reads on “immune-related disorder”) such as cancer (See ¶0014-0015). Rezvani et al. teach that cells expressing antigen receptors can be assessed for efficacy of killing target cells by cytotoxicity assays (which reads on “the cells are expanded prior to… analysis by one or more functional assays”) (See ¶00266).
It would have been obvious to one having ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method of Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., to comprise delivery of nucleic acids encoding hIL-15 and two antigen receptors, as taught by Rezvani et al., for treating cancer. One would be motivated to make this modification because Rezvani et al. teach that expression of IL-15 and multiple antigen receptors can reduce the risk of antigen-negative tumor escape (See ¶0028). There would be a reasonable expectation of success in doing so because Cotta-Ramusino et al. teach that the therapeutic protein-encoding nucleic acids can be delivered to cells before or after CRISPR components (See ¶458) and because Rezvani et al. teach that MSCs are appropriate for expressing such anti-cancer proteins (See ¶0007, 0079, and 0097).
It also would have been obvious to modify the method of Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., to incorporate the cytotoxicity assays taught by Rezvani et al. following modification of the cells. One would be motivated to make this modification because Rezvani et al. teach that the efficacy of killing target cells can be measured this way (See ¶00266), and such a modification could be readily made.
Regarding claim 17: Following the discussion of claims 1-8, 11, 18, 23, 29-30, 32-33, 37-38, 48, and 50, Cotta-Ramusino et al., modified by Kim et al., Davies et al., Ju et al., and Rezvani et al., render obvious the delivery of CRISPR components and nucleic acids for expression of IL-15 and antigen receptors to MSCs but do not expressly teach the origin of the MSCs.
Rezvani et al. teach that MSCs for modification can be obtained from bone marrow (See ¶0007, 0049, and 0079).
It would have been obvious to one having ordinary skill in the art prior to the effective filing date to further modify the method of Cotta-Ramusino et al., modified by Kim et al., Davies et al., Ju et al., and Rezvani et al., to comprise bone marrow-derived MSCs, such as are taught by Rezvani et al. One would be motivated to make this modification because Rezvani et al. teach such cells as suitable for genetic modification (See ¶0007, 0049, and 0079), and such a modification could be readily made.
Claims 1-3, 11, 30-31, 48, and 50 are rejected under 35 U.S.C. 103 as being unpatentable over Cotta-Ramusino et al. (WO 2019152519 A1), of record, in view of Kim et al. (Scientific Reports, 2017), of record, Davies et al. (Stem Cells, 2016), and Ju et al. (Biomaterials, 2019), further in view of Lange et al. (Journal of Cellular Physiology, 2007), of record.
The teachings of Cotta-Ramusino et al., Kim et al., Davies et al., and Ju et al. are set forth in the rejection above and are incorporated herein in their entirety.
Regarding claim 31: Following the discussion of claims 1-3, 11, 30, 48, and 50, Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., render obvious the delivery of PD-L1- and PD-L2-targeting gRNAs and a gene editing nuclease to early-passage MSCs but do not teach the MSCs as expanded in media comprising platelet lysate, L-glutamine, and/or heparin.
Lange et al. teach methods for expanding hMSCs wherein one medium (M3) comprises glutamine (which reads on “L-glutamine”), heparin, and platelet lysate (See page 19, col. 1, ¶5).
It would have been obvious to one having ordinary skill in the art prior to the effective filing date of the claimed invention to further modify the method of Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., to comprise the medium taught by Lange et al. One would be motivated to make this modification because Lange et al. teach this medium as capable of supporting MSC expansion (See fig. 3-4), and such a modification could be readily made.
Claims 1-3, 11, 30, and 48-50 are rejected under 35 U.S.C. 103 as being unpatentable over Cotta-Ramusino et al. (WO 2019152519 A1), of record, in view of Kim et al. (Scientific Reports, 2017), of record, Davies et al. (Stem Cells, 2016), and Ju et al. (Biomaterials, 2019), further in view of Lawson et al. (Biochemical Engineering Journal, 2017).
The teachings of Cotta-Ramusino et al., Kim et al., Davies et al., and Ju et al. are set forth in the rejection above and are incorporated herein in their entirety.
Regarding claim 49: Following the discussion of claims 1-3, 11, 30, 48, and 50, Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., render obvious the delivery of PD-L1- and PD-L2-targeting gRNAs and a gene editing nuclease to early-passage MSCs but do not teach expansion of the MSCs to at least 109 cells.
Lawson et al. teach the expansion of hMSCs in a 50 L bioreactor using microcarriers (See Abstract and page 50, col. 2, full ¶1-2). Thawed hMSCs were cultured for two passages as monolayers prior to bioreactor seeding (See page 51, col. 1, full ¶1). The bioreactor was inoculated with 1.5 × 104 cells/ml in 20 L (See page 51, col. 1, full ¶1), or 3 × 108 cells. Yields up to 1.29 ×1010 total cells (which reads on “large-scale expansion”) could be achieved per run (which reads on “passage) (See fig. 6A).
It would have been obvious to one having ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method of Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., to comprise the use of a bioreactor, as taught by Lawson et al., for expanding MSCs. One would be motivated to make this modification because Lawson et al. teach that hMSC therapies require larger-scale processing in order to generate the doses needed (See page 49, col. 2, ¶1). There would be a reasonable expectation of success in doing so because Lawson et al. teach that large amounts of MSCs can be produced from single runs (See fig. 6A), and because the cells in the method of Cotta-Ramusino et al., modified by Kim et al., Davies et al., and Ju et al., could be readily expanded in such a manner at low passage numbers.
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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/J.S.S./Examiner, Art Unit 1633
/CHRISTOPHER M BABIC/Supervisory Patent Examiner, Art Unit 1633