DETAILED ACTION
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 8/14/2025 has been entered.
Response to Amendments
Applicant's amendments filed 8/14/2025 to claims 1, 15, 17, 19 have been entered. Claims 5, 7-9, 11, 13, 16, 18, 20, 26, 27, and 30 are canceled. Claims 1-4, 6, 10, 12, 14, 15, 17, 19, 21-25, 28, 29, and 31-35 remain pending, of which claims 1-4, 6, 10, 12, 14, 15, 17, 19, 21, 22, and 32 are being considered on their merits. Claims 23-25, 28, 29, 31, and 33-35 remain withdrawn from consideration. References not included with this Office action can be found in a prior action.
The instant amendments to claim 1 have overcome the obviousness rejections of record over Wen, which are withdrawn. New grounds of rejection follow below, necessitated by the instant amendments.
Any other rejections of record not particularly addressed below are withdrawn in light of the claim amendments and/or applicant’s comments.
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 1-4, 6, 10, 12, 14, 15, 17, 19, 21, 22, and 32 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.
The term “like” in claim 1 is a relative term which renders the claim indefinite. The term “like” 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. Clarification and/or correction is required.
In so much that claims 2-4, 6, 10, 12, 14, 15, 17, 19, 21, 22, and 32 depend from claim 1 and do not resolve the point of confusion, these claims must be rejected with claim 1 as indefinite.
Claim Rejections - 35 USC § 103
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.
Claims 1-4, 6, 10, 12, 15, 17 and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Wen et al. (STEM CELLS TRANSLATIONAL MEDICINE 2013;2:118–128; provided in the IDS dated 4/20/2022) in view of Sommer et al. (STEM CELLS 2009;27:543–549; Reference U).
Wen teaches a method of reprogramming human fibroblasts into pluripotent stem cells, the method comprising 1) obtaining and culturing human fibroblasts (p119, subheading “Tissue Collection”), 2) singly transfecting the human fibroblasts with a lentiviral vector encoding for Oct4, Kif4, Sox2, and cMyc to generate induced pluripotent stem cells (p119, subheading “Lentivirus Production and Infection”), 3) differentiating the induced pluripotent stem cells to embryoid bodies and then fibroblast-like cells and such as to positively express the pluripotency markers NANOG, OCT4, SOX2, hTERT, SSEA-3, SSEA-4, Tra-1–60, and Tra-1–80 (p120, paragraph starting “For embryoid body (EB) formation…” through paragraph ending “…were determined by immunocytochemistry; Fig. 2A for pluripotency marker expression and discussion of this figure at p121, paragraph starting “To generate iPSCs…”), reading on claim 1, claim 2, claims 6 and 15, the embodiment of mRNA made in the cells from the lentiviral vector for claim 17, and the embodiment of proteins expressed from mRNA made in the cells from the lentiviral vector for claim 19. Wen teaches that the induced pluripotent stem cells express similar levels of senescence associate genes when obtained from an older human subject (i.e. 78 years old) or a younger human subject (i.e. 47 years old) (Fig. 4 and discussion of that figure at p122, paragraph starting “To test the effect of age…”), reading on claims 2 and the embodiment of senescence marker expression for claim 12. Wen teaches culturing the human fibroblasts transfected with a lentiviral vector encoding for Oct4, Kif4, Sox2, and cMyc for 25-31 days before picking induced pluripotent stem cell colonies for further culture and differentiation (p119, subheading “Lentivirus Production and Infection”), reading on claim 3. Wen teaches that the reprogramming of the human fibroblasts to induced pluripotent stem cells was more efficient in the younger subject than the older subject (i.e. 76% vs 56% complete reprogramming respectively (paragraph starting on p122 and continuing on p124; also Fig. 1 alternatively showing partially reprogrammed colonies), reading on claim 10.
Regarding claim 1, Wen is silent culturing the somatic cells during the maturation phase of reprogramming. However, optimization within prior art conditions or through routine experimentation will generally not support patentability absent a showing of criticality of the claimed range to the contrary. See M.P.E.P. § 2144.05, particularly subsections II and III. In this case, Wen is effective to culture the somatic cell (e.g. the initial fibroblast) with Oct4, Kif4, Sox2, and cMyc and differentiating the induced pluripotent stem cells to embryoid bodies and then fibroblast-like cells and such as to positively express the pluripotency markers NANOG, OCT4, SOX2, hTERT, SSEA-3, SSEA-4, Tra-1–60, and Tra-1–80. As such, any difference in timing in Wen’s method as compared to claim 1 must be held as optimization within prior art conditions or through routine experimentation absent any showing to the contrary.
Regarding claim 4, Wen does not teach wherein the somatic cell is cultured in the presence of said one or more Oct4, Kif4, Sox2, and/or cMyc for a period of no more than 17 days, or no more than 15 days. However, optimization within prior art conditions or through routine experimentation will generally not support patentability absent a showing of criticality of the claimed range to the contrary. See M.P.E.P. § 2144.05, particularly subsections II and III. In this case, Wen clearly teaches that a culture time ranges of 25-31 days is both operable and optimal before picking induced pluripotent stem cell colonies for further culture and differentiation as cited above. Therefore, the claimed range must be held prima facie obvious absent any showing to the contrary.
Regarding claim 1, Wen does not teach removing one or more of Oct4, Kif4, Sox2, and cMyc prior to the stabilization phage of reprogramming. Regarding claim 15, Wen is silent regarding any inducible expression cassette.
Sommer teaches a tetracycline-inducible lentiviral expression cassette comprising Oct4, Kif4, Sox2, and cMyc, which is used to induce pluripotent stem cell generation from fibroblasts (Abstract; Fig. 1, and termed “pHAGE-STEMCCA” or “Tet-STEMCCA”; “Cell culture” on p544), reading on the embodiment of an inducible expression cassette for the generic removal (e.g. removal of the inducer) of claim 1 and on claim 15. Sommer teaches that the inducible lentiviral expression cassette is advantageous to assess the kinetics of reprogramming (the fibroblasts to induced pluripotent stem cells) in MEFs derived from a single iPS clone (p547, subheading “Secondary iPS Generation by Reactivation of STEMCCA” and Fig. 5A).
Regarding claims 1 and 15, it would have been obvious to a person of ordinary skill in the art before the invention was filed to substitute the lentiviral expression cassette comprising Oct4, Kif4, Sox2, and cMyc of Wen with the tetracycline-inducible lentiviral expression cassette comprising Oct4, Kif4, Sox2, and cMyc of Sommer in Wen’s methods. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because both Wen and Sommer are directed towards methods of introducing a lentiviral expression cassette comprising Oct4, Kif4, Sox2, and cMyc into fibroblasts to generate induced pluripotent stem cells. The skilled artisan would have been motivated to do so because Sommer teaches that the substitution would be predictably advantageous to assess the kinetics of reprogramming fibroblasts to induced pluripotent stem cells.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claim 14 is rejected under 35 U.S.C. 103 as being unpatentable over Wen and Sommer as applied to claims 1 and 12 above, and further in view of Weidner et al. (Genome Biology (2014), 15:R24) and Horvath (Genome Biology (2013), 14:R115).
The teachings of Wen and Sommer are relied upon as set forth above.
Regarding claim 14, Wen and Sommer do not teach wherein the molecular signature of the reprogrammed and/or non-reprogrammed somatic cell is the epigenetic signature and is determined using the Horvath epigenetic clock.
Weidner teaches that age-associated methylation changes are reversed in induced pluripotent stem cells as measured by 102 AR-CpG sites (i.e. age-related DNA methylation sites) sites (Abstract; Fig. 1), reading on claim 14.
Horvath teaches that the 353 clock CpGs can be divided into two sets
according to their correlation with age with 193 positively and 160 negatively correlated CpGs get hypermethylated and hypomethylated with age, respectively (page 7, the 1st complete paragraph in the left column), reading on claim 14. Horvath teaches that the multi-tissue age predictor used for defining DNAm (i.e. DNA methylation) age is accurate across multiple tissue and cell types (p2, paragraph starting ”To ensure an unbiased validation…” through paragraph ending “…and buccal epithelium (Fig. 2I; Fig. 1 and 2), reading in claim 14.
It would have been obvious to a person of ordinary skill in the art before the invention was filed to further calculate/determine the epigenetic signature of Wen’s induced pluripotent stem cells utilizing the cellular age predicting methods of Horvath in view of Weidner. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because Horvath teaches that the multi-tissue age predictor used for defining DNAm (i.e. DNA methylation) age and comprising 353 clock CpGs sites is accurate across multiple tissue and cell types, and because Weidner conceptually links utilizing DNA methylation sites as a measurement of induced pluripotent stem cell age of Wen, and because both Wen and Weidner are directed towards induced pluripotent stem cells. The skilled artisan would have been motivated to do so because the addition of the method of Horvath would likely complement the cellular senescence methods of Wen in determining if the somatic cells produced from the induced pluripotent stem cells of Wen are “younger” than the starting somatic cell population.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claim 21 is rejected under 35 U.S.C. 103 as being unpatentable over Wen and Sommer as applied to claim 1 above, and further in view of Weltner et al. (Nature Communications (July 6th, 2018), 9:2463).
The teachings of Wen and Sommer are relied upon as set forth above.
Regarding claim 21, Wen and Sommer do not teach wherein the Oct4, Kif4, Sox2, and/or cMyc are introduced into the somatic cell by CRISPR/Cas-9.
Weltner teaches methods reprogramming skin fibroblasts into induced pluripotent stem cells, the method comprising by targeting the endogenous OCT4, KLF4, SOX2, Myc, and LIN28A promoters utilizing CRISPRa (Abstract; subheading “Fibroblast reprogramming with CRISPRa” on page 3 and continuing on page 6), reading on claim 21. Weltner teaches the reprogramming methods are advantageous due to a high multiplexing capacity and ability to target endogenous loci (Abstract), reading on claim 21
It would have been obvious to a person of ordinary skill in the art before the invention was filed to substitute the lentiviral vector comprising exogenous DNA sequence encoding for OCT4, KLF4, SOX2, Myc of Wen with the CRISPRa/Cas9 system targeting expression of endogenous OCT4, KLF4, SOX2, Myc, and LIN28A promoters of Weltner in Wen’s induced pluripotent stem cells. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because Wen and Weltner are directed towards methods of reprogramming fibroblasts into induced pluripotent stem cells. The skilled artisan would have been motivated to do so because Weltner teaches the reprogramming methods comprising CRIPSR are advantageous due to a high multiplexing capacity and ability to target endogenous loci, thus improving upon the methods and cells of Wen.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claims 19 and 32 is rejected under 35 U.S.C. 103 as being unpatentable over Wen and Sommer as applied to claim 1 above, and further in view of Tammam et. al. (Oncotarget (2016), 7(25), 37728-37739;) and Li et al. (Cell Research (2011), 21:196-204).
The teachings of Wen and Sommer are relied upon as set forth above, and read in part on claim 19. Regarding claims 19 and 32, Wen and Sommer do not optionally teach wherein the targeted delivery system comprises a nanoparticle delivery system.
Tammam teaches a method of introducing recombinant OCT4 protein into human fibroblasts, the method comprising loading chitosan nanoparticles with recombinant OCT4 protein and contacting the human fibroblasts with the OCT4-loaded chitosan nanoparticles sufficient to deliver the recombinant OCT4 protein to the nucleus of the cells (p37737, paragraph starting “Human fibroblasts were seeded…” and Fig. 6; also p37730 for recombinant expression of OCT4 protein in Sf9 insect cells), reading on claims 19 and 32. Tammam teaches that the chitosan nanoparticles are advantageous to stabilize the DNA-binding activity of recombinant OCT4 protein (p37729, paragraph starting “In this study…”; subheading “Chitosan S-NPs stabilize OCT4 DNA-binding activity” on p37730-31). Tammam teaches that there is a need in this art to improve upon methods of generating induced pluripotent stem cells, and the most commonly used method is retroviral transduction but which bears the risk of insertional mutagenesis of the genome-integrating viruses (1st paragraph of the Introduction), reading on claims 19 and 32.
Li teaches a method of reprogramming mouse fibroblasts into pluripotent stem cells, the method comprising infecting the cells with a virus carrying a gene encoding for OCT4 and treating the cells with a combination of 0.5 mM VPA, 5 µM tranylcypromine, 3 µM CHIR99021 and 1 µM 616452 (i.e. VC6T) such as to generate induced pluripotent stem cells (p202, subheading “Lentivirus infection and iPS cell generation”; 1st paragraph of the Discussion on p201; Fig. 1 for GFP+ stem cell generation and Fig. 2 for expression of pluripotency markers), reading on claims 19 and 32.
It would have been obvious to a person of ordinary skill in the art before the invention was filed to substitute the lentiviral vector comprising exogenous DNA sequence encoding for OCT4, KLF4, SOX2, Myc of Wen with chitosan nanoparticle and recombinant OCT4 protein composition and methods of loading of fibroblasts with said composition of Tammam and the small molecule treatment methods of Li in Wen’s methods of making induced pluripotent stem cells A person of ordinary skill in the art would have had a reasonable expectation of success to do so because Wen and Li are directed towards methods of generating induced pluripotent stem cells from fibroblasts, and Li shows that the combination of OCT4 and treatment with the combination of 0.5 mM VPA, 5 µM tranylcypromine, 3 µM CHIR99021 and 1 µM 616452 is sufficient to generate induced pluripotent stem cells. The skilled artisan would have been motivated to do so because ). Tammam teaches that there is a need in this art to improve upon methods of generating induced pluripotent stem cells, and the most commonly used method is retroviral transduction but which bears the risk of insertional mutagenesis of the genome-integrating viruses, and so the substitution would likely improve upon the methods of Wen by removing the lentivirus vector and so remove any possibility mutagenizing the fibroblasts of Wen.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claim 22 is rejected under 35 U.S.C. 103 as being unpatentable over Wen and Sommer as applied to claim 1 above, and further in view of Prieur et al. (WO 2012/136841; provided in the IDS dated 4/20/2022).
The teachings of Wen and Sommer are relied upon as set forth above.
Regarding claim 22, Wen and Sommer do not teach further culturing the cells in the presence of LIN28 and/or NANOG.
Prieur teaches a method for reprogramming cells from aged donors or senescent cells to pluripotent cells that have lost marks of senescence (Abstract). Prieur teaches reprogramming said cells into iPSCs (i.e. induced pluripotent stem cells) with reprogramming factors comprising Oct4, Klf4, Sox2, cMyc, Lin28 and, optionally Nanog (Abstract), reading on claim 22.
Regarding claim 22, "It is prima facie obvious to combine two compositions each of which is taught by the prior art to be useful for the same purpose, in order to form a third composition to be used for the very same purpose.... [T]he idea of combining them flows logically from their having been individually taught in the prior art." In re Kerkhoven, 626 F.2d 846, 850, 205 USPQ 1069, 1072 (CCPA 1980) (citations omitted). See M.P.E.P. § 2144.06. In this case, It would have been obvious to a person of ordinary skill in the art before the invention was filed to add the LIN28 and/or NANOG or Prieur to the composition and methods of Wen because Oct4, Klf4, Sox2, cMyc, Lin28 and Nanog are all taught as useful for the same purpose as reprogramming factors to generate induced pluripotent stem cells from a starting somatic cell type.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Response to Arguments
Applicant's arguments on pages 7-15 of the reply have been fully considered, but not found persuasive of error for the reasons given below.
On pages 7-10 of the reply, Applicant alleges that Wen is deficient regarding culturing the cells during the maturation phase of reprogramming the claimed somatic cells. This is not found persuasive because optimization within prior art conditions or through routine experimentation will generally not support patentability absent a showing of criticality of the claimed range to the contrary. See M.P.E.P. § 2144.05, particularly subsections II and III. Nothing in Applicant’s arguments clearly sets forth any particular unexpected result nor discriminates between expected and unexpected results relative to the cited teachings of Wen. See M.P.E.P. § 716.02.
On pages 10-11 of the reply, Applicant alleges that Wen is deficient by not recognizing the claimed culture time range as a result-effective variable. This is not found persuasive because Wen clearly is effective to culture the somatic cell (e.g. the initial fibroblast) with Oct4, Kif4, Sox2, and cMyc and differentiating the induced pluripotent stem cells to embryoid bodies and then fibroblast-like cells and such as to positively express the pluripotency markers NANOG, OCT4, SOX2, hTERT, SSEA-3, SSEA-4, Tra-1–60, and Tra-1–80. As such, any difference in timing in Wen’s method as compared to claim 1 must be held as optimization within prior art conditions or through routine experimentation absent any persuasive showing to the contrary.
On pages 11-15 of the reply, Applicants rely on arguments traversing the above rejection of claim 1 over Wen and Sommer to traverse this rejection of claim 14 further in view of Weidner and Horvath, the rejection of claim 21 further in view of Weltner, the rejection of claim 22 further in view of Prieur, and the rejection of claims 19 and 32 further in view of Tammam and Li. Therefore, the response set forth above to arguments also applies to these rejections.
Conclusion
No claims are allowed. No claims are free of the art.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to SEAN C BARRON whose telephone number is (571)270-5111. The examiner can normally be reached 7:30am-3:30pm EDT/EST (M-F).
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/Sean C. Barron/Primary Examiner, Art Unit 1653