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 Claims / Response to Amendments
The Amendments and Remarks filed 12/23/2025 in response to the Office Action of 09/24/2025 are acknowledged and have been entered.
Claims 1, 2, 4-8, 12-18 and 20 are amended.
Claims 1-18 and 20 are currently pending and under consideration.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office Action.
This Office Action contains New Objections and Rejections Necessitated by Amendments.
Priority
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55. The U.S. effective filing date of all claims under examination is set at 09/30/2019 based on the CN201910945529.3 application (filed on 09/30/2019).
Objections Withdrawn
The drawing objections are withdrawn.
The specification objections are withdrawn.
The claim objections are withdrawn.
Claim Rejections Withdrawn
The rejection of claims 2, 4, 5, 7, 8, 10, 11, 12, 13-17 and 20 under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AlA), second paragraph has been withdrawn.
The enablement rejection of claim 20 under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph has been withdrawn.
The rejection of claims 1-9 and 20 under 35 U.S.C. 103 as being unpatentable over Li et al. (Science. 2010 Jul 2;329(5987):85-9) has been withdrawn.
The rejection of claims 1-18 and 20 under 35 U.S.C. 103 as being unpatentable over Li et al. (Science. 2010 Jul 2;329(5987):85-9), Berenson et al. (CN 1556854 Date Published 2004-12-22), Hosokawa et al. (Nat Immunol 19, 1427–1440 (2018) and de Jong et al. (Immunology. 1991 Oct;74(2):175-82) has been withdrawn.
The rejection of claims 1-9 and 20 under 35 U.S.C. 103 as being unpatentable over Liu et al. (WO 2011007176A1 Date of Publication 2011-01-20) has been withdrawn.
The provisional nonstatutory double patenting rejection of claims 1-18 and 20 as being unpatentable over claims 1-10, 15-22 and 24 of copending Application No. 17/775,812 has been withdrawn. This is in response to a filed Terminal Disclaimer (12/23/2025).
Rejections Maintained
Claim Rejections 35 U.S.C.112(b) - Maintained
Claim 18 remains rejected because it is unclear what step 3’) is referring to when claim 18 depends on claim 1 wherein claim 1 does not recite a “step 3’) in the claim. In addition, claim 18 recites “…wherein…the T cell medium comprises….”. There is insufficient antecedent basis for “the T cell medium” in the claim.
Response to Arguments
In the Reply of 3/11/26, Applicant indicates this rejection should be withdrawn because amendments clarify the claims.
The amendments to the claims and the arguments found in the Reply of 3/11/26 have been carefully considered, but are not deemed persuasive. In regards to the indication this rejection should be withdrawn because amendments clarify the claims, the examiner disagrees. Claim 18 remains unclear for the reasons stated above.
New Claim Objections Necessitated by Amendments
Claims 1, 10 and 14 are objected to because of the following informalities:
Claims 1, 10 and 14 appear to have the following typographical error where the punctuation “:” is missing for “SEQ ID NO25”. For consistency, it is suggested that this be amended to “SEQ ID NO:25”.
Appropriate correction is required.
New Rejections Necessitated by Amendments
Claim Rejections - 35 USC § 103 (first)– Necessitated by Amendments
Claims 1-9 and 20 are rejected under 35 U.S.C. 103(a) as being unpatentable over Li et al. (Science. 2010 Jul 2;329(5987):85-9) in view of Ha et al. (Leukemia 2017 Feb 24;31(11):2503–2514), Pereira et al. (US20200017832A1 Date Published 2020-01-16; Application US16/342,803, Effective Filing Date 2019-04-17) and Zhang et al. (Bioengineered 2016, 7(3), 166–174).
Li et al. teaches that when Bcl11b gene was deleted in mice through conditional knockout (Bcl11b flox/flox), T cells from all developmental stages acquired NK cell properties (Abstract and Pg. 85 Column right Paragraphs first and second and up to Pg. 86 Column left Paragraph spanning). Li et al. also teaches that these killer cells that were reprogrammed from T cells were named as “induced T-to–natural killer cells”, “ITNK cells” or “ITNKs” (Pg 86 Column left Paragraph spanning). Li et al. further teaches that ITNKs could be produced from mature T cells, including CD4+ T cells, CD8+ T cells, and ϒδ T cells, and many ITNKs (NKp46+) were found growing in CD8+ T cell cultures which retained TCRβ on the cell surface (Pg. 86 Column, middle, Paragraph, first). They teach that these ITNK cells were morphologically and genetically similar to conventional NK cells, were able to kill tumor cells in vitro, and effectively prevented tumor metastasis in vivo (Abstract and Fig. 3E and 3F). Therefore, Li et al. teaches that mouse mature T cells could be reprogrammed, and that they retained functions of T cells and have markers and functions of NK cells.
Li et al. does not specifically teach reprogramming of human T cells that involves deletion of a specific 20 base pair target sequence that is knocked out from the BCL11B gene of human T cells, wherein the target sequence is selected from instant SEQ ID NOs: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46 and 49 as recited in instant claim 1. Li et al. also does not specifically teach that the human BCL11B gene knockout is performed by CRISPR/CAS9.
However, these deficiencies are made up in the teachings of Ha et al., Pereira et al. and Zhang et al.
Ha et al. teaches that human BCL11B is essential for T-lineage commitment, particularly the repression of NK potentials (Abstract and Figure 3). They teach that BCL11B gene expression is induced during the initial stages of T-cell differentiation in the human thymus and upregulation of BL11B is associated with T-lineage commitment and subsequence differentiation (Figure 1). They also teach that increased number of NK cells is seen with BCL11B insufficiency in knockdown experiments (Figures 3c and d. Pg 2505 column right lines 17-19 and Pg 2506 column left lines 7-9).
Pereira et al. teaches compositions, nucleic acid constructs, methods and kits for reprogramming cells to the dendritic cell state or antigen presenting cell state that can be used for immunotherapy applications. (Abstract). They teach that cells to be reprogrammed can be human cells (Paragraphs [0022] and [0042]). They also teach the nucleotide sequence of human BCL11B as set forth in SEQ ID NO: 53 (length 8044 bp, ID: NM_138576.3; Pg 128-132). When comparing SEQ ID NO: 53 of Pereira et al. to the target sequences of instant SEQ ID NOs: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46 and 49, it is noted that all of the instant SEQ ID NOs match fully either in the forward or reverse direction to SEQ ID NO: 53 as taught by Pereira et al., except for instant SEQ ID NO: 10 which matched all but one residue (residues 1-19 matched) to SEQ ID NO: 53 of Pereira et al.
Zhang et al. teaches that through guidance of a 20 nucleotide RNA (gRNA), CRISPR-Cas9 finds and cuts target protospacer DNA precisely 3 base pairs upstream of a PAM (Protospacer Adjacent Motif) (Abstract). They teach that SpCas9 recognizes the 3 nucleotide PAM of NGG, where "N" is any nucleobase (Pg 171 column right paragraph first lines 12-13). They also teach the CRISPR-Cas9 technology is applicable to a variety of human cells for potential utility in medicine (Pg 166 column left paragraph first lines 10-13). Upon close inspection of the human BCL11B gene nucleotide sequence as taught by Pereira et al., it is noted that the target sequences recited in instant claim 1 or the complement reverse sequences are upstream of the NGG 3-nucleotide PAM that is recognized by SpCas9 (see Table 1 below).
Table 1: Target sequences as recited in instant claim 1 (nucleotide sequences and SEQ ID NOs) and their proximity to the 3-nucleotide “NGG” PAM sequence.
PNG
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628
736
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Greyscale
One of ordinary skill in the art would have been motivated, with a reasonable expectation of success, to perform a combined method to generate human immune killer lymphocytes by reprogramming mature T cells through knocking out the Bcl11b gene from mouse T cells as taught by Li et al., replacing mature mouse T cells with mature human T cells as taught by Ha et al., by using the amino acid sequence of human BCL11B gene as taught by Pereira et al. to select a target sequence that is 20 nucleotides in length that can be precisely knocked out by performing CRISPR/Cas9 due to its location that is 3 base pairs upstream of a PAM sequence as taught by Zhang et al. (including just any target sequence listed above in Table 1) to generate human reprogrammed immune killer lymphocytes which retain markers and functions of human T cells and have acquired markers and functions of NK cells, and to administer an effective amount of the generated human reprogrammed immune killer lymphocytes to a human subject with a tumor for the purpose of killing human tumor cells in vivo. One of ordinary skill in the art would have been motivated, with a reasonable expectation of success, to generate said human immune killer lymphocytes that have been reprogrammed from human mature T cells due to the predictable advantages of said cells for an improved cell-based therapy as a cancer therapeutic for human cancer because as taught by Li et al., ITNKs derived from mice mature T cells can be extensively expanded, can effectively kill tumor cells in vitro and can eliminate metastatic cells in mice while sparing normal cells, and as taught by Pereira et al. human immune cells can be reprogrammed for improved immunotherapy applications . (A) Combining prior art elements according to known methods to yield predictable results; (B) Simple substitution of one known element for another to obtain predictable results; and (G) Some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. See MPEP 2143. Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art, absent unexpected results.
Regarding instant claims 3-9, it is acknowledged that the cited references do not teach that immune killer lymphocytes or ITNK cells generated by reprogramming mature human T cells through deletion of BCL11B gene from the mature human T cells express functional CD3 and NKp30 or have expression of other genes, whether these are up- or down-regulated. However, it appears such expression would occur in immune killer lymphocytes and ITNK cells generated by reprogramming mature human T cells by deleting BCL11B gene from the mature human T cells. The examiner takes the position that this is not a property having a significance greater than that of the expected property of a predicted therapeutic effect of the ITNK cells as taught by Li et al. Therefore, recitation that immune killer lymphocytes have the property of having up-regulated or down-regulated genes in the claims is not sufficient to rebut obviousness of using mature human T cells to generate these immune killer lymphocytes or ITNK cells when said cells are expected to have the equal or greater property of expected therapeutic benefit. See MPEP 716.02(c). Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art, absent unexpected results. Further, see In re Baxter Travenol Labs., 952 F.2d 388, 21 USPQ2d 1281 (Fed. Cir. 1991), where the court held that the fact that another advantage would flow naturally from following the suggestion of the prior art cannot be the basis for patentability when the differences would otherwise be obvious.
Claim Rejections - 35 USC § 103 (second)– Necessitated by Amendments
Claims 1-18 and 20 are rejected under 35 U.S.C. 103(a) as being unpatentable over Li et al. (Science. 2010 Jul 2;329(5987):85-9), Ha et al. (Leukemia 2017 Feb 24;31(11):2503–2514), Pereira et al. (US20200017832A1 Date Published 2020-01-16; Application US16/342,803, Effective Filing Date 2019-04-17) and Zhang et al. (Bioengineered 2016, 7(3), 166–174), as applied to claims 1-9and 20, and in further view of Berenson et al. (CN 1556854 Date Published 2004-12-22) and de Jong et al. (Immunology. 1991 Oct;74(2):175-82).
Please note that the citations from Berenson et al. below reference positions in the translation attached as an office action appendix.
The teachings of Li et al., Ha et al., Pereira et al. and Zhang et al. render obvious instant claims 1-9 and 20 as discussed above in the first 103 which is incorporated here in its entirety.
Li et al., Ha et al., Pereira et al. and Zhang et al. do not specifically teach the immune killer lymphocytes according to instant claim 1, wherein the reprogramming of the human T cells comprises: 1) activating mature human T cells; 2) knocking out a target sequence from BCL11B gene of the activated mature human T cells in step 1); and 3) culturing the cells in step 2) with a T cell culture medium. They also do not specifically teach, wherein in step 1), the activation is performed using an anti-human CD3 antibody, an anti-human CD28 antibody, and an anti-human CD2 antibody; wherein in step 2), the BCL11B gene knockout is performed by CRISPR/CAS9; and wherein in step 3), the T cell medium comprises IL-2. They further do not specifically teach a method for preparing the immune killer lymphocytes according to instant claim 1, comprising: 1') activating human T cells; 2') knocking out a target sequence from BCL11B gene of the activated mature human T cells in step 1'); and 3') culturing the cells in step 2') with a T cell culture medium; wherein the human T cells are mature human T cells or a cell population comprising mature human T cells; wherein in step 1'), the activation is performed using an anti-human CD3 antibody, an anti-human CD28 antibody, and an anti-human CD2 antibody; wherein in step 2'), the BCL11B gene knockout is performed by CRISPR/CAS9; wherein in step 3'), the T cell medium comprises IL-2.
However, these deficiencies are made up in the teachings of Berenson et al. and de Jong et al.
Berenson et al. teaches a method for activating and amplifying a population of T cells (Abstract and Pg. 14 Paragraph second “Brief description of the invention”). Berenson et al. also teaches cellular activation enhances cell function (see “Stimulation of cell populations). They teach that T cells are exposed to surfaces that are attached to factors including a first, second and third factor to induce T cell proliferation, where these factors include anti-CD3 antibody, anti-CD2 antibody and anti-CD28 antibody (Pg. 15 Paragraph second). They also teach that suitable T cell culture conditions include cultures comprising interleukin-2 (IL-2) (Pg. 40 Paragraph, first).
de Jong et al. teaches that stimulation of purified T lymphocytes with either graded densities of immobilized anti-CD3 monoclonal antibodies (mAb) or with increasing amounts of anti-CD28 mAb in the presence of a constant concentration of anti-CD2 mAb showed optimal cytolytic T lymphocyte differentiation with TcR/CD3 or CD2/CD28 engagement (Abstract).
One of ordinary skill in the art would have been motivated, with a reasonable expectation of success, to perform a combined method to generate activated mature human immune killer T cell lymphocytes that are reprogrammed from human T cells through deleting a target sequence on BCL11B gene as taught by Li et al., Ha et al., Pereira et al. and Zhang et al. as described in the first 103 rejection above, and by employing the steps of :
1) activating the mature human T cells through contacting the mature T cells with a combination of anti-CD3, anti-CD2 and anti-CD28 antibodies that bind CD3, CD2, and CD28 respectively on mature T cells as taught by Berenson et al. and de Jong et al.;
2) performing target sequence knockout of BCL11B gene on the activated T cells of step 1) by performing the combined method of Li et al., Ha et al., Pereira et al. and Zhang et al. with the activated T cells to reprogram the activated T cells to become activated mature human immune killer T cells.; and
3) culturing the activated killer T cells in a medium comprising IL-2 as taught by Berenson et al. to sustain the cells prior to administering the cells to a subject with a tumor,
because Berenson et al. teaches that the three anti-CD3, anti-CD2 and anti-CD28 antibodies are factors that induce T cell proliferation that can activate and amplify a population of T cells for enhanced cell function, Berenson et al. also teaches that cultures comprising IL-2 are suitable for culturing T cells, de Jong et al. teaches optimal cytolytic T lymphocyte stimulation and differentiation is achieved by exposing said cells to anti-CD3/anti-CD2 or anti-CD28/anti-CD2 monoclonal antibodies, and performing the combined method of Li et al., Ha et al., Pereira et al. and Zhang et al. with the activated T cells precisely targets deletion of BCL11B using the CRISPR/Cas9 system to reprogram the activated T cells to become activated mature human immune killer T cells for immunotherapy applications . This is an example of (A) Combining prior art elements according to known methods (activating T cells with antibodies and knocking-out BCL11B) to yield predictable results of generating cytolytic ITNK cells; and (G) Some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. See MPEP 2143. Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art, absent unexpected results.
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.
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/YIE-CHIA LEE (TONYA)/Examiner, Art Unit 1642
/SEAN E AEDER/Primary Examiner, Art Unit 1642