DETAILED ACTION
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 .
Status of the Claims
Applicant’s submission filed 09 July 2026 has been entered. Claims 1-12, 15, 18-19, 22, and 27 are pending. Claims 1 and 22 have been amended, while claims 17, 25, and 29 have been cancelled without prejudice or disclaimer. Therefore, prosecution on the merits commences for claims 1-12, 15, 18-19, 22, and 27.
Status of Prior Rejections/Response to Arguments
RE: Rejection of claim 25 under 35 USC 112(d)
The cancellation of instant claim 25 renders the rejection moot for that claim. Therefore, the rejection is withdrawn.
RE: Rejection of claims 22, 25, 27, and 29 under 35 USC 101
The cancellation of claims 25 and 29 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 22 requiring the multinucleated megakaryocytes to comprise at least one exogenously expressed protein by at least one gene selected from the group consisting of an oncogene, a Polycomb gene, and an apoptosis suppressor gene obviates the rejection of record.
Therefore, the rejection is withdrawn.
RE: Rejection of claims 22, 25, 27, and 29 under 35 USC 102(a)(1) and 35 USC 102(a)(2) over Wu
The cancellation of claims 25 and 29 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 22 requiring the multinucleated megakaryocytes to comprise at least one exogenously expressed protein by at least one gene selected from the group consisting of an oncogene, a Polycomb gene, and an apoptosis suppressor gene obviates the rejection of record.
Therefore, the rejection is withdrawn.
RE: Rejection of claims 1-2, 4-9, 12, 15, and 17-19 under 35 USC 103 over Eto et al in view of Elagib et al
The cancellation of claim 17 renders the rejection moot for that claim. For the remaining claims, Applicant’s arguments filed 09 July 2026 have been fully considered but they are not persuasive.
Applicant has traversed the rejection, asserting in Pages 7-8 of the Remarks filed 09 July 2026 that the combination of Eto et al and Elagib et al fail to teach that the produced multinucleated megakaryocytes have an enhanced platelet production capability. In response, the Examiner respectfully submits that both Eto et al and Elagib et al teach that the produced multinucleated megakaryocytes have an enhanced platelet production capability. See, for example, Paragraphs [0114]-[0116] of Eto et al and the third paragraph of Elagib et al.
Applicant has further traversed the rejection, asserting in Pages 8-9 of the Remarks filed 09 July 2026 that Elagib et al fail to teach that the harmine is administered to proliferative megakaryocytes, as well as provide a specific protocol for the addition. In response, the Examiner respectfully submits that Elagib et al disclose the treatment of infantile cord blood megakaryocytes with harmine strongly enhanced polyploidization. See, for example, the third paragraph of Elagib et al. Therefore, it would not have been outside the skillset of the ordinary artisan to first recognize that harmine can be administered to pre-multinucleated megakaryocytes that are capable of polyploidization, and then administer an effective amount of harmine to the pre-multinucleated megakaryocytes.
Applicant has further traversed the rejection, asserting in Page 9 of the Remarks filed 09 July 2026 that the combination of Eto et al and Elagib et al fail to teach that the pre-multinucleated megakaryocytes exogenously and inducibly express at least one gene selected from the group consisting of an oncogene, a Polycomb gene, and an apoptosis suppressor gene by gene transfer. In response, the Examiner respectfully submits that Eto et al teach that the pre-multinucleated megakaryocytes inducibly and exogenously express at least one gene selected from the group consisting of an oncogene, a Polycomb gene, and an apoptosis suppressor gene, wherein the gene is introduced into the cell via viral vectors – or by gene transfer. See, for example, Paragraphs [0045], [0078]-[0080], [0085], [0108], [0111], [0129], [0133], [0159], and [0173] of Eto et al.
Therefore, the rejection is maintained and amended to encompass the claims as currently written.
RE: Rejection of claims 1-12, 15, and 17-19 under 35 USC 103 over Eto et al in view of Elagib et al and Ohtori et al
The cancellation of claim 17 renders the rejection moot for that claim. For the remaining claims, Applicant’s arguments filed 09 July 2026 have been fully considered but they are not persuasive.
Applicant has traversed the rejection, citing the same assertions as discussed in Pages 7-9 of the Remarks filed 09 July 2026. In response, the Examiner respectfully directs Applicant to the discussion of the 35 USC 103 rejection over Eto et al in view of Elagib et al above.
Therefore, the rejection is maintained and amended to encompass the claims as currently written.
New/Maintained Grounds of Rejection
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.
Claims 22 and 27 are rejected under 35 U.S.C. 103 as being unpatentable over Eto et al (US 2019/0048317 A1, of record on IDS filed 02 July 2025).
Eto et al is considered prior art under 35 USC 102(a)(1) and 35 USC 102(a)(2), with a publication date of 14 February 2019.
Regarding claims 22 and 27: Eto et al disclose a method for obtaining multinucleated megakaryocytes, wherein megakaryocyte progenitor cells are first engineered to inducibly express a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene, wherein the gene is transferred into the megakaryocyte progenitor cell via a viral vector, and then contacted with a platelet production promoting agent and a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene such that the megakaryocyte progenitor cells become multinucleated megakaryocytes (Paragraphs [0045], [0077]-[0081], [0085], [0108], [0110]-[0111], [0115], [0129], [0133], [0155], [0159], [0166], [0173], [0248]).
Eto et al further disclose that the multinucleated megakaryocytes have a genome that is 16 times to 32 times larger than that of normal cells – or a nuclear phase of 16N to 32N (Paragraph [0078]).
Eto et al fail to teach that the population of multinucleated megakaryocytes comprises at least 20% of multinucleated megakaryocytes having a nuclear phase of 16N or more and 5-50% of multinucleated megakaryocytes having a nuclear phase of 32N or more, as required by instant claims 22 and 27.
However, it would have been prima facie obvious to have modified the method of Eto et al such that the generated population of multinucleated megakaryocytes comprises 20% 16N multinucleated megakaryocytes and 5% 32N multinucleated megakaryocytes. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to ensure the population comprised those percentages, as megakaryocytes having a nuclear phase of 16N and 32N indicates that they have been successfully multinucleated. Furthermore, the ordinary artisan would have had a reasonable expectation of success given that Eto et al disclose that the generated multinucleated megakaryocytes have a nuclear phase of 16N and 32N, and it would not have been outside the skillset of the ordinary artisan to modify the proportion of 16N and 32N multinucleated megakaryocytes within the population via well-known sorting and/or fractionation techniques. See MPEP § 2143(I)(G) and MPEP § 2144.05.
Consequently, Eto et al render obvious a population of multinucleated megakaryocytes, wherein megakaryocyte progenitor cells are first engineered to inducibly express an exogenous protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene via viral vector gene transfer, and wherein the generated population of multinucleated megakaryocytes comprises 20% of multinucleated megakaryocytes having a nuclear phase of 16N (claim 22) and 5% of multinucleated megakaryocytes having a nuclear phase of 32N (claim 27). This therefore renders obvious the method of the instant claims.
Claims 1-2, 4-9, 12, 15, and 18-19 remain rejected under 35 U.S.C. 103 as being unpatentable over Eto et al (US 2019/0048317 A1, of record on IDS filed 02 July 2025) in view of Elagib et al (Blood, 2019, of record on IDS filed 18 December 2023).
Eto et al is considered prior art under 35 USC 102(a)(1) and 35 USC 102(a)(2), with a publication date of 14 February 2019. Elagib et al is considered prior art under 35 USC 102(a)(1).
Regarding claims 1-2, 6, and 8: Eto et al disclose a highly functional platelet production promoting agent comprising AhR inhibitors and ROCK inhibitors (Abstract; Paragraph [0118]).
As such, Eto et al disclose a method for obtaining multinucleated megakaryocytes, wherein the method comprises contacting megakaryocyte progenitor cells with the platelet production promoting agent and a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene, wherein the megakaryocyte progenitor cells become multinucleated megakaryocytes (Paragraphs [0077]-[0081], [0110]-[0111], [0115], [0129], [0155], [0166], [0248]).
Eto et al further disclose that the megakaryocyte progenitor cells are engineered to inducibly express a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene, wherein the gene is transferred into the megakaryocyte progenitor cell via a viral vector (Paragraphs [0045], [0078]-[0080], [0085], [0108], [0111], [0129], [0133], [0159], [0173]).
Eto et al do not disclose that the megakaryocyte progenitor cells are multinucleated in the presence of harmine, as required by instant claim 1.
Elagib et al, however, disclose that the treatment of megakaryocyte progenitors with harmine results in multinucleated megakaryocytes having an enhanced in vitro and in vivo platelet release (Page 2). Elagib et al further disclose that the harmine treatment can be combined with an AhR inhibitor (Page 2).
Therefore, it would have been prima facie obvious to have modified the method of Eto et al such that the megakaryocyte progenitor cells are further contacted with harmine, as detailed in Elagib et al. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to treat the megakaryocyte progenitor cells with harmine, as it independently enhances platelet release in multinucleated megakaryocytes and increases morphogenesis when combined with AhR inhibitors, and would have had a reasonable expectation of success given that the disclosures of Eto et al and Elagib et al are concerned with the production of multinucleated megakaryocytes having an enhanced platelet production. See MPEP § 2143(I)(G).
Consequently, Eto et al as modified by Elagib et al render obvious a method for obtaining multinucleated megakaryocytes having an enhanced platelet production, the method comprising contacting megakaryocyte progenitor cells with an AhR inhibitor (claims 2, 6), a ROCK inhibitor (claim 8), and harmine, wherein the megakaryocyte progenitor cells also exogenously and inducibly express a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene. This therefore renders obvious the method of instant claim 1.
Regarding claims 4-5: Following the discussion of claim 1, Eto et al further disclose that the megakaryocyte progenitor cells are first expanded in culture medium that does not comprise the AhR or ROCK inhibitors (claim 4), and are then switched to a culture medium comprising the AhR inhibitor and ROCK inhibitor (claim 5) (Paragraphs [0111]-[0114], [0129], [0190]-[0203], [0236]-[0248]). This therefore reads on the method of the instant claims.
Regarding claim 7: Following the discussion of claim 2, Eto et al further disclose that the AhR inhibitor is GNF-351 (Paragraphs [0014], [0016], [0039], [0049], [0059], [0063], [0105]-[0107], [0116], [0248]). This therefore reads on the method of the instant claim.
Regarding claim 9: Following the discussion of claim 2, Eto et al further disclose that the ROCK inhibitor is Y-39983 (Paragraphs [0015]-[0016], [0040], [0050], [0063], [0114]-[0116], [0248]). This therefore reads on the method of the instant claim.
Regarding claims 12 and 15: Following the discussion of claim 1, Eto et al further disclose that the multinucleated megakaryocytes have a genome that is 16 times to 32 times larger than that of normal cells – or a nuclear phase of 16N to 32N (Paragraph [0078]).
The combination of Eto et al and Elagib et al fail to teach that the population of multinucleated megakaryocytes comprises at least 20% of multinucleated megakaryocytes having a nuclear phase of 16N or more and 5-50% of multinucleated megakaryocytes having a nuclear phase of 32N or more, as required by instant claims 12 and 15.
However, it would have been prima facie obvious to have modified the method of Eto et al in view of Elagib et al such that the generated population of multinucleated megakaryocytes comprises 20% 16N multinucleated megakaryocytes and 5% 32N multinucleated megakaryocytes. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to ensure the population comprised those percentages, as megakaryocytes having a nuclear phase of 16N and 32N indicates that they have been successfully multinucleated. Furthermore, the ordinary artisan would have had a reasonable expectation of success given that Eto et al disclose that the generated multinucleated megakaryocytes have a nuclear phase of 16N and 32N, and it would not have been outside the skillset of the ordinary artisan to modify the proportion of 16N and 32N multinucleated megakaryocytes within the population via well-known sorting and/or fractionation techniques. See MPEP § 2143(I)(G) and MPEP § 2144.05.
Consequently, Eto et al as modified by Elagib et al render obvious a method for obtaining multinucleated megakaryocytes having an enhanced platelet production, wherein the generated population of multinucleated megakaryocytes comprises 20% of multinucleated megakaryocytes having a nuclear phase of 16N (claim 12) and 5% of multinucleated megakaryocytes having a nuclear phase of 32N (claim 15). This therefore renders obvious the method of the instant claims.
Regarding claim 18: Following the discussion of claim 1, Eto et al further disclose that the megakaryocyte progenitor cells are immortalized megakaryocytes (Paragraph [0079], [0111], [0129], [0150], [0169]-[0203], [0236]). This therefore reads on the method of the instant claim.
Regarding claim 19: Following the discussion of claim 1, Eto et al further disclose that the multinucleated megakaryocytes induced from megakaryocyte progenitor cells produce platelets (Paragraphs [0003], [0053], [0079], [0155]). This therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of instant claim 1.
Claims 1-12, 15, and 18-19 remain rejected under 35 U.S.C. 103 as being unpatentable over Eto et al (US 2019/0048317 A1, of record on IDS filed 02 July 2025) in view of Elagib et al (Blood, 2019, of record on IDS filed 18 December 2023), and further in view of Ohtori et al (US 2023/0030814 A1, of record on IDS filed 18 December 2023).
The discussion of Eto et al as modified by Elagib et al regarding claims 1-2 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Eto et al as modified by Elagib et al render obvious claims 1-2, 4-9, 12, 15, and 18-19. Ohtori et al is considered prior art under 35 USC 102(a)(2), with an effective filing date of 12 December 2019.
Regarding claims 3 and 10-11: As aforementioned in the discussion of claims 1-2 above, Eto et al as modified by Elagib et al render obvious a method for obtaining multinucleated megakaryocytes having an enhanced platelet production, the method comprising contacting megakaryocyte progenitor cells with an AhR inhibitor, a ROCK inhibitor, and harmine, wherein the megakaryocyte progenitor cells also exogenously and inducibly express a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene.
The combination of Eto et al and Elagib et al fail to teach that the megakaryocyte progenitor cells are contacted with a myosin 2 inhibitor, as required by instant claims 3 and 10.
Ohtori et al, however, disclose the multinucleation of megakaryocytes via contacting megakaryocyte progenitor cells with an AhR inhibitor, a ROCK inhibitor, a myosin 2 inhibitor, and a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene (Paragraphs [0049]-[0050], [0080], [0104], [0106]-[0109]). Ohtori et al further disclose that the myosin 2 inhibitor is blebbistatin (Paragraph [0104]).
Therefore, it would have been prima facie obvious to have modified the method of Eto et al in view of Elagib et al such that the megakaryocyte progenitor cells are further contacted with the myosin 2 inhibitor, blebbistatin, as detailed in Ohtori et al. One of ordinary skill before the effective filing date of the invention would have been motivated to contact the megakaryocyte progenitor cells with an inhibitor known to induce multinucleation within the megakaryocyte progenitor cells, and would have had a reasonable expectation of success given that the disclosures of both Eto et al and Ohtori et al are concerned with the generation of multinucleated megakaryocytes utilizing methods that would require minimal adaptation. See MPEP § 2143(I)(G).
Consequently, Eto et al as modified by Elagib et al and Ohtori et al render obvious a method for obtaining multinucleated megakaryocytes having an enhanced platelet production, the method comprising contacting megakaryocyte progenitor cells with an AhR inhibitor, a ROCK inhibitor, a myosin 2 inhibitor (claim 10), and harmine (claim 3), wherein the megakaryocyte progenitor cells also exogenously and inducibly express a protein encoded by a cancer gene, a polycomb gene, or an apoptosis suppressor gene. As the myosin 2 inhibitor is blebbistatin, this therefore renders obvious the method of instant claim 11.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to ALYSSA G WESTON whose telephone number is (571)272-0337. The examiner can normally be reached Monday-Thursday 8AM - 4PM (CT); Friday 8AM - 11AM (CT).
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If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Christopher Babic can be reached at (571) 272-8507. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/ALYSSA G WESTON/Examiner, Art Unit 1633
/CHRISTOPHER M BABIC/Supervisory Patent Examiner, Art Unit 1633
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