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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 11/05/2025 has been entered.
Claim Status
1. The amendment filed 11/05/2025 has been entered. Claims 1, 4, 8, 13, 14, 16, 19, 23, and 39 – 48 remain pending. Claim 22 has been canceled.
Election/Restrictions
2. Applicant's election with traverse of Group I (claims 1, 4, 8, 13 – 16, 19, and 22) in the reply filed on 01/24/2025 is acknowledged. The traversal is on the ground(s) that if search and examination of an entire application can be made without serious burden, the examiner must examine the entire application on the merits, even though the entire application includes claims to independent or distinct inventions and it is Applicant’s position that it would not be unduly burdensome to perform a search on all of the claims together in the present application. This is not found persuasive because Groups I – IV are drawn to distinct inventions that lack unity of invention resulting in a serious search burden.
The requirement is still deemed proper and is therefore made FINAL.
3. Claims 23, 24, and 39 – 48 are withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected invention, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 01/24/2025.
Priority
4. This application claims priority to U.S. Provisional Application No. 62/898,430 filed 09/10/2019.
Withdrawn Drawings Objection and Claim Objection
5. The objection to the disclosure for description of color is withdrawn in view of Applicant’s amendment to the specification at para. 0015 to remove “color”.
6. The objection to claim 8 is withdrawn in view of Applicant’s amendment to the claim.
Withdrawn Claim Rejections
7. The rejection of claim 22 under 35 U.S.C. 101 is rendered moot by Applicant’s cancellation of the claim.
8. The rejection of claim 22 under 35 U.S.C. 103 is rendered moot by Applicant’s cancellation of the claim.
9. The rejection of claims 1, 4, 8, 13 – 14, and 16 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to claim 1.
10. The rejection of claim 19 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to claim 1.
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.
11. Claim(s) 1, 4, 8, 13, 14, and 16 are rejected under 35 U.S.C. 103 as being unpatentable over Lu (Lu, Jianfeng, et al. Nature biotechnology 34.1 (2016): 89-94; previously cited), hereinafter Lu which is cited on the IDS filed 08/16/2024 in view of as evidenced by Neely (Neely MD, et. al. ACS Chem Neurosci. 2012 Jun 20;3(6):482-91; previously cited), hereinafter Neely in view of Pasca (Paşca, Anca M., et al. Nature methods 12.7 (2015): 671-678; previously cited), hereinafter Pasca in view of Trumpp (US2020/0140812-A1; Filed 06/21/2018; Published 05/07/2020), hereinafter Trumpp.
Regarding claim 1(a), Lu teaches culturing human pluripotent stem cells in in SDC (TGF-β inhibitor SB431542, BMP inhibitor DMH1, and GSK3-β inhibitor CHIR99021) media for one week (“up to 7 days”) to form rostral hindbrain neural stem cells (NSCs) (“neural progenitor”) (page 2, paragraph 2 – 3; page 6, last paragraph; Figure 1a). Lu does not teach “in 3D suspension culture”.
Regarding claim 1(b), Lu teaches in Figure 1a differentiating rostral hindbrain NSCs to ventral rostral hindbrain NSCs by culturing in SDC media with SHH for one week followed by adding FGF4 to the SDC + SHH media and culturing for 1 week (Figure 1a; page 2, paragraph 2 – 3; page 6, last paragraph; page 3, para. 3). Lu does not teach “culturing the neural spheroid in suspension culture”.
Regarding claim 1(c), Lu teaches from the fourth week, serotonergic progenitors were cultured in a neuronal differentiation medium (NDM) and the timeline in Figure 1a (and shown below) shows this culturing was for at least one week such that serotonin neurons were produced (page 2, paragraph 2 – 3; page 6, last paragraph; Figure 1a).
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Lu does not teach “glutamatergic neurons” or “oligodendrocytes” of claim 1(c).
Regarding claim 4, Lu teaches culturing human pluripotent stem cells in medium comprising DMH1 (“inhibitor of BMP is dorsomorphin”), SB431542 (“inhibitor of TGFβ is SB-431542”) and CHIR99021 (“inhibitor of GSK-3 is CHIR99021) for one week (page 2, paragraph 2 – 3; page 6, last paragraph; Figure 1a). DMH1 is an analogue of dorsomorphin that is a highly selective small molecule BMP-inhibitor as evidenced by Neely (Abstract; page 484, left col. paragraph 1). Lu does not teach dorsomorphin.
Regarding claim 8, Lu teaches differentiating hPSCs to hindbrain neural stem cells by culturing in medium comprising TGFβ inhibitor SB431542 at 2 µm, BMP inhibitor DMH1 and GSK3-β inhibitor CHIR99021 at 1.4 µM (SDC medium) for one week (page 2, paragraph 2 – 3; page 3, paragraph 1; page 6, last paragraph). Lu does not teach SB431542 “between 5 µm to 20 µm”.
Regarding claim 13, Lu teaches applying FGF4 for one week at the start of the second week of differentiation (page 3, para. 3).
Regarding claim 14, Lu teaches serotonergic progenitors were cultured in neuronal differentiation medium comprising vitamin C (“AA”), DAPT, and BDNF (page 7, paragraph 1).
Regarding claim 16, Lu teaches applying FGF one week after cells were ventralized by SHH, cells enriched with TPH2 suggesting that FGF4 facilitates the serotonergic program of the ventral hindbrain progenitors and serotonergic progenitors were cultured in neuronal differentiation medium comprising vitamin C (“AA”), DAPT, and BDNF (page 3, paragraph 3; page 5, last paragraph; page 6, paragraph 1; page 7, paragraph 1).
Lu does not teach “in 3D suspension culture” of claim 1(a), “culturing the neural spheroid in suspension culture” of claim 1(b), “glutamatergic neurons, and oligodendrocytes” of claim 1(c), dorsomorphin of claim 4, SB431542 (TGFβ inhibitor) “between 5 µm to 20 µm” of claim 8, “the neural spheroid is cultured” of claim 13, “culturing the neural spheroid” of claim 15, or “nuclei spheroid” of claim 22. However, Lu teaches serotonin neurons located in the raphe nucleus of the hindbrain have crucial roles in regulating brain functions and have been implicated in various psychiatric disorders yet functional human serotonin neurons are not available for in vitro studies (Abstract). Lu teaches treating the cells obtained by the method with FDA-approved drugs and the cells release or uptake serotonin in a dose- and time-dependent manner, suggesting the utility of these cells for the evaluation of drug candidates (Abstract; page 4, last paragraph). Lu teaches serotonin neurons are localized to the raphe nucleus of the hindbrain (page 2, paragraph 1). Lu teaches the method facilitates in vitro study of human central serotonin neurons and the development of therapeutics that target the serotonin system (page 2, paragraph 2). Lu teaches the method produces serotonin neurons in large quantities and thus they can be used to validate drugs that affect serotonin release or uptake as an alternative to current assays (page 6, paragraph 2).
Regarding in 3D suspension culture” of claim 1(a), “culturing the neural spheroid in suspension culture” of claim 1(b), dorsomorphin of claim 4, the concentration of TGFβ inhibitor of claim 8, “the neural spheroid is cultured” of claim 13, “culturing the neural spheroid” of claim 16, Pasca teaches a method of 3D suspension culture of human induced pluripotent stem cells (hiPSCs) in exclusively nonadherent conditions and in the absence of extracellular scaffolding comprising transferring hiPSC colonies into low-attachment plates where floating hiPSC colonies folded into spherical structures (page 671, right col. paragraph 3; page 672, left col. last paragraph; Figure 1a and 1d; page 679, Online Methods, left col. paragraph 2). Pasca teaches neural induction of the hiPSC spheroids by addition of dorsomorphin to inhibit BMP and SB-431542 both at 10 µM to inhibit TGFβ (page 672, right col. paragraph 1; Figure 1a and 1d; page 679, Online Methods, left col. paragraph 2). Pasca teaches subsequent differentiation of the neural progenitors into neurons (page 672, right col. paragraph 2; Figure 1a; page 679, Online Methods, left col. paragraph 2). Pasca teaches the spheroids are electrophysiologically mature and display spontaneous activity (Abstract). Pasca teaches the method is simple and does not involve cell (re)-plating, embedding into extracellular matrices or culture in complex environments (page 677, left col. paragraph 3). Pasca teaches differentiation protocols of pluripotent stem cells in monolayers have led to the generation of a variety of neural cell types, but these two-dimensional (2D) methods and unlikely to recapitulate the cytoarchitecture of the developing three-dimensional (3D) nervous system or the complexity and functionality of in vivo neural circuits (page 671, right col. paragraph 2). Pasca teaches 3D approaches for generating organoid cultures display a level of self-organization beyond what is possible in 2D cultures, there remains a need for controlled specification of cell types (page 671, right col. paragraph 2). Pasca teaches the method is simple, scalable, and reproducible and the spheroids have the potential to reveal cellular phenotypes associated with neuropsychiatric disorders, identify biomarkers for early diagnosis and clinical stratification, and provide a platform for drug screening in vitro (page 672, left col. para. 1). Pasca does not teach “glutamatergic neurons and oligodendrocytes” of claim 1(c). One would have been motivated to combine the teachings of Lu and Pasca as both teach method to generate cells for in vitro development of therapeutics to treat neural disorders.
Regarding “glutamatergic neurons and oligodendrocytes” of claim 1(c), Trumpp teaches a method of generating glutamatergic neurons and oligodendrocytes from pluripotent stem cells by the method of Lu where serotonin neurons, glutamatergic neurons, and oligodendrocytes were present in cultures after 10 weeks of differentiation as shown in Figure 3 (page, 3, para. 0035; page 5 – 6, para. 0052; Figure 3; page 14, para. 0180; page 25, para. 0319 -0320; page 29, para. 0355 – 0356; page 30, para. 0364 – 0367; page 31, para. 0376; page 38, para. 0448). Trumpp teaches serotonergic differentiation was initiated by culturing in media containing CHIR99028, SB431542, purmorphamine for one week, followed by culture in CHIR 99028, SB431542, purmorphamine, and FGF4 for another week, followed by culturing in neuronal maturation medium containing dbcAM and BDNF for at least 5 more weeks where oligodendrocytes could be found starting after 5 weeks of differentiation (page 30, para. 0365). Trumpp teaches the method for producing the cells is for development of disease models for studying neurological disorders and diseases and for developing novel treatments based on such models and/or such cells differentiated in vitro (page 1, para. 0013).
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 Lu regarding a method of producing produces cells resembling central serotonin neurons, primarily those located in the rhombomeric segments 2 – 3 of the rostra raphe with the teachings of Pasca regarding a method of 3D suspension culturing of hiPSCs with the teachings of Trumpp regarding using the method of Lu to produce serotonergic neurons, glutamatergic neurons, and oligodendrocytes to arrive at the claimed method and claimed hRNS spheroid where human PSCs in 3D suspension culture are induced to a neural fate to generate a neural spheroid, differentiating the neural spheroid into a hRNS and maintaining the hRNS in neural medium such that the hRNS comprises serotonergic neurons, glutamatergic neurons, and oligodendrocytes. One would have been motivated to combine the teachings of Lu, Pasca, and Trumpp in a method of generating hRNS spheroid for in vitro testing of drugs to treat neural disorders as Lu teaches serotonin neurons located in the raphe nucleus of the hindbrain have crucial roles in regulating brain functions and have been implicated in various psychiatric disorders yet functional human serotonin neurons are not available for in vitro studies and Pasca teaches there remains a need for controlled specification of cell types in 3D approaches for generating organoid cultures and Pasca teaches the method is simple and does not involve cell (re)-plating, embedding into extracellular matrices or culture in complex environments and Trumpp teaches the method for producing the cells is for development of disease models for studying neurological disorders and diseases and for developing novel treatments based on such models and/or such cells differentiated in vitro. One would have a reasonable expectation of success in combining the teachings as Lu teaches the method produces cells that respond to FDA-approved drugs by releasing or uptake of serotonin in a dose- and time-dependent manner, Lu and Pasca teach the same media components for neural induction of hPSCs, Pasca teaches the spheroids are electrophysiologically mature and display spontaneous activity and Trumpp teaches the method of Lu to produce serotonergic neurons, glutamatergic neurons, and oligodendrocytes.
12. Claim(s) 19 is rejected under 35 U.S.C. 103 as being unpatentable over Lu (Lu, Jianfeng, et al. Nature biotechnology 34.1 (2016): 89-94; previously cited), hereinafter Lu which is cited on the IDS filed 08/16/2024 in view of as evidenced by Neely (Neely MD, et. al. ACS Chem Neurosci. 2012 Jun 20;3(6):482-91; previously cited), hereinafter Neely in view of Pasca (Paşca, Anca M., et al. Nature methods 12.7 (2015): 671-678; previously cited), hereinafter Pasca in view of Trumpp (US2020/0140812-A1; Filed 06/21/2018; Published 05/07/2020), hereinafter Trumpp as applied to claims 1, 4, 8, 13, 14, and 16 above, and further in view of Brennand (Brennand, Kristen J., et al. Nature 473.7346 (2011): 221-225; previously cited), hereinafter Brennand.
Lu in view of Pasca and Trump make obvious the limitations of claim 1 as set forth above. Lu and Trumpp do not teach claim 19. However, Lu teaches dysfunction of the serotonin system, especially misregulation of serotonin release or uptake, has been linked to schizophrenia, depression, bipolar disorder, anxiety, obsessive-compulsive disorder, chronic pain syndrome and eating disorders and thus the serotonin system is a major target for the treatment of multiple psychiatric disorders (page 1, paragraph 1). Lu teaches treating serotonergic neuron culture with Ultram to treat pain and Lexapro to treat major depressive disorder and generalized anxiety disorder where serotonin was released in a time-dependent manner (page 4, last paragraph).
Pasca does not teach claim 19. However, Pasca teaches the iPSCs used in the 3D suspension culturing method were derived from subjects (page 672, left col. paragraph 2; page 679, Online Methods, left col. paragraph 1).
Brennand teaches a method of deriving iPSCs from human fibroblasts from patients with schizophrenia (SCXD) and subsequently differentiating the hiPSCs to neural progenitor cells (NPCs) and neurons (page 221, left col. paragraph 1; Abstract; page 222, right col. paragraph 3 – 6; page 225, left col.; page 6, METHODS, left col. paragraph 1 – 2 and 4 – 8; Figure 1 and 4). Brennand teaches gene expression profiles of SCZD hiPSC neurons identified altered expression of components of the cyclic AMP and WNT signaling pathways (Abstract). Brennand teaches key cellular and molecular elements of the SCZD phenotype were ameliorated following treatment of SCZD hiPSC neurons with the antipsychotic loxapine (Abstract; page 222, left col. paragraph 2; Figure 2). Brennand teaches to date, hiPSC neuronal pathology has only been demonstrated in diseases characterized by both the loss of function of a single gene product and rapid disease progression in early childhood (Abstract). Brennand teaches hiPSC neuronal phenotypes and gene expression changes associated with SCZD (Abstract). Brennand teaches SCZD is a debilitating neurological disorder with a world-wide prevalence of 1% and there is a strong genetic component with an estimated heritability of 80 – 85% (Abstract).
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 Lu regarding a method of producing produces cells resembling central serotonin neurons , primarily those located in the rhombomeric segments 2 – 3 of the rostra raphe from hPSCs with the teachings of Pasca regarding a method of 3D suspension culturing of hiPSCs to produce neural spheroids where the hiPSCs were derived from subjects with the teachings of Trumpp regarding using the method of Lu to produce serotonergic neurons, glutamatergic neurons, and oligodendrocytes with the teachings of Brennand regarding a method of deriving hiPSCs from SCDZ patient fibroblasts and differentiation to NPCs and neurons to arrive at the claimed method where hRNS comprise at least one allele associated with schizophrenia. One would have been motivated to combine the teachings of Lu, Pasca, Trumpp, and Brennand in a method of generating hRNS spheroids from subjects with SCZD for in vitro testing of drugs to treat SCZD as Lu teaches serotonin neurons located in the raphe nucleus of the hindbrain have crucial roles in regulating brain functions and have been implicated in various psychiatric disorders yet functional human serotonin neurons are not available for in vitro studies and Brennand teaches SCZD is a debilitating neurological disorder with a world-wide prevalence of 1% and there is a strong genetic component with an estimated heritability of 80 – 85%. One would have a reasonable expectation of success in combining the teachings as Lu teaches the method produces cells that respond to FDA-approved drugs by releasing or uptake of serotonin in a dose- and time-dependent manner and Brennand teaches the hiPSCs neurons had features found in SCZD patients and the hiPSC neurons responded to a SCZD drug.
Applicant’s Arguments/ Response to Arguments
13. Applicant Argues: On page 7, last para. and page 8, para. 1, Applicant asserts that Lu does not disclose glutamatergic neurons or oligodendrocytes as recited in amended claim 1.
Response to Argument: The previous rejections of the claims have been withdrawn in view of Applicant’s amendment to claim 1. New rejections are set forth above in which Trumpp teaches the method of Lu produces serotonergic neurons, glutamatergic neurons, and oligodendrocytes after 5 weeks or 10 weeks of culturing in medium containing dbcAMP and BDNF (page 30, para. 0364; page 25, para. 0319 – 0320). Therefore, upon further consideration and search, Lu in view of Pasca and Trump make obvious the limitations of the limitations of amended claim 1.
Conclusion
No claims allowed.
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/Z.M.B./Examiner, Art Unit 1632
/MARCIA S NOBLE/Primary Examiner, Art Unit 1632