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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 01/26/2026 has been entered.
Claim Status
1. The amendment filed 01/26/2026 has been entered. Claims 1, 2, 4, 5, 7 – 10, 13 – 15, 17, 18, 25, 27, 28, 30, 33 – 37, 40, 41, 43, 44 remain pending.
Election/Restrictions
2. Applicant’s election without traverse of Group I (claims 1, 2, 4, 5, 7 – 10, 13 – 15, and 17 – 18) in the reply filed on 02/19/2025 is acknowledged.
3. Claims 25, 27, 28, 30, 33 – 37, 40, 41, and 43 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 02/19/2025.
4. Claims 1, 2, 4, 5, 7 – 10, 13 – 15, 17 – 18, and 44 are under consideration.
Priority
5. This application is a 371 of PCT/US2020/062180, which claims priority to U.S. provisional application No. 62/939,918, filed in the U.S. on 25 November 2019.
Information Disclosure Statement
6. The information disclosure statement (IDS) submitted on 01/26/2026 is acknowledged. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Withdrawn Claim Objections
7. The objection to claim 5 is withdrawn in view of Applicant’s amendment to the claim to require hiPSCs and not hESCs.
Withdrawn Claim Rejections
8. The rejection of claims 1, 2, and 4 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims to require the hTEPCs or hTECs are derived from hiPSCs.
9. The rejection of claim 5 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims to require the hTEPCs or hTECs are derived from hiPSCs.
10. The rejection of claims 7 – 10 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims to require the hTEPCs or hTECs are derived from hiPSCs.
11. The rejection of claims 13 – 15 and 17 – 18 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims to require the hTEPCs or hTECs are derived from hiPSCs.
12. The rejection of claim 44 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims to require the hTEPCs or hTECs are derived from hiPSCs.
New Rejections
Claim Rejections - 35 USC § 112
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.
13. Claims 7 – 10, 13 – 15, 17, and 18 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.
14. Regarding claim 7, the scope of the claim is not apparent because claim 1 recites “a method of making a bioengineered thymus organoid” but the end-product of the result of claim 7 is “to produce human immune cells”. Further, the claim only requires “the thymus construct” of claim 1 and not the “bioengineered thymus organoid”. Claims 8 – 10 are also rejected as they depend from claim 7 and do not clarify the grounds of the rejection.
15. Regarding claim 13, the scope of the claim is not apparent because claim 1 recites “a method of making a bioengineered thymus organoid” but the end-product of the result of claim 13 is a host animal with the transplanted thymus construct. Further, the claim only requires “the thymus construct” of claim 1 and not the “bioengineered thymus organoid” of claim 1. Claims 14, 15, 17, and 18 are also rejected as they depend from claim 13 and do not clarify the grounds of rejection.
Claim Interpretation
16. Claim 2 is interpreted as descriptive of the hiPSCs or the hHSCs that are derived from one or more donor individuals and not an active step of derivation of the hiPSCs or hHSCs.
17. Claim 5 is interpreted as a product-by-process limitation of the hTEPCs or hTECs because claim 1 does not recite any active steps for deriving the hTEPCs or hTECs. The “wherein” clause of claim 5 is descriptive of the hiPSC and not active method steps.
18. Claims 7 – 10 are interpreted as positively recited intended use/result because the “wherein” clauses are descriptive and not active method steps and because claim 1 does not recite a method of producing human immune cells.
19. Claims 13 – 15, 17, and 18 are interpreted as positively recited intended use/result because the “wherein” clauses are descriptive and not active method steps and because claim 1 does not recite method steps of transplanting the thymus construct.
New Rejections Necessitated By Amendment
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
20. Claim(s) 1, 2, 4, 5, 13 – 15, 17, 18, and 44 is/are rejected under 35 U.S.C. 103 as being unpatentable over Fan (Fan Y, et. al. Mol Ther. 2015 Jul;23(7):1262-1277; previously cited), hereinafter Fan which is cited on the IDS filed 05/24/2022 in view of Chung (Chung B, et. al. Stem Cells. 2014 Sep;32(9):2386-96; previously cited), hereinafter Chung which is cited on the IDS filed 10/04/2024 in view of Vizcardo (US-11898166-B2; Filed 09/19/2018; Published 02/13/2024), hereinafter Vizcardo.
Regarding claim 1, Fan teaches a method of obtaining mouse TECs (“thymic epithelial cells”) and combining the mouse TECs with mouse Lin- progenitors (“hematopoietic stem cells”) at a 1:1 ratio (“in a defined ratio to form a combination”) and injecting the combination into decellularized thymus scaffolds (“seeding the combination into an extracellular matrix of a de-cellularized thymus scaffold”) to form reconstructed thymus organoids that were cultured (“culturing the thymus construct”) (page 1263, left col. last para. and right col. para. 1; page 1274, right col. last para.; page 1275, left col. para. 1 – 3; page 1265). Fan teaches 7 days after thymus reconstruction, some of the injected cells began to assume a fibroblast-like morphology, suggesting that these cells successfully colonized the 3D ECM (page 1262, right col. para. 1). Fan teaches the TECs remained viable for > 3 weeks in the thymic scaffolds (page 1262, right col. para. 1). Fan does not teach “hTECs” or “wherein the hTECs are derived from hiPSCs”.
Regarding claim 4, Fan teaches the decellularized thymus scaffold is from a mouse (page 1274, right col. para. 4).
Regarding claim 13, Fan teaches the thymus organoid was transplanted to a mouse (page 1275, left col. para. 2).
Regarding claim 18, Fan teaches to examine whether T cells from the bioengineered thymus can provide helper function to humoral responses, the thymus recipients were immunized with OVA and high titers of anti-OVA IgG Isotypes IgG2b and IgG3 were found in serum samples harvested from the nude mice (page 1266, right col. last para.; Figure 4f). Fan does not teach the IgG is fully human. However, Fan teaches one of the essential roles of T cells in adaptive immunity is their helper function for humoral immunity, namely to mediate Ig class switch in antibody-producing B cells (page 1266, right col. last para.).
Regarding claim 44, Fan teaches the TECs and HSCs were both harvested from B6.CD45.1 mice to make the thymus organoids (page 1265, right col. last para.; page 1266, left col. para. 1).
Fan does not teach “hTECs” or “wherein the hTECs are derived from hiPSCs” of claim 1 and 5 or “the hiPSCs or the hHSCs are derived from one or more donor individuals” of claim 2 or “the host animal is a preconditioned humanized immune-deficient animal” of claim 14 or “mouse” of claim 15 or “provided hHSCs and produces human immune cells” of claim 17 or “fully human Immunoglobulin G” of claim 18. However, Fan teaches a study by Chung in which human TECs and thymic mesenchymal cells were co-cultured and the resulting engineered human thymic aggregates can support thymopoiesis both in vitro and in vivo (page 1274, left col. para. 1). Fan teaches the mouse thymus scaffolds can be stored at 4 °C for up to 1 month before use and it is likely that human thymus scaffolds can be preserved in a similar (page 1274, right col. para. 1). Fan teaches because there is no cellular component in the decellularized thymus scaffold, it is believed that allogeneic or even xenogeneic rejection will not be a concern (page 1274, right col. para. 1). Fan teaches one of the major obstacles in organ transplantation is to establish immune tolerance of allografts and the efficacies of immunosuppressive drugs are limited, transient, and associated with severe side effects (Abstract). Fan teaches induction of thymic central tolerance to allografts remains challenging largely because of the difficulty of maintaining donor TECs in vitro to allow successful bioengineering (Abstract; page 1262, right col. para. 3). Fan teaches the 3D scaffold environment of the decellularized thymus can support the long-term survival of TECs in vitro and enable them to retain the thymic specific patterns of molecule expression that is essential for T-cell development (page 1264, right col.). Fan teaches the thymus organoids when transplanted into athymic nude mice effectively promoted homing of lymphocyte progenitors and supported thymopoiesis and tolerance to skin allografts was achieved by transplanting thymus organoids constructed with either TECs co-expressing both syngeneic and allogenic major histocompatibility complexes, or mixtures of donor and recipient TECs (Abstract). Fan teaches the thymus organoids can support the development of T cells in vitro and can support lymphopoiesis in vivo (page 1265, right col.; page 1266, left col.). Fan teaches the thymus-reconstructed mice did not display any pathological sign of autoimmunity (page 1266, left col. para. 3).
Regarding “hTECs” of claim 1, Chung teaches hTECs were generated from post-natal thymic samples removed as waste tissue from patients during cardiothoracic surgery and cultured (page 2387, left col. last para. and right col. para. 1 – 2; page 2389, left col. para. 2 and right col.; Figure 2A; page 2390, left col.). Chung teaches generation of human thymic aggregates by mixing hTECs with human thymic mesenchyme (TM) (page 2387, right col. last para.; page 2390, right col.). Chung teaches implantation of the human thymic aggregates into NSG mice (page 2388, left col. para. 2; Figure 2A and C; page 2391, left col. para. 1).
Regarding claim 14 and 15, Chung teaches sublethally irradiating (“preconditioned” of claims 14 and 15) NSG mice (“immune-deficient animal” of claim 14 and “mouse” of claim 15) and transplanted with CB CD34+ HSPC from a HLA-A2+ donor to establish stable marrow engraftment (“preconditioned humanized immune-deficient animal” of claim 14 and “mouse” of claim 15) (page 2393, right col. para. 1). Chung teaches at 4 weeks post-HSPC transplantation, hTECs and CB CD34+ cells from HLA-A2 negative donors were implanted into the inguinal region of the mice (page 2393, right col. para. 1; Figure 6A).
Regarding claim 17, Chung teaches human CB CD34+ HSPCs are transplanted into the NSG mice and the mice produced T cells (page 2393, left col. para. 1).
Regarding “fully human Immunoglobulin G” of claim 18, Chung teaches human HSPCs were detected in murine bone marrow and human T cells were detected in human thymic aggregates
(page 2393, right col. para. 2; Figure 6C – E). As Fan teaches the mouse T cells produced IgGs, the human T cells produced by the hTECs would produce human IgGs.
Chung does not teach “wherein the hTECs are derived from hiPSCs” of claim 1 and claim 5. However, Chung teaches a system that allows manipulation of the human thymic microenvironment is needed both toe elucidate the extrinsic mechanisms that control human thymopoiesis and to develop potential cell therapies for thymic insufficiency (Abstract). Chung teaches the T cell defects that result from thymic dysfunction render patients highly susceptible to infections, malignancies, and/or autoimmune disease (page 2386, left col.). Chung teaches in addition to thymic involution that occurs with aging, damage to TECs can be caused by many external insults including infection, radiation, immunosuppressant therapy, and graft versus host disease after HSCT (page 2386, left col.). Chung teaches an experimental model to study how human T cell development is regulated within the human thymic microenvironment is needed to advance our understanding of thymic insufficiency (page 2386, left col.). Chung teaches progress toward thymic transplantation as a therapeutic option will require methods by which the thymic tissue can be manipulated ex vivo to remove endogenous T cells and to maintain the stromal compartments required to control normal thymopoiesis (page 2386). Chung teaches the frequency of TM and TEC in the thymus is extremely low and fragments of the human fetal thymus likely consist of variable and sometimes inadequate components of TM and TEC (page 2395, left col.). Chung teaches the method of culturing hTECs and TM provides the benefit of depleting contaminating thymocytes (page 2395, left col. para. 1). Chung teaches thymic implants have been used clinically but the risk of graft versus host disease from allo-reactive fetal thymocytes and the logistical difficulties of obtaining fetal thymus for transplantation has led to the use of postnatal thymic tissue cultured as intact chunks to remove contaminating thymocytes and surgically implanted in the muscle sheath (page 2395, left col. para. 2). Chung teaches that patients have remained severely T lymphopenic and autoimmune disease has developed likely because of suboptimal survival and function of TECs during culture and because of difficulty establishing vascularization throughout the relatively thick thymic chunks (page 2395, left col. para. 2). Chung teaches TEC cultures were not passaged and were typically used after approximately 3 weeks because longer cultures resulted in a lower percentage of TECs (page 2387, right col. para. 2). One would have been motivated to combine the teachings of Fan and Chung and substitute the mouse TECs with human TECS because Fan teaches the 3D scaffold environment of the decellularized thymus can support the long-term survival of TECs in vitro where the TECs remained viable for > 3 weeks in the thymic scaffolds and enable them to retain the thymic specific patterns of molecule expression that is essential for T-cell development, whereas Chung teaches culturing hTECs beyond 3 weeks resulted in a lower percentage of hTECs and Fan teaching that because there is no cellular component in the decellularized thymus scaffold, it is believed that allogeneic or even xenogeneic rejection will not be a concern.
Regarding “wherein the hTECs are derived from hiPSCs” of claim 1 and claim 5, Vizcardo teaches thymus organoids comprising hTECs and hTEPCs derived from hiPSCs (col. 16, lines 24 – 67; col. 17, lines 1 – 15 and 52 – 67; col. 18 – 21; Figure 1; col. 1, lines 35 – 52). Vizcardo teaches a method of deriving hTECs and hTEPCs from hiPSCs in Figure 1, 4, 5A, 6, and 8 which meets the product-by-process limitation of claim 5.
Regarding claim 2, Vizcardo teaches the hiPSCs are derived from human cord blood cells or T cells (col. 16, lines 61 – 65; col. 17, lines 55 – 60; Figure 9).
Vizcardo teaches there exists a need for improved materials and methods useful for preparing adoptive cell therapy (col. 1, lines 29 – 31). Vizcardo teaches the hTECs can be administered to a mammal to treat a condition in the mammal (col. 1, lines 62 – 67; Figure 9).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Fan regarding a method for making a bioengineered thymus organoid comprising mouse TECs with the teachings of Chung regarding a method of making thymic aggregates with human TECs with the teachings of Vizcardo regarding deriving human TECs from hiPSCs to arrive at the claimed method for making a bioengineered thymus organoid, comprising: obtaining a cell population comprising human thymic epithelial progenitor cells (hTEPCs)
or human thymic epithelial cells (hTECs) or both, wherein the hTEPCs or hTECs are derived from
human induced pluripotent stem cells (hiPSCs); combining the cell population with human hematopoietic stem cells (hHSCs) in a defined ratio to form a combination; seeding the combination into an extracellular matrix of a de-cellularized thymus scaffold to generate a thymus construct; and
culturing the thymus construct under conditions permitting cellular attachment onto the
extracellular matrix thereby making the bioengineered thymus organoid. One would have been motivated to combine the teachings of Fan, Chung, and Vizcardo in a method of making a thymus construct containing human TECs derived from hiPSCs to study and treat disorders related to thymic dysfunction and graft rejection as Fan teaches one of the major obstacles in organ transplantation is to establish immune tolerance of allografts and the efficacies of immunosuppressive drugs are limited, transient, and associated with severe side effects and Fan teaches induction of thymic central tolerance to allografts remains challenging largely because of the difficulty of maintaining donor TECs in vitro to allow successful bioengineering and Chung teaches the T cell defects that result from thymic dysfunction render patients highly susceptible to infections, malignancies, and/or autoimmune disease and Chung teaches an experimental model to study how human T cell development is regulated within the human thymic microenvironment is needed to advance our understanding of thymic insufficiency and Chung teaches that patients have remained severely T lymphopenic and autoimmune disease has developed with transplantation of postnatal thymic tissue likely because of suboptimal survival and function of TECs during culture and because of difficulty establishing vascularization throughout the relatively thick thymic chunks and Vizcardo teaches there exists a need for improved materials and methods useful for preparing adoptive cell therapy. One would have a reasonable expectation of success in combining the teachings as Fan teaches the thymus organoids can support the development of T cells in vitro and can support lymphopoiesis in vivo and Fan teaches the thymus-reconstructed mice did not display any pathological sign of autoimmunity and Chung teaches mice transplanted with the hTEC aggregates produced T cells and Vizcardo teaches human thymus organoids can be generated from hiPSCs.
21. Claim(s) 7 – 10 is/are rejected under 35 U.S.C. 103 as being unpatentable over Fan (Fan Y, et. al. Mol Ther. 2015 Jul;23(7):1262-1277; previously cited), hereinafter Fan which is cited on the IDS filed 05/24/2022 in view of Chung (Chung B, et. al. Stem Cells. 2014 Sep;32(9):2386-96; previously cited), hereinafter Chung which is cited on the IDS filed 10/04/2024 in view of Vizcardo (US-11898166-B2; Filed 09/19/2018; Published 02/13/2024), hereinafter Vizcardo as applied to claims 1, 2, 4, 5, 13 – 15, 17, 18, and 44 above, and further in view of Hamilton (US11371000B2; Filed 03/30/2017; Published 06/28/2022; previously cited), hereinafter Hamilton in view of Beaudette (Beaudette-Zlatanova, Britte C., et al. Experimental hematology 39.5 (2011): 570-579; previously cited), hereinafter Beaudette in view of Mamonkin (Mamonkin, Maksim, et al. Blood, The Journal of the American Society of Hematology 126.8 (2015): 983-992), hereinafter Mamonkin.
Fan in view of Chung and Vizcardo make obvious the limitations of claim 1 as set forth above but do not teach “the thymus construct is placed into a flow cell with a continuous feed of nutrients and human cells to produce human immune cells” of claim 7. However, Fan teaches the thymus is a vulnerable organ where anticancer drug treatments can irreversibly compromise its function (page 1262, right col. para. 1).
Regarding claim 8 and 9, Fan teaches culturing the thymus organoid in vitro produces T cells (“immune cells” of claim 8 and “T-cells” of claim 9) (page 1265, left col. and right col. para. 1) but Fan does not teach “B-cells” of claim 9. Chung teaches the human thymic aggregates produced T cells (page 2391, right col, para. 1) but does not teach “B-cells” of claim 9. However, Chung teaches Notch signaling is critical for the earliest stages of T cell commitment from HSPC (page 2394, right col. para. 1). Chung teaches expression of notch ligands in TECs is downregulated during monolayer culture and therefore retroviral expression of DLL1 or DLL4 in murine stromal cells has been used as an in vitro method to induce T cell differentiation from HSPC (page 2394, right col. para. 1). Vizcardo teaches the hiPSC thymus organoid expresses notch ligand DLL4 (Figure 15B; col. 20, lines 1 – 13).
Regarding “flow cell with continuous feed of nutrients and human cells” of claim 7, Hamilton teaches Thymus-on-Chip that is a microfluidic platform that is in fluid communication with Cancer-on-Chips (col. 25, lines 35; col. 35, lines34 – 41; col. 38, lines 37 – 54; Figure 17). Hamilton teaches the microfluidic device can comprise an inlet channel connecting an inlet fluid port to a first chamber where the inlet channels and inlet ports can be used to introduce cells and culture media into the first chamber (col. 44, lines 32 – 37). Hamilton teaches the tumor cells in the microfluidic device are in close proximity with at least one type of immune cells including T cells (col. 2, lines 23 – 48). Hamilton teaches in Example 3 studying immune cell migration in relation to cancer cell/tumor growth (col. 52, lines 30 – 50). Hamilton teaches microfluidic platforms or chips for testing and understanding cancer where the interaction between cancer cells and immune cells can be tested by linking a cancer chip to another chip (col. 1, lines 59 – 66). Hamilton teaches the interaction with circulating immune cells recruited to the tumor site will be enabled to allow testing of immunomodulatory agents, confirm immune surveillance (or lack thereof), and provide a platform for testing of immunotherapeutics (col. 2, lines 1 – 6). Hamilton teaches microfluidic platforms can increase our understanding of tumor growth and all other aspects of cancer including the role of ECM on this process, resistance to immune surveillance, and development of metastatic disease (col. 2, lines 6 – 12). Hamilton teaches resident immune cells (B cells, T cells, dendritic cells, macrophages, and innate lymphoid cells) may be isolated from cancer patients and incorporated in the chip (col. 29, lines 45 – 63).
Hamilton does not teach “B cells” of claim 9 or CAR of claim 10. However, Hamilton teaches CAR T cell therapy has dramatically improved the outcomes of blood cancer patients with advanced forms of leukemia and lymphoma but the full potential of CARs for treating solid tumors has not been reached and many challenges remain (col. 42, lines 40 – 45). Hamilton teaches having more predictive, human relevant systems to study human tumor biology and the interactions of the human immune system with the tumor would advance our knowledge and help to provide the most robust and precise preclinical platforms for drug discovery and enable the advancement of immunotherapies (col. 42, lines 45 – 52). One would have been motivated to combine the teachings of Fan and Hamilton in a method of making a thymus organoid in a flow cell to study the interaction of immune cells produced by the organoid with cancer cells as Fan teaches anticancer drug treatments can irreversibly compromise thymus function and Hamilton teaches microfluidic platforms can increase our understanding of tumor growth and all other aspects of cancer including the role of ECM on this process, resistance to immune surveillance, and development of metastatic disease.
Regarding “B cells” of claim 9, Beaudette teaches co-culture of human Lin-CD34+HPCs with human TECs expressing Notch ligands resulted in formation of T-cells and B-lineage cells (page 571, right col. para. 2; page 573, left col. and right col. para. 1; page 575, right col. para. 2). Beaudette teaches the ability to form these cells from co-culture of TECs and HPCs represent a novel tool for studying human lymphopoiesis and have the potential to generate lymphocytes for in vivo therapy in patients with hematological malignancies and immunodeficiency disorders (page 578, right col. para. 3). Beaudette does not teach “CAR” of claim 10. One would have been motivated to combine the teachings of Fan, Hamilton, and Beaudette because Fan and Beaudette teach methods of generating immune cells for treating patients with immunodeficiency disorders and Hamilton teaches the use of immune cells including B cells and T cells for studying immunotherapeutics.
Regarding “a chimeric antigen receptor” of claim 10, Mamonkin teaches transduction of T cells with a CD5 CAR (page 983, right col. para. 3). Mamonkin teaches CD5 CAR T cells recognize and eliminate malignant T cells in vitro (page 984, right col. para. 3 – 5). Mamonkin teaches CD5 CAR T cells kill primary T-ALL cells from patients (page 985, right col. para. 4 – 5; page 986, left col. para. 1). Mamonkin teaches prognosis of patients with primary chemotherapy-refractory or relapsed lymphoid malignancies remains poor (page 983, left col. para. 1). Mamonkin teaches CAR T cell therapy that targeted a tumor antigen shared between normal and malignant T cells might lead to fratricide of the CAR T cells thus jeopardizing their therapeutic efficacy (page 983, left col. para. 1). Mamonkin teaches CD5 CAR can effectively redirect human T cells to target malignant CD5+ cells and this may provide novel treatment options for patients with refractory or relapsed CD5+ T-cell neoplasms (page 991, right col. 3).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Fan regarding a method for making a bioengineered thymus organoid that produces T cells when cultured in vitro with the teachings of Chung regarding a method of making thymic aggregates with human TECs that produces T cells when cultured in vitro with the teachings of Vizcardo regarding deriving human thymus organoids from hiPSCs that express notch ligands with the teachings of Hamilton regarding Thymus-on-Chip with the teachings of Beaudette regarding in vitro co-culture of TECs expressing notch ligands and HPCs produces T cells and B cells with the teachings of Mamonkin regarding transducing T cells with a CAR to treat cancer to arrive at the claimed method where the thymus construct is placed into a flow cell with a continuous feed of nutrients and human cells to produce human cells. One would have been motivated to combine the teachings of Fan, Chung, Vizcardo, Hamilton, Beaudette, and Mamonkin in a method for making a thymus organoid to test immunotherapeutics on immune cell function and thymus function as Fan teaches the thymus is a vulnerable organ where anticancer drug treatments can irreversibly compromise its function and Beaudette teaches the ability to form B and T cells from co-culture of TECs and HPCs represent a novel tool for studying human lymphopoiesis and has the potential to generate lymphocytes for in vivo therapy in patients with hematological malignancies and immunodeficiency disorders and Hamilton teaches CAR T cell therapy has dramatically improved the outcomes of blood cancer patients with advanced forms of leukemia and lymphoma but the full potential of CARs for treating solid tumors has not been reached and many challenges remain, and Hamilton teaches having more predictive, human relevant systems to study human tumor biology and the interactions of the human immune system with the tumor would advance our knowledge and help to provide the most robust and precise preclinical platforms for drug discovery and enable the advancement of immunotherapies. One would have a reasonable expectation of success in combining the teachings as Fan and Chung teach in vitro culture of TECs with HPCs produces immune cells and Hamilton teaches studying immune cell migration in relation to cancer cell/tumor growth with the chips and Mamonkin teaches the T cells express the CAR and can target and kill malignant cells.
Applicant’s Arguments/ Response to Arguments
22. The affidavit under 37 CFR 1.132 filed 01/26/2026 is insufficient to overcome the new rejection of claims 1 – 2, 4 – 5, 7 – 10, 13 – 15, and 17 – 18, which was necessitated by the amendment to claim 1 requiring the hTEPCs or hTECs be derived from hiPSCs, based upon the Parent reference applied under 35 U.S.C. 103 as set forth in the last Office action because: the previous rejection of the claims using the teachings of Parent have been withdrawn in view of the amendments to the claims and a new rejection of the claims is set forth above using the combined teachings of Fan, Chung, and Vizcardo and because Applicant’s specification teaches that iPS-derived TECs were prepared according to the adopted protocol of Parent at page 37 – 38, para. 00144. The evidence presented in the affidavit regarding the teachings of Fan and Beaudette is rebutted as follows because the new rejection set forth above cites the teachings of Fan and Beaudette:
Affidavit States: On page 4, paragraph 14, Applicant states that a POSITA would not have expected that iPSCs could be successfully used to produce mature TECs capable of supporting T cell lineage-specification from HSPCs in vitro without the need to artificially overexpress important regulators such as Foxn1 or Dl1. On page 6, last para., Applicant states that the teachings of Beaudette-Zlatanova do not provide any evidence or suggestion that hTEPCs and hTECs generated from iPSCs can mature enough to support T cell development in a bioengineered system either. On page 7, para. 24, Applicant states the data from Beaudette suggest that even with cells derived from mature TEPCs/TECs, it is not predictable whether the TEC lines will be able to support mature T cell development and a POSITA would infer that genetic modification of the TECs to overexpress Dl1 is required to steer HSCs towards T cell, opposed to B cell development.
Response: In the new rejection set forth above, Chung teaches human thymic aggregates prepared from human TECs, human TM, and CB-derived CD34+ cells mixed at a ratio of 2:1:4 could produce T cells in vitro (page 2387, left col. last para. and right col.; page 2390, right col.). Vizcardo teaches hTECs and human thymus organoids derived from hiPSCs that express Foxn1 and Dl4 (col. 18, lines 13 – 26; col. 21, lines 9 – 17 and lines 50 – 62). Therefore, a POSITA would have expected that iPSCs could be successfully used to produce mature TECs capable of supporting T cell-lineage specification from HSPCs in vitro because the hTECs of Vizcardo derived from iPSCs express Foxn1 and Dl4.
Affidavit States: On page 9, para. 31, Applicant states that TECs of the instant application expressed maturity markers like PAX9 and FOXN1 and also expressed cytokeratin genes. On page 11, para. 35, Applicant states that the mature TECs express AIRE.
Response: In response, Vizcardo teaches hTECs and human thymus organoids derived from hiPSCs that express Foxn1 and cytokeratins (col. 18, lines 13 – 26; col. 21; Figure 1, 5A, 6). Chung teaches the hTECs expressed DLL4 and AIRE (page 2392, right col. last para.; Figure 5). Therefore, the hTECs of Chung and Vizcardo show markers of mature TECs.
Affidavit States: On page 17, paragraph 44, Applicant states that due to the significant differences in cells used to generate the stromal compartment of a bioengineered thymus in Fan vs. the claimed invention, a POSITA would not have reasonably expected that hTEPCs or hTECs derived from hiPSCs could be successfully used to create a functional, human thymus organoid capable of supporting human T cell development.
Response: In response, the previous rejection has been withdrawn in view of the amendment to claim 1. Therefore, the arguments stated in the Affidavit regarding the teachings of Parent are moot. However, it is noted that Applicant’s specification teaches that iPS-derived TECs were prepared according to the adopted protocol of Parent at page 37 – 38, para. 00144.
In the new rejection necessitated by the amendment set forth above, the teachings of Fan are cited and therefore Applicant’s arguments addressing the Fan reference are addressed. Fan does not teach human TECs or human TEPCs and instead teaches mouse TECs. Chung teaches human thymic aggregates formed from human TECs. One would have been motivated to combine the teachings of Fan and Chung and substitute the mouse TECs with human TECs because Fan teaches the 3D scaffold environment of the decellularized thymus can support the long-term survival of TECs in vitro where the TECs remained viable for > 3 weeks in the thymic scaffolds and enable them to retain the thymic specific patterns of molecule expression that is essential for T-cell development, whereas Chung teaches culturing hTECs beyond 3 weeks resulted in a lower percentage of hTECs and Fan teaching that because there is no cellular component in the decellularized thymus scaffold, it is believed that allogeneic or even xenogeneic rejection will not be a concern. Chung teaches generation of human thymic aggregates by mixing hTECs with human thymic mesenchyme (TM) (page 2387, right col. last para.; page 2390, right col.). Chung teaches the thymic aggregates support the development of human T cells in vitro and in vivo (page 2388, left col. para. 2; Figure 2A and C; page 2391, left col. para. 1; page 2393, left col. para. 1 and right col. para. 1; Figure 6A). Vizcardo teaches thymus organoids comprising hTECs and hTEPCs derived from hiPSCs and as stated previously, these cells express markers of mature TECs (col. 16, lines 24 – 67; col. 17, lines 1 – 15 and 52 – 67; col. 18 – 21; Figure 1, 4, 5A, 6, and 8; col. 1, lines 35 – 52). One would have been motivated to substitute the hTECs of Chung with hTECs derived from hiPSCs of Vizcardo because Chung teaches the frequency of TM and TEC in the thymus is extremely low and fragments of the human fetal thymus likely consist of variable and sometimes inadequate components of TM and TEC (page 2395, left col.) and to eliminate the need for using human fetal thymus because Chung teaches thymic implants have been used clinically but there is a risk of graft versus host disease from allo-reactive fetal thymocytes and logistical difficulties of obtaining fetal thymus for transplantation (page 2395, left col. para. 2). Therefore, the limitations of claim 1 are obvious over the teachings of Fan, Chung, and Vizcardo and a POSITA would have expected that using the hTECs of Chung or the hTECs derived from hiPSCs of Vizcardo would successfully result in a thymus organoid capable of supporting human T cell development.
23. Applicant Asserts: On page 10 of Applicant’s Remarks, para. 1 and page 11, para. 2, Applicant asserts that a POSITA would have been required at the priority date of the instant application to modify the teaching of Parent to use hiPSCs instead of hESCs to arrive at the claimed invention. On page 12 – page 15, para. 1, Applicant asserts that Beaudette highlights the challenges and unpredictability of in vitro systems seeking to recapitulate thymus function to generate T cells from HSPCs.
Response to Argument: The previous rejections of the claims has been withdrawn and new rejections set forth in view of the amendment to the claims to require hiPSCs that do not cite the teachings of Parent. Therefore, the arguments addressing Parent are moot. In the new rejection, Chung teaches hTECs can generate T cells in vitro and Vizcardo teaches hTECs that are not genetically modified to express a notch ligand can be derived from hiPSCs and that these hTECs have markers of mature TECs as stated above. Therefore, a POSITA would have expected that hTECs derived from hiPSCs would result in the generation of mature TECs that would be sufficiently mature to promote T cell lineage specification in developing T cells in vitro.
Applicant Argues: On page 15 – 16, para. 1 – 3 and page 20, para. 2 of Applicant’s Remarks, Applicant asserts that the organoids and methods disclosed in the instant application exhibit unexpected and superior properties as compared to the art as they are significantly more mature than the TECs produced by Parent and Chhatta and the organoids allow for the generation of mature TECs from hiPSCs wherein the TECs are capable of supporting T cell lineage specification in vitro.
Response to Argument: This is not found persuasive because claim 1 requires broadly obtaining hTECs or hTEPCs and does not recite any special features of the hTECs or hTEPCs other than they are derived from hiPSCs. As Vizcardo teaches hTECs and hTEPCs can be derived from hiPSCs, the combined teachings of Fan, Chung, and Vizcardo make obvious the limitations of claim 1. The scope of claim 1 is a method for making a bioengineered thymus organoid and not a method of supporting T cell lineage specification in vitro. Nevertheless, Chung teaches that hTECs can support T cell generation in vitro as previously stated. Therefore, the intended use of hTECs derived from hiPSCs to support T cell generation in vitro would be expected.
Applicant Argues: On page 17, paragraph 1 – 2 of Applicant’s Remarks, Applicant disagrees with the obviousness rejection set forth in the previous Office action citing the teachings of Mantalaris and Randle.
Response to Argument: The previous rejection of the claims citing the teachings of Mantalaris and Randle have been withdrawn and therefore the arguments are moot.
Applicant Argues: On page 18, para. 1 – 2 of Applicant’s Remarks, Applicant asserts that the device of Hamilton is for testing and understanding cancer and neither Hamilton nor Mamonkin provide any teaching or motivation for a POSITA to arrive at amended claim 1.
Response to Argument: This is not found persuasive because the combined teachings of Fan, Chung, and Vizcardo make obvious the limitations of claim 1. Hamilton teaches in Example 3 studying immune cell migration in relation to cancer cell/tumor growth (col. 52, lines 30 – 50). Hamilton teaches microfluidic platforms or chips for testing and understanding cancer where the interaction between cancer cells and immune cells can be tested by linking a cancer chip to another chip (col. 1, lines 59 – 66). Hamilton teaches the interaction with circulating immune cells recruited to the tumor site will be enabled to allow testing of immunomodulatory agents, confirm immune surveillance (or lack thereof), and provide a platform for testing of immunotherapeutics (col. 2, lines 1 – 6). Hamilton teaches microfluidic platforms can increase our understanding of tumor growth and all other aspects of cancer including the role of ECM on this process, resistance to immune surveillance, and development of metastatic disease (col. 2, lines 6 – 12). Hamilton teaches resident immune cells (B cells, T cells, dendritic cells, macrophages, and innate lymphoid cells) may be isolated from cancer patients and incorporated in the chip (col. 29, lines 45 – 63). Hamilton teaches CAR T cell therapy has dramatically improved the outcomes of blood cancer patients with advanced forms of leukemia and lymphoma but the full potential of CARs for treating solid tumors has not been reached and many challenges remain (col. 42, lines 40 – 45). Hamilton teaches having more predictive, human relevant systems to study human tumor biology and the interactions of the human immune system with the tumor would advance our knowledge and help to provide the most robust and precise preclinical platforms for drug discovery and enable the advancement of immunotherapies (col. 42, lines 45 – 52). Therefore, one would have been motivated to combine the teachings of Fan, Chung, Vizcardo, and Hamilton in a method for making a thymus organoid in a flow cell to study the interaction of immune cells produced by the organoid with cancer cells as Fan teaches anticancer drug treatments can irreversibly compromise thymus function and Hamilton teaches microfluidic platforms can increase our understanding of tumor growth and all other aspects of cancer including the role of ECM on this process, resistance to immune surveillance, and development of metastatic disease.
Applicant Argues: On page 18, last para. of Applicant’s Remarks, Applicant asserts that a POSITA would have understood that the development of T cells requires a functional stromal compartment containing mature TECs and Chung provides no teaching or motivation for a POSITA to arrive at the amended claim 1.
Response to Argument: This is not found persuasive because one would have been motivated to combine the teachings of Fan and Chung and substitute the mouse TECs with human TECS because Fan teaches the 3D scaffold environment of the decellularized thymus can support the long-term survival of TECs in vitro where the TECs remained viable for > 3 weeks in the thymic scaffolds and enable them to retain the thymic specific patterns of molecule expression that is essential for T-cell development, whereas Chung teaches culturing hTECs beyond 3 weeks resulted in a lower percentage of hTECs and Fan teaching that because there is no cellular component in the decellularized thymus scaffold, it is believed that allogeneic or even xenogeneic rejection will not be a concern.
Applicant Argues: On page 19, paragraph 2 – 3 – page 20, para. 1 of Applicant’s Remarks, Applicant states that Danner, Gragert, Tan, and Inami provide no teaching or motivation for a POSITA to arrive at the amended claim 1.
Response to Argument: The previous rejection of the claims citing the teachings these references have been withdrawn and therefore the arguments are moot.
Conclusion
No claims allowed.
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/ZANNA MARIA BEHARRY/Examiner, Art Unit 1632