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 .
DETAILED ACTION
This action is in response to papers filed on September 10, 2024. Claims 1-2, and 8-25 are currently pending. Currently, claims 1, 11, 17, 18, 22, 23 are independent claims.
Therefore, claims 1, 2 and 8-25 are currently under examination as to which the following grounds of rejection are applicable.
Priority
The instant applications claims priority of International Application PCT/CN2023/070817 filed January 6 2023, which claims priority to Chinese Application PCTCN2022080149 filed March 10, 2022.
The Examiner notes that on the Bibliographic Data Sheet (BIB), that the Foreign Priority is claimed under Application PCTCN2022080149, but is pertaining to Chinese Application CN2022080149.
Therefore, the earliest effective filing date for the instant application is March 10, 2022.
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.
Claims 10-16, 18-25 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 claim 10, it is indefinite in its recitation of the phrase “wherein the CDTs are present in relative amounts”. The term “relative amounts ” in claim 10 is a relative term which renders the claim indefinite. The term “relative” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention.
Additionally claim 10 is indefinite in its recitation of “to put into cell culture media for pluripotent cells to induce totipotency”. Claim 10 depends on claim 1 which recites “to induce an untreated cell into a totipotent potential stem (TPS) cell”. Thus, it is unclear whether there is stage of differentiation between untreated cells and TPS cells. As such the metes and bounds of the claim are indefinite.
Regarding claim 11, it is indefinite in its recitation of the term period of time effective to induce pluripotency. The term “effective ” in claim 10 is a relative term which renders the claim indefinite. The term “effective” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention.
Regarding claims 19-21, it is indefinite in its recitation of the term “the cell” as it lacks proper antecedent basis. Claim 18 recites an isolated ciTPSC.
Claims 12-16, 18, and 22-25 are rendered indefinite as they depend on claims 18 and 11.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
Claims 17-22 are rejected under 35 U.S.C. 101 because the claimed invention is directed to a product of nature without significantly more. The claims recite the nature-based product drawn to an isolated chemically induced totipotent potential stem cell (ciTPSC) with totipotency. MPEP 2106 sets forth the multistep process for determining subject matter eligibility.
In accordance with the MPEP § 2016, claimed found to recite a statutory subject matter (e.g., compositions of matter) (Step 1: YES) are the analyzed to determine if the claims recite any steps that equate to an abstract idea, law of nature, or natural phenomenon. Claims 17-22 are drawn to an isolated population of ciTPSCs that occur in nature (2-cell embryos) within mammalian early embryonic development. Specifically, the claims recite naturally occurring cell populations isolated from mouse embryos, as discussed in Baker et al. (Published 2018. Capturing Totipotent Stem Cells Cell Stem Cell, 22, 25-34) and Yang et al. (Published March 3 2022. Cited in IDS filed: 12/12/2024, Cite No 42. Cell Press, Volume 29, Issue 3, 3 March 2022, Pages 400-418.e13). As such, the claims and art merely recite the isolation and maintenance of such naturally occurring cell populations outside of the body, and the process of culturing does not impart any properties to the product of ciTPSC by the claimed steps. The claimed isolated ciTPSCs have the same structure as the naturally occurring 2 cell mouse embryos. They have merely being isolated and placed into culture.
MPEP 2106.04(c) states that if a claim includes a nature-based product that does not exhibit markedly different characteristics from its naturally occurring counterpart in its natural state, then the claim recites a “product of nature” exception, and requires further analysis in Step 2A Prong Two to determine whether the claim as a whole integrates the exception into practical application (Step 2A, Prong One: YES)
The judicial exception is not integrated into a practical application because they do not recite any elements in addition to the recited judicial exception. MPEP 2106.04(d), subsection III states that a judicial exception alone is not eligible subject matter; therefore, if there are no additional claim elements besides the judicial, or if the additional claim elements merely recite another judicial exception, that is insufficient to integrate the judicial exception into a practical application. As such, claims 17-22 are directed to a natural product with no additional elements to demonstrate the claims as a whole integrate the exception into practical application. (Step 2A, Prong 2: NO).
The claims do not include additional elements that are sufficient to amount to significantly more than the judicial exception. MPEP 2106.05, subsection I states that additional elements in the claims must be evaluated to determine whether they amount to an inventive concept, which requires considering them both individually and in combination to ensure that they amount to significantly more than the judicial exception itself. Any additional steps, such as isolating the cell populations from embryo, culturing such donor cells with a chemically induced culture medium, are no more than routine in the art and do not transform the nature of the claim to be something distinct from their naturally occurring counterpart. Hence, because claims 17-22 do not contain additional elements to demonstrate the claims as a whole integrate the exception into a practical application, the claim simultaneously does not recite significantly more than the exception itself and there is no meaningful limitation in the claim that transforms the exception into a patent- eligible application. Therefore, claims 17-22 are directed to a judicial exception without significantly more and does not have an eligible subject matter under 35 U.S.C. § 101 (Step 2B: NO).
Note that claims 23-25 are not included in this rejection because the cell culture medium of claim 23 maintains the isolated ciTPSC in a totipotent cells for various passages.
Claim Rejections - 35 USC § 112: Written Description
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-2, and 8-25 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
In making a determination of whether the application complies with the written description requirement of 35 U.S.C. 112, first paragraph, it is necessary to understand what Applicant has possession of and what Applicant is claiming. Claim 1 recites “a cell culture comprising chemical derivers of totipotency: an HDAC inhibitor, a DOT1L inhibitor and a RARy agonist” driving untreated cells to totipotency in cells, which encompasses a genus of inhibitors, e.g, an HDAC inhibitor and a DotlL inhibitor and a genus of agonists, e.g., an RARy agonist. There is not structure / function correlation for the claimed combination of CDTs and their ability to induce an untreated cell into a totipotent potential stem (TPS) cell in an effective amount.. Therefore, the claims are drawn to a genus of combination of an HDAC inhibitor, a DOT1L inhibitor and a RARy agonist for which there is inadequate written description. Note that a GSK inhibitor is recited as an optional CDT.
The written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice (see MPEP 2163(II)(3)(a)(i)(A), reduction to drawings MPEP 2163(II)(3)(a)(i)(B), or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus MPEP 2163(II)(3)(a)(i)(C).
The Specification teaches that the combination of R one species for the claimed combination that fall within the genus claimed (para 0068, “Scale bar, 200 μm. V, VPA. E, EPZ004777. CD, CD1530. CTR, N2B27 medium supplemented with CHIR 99021”; para 0155, “ Mouse middle 2-cell embryos were collected and treated using the 4S chemical cocktail (VPA 500 μM, EPZ004777 1 μM, CD1530 0.5 μM, CHIR 99021 3 μM), which was added into the KSOM medium.”). Moreover, the Specification only discloses examples of deriving totipotent stem cells (TPS) from 2-cell embryos (para 0126) and mouse embryonic stem cells (para 0127-129), and TPS cell selected based pm their expression of ZSCAN4 (para 0152) or MERVL (para 0179, “ To identify small molecules that can induce totipotent features in mouse EPS cells, we used cell lines carrying a MERVL-TdTomato or a Zscan4-Emerald GFP reporter.”)
In the instant case, claim 1 requires the combination of the claimed genus of CDTs has an activity of “deriving cell totipotency in vitro of isolated cells”. However, the specification is silent about what structure of each of EPZ004777, VPA, CD1530 as disclosed in the examples discussed in paragraphs 0126-0129 is sufficient for the claimed activity when in effective amounts other than the cocktail of EPZ004777, VPA, CD1530 to induce 2-cell embryos into TPS.
Hu et al. (Published: 2020. US Patent Publication: US 20200277567 A1) teaches chemically induced lineage reprogramming of cells using synthase kinase inhibitors, TGFβ receptor inhibitors, cyclic AMP agonists or histone acetylators (Abstract; Claim 1, “A kit or cell culture media composition for inducing lineage reprogramming in eukaryotic cells, the composition comprising chemical inducers of lineage reprogramming (CiLRs) from each of the following groups (1) glycogen synthase kinase (GSK) inhibitors, (2) TGFβ receptor inhibitors, (3) cyclic AMP (cAMP) agonists, (4) histone acetylators, (5) DOT1L methyltransferase inhibitors, (6) Retinoic acid receptor (RAR) agonist, and (7) inhibitors of histone demethylation in amounts effective to induce reprograming a cell of a first type/lineage to a cell of a second type/lineage.”). Moreover, Hu teaches that regulation of JAK-STAT signaling pathway helps reprogram dedifferentiated cells towards a mesenchymal lineage, one of those regulators being HDAC (para 0013; claim 2, “…a combination thereof; a gene or protein target associated with the inhibition of HDAC”) (para 0005, “An object of this application is to provide a method of inducing dedifferentiation of somatic cells with small molecules to prepare rejuvenated mesenchymal stem cells and uses thereof, where the method involves the regulation of JAK-STAT (Janus kinase-signal transducer and activator of transcription) signaling pathway tp regulate cell differentiation, dedifferentiation, transdifferentiation, rejuvenation, aging and apoptosis, reversing the aging process and prolonging the lifespan”).
Therefore, it is unclear if certain small chemical drivers of totipotency (CDTs) such as HDAC inhibitor promote all cell types towards a totipotent state, and that HDAC inhibitor, RARy agonist and DOT1L inhibitor alone can cause all cell types to reprogram into a totipotent state, especially as some CDTs are found to promote mesenchymal characteristics rather than totipotent. The art and Specification are silent on whether cell populations outside of 2-cell embryos, embryonic stem cells, etc. can derive totipotent stem cells with the claimed CDTs with reasonable expectations of success.
The specification fails to provide a representative number of species within the recited genus. Accordingly, in the absence of sufficient recitation of distinguishing identifying characteristics of the genus as a whole, or representative number of species within the genus, the specification does not provide adequate written description of the claimed genus of combined CDTs.
Claim Rejections - 35 USC § 112: Scope of Enablement
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1,2, 8-25 rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph,
Claims 1-2, and 8-25 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for
a cell culture media comprising the combination of EPZ004777, VPA, CD1530, and induction of 2-cell embryos to totipotent potential stem (TPS) cells,
does not reasonably provide enablement for cell culture media comprising any combination of a genus of HDAC inhibitors, a genus of DOT1L inhibitors, and a genus of RARγ agonists and any type of untreated cell. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims.
The criteria for enablement set out In re Wands, MPEP 2162.01(a), considers the following factors:
Breadth of the Claims
The claims are broadly directed to a cell culture composition comprising cocktails of any HDAC inhibitor, any DOT1L inhibitor and any RARy agonist. The claims also are broadly but reasonably interpreted as any untreated cell that can be induced to be a TPS cell when culture with said cocktail. As such, the breadth of the claims is great.
State of Prior Art
Li et al. (Published: 2014. Cell stem cell, 13(3), 270–283.) teaches various chemicals that influence stem cell reprogramming, of those chemicals, them including HDAC inhibitor, EPZ004777 inhibitor and RARy agonist (FIG 2; pp. 274, col 2, “Acetylated histones are associated with active gene expression. Similarly, histone deacetylase inhibitors (including valproic acid [VPA] and sodium butyrate)”; pp. 275 col 2, “Notably, it was recently found that ectopic expression of retinoic acid receptor (RAR) a/g and Nr5a2 greatly enhanced reprogramming efficiency and kinetics”)
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Therefore, Li is silent on whether the specific combination of VPA, EPZ004777, and CD1530 would reasonably result in cells being totipotent.
Hu et al. (Published: 2020. US Patent Publication: US 20200277567 A1) teaches chemically induced lineage reprogramming of cells using synthase kinase inhibitors, TGFβ receptor inhibitors, cyclic AMP agonists or histone acetylators (Abstract; Claim 1, “A kit or cell culture media composition for inducing lineage reprogramming in eukaryotic cells, the composition comprising chemical inducers of lineage reprogramming (CiLRs) from each of the following groups (1) glycogen synthase kinase (GSK) inhibitors, (2) TGFβ receptor inhibitors, (3) cyclic AMP (cAMP) agonists, (4) histone acetylators, (5) DOT1L methyltransferase inhibitors, (6) Retinoic acid receptor (RAR) agonist, and (7) inhibitors of histone demethylation in amounts effective to induce reprograming a cell of a first type/lineage to a cell of a second type/lineage.”). Moreover, Hu teaches that regulation of JAK-STAT signaling pathway helps reprogram dedifferentiated cells towards a mesenchymal lineage, one of those regulators being HDAC (para 0013; claim 2, “…a combination thereof; a gene or protein target associated with the inhibition of HDAC”) (para 0005, “An object of this application is to provide a method of inducing dedifferentiation of somatic cells with small molecules to prepare rejuvenated mesenchymal stem cells and uses thereof, where the method involves the regulation of JAK-STAT (Janus kinase-signal transducer and activator of transcription) signaling pathway tp regulate cell differentiation, dedifferentiation, transdifferentiation, rejuvenation, aging and apoptosis, reversing the aging process and prolonging the lifespan”).
Therefore, it is unclear if certain small compounds such as HDAC inhibitor promote all cell types towards a totipotent state, and that HDAC inhibitor, RARy agonist and DOT1L inhibitor alone can cause all cell types to reprogram into a totipotent state. The art and Specification is silent on whether cell populations outside of 2-cell embryos, embryonic stem cells, etc. can derive totipotent stem cells with the claimed CDTs with reasonable expectations of success.
Amount of Direction Provided by the Inventor
The Specification provides insufficient direction and guidance to enable a person of ordinary skill in the art to practice the full scope of the claimed invention. Although the claims broadly encompass a cell culture composition comprising combinations of any HDAC inhibitor, any DOT1L inhibitor and any RARy agonist, the Specification only provides a single working example for VPA, EPZ004777 and CD1530 (para 0068, “Scale bar, 200 μm. V, VPA. E, EPZ004777. CD, CD1530. CTR, N2B27 medium supplemented with CHIR 99021”; para 0155, “ Mouse middle 2-cell embryos were collected and treated using the 4S chemical cocktail (VPA 500 μM, EPZ004777 1 μM, CD1530 0.5 μM, CHIR 99021 3 μM), which was added into the KSOM medium.”). The Specification only provides one example of untreated cells that are induced into stem cell reprogramming.
The Specification is silent on whether other cocktails of inhibitors of HDAC and a DotlL inhibitor and a agonists of an RARy used within the culture medium will successfully result in deriving 2-cell embryos to totipotent potential stem (TPS) cells totipotent stem cells, let alone any untreated cell into a TPS cell .
Presence or Absence of Working Examples
The specification only provides working examples for VPA, EPZ004777 and CD1530 CD1530 (para 0068, “Scale bar, 200 μm. V, VPA. E, EPZ004777. CD, CD1530. CTR, N2B27 medium supplemented with CHIR 99021”; para 0155, “ Mouse middle 2-cell embryos were collected and treated using the 4S chemical cocktail (VPA 500 μM, EPZ004777 1 μM, CD1530 0.5 μM, CHIR 99021 3 μM), which was added into the KSOM medium.”). Moreover, the Specification only discloses examples of deriving totipotent stem cells (TPS) from 2-cell embryos (para 0126) and mouse embryonic stem cells (para 0127-129), and TPS cell selected based pm their expression of ZSCAN4 (para 0152) or MERVL (para 0179, “ To identify small molecules that can induce totipotent features in mouse EPS cells, we used cell lines carrying a MERVL-TdTomato or a Zscan4-Emerald GFP reporter.”)
The Specification is silent on whether other cocktails of inhibitors or agonists used within the culture medium will successfully result in deriving totipotent stem cells. The examples recited in the Specification are not representative of the full scope of the claims.
Quantity of Experimentation Necessary
A person with ordinary skill in the art would not reasonably expect that other combinations of HDAC inhibitor, RARy agonist and DOT1L inhibitor, would result in the derivation and isolation of chemically induced totipotent stem cells. A person with ordinary skill in the art would not reasonably expect that other untreated cells were induced into a totipotent potential stem (TPS) cells
A person with ordinary skill in the art would have to perform undue, substantial, experimentation to fully encompass the scope of the claims.
Conclusion
In light of the unpredictability surrounding the claimed subject matter in the art and lack of adequate guidance in the Specification, one wishing to practice the presently claimed invention would be unable to do so without engaging in undue experimentation. It is especially noted that applicants provide no data, examples, figures, etc. demonstrating that other cocktails of the claimed inhibitors and agonists would reasonably result in chemically induced totipotent stem cells from untreated cells, and if this can be applied to the full scope of the claims. In the absence of such information, a person of ordinary skill in the art would reasonably require an undue quantity of experimentation.
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.
Claim(s) 1, 2, 10-25 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yang et al. (Published March 3 2022. Cited in IDS filed: 12/12/2024, Cite No 42. Cell Press, Volume 29, Issue 3, 3 March 2022, Pages 400-418.e13) in view of Iturbide et al. (Published: 2021. Cited in IDS filed: 12/12/2024, Cite No 14. Nature structural & molecular biology, 28(6), 521–532).
Regarding claim 1 and 2 Yang teaches a media composition for deriving totipotency within isolated cells (pp. 402 col 2, “Collectively, these analyses suggest that 2 cell-like cell (2CLCs) are partially reprogrammed mimics of the totipotent state, and the presence of chromocenters is likely associated with the semi-totipotent state that is distinctive from in vivo totipotency”; pp. e6, “For TLSC derivation from late 2C, 4C, 8C embryos, morula or blastocysts, these embryos were dissociated into single blastomeres, and then plated onto feeder/gelatin-coated 48-well plates in the medium comprising KnockOut DMEM, 20% KnockOut serum replacement, 13 non-essential amino acids solution, 2 mM L-glutamine, 50 mg/ml BSA, 100 mg/ml L-ascorbic acid, 100 mM2-mercaptoethanol, 10ng/ml IL6, 10 ng/ml sIL-6R, 2 mM SGC0946,2mMAS8351(orplusA366[2mM]).”), wherein the composition comprises an HADC inhibitor, (pp. e6, para 1, "we observed that the inhibitors of HDAC (NaB) and SUMO2 (ML-792) promoted the transition of 2C::GFP- mESCs to 2C::GFP+ cells under treatment with SGC0946 (Figure 3B)"), and a DOT1L inhibitor (Abstract, "DOT1L inhibition collapses chromocenters to induce an alternative 2C-like state"; Figure 4A; pp. e6 para 3, “Note that other selective Dot1l inhibitors (e.g. EPZ004777) can be substituted for SGC0946 in the above medium.”).
However, Yang does not teach the composition further comprising an RARγ agonist.
Iturbide teaches a media composition comprising retinoic acid (RA), which is a known RAR agonist (pp. 523 col 2, “RA induces 2CLC reprogramming without inducing differentiation. 2CLCs arise preferentially from naive ES cells10. Because RA promotes ES cell differentiation”; pp. 518, col 1, “Finally, we investigated a potential role of RA signaling during the totipotency transition in embryos”; pp. 22, “This suggests that the RA pathway might be active in 2CLCs, and possibly also in totipotent cells in vivo”) .
It would have been obvious to modify the media composition of Yang, to further comprise the RARγ agonist, retinoic acid, as it as a role in programming embryos towards totipotency. There would have been reasonable expectations of success in combining these teachings as one of ordinary skill in the art would recognize to combine known elements in the art to give predictable results.
Regarding claim 10, a kit is a collection of components. The combined teachings of Yang and Iturbide teach a culture media containing an HDAC inhibitor, a DOT1L inhibitor, and an RARγ agonist, which are the only components recited in the kit. Therefore, the prior art fulfills the limitations of the kit comprising the composition, by teaching the components of the kit. See In re Venezia 189 USPQ 49 (CCPA 1976), where kits are drawn to the structural attributes of interrelated component parts and not to activities that may or may not occur.
Regarding claim 11, the combined teachings of Yang and Iturbide render obvious the claimed composition of claim 1. Moreover, Yang teaches culturing the mESCs until they were totipotent stem cells (pp. 408, “Of note, the TLSCs still retained self-renewal and high expression of totipotent genes for at least two passages, when IL6 was withdrawn from the medium (Figures S4D and S4E).”; Figure 4A and 4B, )
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Regarding claim 12 and 13, the combined teachings of Yang and Iturbide render obvious the claimed composition of claim 1, and the claimed method of claim 11. Moreover, Yang teaches that the donor cells are mouse embryonic stem cells (pp. e6, “Derivation of TLSCs (totipotent-like stem cells) from mouse ES cells”; pp. e6, “For TLSC derivation from late 2C, 4C, 8C embryos, morula or blastocysts, these embryos were dissociated into single blastomeres, and then plated onto feeder/gelatin-coated 48-well plates in the medium comprising KnockOut DMEM, 20% KnockOut serum replacement, 13 non-essential amino acids solution, 2 mM L-glutamine, 50 mg/ml BSA, 100 mg/ml L-ascorbic acid, 100 mM2-mercaptoethanol, 10ng/ml IL6, 10 ng/ml sIL-6R, 2 mM SGC0946,2mMAS8351(orplusA366[2mM])”).
Regarding claim 14, the combined teachings of Yang and Iturbide render obvious the claimed composition of claim 1, and the claimed method of claim 11. Moreover, Yang teaches donor cells isolated from 2-cell (2c) mouse embryos cells (pp. e6, “Derivation of TLSCs (totipotent-like stem cells) from mouse ES cells”; pp. e6, “For TLSC derivation from late 2C, 4C, 8C embryos, morula or blastocysts, these embryos were dissociated into single blastomeres, and then plated onto feeder/gelatin-coated 48-well plates in the medium comprising KnockOut DMEM, 20% KnockOut serum replacement, 13 non-essential amino acids solution, 2 mM L-glutamine, 50 mg/ml BSA, 100 mg/ml L-ascorbic acid, 100 mM2-mercaptoethanol, 10ng/ml IL6, 10 ng/ml sIL-6R, 2 mM SGC0946,2mMAS8351(orplusA366[2mM])”).
Regarding claim 15, the combined teachings of Yang and Inturbide render obvious the claimed method of claim 11 and the claimed composition of claim 1. Moreover, Yang teaches that mouse embryonic stem cells can be converted to TLSCs (pp. 406, col 1, “We also successfully converted mESCC57/SPR (derived from blastocysts of mixed background C57BL/6J × SPRET/EiJ) to TLSCs (hereinafter named TLSCC57/SPR), which expressed multiple totipotent genes at high levels comparable to 2C embryos (Figure S4F), suggesting that our condition supports the transition of mESCs of multiple genetic backgrounds into TLSCs”).
Regarding claim 16, the combined teachings of Yang and Iturbide render obvious the claimed method of claim 11 and the claimed product of claim 1. Moreover, Yang teaches that the donor cells are seeded single-celled (pp. e5, para 4, “When passaging, cells were dissociated by 0.05% Trypsin-EDTA into single cells and re-plated every three days at a ratio of 1:10- 1:12 (for mESC) or 1:4- 1:6 (for TLSCs). Mouse ESCs and TLSCs were maintained in feeder cells derived from mouse embry onic fibroblasts.”).
Regarding claim 17, the combined teachings of Yang and Iturbide render obvious the claimed methodology and product. Moreover, Yang teaches a chemically induced totipotent stem cell (pp. 408 col 1, “We also successfully converted mESCC57/SPR (derived from blastocysts of mixed background C57BL/6J × SPRET/EiJ) to TLSCs (hereinafter named TLSCC57/SPR), which expressed multiple totipotent genes at high levels comparable to 2C embryos (Figure S4F), suggesting that our condition supports the transition of mESCs of multiple genetic backgrounds into TLSCs”).
Regarding claim 18, the combined teachings of Yang and Iturbide render obvious the claimed methodology of claim 11 and the composition of claim 1. Moreover, Yang teaches the chemically induced totipotent stem cells (Figure 4A and 4B). The structure implied by the process steps should be considered when assessing the patentability of product-by-process claims over the prior art, especially where the product can only be defined by the process steps by which the product is made, or where the manufacturing process steps would be expected to impart distinctive structural characteristics to the final product. See, e.g., In re Garnero, 412 F.2d 276, 279, 162 USPQ 221, 223 (CCPA 1979).
Regarding claim 19, the combined teachings of Yang and Iturbide render obvious claims 1, 11, and 18. Moreover, Yang taught that the cells express ZSCAN4 and pluripotent genes were downregulated (pp. 408, col 1, “These cells stably expressed key totipotent markers (ZSCAN4 and MERVL-GAG) but lacked OCT4 protein (Figures 4A–4D)… high expression of multiple totipotent genes in the TLSCs comparable to the 2C embryo, whereas the transcription of pluripotent genes was reduced”).
Regarding claim 20, the combined teachings of Yang and Iturbide render obvious claims 1, 11 and 18. Moreover, Yang teaches that the cell can generate both embryonic and extraembryonic lineages (pp. 412, col 2, “We observed the incorporation of single-TLSC derived cells in both embryonic and extraembryonic tissues in E13.5 conceptuses with relatively high frequency (Figures 7C andS6D)”; pp. 414 col 2, “The TLSCs have the embryonic and extraembryonic developmental potency and unique molecular features of totipotent blastomeres, in line with the stringent criteria of totipotency (Posfai et al., 2021).”).
Regarding claim 21, the combined teachings of Yang and Iturbide render obvious claims 1, 11 and 18. Moreover, Yang teaches that the totipotent-like stem cells (TLSCs) can generate blastocyst like structures (pp. 412, col 2, “Blastocyst-like structures generated solely from TLSCs”).
Regarding independent claims 22 and 23 directed a therapeutic composition and a cell culture, respectively, the combined teachings of Yang and Iturbide render obvious claims 1, 11 and 18 as stated above to render obvious claims 1, 11 and 18 . In particular , Yang teaches chemically induced totipotent like stem cells (TLSCs) (pp. 402 col 2, “Collectively, these analyses suggest that 2CLCs are partially reprogrammed mimics of the totipotent state, and the presence of chromocenters is likely associated with the semi-totipotent state that is distinctive from in vivo totipotency”; pp. e6, “For TLSC derivation from late 2C, 4C, 8C embryos, morula or blastocysts, these embryos were dissociated into single blastomeres, and then plated onto feeder/gelatin-coated 48-well plates in the medium comprising KnockOut DMEM, 20% KnockOut serum replacement, 13 non-essential amino acids solution, 2 mM L-glutamine, 50 mg/ml BSA, 100 mg/ml L-ascorbic acid, 100 mM2-mercaptoethanol, 10ng/ml IL6, 10 ng/ml sIL-6R, 2 mM SGC0946,2mMAS8351(orplusA366[2mM]).”), wherein the composition comprises an HADC inhibitor, (pp. e6, para 1, "we observed that the inhibitors of HDAC (NaB) and SUMO2 (ML-792) promoted the transition of 2C::GFP- mESCs to 2C::GFP+ cells under treatment with SGC0946 (Figure 3B)"), and a DOT1L inhibitor (Abstract, "DOT1L inhibition collapses chromocenters to induce an alternative 2C-like state"; Figure 4A; pp. e6 para 3, “Note that other selective Dot1l inhibitors (e.g. EPZ004777) can be substituted for SGC0946 in the above medium.”) and Iturbide teaches a media composition comprising retinoic acid (RA), which is a known RAR agonist (pp. 523 col 2, “RA induces 2CLC reprogramming without inducing differentiation. 2CLCs arise preferentially from naive ES cells10. Because RA promotes ES cell differentiation”; pp. 518, col 1, “Finally, we investigated a potential role of RA signaling during the totipotency transition in embryos”; pp. 22, “This suggests that the RA pathway might be active in 2CLCs, and possibly also in totipotent cells in vivo”) .
Regarding claim 24, the combined teachings of Yang and Inturbide render obvious claims 1, 11, 18, and 23. Moreover, Yang teaches that the TLSCs still remained totipotent after being passaged twice within the medium without IL-6 (pp. 406, col 1, “Of note, the TLSCs still retained self-renewal and high expression of totipotent genes for at least two passages, when IL6 was withdrawn from the medium”).
Regarding claim 25, the combined teachings of Yang and Inturbide render obvious claims 1, 11, 18, 23 and 24. Moreover, Yang taught that the cells express ZSCAN4 and pluripotent genes were downregulated (pp. 408, col 1, “These cells stably expressed key totipotent markers (ZSCAN4 and MERVL-GAG) but lacked OCT4 protein (Figures 4A–4D)… high expression of multiple totipotent genes in the TLSCs comparable to the 2C embryo, whereas the transcription of pluripotent genes was reduced”).
***
Claim(s) 1, 8 and 9 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yang et al. (Published March 3 2022. Cited in IDS filed: 12/12/2024, Cite No 42. Cell Press, Volume 29, Issue 3, 3 March 2022, Pages 400-418.e13) in view of Iturbide et al. (Published: 2021. Cited in IDS filed: 12/12/2024, Cite No 14. Nature structural & molecular biology, 28(6), 521–532), and in further view of Briggs et al. (Published: 2011. US Patent Publication: US 20110318830 A1) and R&D Systems (Wayback Machine Date: 2019. R&D Systems, CD1530, Tocris Bioscience.)
With regard to claim 1, the combined teachings Yang and Iturbide render obvious the claimed composition, as iterated above in the 103 rejection the content of which is incorporated in claim 1. Moreover, Yang teaches that the DOT1L inhibitor can be EPZ004777 (pp. e6, para 3, "Note that other selective Dot1l inhibitors (e.g. EPZ004777) can be substituted for SGC0946 in the above medium").
However, Yang and Iturbide do not teach VPA or CD1530 as required by claim 8.
R&D Systems teaches that CD1530 is a selective RARγ receptor agonist (“CD 1530 is a potent and selective RARγ receptor agonist (Ki values are 150, 1500 and 2750 nM for RARγ, RARβ and RARα receptors respectively).”)
Further, Briggs teaches that valproic acid (VPA) improves reprogramming efficiency and is a histone deacetylase inhibitor (Claim 23, para 0229, “In one embodiment, valproic acid (VPA), a histone deacetylase inhibitor, is added to improve reprogramming efficiency; which can be by more than 100-fold without introduction of the oncogene c-Myc”)
It would have been obvious to modify the culture composition Yang to further comprise both CD1530 and valproic acid as they were known RARγ receptor agonist and HDAC inhibitor respectively. Moreover, a skilled artisan would have been motivated to use VPA as it has been shown to improve reprogramming efficiency. Further, a skilled artisan would have been motivated to modify the concentrations of EPZ004777, CD1530 and VPA to reflect the claimed concentrations of claim 8, as the present result effective variables that one would routinely optimize in order to achieve pluripotent stem cells. There would have been reasonable expectations of success in combining these teachings as one of ordinary skill in the art would recognize to combine known elements in the art to give predictable results.
Regarding claim 9, the combined teachings of Yang, Iturbide, R&D Systems, and Briggs render obvious the claimed composition of claim 1. Moreover, Iturbide teaches CHIR99021 at a concentration of 3µM (pp. 533, col 1 “Medium was supplemented with 2i (3 µM CHIR99021 and 1 µM PD0324901, Miltenyi Biotec) for maintenance and expansion. The 2i was removed 24 h before starting experiments”)
Conclusion
No Claims allowed.
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/KATRIEL BARCELLANO KASAYAN/Examiner, Art Unit 1634
/MARIA G LEAVITT/Supervisory Patent Examiner, Art Unit 1634
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