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 1-3, 5-8, and 10-14 are pending and examined on the merits herein.
Power of Attorney
It is noted that a Power of Attorney is not on record for the instant application. The Applicant is encouraged to file a Power of Attorney in the event that the Examiner needs to communicate with an authorized representative for the Applicant during the prosecution of the case.
Grounds of Rejection Withdrawn
Previous objection to the specification is withdrawn in view of amendment.
Previous objections to claims 1-3 are withdrawn in view of claim amendments.
All previous objections and rejections of claims 4 and 9 are rendered moot by claim cancellation.
Previous rejection of claims 1 and 8 under 35 U.S.C. 112(b) are withdrawn in view of claim amendments.
Claim Objections
Objection Maintained
Claims 10-13 are objected to because of the following informalities:
Claims 10-13 all recite “A method of claim 8” these should read “The method of claim 8”.
Appropriate correction is required.
Claim Rejections - 35 USC § 103
New 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 1-2, 8 and 14 are rejected under 35 U.S.C. 103 as being unpatentable over Morsut (Cell. 2016 Feb 11;164(4):780-91; cited in OA 11/25/2025), Lupo (J Hematol Oncol. 2020 Jun 12;13(1):76; cited in OA 11/25/2025), Roybal (Cell. 2016 Oct 6;167(2):419-432.e16; cited in OA 11/25/2025) and Chang (Curr Opin Chemical Engineering, 2020, 30: 69-74; cited in OA 11/25/2025).
Regarding claims 1-2, 8, and 14, Morsut teaches synthetic notch receptors that use surface antigen binding to regulate cleavage through a small regulatory region of Notch and regulate transcription through an intracellular GAL4-VP64 to generate a novel transcriptional output (Fig 1a). Morsut further teaches that the notch core is comprised of 3 Lin-12 Notch repeats (LNRs) which in the basal state bind and shield ADAM/ TACE protease cleavage site, but upon binding of the extracellular domain to a ligand releases the LNR domains exposing the ADAM/TACE protease cleavage site which can then initiate cleavage (page 797; Fig 7). Morsut further teaches the intracellular domain of Notch can be replaced with an artificial transcription factor such as GAL4-VP64 to create a reporter of Notch activity (page 780, col 2, para 4). Morsut further teaches that all synNotch receptors contain an N-terminal CD8a signal peptide that can be fused to an scFv fused to a Notch core domain fused to a GAL4-VP64 to activate a reporter construct (page 789, col 1, para 4). Morsut further teaches generation of cell lines with the synNotch construct using lentiviral transduction (experimental procedures). Morsut further teaches that synNotch receptors may prove to be very useful in therapeutic cells, such as engineered immune cells and in regenerative medicine, both of which require specific sensing of environmental signals linked to precise cellular responses (page 789, col 1, para 3) and synNotch receptors are therefore a powerful tool due to their customizable input/ output function (page 781, col 2, para 2). This results in a construct that contains 5’ to 3’: signal peptide- scFv- Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- reporter protein. Morsut further teaches use of a UAS upstream of the reporter BPS in an anti-GFP, synNotch, GAL-VP64, UAS-BFP construct (Figure 4).
The synNotch is interpreted as comprising a notch core domain.
Regarding claim 2, Morsut teaches that the synNotch constructs were engineered into a vector and then transfected into lentivirus, that was then used for transduction into cell lines for testing (experimental procedures).
Morsut does not teach a CD155/TIGIT-binding protein or use of NK cells that are derived from pluripotent stem cells.
Regarding claims 1-2, and 8, Lupo teaches that natural killer (NK) cells are powerful immune effectors that modulate their anti-tumor function through a balance of activating and inhibiting ligands on their cell surface. Lugo further teaches that the tumor microenvironment of glioblastoma (GBM) overexpresses CD155 which has emerged as a pro-tumorigenic antigen and contributes to GBM migration and aggressiveness. Lupo further teaches that CD155 activates NK cells through CD226 or CD96 but inhibits them through TIGIT, but TIGIT is upregulated in cancer so that CD155 is a predominantly inhibitory receptor in the context of solid tumors (abstract; Fig 1). Lupo further teaches that the CD155/TIGIT axis has been shown to be involved in the regulation of immune cells and tumor progression, proliferation and invasion (page 7, col 2, para 3). Lupo further teaches that clinically, significant responses of autologous blood-derived NK cells to treat malignant gliomas have been reported (page 5, col 1, para 3). Lupo further teaches that there are numerous immunotherapies targeting the CD155/TIGIT axis being tested in clinical trials for GBM as well as other tumor types (Table 1), and that targeting CD155 in GBM has already demonstrated efficacy (page 7, col 2, para 3). Lupo further teaches that targeting CD155/TIGIT axis with engineered NK cells holds tremendous potential to treat GBM to improve clinical outcomes (page 7, col 2, para 3) especially as there have been novel advances in activation strategies for NK cells,
the development of new NK cell sources, as well as advancements in genetic engineering toward improved targeting approaches for various cancers (page 5, col 1, para 2).
Regarding claim 8, Lupo teaches that inhibition of CD155 has been shown to result in a reduction in tumor invasiveness as well as a reduction in secretion of MMP-2 (page 5, col 2, para 4). This is interpreted as an alteration of target cell function with the target cell being a tumor cell.
Regarding claim 14, Lupo teaches that inhibition of TIGIT strongly enhances NK cell function including proliferation and production of pro-inflammatory cytokines (page 7, col 1, para 1). This is interpreted as an alteration of NK cell activity.
Regarding claim 1, Roybal teaches use of synthetic Notch receptors with extracellular domains targeting cancer cells to deliver custom response such as cytokines, cytotoxic agents, or therapeutic antibodies (graphical abstract). Roybal further teaches that synNotch cells drive customized therapeutic responses for cancer or other diseases by sensing tumor antigens and locally delivering therapeutic modalities such as cytokines or antibodies (highlights). Roybal further teaches use of the synthetic Notch receptor comprising an anti-CD19-synNotch-Gal4VP64-UAS-IL2 with demonstrated antigen induced cytokine production (Figure 2c and Figure S2 and legend). Roybal further teaches that a potential way to increase the effectiveness of the therapeutic antibodies and reduce adverse events is to have T cells locally produce the antibodies in tumors (page 425, col 2, para2) synNotch can be used to this effect which was demonstrated using an anti-GFP – synNotch – GAL4 – anti-PD-1 or anti-CTLA output (Figure 5). Roybal further demonstrated in vivo efficacy with this model in a mouse GFP tumor model using a synNotch construct driving expression of a cytokine or a BITE targeting CD19 and CD3 in T cells (Figure 6). Roybal further teaches using the synNotch receptor with a cancer antigen in an immune cell can be used to disrupt the disease microenvironment, lower systemic toxicity and effectively overcome immunosuppression put up by many tumors (discussion).
Regarding claims 1-2 and 14, Chang teaches methods of differentiating NK from pluripotent stem cells (PSC) (figure 1). Chang further teaches that adoptive cell therapy with NK cells is attractive because of their unique innate ability to kill tumor cells without prior sensitization or antigen presentation and lack of graft versus host disease but there are major drawbacks including lack of in vivo persistence, challenge in obtaining sufficient healthy NK cells, resistance to genome editing, and suboptimal cytotoxicity against solid tumor (introduction). Chang further teaches that the ability to easily generate large numbers of universally histocompatible NK cells and ease of genome editing enable the continuously renewing human pluripotent stem cells (hPSCs) as a promising source to develop a truly off-the-shelf cellular immunotherapy (introduction). Chang further teaches the genetic modification of the NK cells derived human PSC for improved cellular function (abstract), and further various PSC derived NK cells that were genetically altered through different constructs to enhance cytotoxicity (table 2). Chang further teaches that recent advances in the differentiation and genome editing of hPSCs for NK cell-based immunotherapy and particularly, the combination of genome editing and hPSCs have significantly enhanced the persistence and cytotoxicity of engineered NK cells (conclusion).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the instant application to use a signal peptide- scFv- Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- reporter protein construct as taught by Morsut in the engineered NK cells targeting CD155/TIGIT as taught by Lupo and to substitute an effector protein as taught by Roybal, derived from pluripotent stem cells as taught by Chang. This results in PSC derived NK cells genetically engineered to express signal peptide- CD155/TIGIT binding protein- Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- effector protein. The ordinary artisan would have been motivated to do so because Morsut teaches that synNotch receptors would be very useful in engineered immune cells which require specific sensing of environmental signals linked to precise cellular responses. Roybal teaches that synthetic Notch receptors with extracellular domains targeting cancer cells to deliver custom responses, such as cytokines, cytotoxic agents, or therapeutic antibodies in an immune cell can be used to disrupt the disease microenvironment, lower systemic toxicity and effectively overcome immunosuppression put up by many tumors. Lupo teaches that there are numerous immunotherapies targeting the CD155/TIGIT axis and that targeting CD155 in GBM has already demonstrated efficacy as well as adoptive cell therapy using autologous NK cells. Lupo further teaches that TIGIT is upregulated on NK cells in cancer and is an inhibitory signal and that inhibition of TIGIT strongly enhances NK cell function including proliferation and production of pro-inflammatory cytokines. Chang teaches the ability to easily generate large numbers of universally histocompatible NK cells and ease of genome editing enable the continuously renewing human pluripotent stem cells (hPSCs) as a promising source to develop a truly off-the-shelf cellular immunotherapy and genetic engineering enhances the cytotoxicity for adoptive cell therapy. The ordinary artisan has a reasonable expectation of success to substitute an effector protein for the reporter such as a cytokine or antibody in the NK cells derived from PSC in the known synthetic Notch receptor construct comprising a signal peptide- CD155/TIGIT binding protein - Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- reporter protein that has the ability to overcome the immunosuppressive tumor microenvironment and increase efficacy of cancer immunotherapy.
Claims 5 and 10 are rejected under 35 U.S.C. 103 as being unpatentable over Morsut (Cell. 2016 Feb 11;164(4):780-91; cited in OA 11/25/2025), Lupo (J Hematol Oncol. 2020 Jun 12;13(1):76; cited in OA 11/25/2025), Roybal (Cell. 2016 Oct 6;167(2):419-432.e16; cited in OA 11/25/2025) and Chang (Curr Opin Chemical Engineering, 2020, 30: 69-74; cited in OA 11/25/2025) as applied to claims 1-2, 8, and 14 above, and further in view of Chambers ( Front Mol Biosci. 2019 Jul 24;6:60; cited in OA 11/25/2025).
The teachings of Morsut, Lupo, Roybal, and Chang and regarding claims 1-2, 8, and 14 are detailed above.
Morsut, Lupo, Roybal, and Chang do not teach wherein the effector molecule is an anti-CD73 binding protein.
Chambers teaches that immunosuppressive signaling via CD73 has also been reported to occur from exosomes secreted from tumors that are often involved in malignant reprogramming of immune cells (page 3, col 1, para 3). Chambers further teaches that when exosomes expressing CD73 and CD39 were incubated with NK cells the lytic functions and survival of NK cells were inhibited (page 3 col 2, para 1). Chambers further teaches that Multiple studies have shown that CD73 expression is rapidly enhanced under hypoxic conditions (page 3, col 2, last para) and that NK cells are sensitive to hypoxia (page 4, col 2, para 1). Chambers further teaches that antibody-mediated blockade of CD73 enhanced anti-metastatic function of NK cells by
blocking tumor growth, as well as enhanced cytotoxicity and intra-tumoral infiltration of chimeric antigen receptor-NK cells in vivo (table 2). Chambers further teaches that despite successful targeting of CD73 alone, more profound responses could be obtained when CD73 targeting is used as part of combination treatments (page 8, col 1, para 3).
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 anti-CD73 antibody as the effector protein as taught by Chambers in the signal peptide- CD155/TIGIT binding protein - Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- effector protein expressing NK cells derived from PSCs as taught by Morsut, Lupo, Chang, and Roybal. This results in PSC derived NK cells genetically engineered to express signal peptide- CD155/TIGIT binding protein- Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- anti-CD73. The ordinary artisan would have been motivated to do so because Chambers teaches that CD73 is expressed by exosomes secreted from tumors and has an inhibitory effect on NK cell function and survival. Chambers also teaches that antibody-mediated blockade of CD73 enhanced anti-metastatic function, cytotoxicity, and intra-tumoral infiltration of NK cells and that despite successful targeting of CD73 alone, more profound responses could be obtained when CD73 targeting is used as part of combination treatments. Roybal teaches that use of synthetic Notch receptors with extracellular domains targeting cancer cells to deliver custom response such as cytokines, cytotoxic agents, or therapeutic antibodies. Roybal further teaches that a potential way to increase the effectiveness of the therapeutic antibodies and reduce adverse events is to have cells locally produce the antibodies in tumors and that synNotch can be used to this effect. The ordinary artisan has a reasonable expectation of success to generate NK cells derived from PSC genetically engineered to express a synthetic Notch receptor construct comprising a signal peptide- CD155/TIGIT binding protein - Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- anti-CD73 to overcome the immunosuppressive tumor microenvironment, increase efficacy of cancer immunotherapy and lower adverse events by local delivery of the anti-CD137 antibody.
Claims 7 and 11-13 are rejected under 35 U.S.C. 103 as being unpatentable over Morsut (Cell. 2016 Feb 11;164(4):780-91; cited in OA 11/25/2025), Lupo (J Hematol Oncol. 2020 Jun 12;13(1):76; cited in OA 11/25/2025), Roybal (Cell. 2016 Oct 6;167(2):419-432.e16; cited in OA 11/25/2025) and Chang (Curr Opin Chemical Engineering, 2020, 30: 69-74; cited in OA 11/25/2025) as applied to claims 1-2, 8, and 14 above, and further in view of Makkouk (Eur J Cancer. 2016 Feb;54:112-119; cited in OA 11/25/2025).
The teachings of Morsut, Lupo, Roybal, and Chang regarding claims 1-2, 4, 8-9, and 14 are detailed above.
Morsut, Lupo, Roybal, and Chang do not teach wherein the effector molecule is an anti-CD137 binding protein.
Makkouk teaches that targeting of CD137 has promising clinical applications as it harnesses co-stimulatory receptors to activate innate and adaptive immune cells and to modulate the tumor microenvironment (TME) (abstract). Makkouk further teaches that preferential expression of CD137 on tumor reactive T cells and its ability to increase T cell recruitment into tumor sites, enhance their cytotoxicity as well as protect T cells from activation induced cell death plays a role in anti-CD137 monoclonal antibody response (page 113, col 2, para 4). Makkouk further teaches that several reports have shown a critical role for natural killer cells in mediating the therapeutic effects of CD137 through perforin and granzyme (page 114, col 1, para 2; Figure 1). Makkouk further teaches that there are numerous anti-CD137 antibodies that have entered clinical testing including agonistic antibodies, CARs, and bispecifics (Table 1). Makkouk further teaches that immunosuppression in the TME makes checkpoint inhibitors a rational combination partner with CD137 therapeutic treatment (page 116, col 1, para 5). Makkouk further teaches that clinical CD137 antibodies have met with difficulty due to on target off tumor toxicity and associated adverse events (page 116, col 2, para 1). Makkouk further teaches that while early success of CD137-mediated immunotherapy using anti-CD137 monotherapy was limited, we believe that combination therapies and novel modalities that are designed to positively influence the TME offer a unique opportunity for achieving durable complete responses against multiple types of cancer (page 117, col 2, para 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 an anti-CD137 antibody as the effector protein as taught by Makkouk in the signal peptide- CD155/TIGIT binding protein - Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- effector protein expressing NK cells derived from PSCs as taught by Morsut, Lupo, Roybal, and Chang. This results in PSC derived NK cells genetically engineered to express signal peptide- CD155/TIGIT binding protein- Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- anti-CD137. The ordinary artisan would have been motivated to do so because Makkouk teaches that there is a critical role for natural killer cells in mediating the therapeutic effects of CD137 through perforin and granzyme and further that combination therapies and novel modalities with anti-CD137 that are designed to positively influence the TME offer a unique opportunity for achieving durable complete responses against multiple types of cancer, while overcoming the on target off tumor toxicity seen with initial clinical CD137 testing. Roybal teaches that use of synthetic Notch receptors with extracellular domains targeting cancer cells to deliver custom response such as cytokines, cytotoxic agents, or therapeutic antibodies. The ordinary artisan has a reasonable expectation of success to generate NK cells derived from PSC genetically engineered to express a synthetic Notch receptor construct comprising a signal peptide- CD155/TIGIT binding protein - Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- anti-CD137 to overcome the immunosuppressive tumor microenvironment to activate cytotoxicity of NK cells and increase the efficacy of cancer immunotherapy.
Response to Arguments
Applicant's arguments filed 04/25/2026 have been fully considered but they are not persuasive.
Applicant submits: Applicant respectfully traverse the rejection. As now amended, claim 1 and 14 recites to a pluripotent NK cell stably transformed with an expression construct, wherein said synthetic notch receptor protein construct is further linked to a secondary effector molecule construct which encodes for a UAS, signal peptide and effector molecule.
The asserted art, either alone or in combination, do not teach the creation of such a pluripotent NK cell. There has been no suggestion or teaching in the cited art of an NK cell transformed with the expression construct as encompassed by the claimed invention. The expression of a secondary effector component in addition to, and not linked to the reporter molecule has not been taught or suggested in the asserted art.
In Response: The updated 103 rejection above addresses the amendment which incorporated limitations previously addressed in the OA mailed 11/25/2025. Specifically, the combined teachings of Morsut, Lupo, Roybal, and Chang teach PSC derived NK cells genetically engineered to express signal peptide- CD155/TIGIT binding protein- Notch core domain (which contains 3 LNR elements and an ADAM/TACE element)- GAL4-VP64- effector protein as detailed above.
Allowable Subject Matter
Claim 3 is allowed.
Claim 6 is objected to as being dependent upon a rejected base claim but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening claims.
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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/AMBER K FAUST/Examiner, Art Unit 1643
/GARY B NICKOL/Primary Examiner, Art Unit 1643
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