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 .
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 12/09/2025 has been entered.
Claims 19-32 are pending. Claims 19, 27, and 28 are independent claims. Applicant cancelled claims 1-18 by amendment filed on 11/12/2025.
Therefore, claims 19-32 are currently under examination to which the following grounds of rejection are applicable.
Priority
The instant application is a national stage entry under 35 USC 371 of PCT/JP2020/040732 (filed 10/30/2020), which claims benefit of JAPAN 2019-199781 filed 11/01/2019.
Acknowledgment is made of applicant’s claim for foreign priority under 35 U.S.C. 119 (a)-(d). Thus, the earliest possible priority date is 11/01/2019.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on 6/08/2022, 9/12/2023, and 6/24/2024, were filed before the mailing date of the Non-final office action. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Withdrawn Rejections in response to Applicants’ arguments or amendments
Claim Rejections - 35 USC § 101
The rejection of claims 13-18 under 35 USC § 101 is rendered moot in view of the amendments in the response filed on 11/12/2025. The claims have been cancelled and the new claims include clarifications to the previously presented claim. The amended claim 27-32 (which correlate to the cancelled claims 13-18) now recite that a knockout gene is present in the cell, and this engineered cell type is not a cell that would not be a naturally occurring product.
Claim Rejections - 35 USC § 112(b)
The rejection of claims 13-18 under 35 USC § 112(b) is withdrawn in view of the amendments in the response filed on 11/12/2025. The claims have been cancelled and the new claims include clarifications to the previously presented claim. The amended claim 27-32 (which correlate to the cancelled claims 13-18) have deleted the term “substantially”.
Claim Rejections - 35 USC § 103
The rejection of claims 1-3 and 8-9 under 35 USC § 103 as being unpatentable over Vodyanyk et al., (AU 2016342183 Al) in view of Fowler et al., (US 2004/0241153 Al) as
evidenced by Leoncini et al. (Oxidative medicine and cellular longevity, 2015)) is rendered mooot in view of the amendments in the response filed on 11/12/2025. The claims have been cancelled and the new claims include the limitations of claims 6 and 7, which were not rejected by the prior art.
Applicants’ arguments with regard to any withdrawn objection/rejection are moot.
New objections and rejections in response to Applicants’ arguments or amendments
Claim Objections
Claims 19 and 28 are objected to because of the following informalities: the recitation of both TBX21 in the same Markush group as T-bet is improper. TBX21 is the gene encoding the protein T-bet. As the claims are drawn to a gene knockout, T-bet should be deleted. Appropriate correction is required.
Claim Rejections - 35 USC § 112(a)
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 19-32 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.
The independent claims are claims 19, 27, and 28. Claims 27 and 28 are product claims.
Claim 19 recites, “A method for producing a Thl-type or Th2-type CD4 single-positive T cell, comprising a step of inducing a CD4 single-positive T cell from a hematopoietic stem cell (HSC) and/or a hematopoietic progenitor cell (HPC): wherein a gene involved in IL-4 secretion is knocked out, wherein the gene involved in IL-4 secretion is selected from the group consisting of IL-4, IL-4R, Stat6, GATA3, mel-18, IL-5 and IL-13 genes; or wherein a gene involved in IFN-y secretion is knocked out, wherein the gene involved in IFN-y secretion is selected from the group consisting of TBX21, IL-12, IFN-y, IL-12R~2, IFNyR, Stat4, T-bet and HLX genes.
Claim 27 recites, “A Thl-type CD4 single-positive T cell, wherein a gene involved in IL-4 secretion is knocked out, wherein the gene involved in IL-4 secretion is selected from the group consisting of IL4, IL-4R, Stat6, GATA3, mel-18, IL-5 and IL-13 genes.”
Claim 28 recites, “A Th2-type CD4 single-positive T cell, wherein a gene involved in IFN-y secretion is knocked out, wherein the gene involved in IFN-y secretion is selected from the group consisting of TBX21, IFN-y, IL-12RB2, IFNyR, Stat4, T-bet and HLX genes.”
The instant disclosure defines a Th-1 type or Th2-type T cell as follows: IL-4 non-secreting and IFNy secreting (Thl-type) or IFN-y non-secreting and IL-4 secreting (Th2-type) CD4 single-positive T cell (CD4SP T cell) (Pg. 7, Paragraph [0012]).
The instant disclosure fails to provide sufficient support pertaining to a Th1-type or Th2-type T cell wherein knockout of any of the genes in the instant claim would retain the functionalities of a Th1-type or Th2-type T cell. The specification does not provide sufficient support for all claimed knockouts that would be encompassed by the structural limitations of the independent claims and still be able to produce functional requirements of either an IL-4 non-secreting and IFNy secreting (Thl-type) or IFN-y non-secreting and IL-4 secreting (Th2-type) CD4 single-positive T cell (CD4SP T cell). The instant specification does not provide a sufficient representative sampling of cells that harbor a knockout of genes as claimed which includes, IL-4R, Stat6, GATA3, mel-18, IL-5 and IL-13 genes to produce a Th1 type T cell or TBX21, IL-12, IFN-y, IL-12R~2, IFNyR, Stat4, T-bet and HLX genes to produce a Th2 type T cell. Dependent claims 20-26 and 29-32 do not serve to overcome the written description deficiencies.
Nature of the Invention
The instant claims are drawn to a method for producing an IL-4 non-secreting and IFNy secreting (Thl-type) or IFN-y non-secreting and IL-4 secreting (Th2-type) CD4 single-positive T cell (CD4SP T cell) and the cells that are produced.
Specification Disclosure
The specification describes only the following KO cell lines at Pg. 62 (Table 2, provided below where specific gRNAs have been used to knock out various nucleotide mutations) and the describes the KO cell line production workflow (essentially iPSC -> HPC / HPC -> Thl cell or Th2 cell) at example 2, Pg. 63-64.
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The specification does not provide sufficient support for cells that harbor a knockout of genes as claimed which includes, IL-4R, Stat6, GATA3, mel-18, IL-5 and IL-13 genes to produce a Th1 type T cell or TBX21, IL-12, IFN-y, IL-12R~2, IFNyR, Stat4, T-bet and HLX genes to produce a Th2 type T cell.
The specification states that “The gene involved in IL-4 secretion is typically the IL4 gene, but it may be another gene as long as it can suppress IL-4 secretion by substantial defectiveness and neither interferes with the production of a Thl-type CD4SP T cell nor suppresses IFN-y secretion. More specifically, the gene is involved in IL-4 secretion can be selected from genes encoding the proteins shown in the right figure of FIG. 1 (Th2 signals), that is, IL-4, IL-4R, Stat6, GATA3, mel-18, IL-5 and IL-13 (at least a portion of the protein if it is a
complex).” and “The "gene involved in IFN-y secretion" is typically the "TBX21 gene" that encodes T-bet (see FIG. 1), but it may be another gene as long as it can suppress IFN-y secretion by substantial defectiveness and neither interferes with the production of a Th2-type CD4SP T cell nor suppresses IL-4 secretion. More specifically, the "gene involved in IFN-y secretion" can be selected from genes encoding the proteins shown in the left figure of FIG. 1 (Thl signals), that is, IL-12, IFN-y, IL-12R~2, IFNyR, Stat4, T-bet and HLX (at least a portion of the protein if it is a complex).” (Pg. 27 and 28, Paragraph [0030] – [0031]).
However, the caveats provided in the instant specific that “it may be another gene as long as it can suppress IFN-y secretion by substantial defectiveness and neither interferes with the production of a Th2-type CD4SP T cell nor suppresses IL-4 secretion” would mean that the applicant is aware that the knockout of these genes may have an undesired effect and the specification is silent on the effects of knocking out these genes, save for the IL-4 knockout to produce a TH1-type Tcell and the TBX21 knockout to produce a TH2-type Tcell.
In summary, the specification provides only a narrow subset (IL-4 and TBX21 KO), while the claims encompass a broader and heterogenous set of gene knockouts, of which some may not predictably produce the claimed TH1-type/Th2-type cells. The specification does not provide sufficient support to identify whether the structural limitations (knockouts encompassed by the independent claims) would be able to produce a Tcell with the functional requirements as claimed.
Guidance Provided by the Art
A thorough search of the art revealed the genes recited in the instant claims may play multifaceted and complex roles within Tcell biology and that knockout/disruption of these genes may not necessarily produce the claimed TH1-type/Th2-type cells.
For example, the assertion by the applicant that IL4R KO would necessarily produce an IL-4 non-secreting cell is also not certain. Noben-Trauth et al. (Noben-Trauth, N et al., Proceedings of the National Academy of Sciences of the United States of America, 1997) demonstrates that IL4R disruption does not necessarily produce a Tcell with loss of IL-4 production, “To more definitively investigate the role of IL-4 in TH2 development, we disrupted the IL-4Rα gene by homologous recombination in BALB/c-I embryonic stem (ES) cells. Because these mice lack sensitivity to IL-4, we could evaluate the relative importance of IL-4 in development of TH2 responses and could determine what cell populations retain the capacity to produce IL-4 even when they differentiate in the total absence of IL-4. Here we describe results from these studies in which we tested the relative ability of CD4+ T cells from wild-type and IL-4Rα-deficient mice to develop into IL-4 producers.” (Pg. 10838, 2nd column, 1st paragraph).
With respect to GATA3, Bach et al. (Bacha R et al., Biochim Biophys Acta Mol Cell Res., 2025) teaches “GATA3 is a significant regulator of the specification, survival, and differentiation of T-cells into T-helper 2 cells (Th2 cells). It inhibits the differentiation of T-helper 1 cells (Th1) and T-helper 17 cells (Th17 cells) while enhances the expression of Th2 cytokines (Interleukin-4 (IL-4), Interleukin-5 (IL-5), Interleukin-13 (IL-13)). Strikingly, GATA3 plays a critical role not only in the development of innate lymphoid cells (ILCs), particularly ILC2s, but also in the regulation of adaptive immune responses, such as Th2 cell differentiation. The distinction between the gene expression profiles and regulatory networks of Th1 and Th2 cells, identified through cDNA representational difference analysis, highlights the specific influence of GATA3 in driving the Th2 cell lineage, further underscoring its role in coordinating immune cell development across both innate and adaptive systems.” (Pg. 3, Section 3.1; Figure 2 provided
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below).
Zhu et al. (Jinfang Zhu et al., The Journal of Immunology, 2001, Pages 7276–7281) teaches Stat6 has been shown to be required for Th2 differentiation (Abstract) and further teaches with respect to STAT6, “The differentiation of naive T cells into Th2 cells requires both TCR- and IL-4-mediated signals. IL-4 activation of Stat6 has been shown to be a critical step in driving Th2 differentiation; the role of IRS2 and other PTBD proteins during Th2 differentiation is unclear. To examine a role for the PTBD proteins in IL-4-mediated Th2 differentiation and IL-4-driven proliferation in CD4 cells, we used a retrovirus (RV) system to introduce h wild-type (WT) and mutant IL-4Rα into CD4 T cells. It was found that Y497, the binding site for PTBD protein, is dispensable for both IL-4-driven Th2 differentiation and for cell expansion. Using Stat6-knockout CD4 T cells, we found that Stat6 is necessary for both of these IL-4-induced functions in CD4 T cells. Moreover, using a RV containing constitutively activated Stat6, it was found that Stat6 is not only necessary but also sufficient for the IL-4 effects in Th2 differentiation and cell expansion.” (Pg. 7276, 2nd column, 2nd full paragraph).
Taken together, the prior art provides clear evidence that ability to produce the recited cell types with the function of IL-4 non-secreting and IFNy secreting (Thl-type) or IFN-y non-secreting and IL-4 secreting (Th2-type) CD4 single-positive T cell (CD4SP T cell) is not readily established. As such, it is clear that many, if not all, of the recited genes that are not supported by the specification are obviously critical and knockout may not necessarily produce the claimed cells.
Conclusion
Considering the broadness in the genus encompassed by the instant claims, the singular species described in the specification for each cell type, and the lack of predictability provided by the specification and art for the full scope of the claimed genus, it is reasonable to conclude that applicant did not possess the invention as claimed at the time of filing.
Claim Rejections - 35 USC § 112 (b)
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 19-26, 28, and 31-32 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.
Claim 19 recites the following, “…wherein a gene involved in IFN-y secretion is knocked out, wherein the gene involved in IFN-y secretion is selected from the group consisting of TBX21, IL-12, IFN-y, IL-12RB2, IFNyR, Stat4, T-bet and HLX genes”.
IL-12 is a cytokine made up of two subunits, each encoded by a separate gene. As written, it is unclear to which gene the applicant is intending to be knocked out, therefore the claim is rendered indefinite.
IFNyR is a heterodimeric receptor. The gene IFNGR1 encodes IFN-γR1, which is the ligand-binding chain (alpha) of the heterodimeric gamma interferon receptor and IFNGR2, encodes IFN-γR2, the non-ligand-binding partner of the heterodimeric receptor. As written, it is unclear to which gene the applicant is intending to be knocked out, therefore the claim is rendered indefinite.
Claims 20-26 are rejected insofar as they depend from claim 19.
Claim 26 recites the following, “The method according to claim 25, wherein the T cell is a CD4 single-positive T cell.”. It is unclear what is being referred to by “the T cell”. As claim 25 recites a T-cell “the iPSC is a T cell-derived iPS cell” and claim 19, from which claims 25-26 ultimately depend, recite multiple T cells. Therefore, it is unclear to which T cell is being referenced, thus the claim is rendered indefinite. If applicant intends the T-cell is referring to the claim 25 recitation “the iPSC is a T cell-derived iPSC cell”, then applicant should amend the claim 26 to recite, wherein T cell from which the iPSC is derived is a CD4 single-positive T cell” or similar.
Claim 28 recites the following, “…wherein a gene involved in IFN-y secretion is knocked out, wherein the gene involved in IFN-y secretion is selected from the group consisting of TBX21, IFN-y, IL-12RB2, IFNyR, Stat4, T-bet and HLX genes”.
IFNyR is a heterodimeric receptor. The gene IFNGR1 encodes IFN-γR1, which is the ligand-binding chain (alpha) of the heterodimeric gamma interferon receptor and IFNGR2, encodes IFN-γR2, the non-ligand-binding partner of the heterodimeric receptor. As written, it is unclear to which gene the applicant is intending to be knocked out, therefore the claim is rendered indefinite.
Claims 31-32 are rejected insofar as they depend from claim 28.
Conclusion
Claims 19-32 are rejected. No claim is allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to KODYE LEE ABBOTT whose telephone number is (703)756-1111. The examiner can normally be reached M-F 8-5.
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/KODYE LEE ABBOTT/Examiner, Art Unit 1634
/MARIA G LEAVITT/Supervisory Patent Examiner, Art Unit 1634
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