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 .
Application Status
Claims 48-54, 56-58, 60-61, 63, 65-67, and 69-73 are pending and examined on the merits herein.
Grounds of Rejection Withdrawn
Previous objections to the specification are withdrawn in view of amendment.
Previous objections to the claims are withdrawn in view of claim amendments.
All previous rejections of claims 55, 59, 62, 64, and 68 are rendered moot by claim cancellation.
Previous rejection of claims 53-54, 56-57, 64 and 65 under 35 U.S.C. 112(a) are withdrawn in view of claim amendments.
Previous rejection of claims 48-52 and 61 under 35 U.S.C. 102 are withdrawn in view of claim amendments.
Claim Rejections - 35 USC § 112(b)
New Rejection Necessitated by Amendment
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 50, 52-54, 56-58, 60-61, 63, 65-67, 69, and 71-73 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claims 50, 52-54, 56-58, 60-61, 63, 65-67, 69, and 71-73, the phrase "preferably" renders the claim indefinite because it is unclear whether the limitation(s) following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Claim Rejections - 35 USC § 103New Rejection Necessitated by Amendment
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.
Claims 48-54, 56-58, 60-61, 63, 65-67, and 69 are rejected under 35 U.S.C. 103 as being unpatentable over Tao (CN 111440801A; IDS entered February 14, 2023; Espacenet English translation used in this rejection; IDS entered February 14, 2023), Wang (WO 2020/088631 A1; cited in OA 11/20/2025) and Ureña-Bailén (Brief Funct Genomics, 2020 May 20;19(3):191-200; cited in OA 11/20/2025).
Regarding claims 48-51, 61, and 69, Tao teaches a sgRNA for targeted knockout of human NKG2A/ KLRC1 gene, characterized in that the nucleotide sequence of the sgRNA is as shown in any one of SEQ ID NO: 1-56 (claim 1) and the present invention can directly edit T cells (page 6, line 25); SEQ ID NOs: 2, 51, 4, 21, 1, 20, 23, 39, and 32 have 100% sequence identity to the instant claimed SEQ ID NOs: 12-13, 15, 61-64, 66, and 68 respectively. Tao further teaches that using plasmid pGL3-2U6-NKG2A/KLRC1-sg(23)-sg(20) the expression reduction rate of NKG2A on the surface was 76.4%-91.8 % (page 11, lines 14-17; Figure 3).
Regarding claims 50 and 61, Tao teaches a CRISPR/Cas9 system comprising the sgRNA targeted to knock out the human NKG2A/KLRC1 gene according to any one of claims 1 to 4, characterized in that it further comprises a Cas9 protein expression plasmid (claim 6).
Regarding claim 52, Tao teaches that the edited cells are PBMCs which include T cells (page 11, lines 23-24), and are therefore primary T cells.
Tao does not teach the engineered T cell further comprising an exogenous protein; wherein the exogenous protein comprises a CAR, TCR, TFP, TAC, YCR-T or combination thereof; the targets of the CAR; or endogenous TCR knockdown.
Regarding claim 53-54, 56, 58, 60, 63, and 65-66, Wang teaches a cell, comprising: a functionally inactive T cell receptor (TCR) , and one or more chimeric antigen receptors (CARs), wherein each individual CAR of said one or more CARS comprising a binding moiety, which binding moiety comprises (i) a first antigen binding domain, which first antigen binding domain suppresses or reduces a subject’s immune response toward said engineered immune cell when administered into said subject and (ii) a second antigen binding domain that binds to a disease-associated antigen, and wherein each CAR of said one or more CARS further comprises a transmembrane domain and an intracellular signaling domain (claim 111), wherein said first antigen binding domain binds to an immune cell antigen (claim 118), wherein said immune cell is a T cell (claim 120), wherein said immune cell antigen is CD159a (NKG2A) (claim 121), wherein a gene encoding an endogenous surface marker of said cell is inactivated, wherein said endogenous surface marker is capable of binding to said first antigen binding domain when expressed (claim 139), wherein said endogenous surface marker is CD159a (NKG2A) (claim 140), wherein said disease-associated antigen is a tumor associated antigen (claim 122), wherein said tumor-associated antigen is BCMA (claim 123). This results in a T cell with a BCMA-CAR and a NKG2A-CAR with endogenous TCR and NKG2A knockdown.
Regarding claim 57, Wang teaches wherein said first antigen binding domain and said second antigen binding domain is arranged, from amino terminus to carboxyl terminus, as: (iii) VL1-VH2-VL2-VH1; wherein VH1 is heavy chain variable domain of said first antigen binding domain, VL1 is a light chain variable domain of said first antigen binding domain, VH2 is a heavy chain variable domain of said second antigen binding domain, and VL2 is a light chain variable domain of said second antigen binding domain (claim 114), which corresponds to the instant claimed option (2).
Regarding claims 58 and 63, Wang teaches the CAR comprising two antigen binding domains can have a structure of L8-VL1-VH2-I-VL2-VH1-H8-TM8-C8-CD3ζ, wherein I is a flexible joint; H8 is an optional hinge region; TM8 is a transmembrane domain; C8 is a costimulatory domain; CD3 is a cytoplasmic signaling sequence derived from CD3 (para 00139).
Regarding claims 60, 66, and 73, Wang teaches knockdown of beta-2-microglobulin to prevent donor CAR-T cells from being attacked by host T cells (para 00221). Wang further teaches TCR/ B2M dual knockout cells (section 1.8; para 00338). Wang further teaches use of gRNA to target TRAC using CRISPR (section 1.2; paras 00310).
Regarding claim 67, Wang teaches the present disclosure provides a method of treating or diagnosing a disease in a subject, comprising administering the pharmaceutical composition to the subject comprising the engineered immune cell which is derived from an allogeneic immune cell. Wang teaches in some embodiments the disease is cancer and the cancer is lymphoma or leukemia. Wang further teaches the terms “subject,” “individual,” and “patient” are used interchangeably herein refer to a vertebrate, preferably a mammal such as a human.
Ureña-Bailén teaches that problems such as targeting healthy cells and committing fratricide (on-target off-tumour effect) can be prevented by a rational selection of the CAR such as avoiding self-destruction by disrupting the expression of T-cell antigens of broad expression in CAR-T cells through CRISPR/Cas9 technology (page 192, col 2, para 3; Fig 1). Ureña-Bailén further teaches that CRISPR/ Cas9 has emerged in the last few years as one of the best options for gene editing that is a very powerful editing system with several positive assets, including simplicity, high efficiency and wide applicability (page 196, col 2, para 2). Ureña-Bailén further teaches that graft versus-host disease (GvHD) and host-versus-graft effect (HvG) are susceptible to occur with CAR-T cell therapy, but can be prevented by knocking out genes such as TRAC and B2M (page 194, last para-page 195 first para; table 1) which can be achieved using CRISPR/Cas9 technology (page 195, col 1, para 2; figure 2).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the instant application to use the CRISPR/Cas NKG2A knockdown cell method of Tao to generate the BCMA CAR and NKG2A CAR with TCR and NKG2A knockdown as taught by Wang, with the reasoning as taught by Ureña-Bailén. This results in a T cell with a BCMA-CAR, a NKG2A-CAR with TCR and NKG2A knockdown achieved through CRISPR/ Cas. The ordinary artisan would have been motivated to do so because Wang teaches bringing an immune cell and a disease antigen expressing cell together through this dual CAR system targeting BCMA and NKG2A. Wang also teaches knockdown of NKG2A expression in the T cells expressing the CAR with the same target as well as knockdown of B2M and TRAC to block endogenous TCR expression and Tao teaches the specific gRNA target sequences to use for the CRISPR/Cas targeting of NKG2A. Ureña-Bailén teaches that CRISPR/ Cas9 technology can be used to knock out genes such as TRAC and B2M, as well as the endogenously expressed CAR target to prevent fratricide as well as graft versus host disease or host versus graft disease in the subject. Ureña-Bailén further teaches that CRISPR/ Cas is a very powerful editing system with several positive assets, including simplicity, high efficiency and wide applicability. The ordinary artisan has a reasonable expectation of success to apply the gRNA sequences and CRISPR/Cas method of Tao to knockdown NKG2A in T cells with high efficiency as taught by Ureña-Bailén to improve the CAR-T therapy in the dual BCMA and NKG2A-CAR-T cells with endogenous NKG2A and TCR knockdown as taught by Wang.
The rationale to apply a technique taught by the prior art as improving the therapeutic and production characteristics of a similar construct is to predictably obtain an improvement to the second construct and is consistent with the exemplary rationales provided by the Supreme Court in KSR International Co. v. Teleflex Inc., 82 USPQ2d 1385, 1395-97 (2007) and discussed in M.P.E.P. § 2143. For these reasons, the invention as a whole would have been prima facie obvious to one ordinary skill in the art before the effective filing date of the claimed invention.
Claim 70 is rejected under 35 U.S.C. 103 as being unpatentable over Tao (CN 111440801A; IDS entered February 14, 2023; Espacenet English translation used in this rejection; IDS entered February 14, 2023), Wang (WO 2020/088631 A1; cited in OA 11/20/2025), and Ureña-Bailén (Brief Funct Genomics, 2020 May 20;19(3):191-200; cited in OA 11/20/2025), as applied to claims 48-54, 56-58, 60-61, 63, 65-67, and 69 above, and further in view of Andre (WO 2016/041945 A1; PTO-892).
The teachings of Tao, Wang and Ureña-Bailén regarding claims 48-54, 56-58, 60-61, 63, 65-67, and 69 are detailed above.
Tao, Wang and Ureña-Bailén do not teach the sequences of the NKG2A binding moiety.
Andre teaches an anti-NKG2A antibody that neutralizes inhibitory activity of human NKG2A for use in treating cancer in a human patient (claim 1), wherein an antibody that neutralizes the inhibitory activity of NKG2A comprises the CDR1 , CDR2 and CDR3 domains of a heavy chain having the sequence set forth in any one of SEQ ID NOS: 4-8, and the CDR1 , CDR2 and CDR3 domains of a light chain having the sequence set forth in SEQ ID NO: 9. SEQ ID NO: 9 has 100% sequence identity to the instant claimed SEQ ID NO: 2 and 6-8 and SEQ ID NO: 5 has 100% sequence identity to the instant claimed SEQ ID NO: 1 and 3-5.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the instant application to use an established NGK2A antibody in the BCMA CAR/ NKG2A CAR with TCR and NKG2A knockdown achieved by CRISPR/Cas as taught by Tao, Wang, and Ureña-Bailén. This results in a T cell with a BCMA-CAR, a NKG2A-CAR with TCR and NKG2A knockdown achieved through CRISPR/ Cas. The ordinary artisan would have been motivated to do so because this is a simple substitution of an established sequence into the same element of a structure. The ordinary artisan has a reasonable expectation of success at using an established sequence for a BCMA antibody to generate the BCMA CAR portion of the BCMA CAR/ NKG2A CAR with TCR and NKG2A knockdown.
Claims 71-72 are rejected under 35 U.S.C. 103 as being unpatentable over Tao (CN 111440801A; IDS entered February 14, 2023; Espacenet English translation used in this rejection; IDS entered February 14, 2023), Wang (WO 2020/088631 A1; cited in OA 11/20/2025), and Ureña-Bailén (Brief Funct Genomics, 2020 May 20;19(3):191-200; cited in OA 11/20/2025), as applied to claims 48-54, 56-58, 60-61, 63, 65-67, and 69 above, and further in view of Peng (WO 2018/133877 A1; cited in OA 11/20/2025; Machine English translation used in this rejection attached as NPL; cited in OA 11/20/2025).
The teachings of Tao, Wang and Ureña-Bailén regarding claims 48-54, 56-58, 60-61, 63, 65-67, and 69 are detailed above.
Tao, Wang and Ureña-Bailén do not teach the sequences of the BCMA binding moiety.
Regarding claims 71-72, Peng teaches an antibody that targets BCMA wherein the heavy chain variable region of the antibody has the amino acid sequence of SEQ ID NO: 56 (claim 1-1) and wherein the light chain variable region of the antibody has the amino acid sequence of SEQ ID NO: 19 (claim 1-2), SEQ ID NO: 56 has 100% sequence identity to the instant claimed SEQ ID NO: 22 which comprises the instant claimed SEQ ID NOs: 16, 17 and 18; SEQ ID NO: 19 has 100% sequence identity to instant claimed SEQ ID NO: 23. SEQ ID NOs 19 and 56 are both in instant claimed Seq ID NO: 24 with a linker as seen in the alignment below. Peng further teaches a linker consisting of GGGGSGGGGSGGGGS, to form a scFv (example 4), which is the exact linker used in instant SEQ ID NO: 24. Peng further teaches use of the antibody for preparing an immune cell expressing a chimeric antigen receptor modification; preferably, the immune cell comprises T lymphocytes (page 3, lines 27-28) the chimeric antigen receptor wherein most preferably, the transmembrane domain is selected from the group consisting of CD8α or CD28, and the costimulatory signal domain is selected from the intracellular signal domain of CD137 or CD28, and the primary signal domain is selected from the group consisting of CD3 (page 4, lines 29-31).
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It would have been obvious to one of ordinary skill in the art before the effective filing date of the instant application to use an established BCMA antibody CAR in the BCMA CAR/ NKG2A CAR with TCR and NKG2A knockdown achieved by CRISPR/Cas as taught by Tao, Wang, and Ureña-Bailén. This results in a T cell with a BCMA-CAR, a NKG2A-CAR with TCR and NKG2A knockdown achieved through CRISPR/ Cas. The ordinary artisan would have been motivated to do so because this is a simple substitution of an established sequence into the same element of a structure. The ordinary artisan has a reasonable expectation of success at using an established sequence for a BCMA antibody to generate the BCMA CAR portion of the BCMA CAR/ NKG2A CAR with TCR and NKG2A knockdown.
Response to Arguments
Applicant's arguments filed 02/12/2026 have been fully considered but they are not persuasive.
Applicant submits: At the time of the invention, NKG2A was well understood as an inhibitory receptor predominantly expressed on NK cells, and was hardly expressed in normal cultured T cells (see, e.g., Example 3 of the instant application). As such, while Tao discloses tools to reduce NKG2A expression in immune cells generally, it provides no motivation for a POSA to select T cells from the mixture for NKG2A knockout, much less such cells that also express an NK-targeting exogenous protein. Nor does Tao provide a POSA with any motivation to combine its teaching with Wang-a reference on T cell engineering.
Wang is cited for its disclosure of engineered immune cells comprising a CAR or TCR targeting various antigens, in which "a gene encoding an endogenous surface marker of said cell is inactivated, wherein said endogenous surface marker is capable of binding to said first antigen binding domain when expressed." Wang, however, does not provide the specific motivation for generating NK-targeting engineered T cells with inactivated NKG2A for at least the following reasons.
First, a close review of Wang reveals that it provides "laundry lists" of potential targets, including immune cell targets (e.g., claims 121 and 140) and tumor-associated antigens (e.g., claim 123).
Second, while Wang mentions generally that "an endogenous surface marker of the engineered immune cell can be inactivated," and includes NKG2A in the laundry list of such markers, Wang does not teach or suggest inactivating NKG2A for T cells.
In Response: Contrary to the assertion that NGK2A is “hardly expressed on T cells” Andre (Cell. 2018 Dec 13;175(7):1731-1743; IDS entered 02/142023) teaches that NGK2A is an ITIM bearing receptor expressed on both T and NK cells (page 1732, col 1, para 1). Andre further teaches that in healthy individuals 5% of human peripheral blood CD8+ T cells express cell surface NKG2A but this is upregulated by chronic antigen stimulation (page 1732, col 1, para 1). Andre further demonstrates that blockade of NGK2A enhances anti0tumor immunity by both NK and CD8+ T cells (page 1732, col 1, para 2).
It is not necessary for a reference to reduce to practice a limitation to be valid prior art please see MPEP § 2121: When the reference relied on expressly anticipates or makes obvious all of the elements of the claimed invention, the reference is presumed to be operable. Once such a reference is found, the burden is on applicant to rebut the presumption of operability. In re Sasse, 629 F.2d 675, 207 USPQ 107 (CCPA 1980).
Targeting of NKG2A is identified by Tao and Wang provides motivation to knockdown a target that is being targeted by an engineered CAR in the same cell. Further the entire application of Wang is directed to engineered T cells (title) and this is exemplified in the inactivation of an endogenous TCR receptor (claim 6 and 63) which would not be relevant in other types of immune cells.
Ureña-Bailén was cited for the teaching that CRISPR/ Cas9 technology can be used to knock out genes such as TRAC and B2M, as well as the endogenously expressed CAR target to prevent fratricide. It was for the rationale and not the target
The motivation to combine is detailed in the new 103 rejection above and a POSA has a reasonable expectation of success as all of the elements were known in the prior art.
Double Patenting
New Rejection Necessitated by Amendment
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.
Claims 48-54, 56, 58, 60-61, 63, 65-67, and 69-73 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 46-49, 51-52, 54-55, 59, 61, 63-64, and 69-70 of copending Application No. 17/623,481 in view of Tao (CN111440801A; IDS entered February 14, 2023; Machine English translation used in this rejection; IDS entered February 14, 2023) and Ureña-Bailén (Brief Funct Genomics, 2020 May 20;19(3):191-200; cited in OA 11/20/2025).
This is a provisional nonstatutory double patenting rejection.
Regarding claims 48, 58, 60-61, 63, and 66, the copending claims teach an isolated T cell resistant to transplantation immune rejection, characterized in that the cell expresses a first protein and has the function of inhibiting or killing Natural Killer (NK) cells of the host; wherein the first protein is a chimeric antigen receptor (CAR) comprising an extracellular domain, a transmembrane domain, and an intracellular signaling domain; wherein the extracellular domain comprises an antigen binding region that specifically recognizes one or more antigens of the NK cells, and the intracellular signaling domain comprises a functional signaling domain of a stimulatory molecule or a costimulatory molecule; wherein the antigen of the NK cells is selected from the group consisting of NKG2-A/NKG2-B type II integral membrane protein (NKG2A)… wherein the transmembrane domain is CD28 transmembrane domain or CD8a transmembrane domain; and wherein the intracellular signaling domain comprises the functional signaling domain of CD3ζ,: and wherein the endogenous β2 microglobulin (B2M) gene is silenced or knocked out (claim 46 and 72), characterized in that an endogenous T cell receptor (TCR) gene is silenced or knocked out (claim 55), wherein the intracellular signaling domain further comprises the functional signaling domain of 4-lBB, CD28, or CD27 (claim 54).
Regarding claim 52, the copending claims teach wherein the cell is an allogeneic cell (claim 49).
Regarding claims 53, 65, and 70, the copending claims teach wherein the antigen binding region that specifically recognizes NKG2A comprises HCDRl having the amino acid sequence of SEQ ID NO: 10, HCDR2 having the amino acid sequence of SEQ ID NO: 11, and HCDR3 having the amino acid sequence of SEQ ID NO: 12; and LCDRl having the amino acid sequence of SEQ ID NO: 13, LCDR2 having the amino acid sequence of SEQ ID NO: 14, and LCDR3 having the amino acid sequence of SEQ ID NO: 15 (claim 63), wherein the antigen binding region that specifically recognizes NKG2A comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 1, and a light chain variable region having the amino acid sequence of SEQ ID NO:2 (claim 64). SEQ ID NOs: 1-2 have 100% sequence identity to the instant claimed same SEQ ID NOs. SEQ ID NOs: 10-15 have 100% sequence identity to the instant claimed SEQ ID NOs: 3-8.
Regarding claims 54, 56, and 63, the copending claims teach wherein the cell further expresses a second protein, wherein the second protein is a CAR comprising an extracellular domain, a transmembrane domain, and an intracellular signal domain; wherein the extracellular domain comprises an antigen binding region that specifically recognizes a tumor antigen; wherein the tumor antigen is BCMA (claims 59 and 61).
Regarding claims 71-72, the copending claims teach wherein the antigen binding region that specifically recognizes BMCA comprises HCDRl having the amino acid sequence of SEQ ID NO: 16, HCDR2 having the amino acid sequence of SEQ ID NO: 17, and HCDR3 having the amino acid sequence of SEQ ID NO: 18; and LCDRl having the amino acid sequence of SEQ ID NO: 19, LCDR2 having the amino acid sequence of SEQ ID NO:20, and LCDR3 having the amino acid sequence of SEQ ID NO:21 (claim 69), herein antigen binding region that specifically recognizes BMCA comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO:22, and a light chain variable region having the amino acid sequence of SEQ ID NO:23 (claim 70). SEQ ID NOs: 16-23 have 100% sequence identity to the instant claimed same SEQ ID NOs.
The copending claims do not teach knockdown of NKG2A; the sequences or method of achieving the knockdown
Regarding claims 48-51, 61, and 69, Tao teaches a sgRNA for targeted knockout of human NKG2A/ KLRC1 gene, characterized in that the nucleotide sequence of the sgRNA is as shown in any one of SEQ ID NO: 1-56 (claim 1) and the present invention can directly edit T cells (page 6, line 25); SEQ ID NOs: 2, 51, 4, 21, 1, 20, 23, 39, and 32 have 100% sequence identity to the instant claimed SEQ ID NOs: 12-13, 15, 61-64, 66, and 68 respectively. Tao further teaches that using plasmid pGL3-2U6-NKG2A/KLRC1-sg(23)-sg(20) the expression reduction rate of NKG2A on the surface was 76.4%-91.8 % (page 11, lines 14-17; Figure 3).
Regarding claims 50 and 61, Tao teaches a CRISPR/Cas9 system comprising the sgRNA targeted to knock out the human NKG2A/KLRC1 gene according to any one of claims 1 to 4, characterized in that it further comprises a Cas9 protein expression plasmid (claim 6).
Regarding claim 52, Tao teaches that the edited cells are PBMCs which include T cells (page 11, lines 23-24), and are therefore primary T cells.
Regarding claims 48-50, 53-54, 56, 60-61, 63, and 65-66, Ureña-Bailén teaches that problems such as targeting healthy cells and committing fratricide (on-target off-tumour effect) can be prevented by a rational selection of the CAR such as avoiding self-destruction by disrupting the expression of T-cell antigens of broad expression in CAR-T cells through CRISPR/Cas9 technology (page 192, col 2, para 3; Fig 1). Ureña-Bailén further teaches that CRISPR/ Cas9 has emerged in the last few years as one of the best options for gene editing that is a very powerful editing system with several positive assets, including simplicity, high efficiency and wide applicability (page 196, col 2, para 2). Ureña-Bailén further teaches that graft versus-host disease (GvHD) and host-versus-graft effect (HvG) are susceptible to occur with CAR-T cell therapy, but can be prevented by knocking out genes such as TRAC and B2M (page 194, last para-page 195 first para; table 1) which can be achieved using CRISPR/Cas9 technology (page 195, col 1, para 2; figure 2).
It would have been obvious to one of ordinary skill in the art to add the CRISPR/Cas9 NKG2A knockdown as taught by Tao to the BCMA CAR and NKG2A CAR with B2M/TCR knockdown as taught by the copending claims, with the reasoning as taught by Ureña-Bailén. This results in a T cell with a BCMA-CAR, a NKG2A-CAR with B2M/TCR and NKG2A knockdown achieved through CRISPR/ Cas. The ordinary artisan would have been motivated to do so because Tao teaches how to knockdown NKG2A expression in the T cells and the specific gRNA target sequences to use for the CRISPR/Cas targeting of NKG2A. Ureña-Bailén teaches that CRISPR/ Cas9 technology can be used to knock out genes such as TRAC and B2M, as well as the endogenously expressed CAR target to prevent fratricide as well as graft versus host disease or host versus graft disease in the subject. Ureña-Bailén further teaches that CRISPR/ Cas is a very powerful editing system with several positive assets, including simplicity, high efficiency and wide applicability. The ordinary artisan has a reasonable expectation of success to apply the gRNA sequences and CRISPR/Cas method of Tao to knockdown NKG2A in T cells with high efficiency as taught by Ureña-Bailén to improve the CAR-T therapy in the dual BCMA and NKG2A-CAR-T cells with NKG2A and B2M/ TCR knockdown as taught by the copending claims.
The rationale to apply a technique taught by the prior art as improving the therapeutic and production characteristics of a similar construct is to predictably obtain an improvement to the second construct and is consistent with the exemplary rationales provided by the Supreme Court in KSR International Co. v. Teleflex Inc., 82 USPQ2d 1385, 1395-97 (2007) and discussed in M.P.E.P. § 2143. For these reasons, the invention as a whole would have been prima facie obvious to one ordinary skill in the art before the effective filing date of the claimed invention.
Claims 48-54, 56, 58, 60-61, 63, 65-66 and 69-73 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 22, 25, 30-31, 35, 39, 41, 43, and 48 of copending Application No. 18/837,379 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Regarding claims 48-49, 60, 66, 69, and 73, the copending claims teach a cell, wherein the cell comprises: b) low or no expression of endogenous TCR/B2M/NKG2A molecules preferably, wherein the endogenous TCR, B2M, CIITA and/or NKG2A molecules are knocked out by CRISPR/Cas9 technology (claim 39). the copending claims teach further comprising a third gRNA targeting TRAC gene and/or a fourth gRNA targeting B2M gene: preferably, wherein the third gRNA comprises a sequence represented by SEQ ID NOs: 24, 64 and/or 65; and/or the fourth gRNA comprises a sequence represented by SEQ ID NOs: 25, 66 and/or 67 (claim 25). SEQ ID NO: 24 has 100% sequence identity to the instant claimed SEQ ID NO: 53 and SEQ ID NO: 25 has 100% sequence identity to the instant claimed SEQ ID NO: 54.
Regarding claims 51 and 61, the copending claims teach a gRNA construct comprising a second gRNA targeting NKG2A, wherein the second gRNA comprises a sequence represented by SEQ ID NO: 14, 15 or 21 (claims 22 and 41). SEQ ID NO: 15 has 100% sequence identity to the instant claimed same SEQ ID NO.
Regarding claims 50, 52, and 61, the copending claims teach a method for gene editing of CIITA or NKG2A in cells by the CRISPR/Cas system, which comprises conducting gene editing of the cells by using the construct according to claim 22 (claims 30-31), wherein the cells are selected from: T cells, preferably, wherein the cells are selected from: autologous T cells or allogeneic T cells, stem cell-derived T cells, primary T cells or autologous T cells derived from human (claim 35).
Regarding claims 48, 53, 58, and 63, the copending claims teach wherein the cell further expresses an exogenous receptor that recognizes an NK cell receptor, a tumor antigen and/or a pathogen antigen, preferably expresses an exogenous receptor that recognizes a NKG2A., preferably, wherein the exogenous receptor comprises a chimeric antigen receptor (CAR) or a recombinant TCR receptor; more preferably, wherein the CAR comprises: more preferably, wherein the CAR comprises: a) an antibody that recognizes a NKG2A polypeptide, a tumor and/or a pathogen antigen, a transmembrane region of CD28 or CD8, a co-stimulatory signaling domain of CD28, and CD38; and/or c) an antibody that recognizes a NKG2A polypeptide, a tumor and/or a pathogen antigen, a transmembrane region of CD28 or CD8, a costimulatory signaling domain of CD28, a costimulatory signaling domain of CD137, and CD3ζ; preferably, wherein the cell is selected from the group consisting of: T cells, ore preferably, wherein the cell is selected from: autologous T cells or allogeneic T cells, stem cell-derived T cells, primary T cells, or autologous T cells derived from human (claim 43)
Regarding claims 53-54, 56, 63, 65, and 70-72, the copending claims teach the cell according to claim 43, wherein the tumor antigen is selected from: CD19, GPC3, Claudinl8.2, WTI, HER2, EGFR, BCMA, or a combination thereof, preferably, the antibody recognizing the NKG2A polypeptide comprises: the heavy chain variable region and the light chain variable region respectively represented by SEQ ID NO: 34 and SEQ ID NO: 35,; or a tandem antibody sequence represented by SEO ID NO: 46, 47, 48, 49 or 50, more preferably, the antibody recognizing a tumor antigen comprises: a heavy chain variable region and a light chain variable region respectively represented by SEQ ID NO: 27 and SEQ ID NO: 28; or a scFv represented by SEO ID NO: 29, 30, 31, 32 or 33; or a tandem antibody sequence represented by SEO ID NO: 46, 47, 48, 49 or 50 (claim 48). SEQ ID NOs: 34-35 have 100% sequence identity to the instant claimed SEQ ID NOs: 1-2 respectively. SEQ ID NO: 46 contains 100% of instant claimed SEQ ID NOs: 22-23.
Response to Arguments
Applicant's arguments filed 02/12/2026 have been fully considered but they are not persuasive.
Applicant submits: At the time of the invention, NKG2A was well understood as an inhibitory receptor predominantly expressed on NK cells, and was hardly expressed in normal cultured T cells (see, e.g., Example 3 of the instant application). As such, while Tao discloses tools to reduce NKG2A expression in immune cells generally, it provides no motivation for a POSA to select T cells from the mixture for NKG2A knockout, much less such cells that also express an NK-targeting exogenous protein. Nor does Tao provide a POSA with any motivation to combine its teaching with Wang-a reference on T cell engineering.
In Response: In the NSDP rejection this argument is moot as the copending claims already select the T cell population. Further as detailed above there is prior art teaching the expression of NKG2A on NK and T cells.
Conclusion
No claims are allowed.
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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/AMBER K FAUST/Examiner, Art Unit 1643
/JULIE WU/Supervisory Patent Examiner, Art Unit 1643
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