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 19 May 2026 has been entered. Claims 1-29, 32, 36, and 39-43 are pending. Claims 1-7, 12-14, 28, 32, 36, and 40 have been amended, while claims 30-31 and 35 have been cancelled without prejudice or disclaimer and claims 41-43 have been newly added. Therefore, prosecution on the merits continues for claims 1-29, 32, 36, and 39-43. All arguments have been fully considered with the status of each prior ground of rejection set forth below.
Status of Prior Rejections/Response to Arguments
RE: Objection to the Drawings
The replacement drawing sheets filed 19 May 2026 are acknowledged and entered into the application file. Accordingly, the replacement drawing sheets remove the figure templates and thus obviate the objection of record.
Therefore, the objection is withdrawn.
RE: Objection to claim 28
Applicant’s amendments to instant claim 28 correct the minor informality and thus obviate the objection of record.
Therefore, the objection is withdrawn.
RE: Rejection of claims 12-14 under 35 USC 112(b)
Applicant’s amendments to each of instant claims 12-14 correcting the antecedent basis obviate the rejections of record.
Therefore, the rejections are withdrawn.
RE: Rejection of claims 30-31, 35, and 40 under 35 USC 101
The cancellation of instant claims 30-31 and 35 renders the rejection of those claims moot. For the remaining claim, Applicant’s arguments filed 19 May 2026 have been fully considered but they are not persuasive.
Applicant has traversed the rejection, asserting in Page 9 of the Remarks filed 19 May 2026 that the population of cells recited in instant claim 40 is enriched in CD4 single positive cells relative to the population of T cells found in nature. Applicant has further traversed the rejection, asserting that this structural difference imparts functionalities to the claimed invention that are not found in the naturally occurring T cell population, namely that the single positive cells are partially matured in the direction of becoming effector T cell. In response, the Examiner respectfully submits that altering the number of CD4 single positive cells within a composition does not structurally change the CD4 single positive cells, per se. Therefore, there are no structural differences between the instantly claimed CD4 single positive cells and those that are naturally occurring. With that, as stated in Pages 8-9 of the Office action filed 19 November 2025, the incorporation of the CD4 single positive cells into a pharmaceutical composition does not modify or transform the CD4 single positive cells, nor apply the cells for a particular treatment of medical condition, nor impose a meaningful limit on the cells recited therein.
Therefore, the rejection is maintained.
RE: Rejection of claims 1-7, 12-14, and 30 under 35 USC 102(a)(1) over Takahama et al
The cancellation of claim 30 renders the rejection moot for that claim. For the remaining claims, Applicant’s amendments to independent claim 1 requiring the concentration of PMA within the culture medium to be between 0.00625 to 0.0125 μg/mL and the concentration of ionomycin to be between 0.125 to 0.250 μg/mL obviates the rejection of record.
Therefore, the rejection is withdrawn.
However, Applicant’s remarks are addressed in so far as the are applicable to the claims as currently written:
Applicant has traversed the rejection, asserting in Pages 9-10 of the Remarks filed 19 May 2026 that the disclosure of Takahama et al fails to recite the specific concentrations of PMA and ionomycin in culture with CD4+CD8+ T cells. In response, the Examiner respectfully submits that while Takahama et al fail to reduce to practice the culture of CD4+CD8+ T cells in the recited concentration ranges, Takahama et al do disclose that concentrations of PMA between 0.016 to 0.16 μM and ionomycin between 0.13 to 1.3 μM allow for positive enrichment of CD4+ T cells from CD4+CD8+ T cells. See Page 1509-1510 of Takahama et al. Therefore, the ordinary artisan would have reasonably arrived at the claimed ranges through routine experimentation. See MPEP § 2144.05.
RE: Rejection of claims 1, 3, 6-7, 12-16, 27-28, 30-32, and 40 under 35 USC 102(a)(2) over Kaneko et al
The cancellation of claims 30-31 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 1 requiring the concentration of PMA within the culture medium to be between 0.00625 to 0.0125 μg/mL and the concentration of ionomycin to be between 0.125 to 0.250 μg/mL obviates the rejection of record.
It is of note, however, that independent claim 40 as currently written comprises a product-by-process limitation that is not overcome by the amendments to independent claim 1 as incorporated. See Claim Interpretation below. Furthermore, Applicant has not particularly argued the merits of the product-by-process limitation in regards to independent claim 40.
Therefore, the rejection is withdrawn for claims 1, 3, 6-7, 12-16, 27-28, 30-32 and maintained for claim 40.
RE: Rejection of claims 35-36 and 39 under 35 USC 102(a)(1) and 35 USC 102(a)(2) over Gorochov et al
The cancellation of claim 35 renders the rejection moot for that claim. For the remaining claims, Applicant’s amendments to independent claim 36 requiring the method of treating a patient in need of immunosuppression to comprise administering a population enriched in CD4+ Treg cells obtained by the method of claim 1 obviates the rejection of record.
Therefore, the rejection is withdrawn.
RE: Rejection of claims 1, 3, 6-8, 10-16, 27-28, 30-32, 35-36 and 39-40 under 35 USC 103 over Kaneko et al in view of Schmidt et al
The cancellation of claims 30-31 and 35 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 1 requiring the concentration of PMA within the culture medium to be between 0.00625 to 0.0125 μg/mL and the concentration of ionomycin to be between 0.125 to 0.250 μg/mL obviates the rejection of record.
Therefore, the rejection is withdrawn.
RE: Rejection of claims 1, 3, 6-7, 12-18, 25, 27-32, and 40 under 35 USC 103 over Kaneko et al in view of Gschweng et al
The cancellation of claims 30-31 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 1 requiring the concentration of PMA within the culture medium to be between 0.00625 to 0.0125 μg/mL and the concentration of ionomycin to be between 0.125 to 0.250 μg/mL obviates the rejection of record.
Therefore, the rejection is withdrawn.
RE: Rejection of claims 1, 3, 6-7, 12-32, and 40 under 35 USC 103 over Kaneko et al in view of Gschweng et al, and further in view of Burleigh et al
The cancellation of claims 30-31 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 1 requiring the concentration of PMA within the culture medium to be between 0.00625 to 0.0125 μg/mL and the concentration of ionomycin to be between 0.125 to 0.250 μg/mL obviates the rejection of record.
Therefore, the rejection is withdrawn.
RE: Rejection of claims 1, 3, 6-7, 12-32, 35-36, and 39-40 under 35 USC 103 over Kaneko et al in view of Conway et al
The cancellation of claims 30-31 and 35 renders the rejection moot for those claims. For the remaining claims, Applicant’s amendments to independent claim 1 requiring the concentration of PMA within the culture medium to be between 0.00625 to 0.0125 μg/mL and the concentration of ionomycin to be between 0.125 to 0.250 μg/mL obviates the rejection of record.
Therefore, the rejection is withdrawn.
New/Maintained Grounds of Rejection
Claim Interpretation
Under the broadest reasonable interpretation of each claim, all of the “optional” limitations are not required.
Furthermore, instant claim 40 is directed to a pharmaceutical composition comprising a population of cells enriched for CD4 single positive cells and a pharmaceutically acceptable carrier, wherein the population of cells is obtained by the method of claim 1. This is a product-by-process limitation. Product-by-process limitations are considered only in so far as the method of production affects the structure of the final product. In the instant case, there is no evidence that the production method of instant claim 1 imparts any particular structure or significance to the enriched CD4 single positive cells. Thus, the claim will be interpreted as if a pharmaceutical composition comprising CD4+ cells derived from any source or production method fulfills the recited claim limitations. See MPEP § 2113.
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.
Claim 40 is 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 40: The instant claim recites the limitation "the population of cells enriched in CD4 single positive cells" in Lines 1-2. There is insufficient antecedent basis for this limitation in the claim, as there is no prior recitation of a population of cells within the instant claim.
Appropriate correction is required.
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.
Claim 40 remains rejected under 35 U.S.C. 101 because the claimed invention is directed to a nature-based product without significantly more.
Claim 40 is directed to a pharmaceutical composition comprising a population of cells enriched for CD4 single positive cells and a pharmaceutically acceptable carrier, wherein the population of cells enriched for CD4 single positive cells is obtained by the method of claim 1. Instant claim 40 comprises a product-by-process limitation, as can be observed in the Claim Interpretation section above and is incorporated in its entirety herein.
The test for 101 eligibility of judicial exceptions can be found at MPEP § 2106:
“First, the claimed invention must be to one of the four statutory categories. 35 U.S.C. 101 defines the four categories of invention that Congress deemed to be the appropriate subject matter of a patent: processes, machines, manufactures and compositions of matter.”
“Second, the claimed invention also must qualify as patent-eligible subject matter, i.e., the claim must not be directed to a judicial exception unless the claim as a whole includes additional limitations amounting to significantly more than the exception. The judicial exceptions (also called "judicially recognized exceptions" or simply "exceptions") are subject matter that the courts have found to be outside of, or exceptions to, the four statutory categories of invention, and are limited to abstract ideas, laws of nature and natural phenomena (including products of nature). Alice Corp. Pty. Ltd. v. CLS Bank Int'l, 573 U.S. 208, 216, 110 USPQ2d 1976, 1980 (2014) (citing Ass'n for Molecular Pathology v. Myriad Genetics, Inc., 569 U.S. 576, 589, 106 USPQ2d 1972, 1979 (2013).”
“The Supreme Court in Mayo laid out a framework for determining whether an applicant is seeking to patent a judicial exception itself, or a patent-eligible application of the judicial exception. See Alice Corp., 573 U.S. at 217-18, 110 USPQ2d at 1981 (citing Mayo, 566 U.S. 66, 101 USPQ2d 1961). This framework, which is referred to as the Mayo test or the Alice/Mayo test, is discussed in further detail in subsection III, below. The first part of the Mayo test is to determine whether the claims are directed to an abstract idea, a law of nature or a natural phenomenon (i.e., a judicial exception). Id. If the claims are directed to a judicial exception, the second part of the Mayo test is to determine whether the claim recites additional elements that amount to significantly more than the judicial exception. Id. citing Mayo, 566 U.S. at 72-73, 101 USPQ2d at 1966). The Supreme Court has described the second part of the test as the "search for an 'inventive concept'". Alice Corp., 573 U.S. at 217-18, 110 USPQ2d at 1981 (citing Mayo, 566 U.S. at 72-73, 101 USPQ2d at 1966).”
Examiners should determine whether a claim satisfies the criteria for subject matter eligibility by evaluating the following steps outlined in a flow chart at MPEP 2106(III) and 2106.04(II)(A):
Step 1: is the Claim to a process, machine, manufacture or composition of matter? If Yes, proceed to Step 2A;
Step 2A, prong one: Is the Claim directed to a law of nature, a natural phenomenon (product of nature), or an abstract idea? If Yes, proceed to Step 2A, prong two;
Step 2A, prong two: Does the claim recite additional elements that integrate the judicial exception into a practical application? If No, proceed to Step 2B;
Step 2B: Does the claim recite additional elements that amount to significantly more (an inventive concept) than the judicial exception? If No, the claim is not eligible subject matter under 35 USC 101.
With regard to Step 1: YES, the claims are each directed to a composition of matter, or product.
With regard to Step 2A, prong one: YES, the claims are directed to pharmaceutical composition comprising an enriched population of CD4 single positive cells, which are nature-based products. More specifically, since the instantly claimed CD4+ cells comprise a product-by-process limitation – see Claim Interpretation section above – the claimed CD4+ cells are compared to the closest naturally occurring counterpart, which are CD4 single positive cells, per se. Thus, there is no marked different between the claim and product of nature. See, for example, Paragraphs [0002]-[0003] of Conway et al (WO 2021/092581 A1, of record on IDS filed 23 July 2024).
With regard to Step 2A, prong two: No, the claims do not include any additional elements that integrate the judicial exceptions into a practical application. Integration into a practical application requires additional elements in the claim to apply, rely on, or use the judicial exception in a manner that imposes a meaningful limit on the judicial exception, such that the claim is more than a drafting effort designed to monopolize the exception.
The incorporation of naturally occurring CD4+ cells in a pharmaceutical composition does not modify or transform the naturally occurring CD4+ cells, nor apply the cells for a particular treatment or medical condition, nor impose a meaningful limit on the cells recited therein.
With regard to Step 2B: No, the claim does not provide any additional elements that amount to significantly more (an inventive concept) than the judicial exception.
Taken together, the claim encompasses a natural product. The judicial exception is not integrated into practical applications as iterated above. The claim does not include additional elements
that are sufficient to amount to significantly more than the judicial exception as iterated above.
Claim Rejections - 35 USC § 102
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 the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claim 40 remains rejected under 35 U.S.C. 102(a)(2) as being anticipated by Kaneko et al (US 2022/0411752 A1). Kaneko et al has an effective filing date of 01 November 2019.
Kaneko et al disclose methods for producing an IL-4 non-secreting and IFN-γ secreting (Th1-type)
or IFN-γ non-secreting and IL-4 secreting (Th2-type) CD4 single-positive T cell (CD4SP T cell) (Abstract).
As such, Kaneko et al disclose the generation of an artificial thymic organoid (ATO) from T-iPSC-derived HSPCs, wherein the ATOs produce CD4+CD8+ T cells that are cultured in medium comprising PMA and ionomycin (Paragraphs [0003], [0005], [0013], [0064], [0066], [0078], [0080], [0133]-[0147], [0168]-[0169], [0187]-[0199]). Kaneko et al further disclose that the iPSC is a universalized iPSC wherein an HLA gene has been knocked out (Paragraph [0133]).
Kaneko et al further disclose that CD4+ T cells are isolated from culture via FACS (Paragraphs [0169], [0199]).
Kaneko et al further disclose a pharmaceutical composition comprising the isolated Th1-type or Th2-type CD4+ T cells and a pharmaceutically acceptable carrier (Paragraphs [0002], [0016], [0097], [0179]-[0180], [0184]-[0185]).
Accordingly, Kaneko et al anticipate the claims as follows:
Regarding claim 40: Instant claim 40 comprises product-by-process. The effect of the product-by-process language is discussed above – see Claim Interpretation – and is incorporated herein in its entirety.
Accordingly, Kaneko et al disclose a pharmaceutical composition comprising the isolated Th1-type or Th2-type CD4+ T cells and a pharmaceutically acceptable carrier. This therefore reads on the pharmaceutical composition of the instant claim.
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 1-5 are rejected under 35 U.S.C. 103 as being unpatentable over Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025).
Takahama et al is considered prior art under 35 USC 102(a)(1).
Regarding claims 1 and 5: Takahama et al disclose the culture of CD4+CD8+ T cells in a culture medium comprising PMA and ionomycin for five days, wherein CD4+ T cells are subsequently isolated from culture using FACS (Pages 1508-1511; Figures 1-2, 4; Table 1). Takahama et al further disclose that the PMA is at a concentration of 0.0016 to 1.6 μM, and ionomycin is at a concentration of 0.13 to 1.3 μM (Pages 1509-1510; Figure 4). It is of note that the PMA concentration range equates to 0.000987 μg/mL to 0.987 μg/mL, while the ionomycin concentration range equates to 0.0971 μg/mL to 0.971 μg/mL (see calculations provided on Page 2 of third party observation for application number EP 2021810847.0, of record on IDS filed 10 July 2025).
However, Takahama et al do not exemplify or reduce to practice the culture of a starting population of CD4+CD8+ T cells in a medium comprising 0.00625 μg/mL to 0.0125 μg/mL PMA and 0.125 μg/mL to 0.250 μg/mL ionomycin, as required by instant claim 1.
Therefore, it would have been prima facie obvious to have modified the culture method of Takahama et al such that the CD4+CD8+ T cells are cultured in a medium comprising 0.00625 μg/mL to 0.0125 μg/mL PMA and 0.125 μg/mL to 0.250 μg/mL ionomycin. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to culture the CD4+CD8+ T cells in a medium having a PMA concentration between 0.00625 μg/mL to 0.0125 μg/mL and an ionomycin concentration between 0.125 μg/mL to 0.250 μg/mL, as it would have been a matter of routine optimization in determining respective PMA and ionomycin concentrations that allow for the positive selection of CD4+CD8- T cells (Takahama et al: Pages 1512-1513). See MPEP § 2144.05(II). Furthermore, the ordinary artisan would have had a reasonable expectation of success given that the disclosure of Takahama et al recites and utilizes different concentrations of PMA and ionomycin that fall within the claimed concentration ranges. See MPEP § 2143(I)(G).
Consequently, Takahama et al render obvious the culture of CD4+CD8+ T cells in a culture medium comprising 0.00625 μg/mL to 0.0125 μg/mL PMA and 0.125 μg/mL to 0.250 μg/mL ionomycin for five days (claim 5). This therefore renders obvious the method of obtaining a population of cells enriched for CD4 single positive T cells of instant claim 1.
Regarding claims 2-4: As aforementioned in the discussion of claim 1, Takahama et al disclose that the PMA is at a concentration of 0.0016 to 1.6 μM, and ionomycin is at a concentration of 0.13 to 1.3 μM. As these molarity ranges encompass a PMA concentration of 0.00625 μg/ml and an ionomycin concentration of 0.125 μg/ml (see calculations provided on Page 2 of third party observation for application number EP 2021810847.0, of record on IDS filed 10 July 2025), this therefore has a weight ratio of PMA to ionomycin of 1:20 (claims 3-4) and renders obvious the method of instant claim 2. See MPEP § 2144.05(I).
Claims 1-7, 12-15, 27, 32, 36, 39, and 42 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025).
Crooks et al is considered prior art under 35 USC 102(a)(1) and 35 USC 102(a)(2). Takahama et al is considered prior art under 35 USC 102(a)(1).
Regarding claims 1 and 5: Crooks et al disclose methods and compositions for the production of T cells (Abstract).
As such, Crooks et al disclose the generation of an artificial thymic organoid (ATO) from iPSC-derived HSPCs, wherein the ATOs produce CD4+CD8+ T cells that are cultured in medium comprising PMA and ionomycin (Paragraphs [0005]-[0007], [0025], [0029]-[0030], [0037], [0041]-[0042], [0054], [0064], [0069], [0083], [0091], [0102], [0111], [0153], [0208], [0336], [0338], [0350], [0354]-[0355], [0359], [0373], [0379]-[0380], [0416]; Figures 6B, 25). Crooks et al further disclose that CD4 single positive T cells are isolated from the ATOs (Paragraphs [0081], [0083], [0089], [0379]; Figure 12).
Crooks et al do not disclose that the concentration of PMA and ionomycin within the culture medium is 0.00625 μg/mL to 0.0125 μg/mL and 0.125 μg/mL to 0.250 μg/mL, respectively, as required by instant claim 1.
Takahama et al, however, disclose the culture of CD4+CD8+ T cells in a culture medium comprising PMA and ionomycin for five days, wherein CD4+ T cells are subsequently isolated from culture using FACS (Pages 1508-1511; Figures 1-2, 4; Table 1). Takahama et al further disclose that the PMA is at a concentration of 0.0016 to 1.6 μM, and ionomycin is at a concentration of 0.13 to 1.3 μM (Pages 1509-1510; Figure 4). It is of note that the PMA concentration range equates to 0.000987 μg/mL to 0.987 μg/mL, while the ionomycin concentration range equates to 0.0971 μg/mL to 0.971 μg/mL (see calculations provided on Page 2 of third party observation for application number EP 2021810847.0, of record on IDS filed 10 July 2025).
Therefore, it would have been prima facie obvious to have modified the method of Crooks et al such that the CD4+CD8+ T cells comprised within the ATO are cultured in medium comprising 0.00625 μg/mL to 0.0125 μg/mL PMA and 0.125 μg/mL to 0.250 μg/mL ionomycin, as detailed in Takahama et al. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to culture the CD4+CD8+ ATO cells in a concentration of PMA and ionomycin known to enhance CD4 single positive cell production, as the ATOs exhibited proportionately fewer CD4 single positive T cells relative to CD8 single positive T cells, which contrasts with the proportion naturally exhibited in the thymus (Crooks et al: Paragraph [0354]). Furthermore, the ordinary artisan would have had a reasonable expectation of success given that the disclosures of Crooks et al and Takahama et al both concern the culture of a population of cells comprising CD4+CD8+ T cells in medium comprising PMA and ionomycin for the eventual sorting of produced CD4+ T cells. See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al render obvious a method wherein iPSC-derived HSPCs form an ATO that produces CD4+CD8+ T cells, which are then cultured in a medium comprising 0.00625 μg/mL to 0.0125 μg/mL PMA and 0.125 μg/mL to 0.250 μg/mL ionomycin for five days (claim 5), and then sorted for CD4+ T cells. This therefore renders obvious the method of obtaining a population of cells enriched for CD4 single positive T cells of instant claim 1.
Regarding claims 2-4: As aforementioned in the discussion of claim 1, Takahama et al disclose that the PMA is at a concentration of 0.0016 to 1.6 μM, and ionomycin is at a concentration of 0.13 to 1.3 μM. As these molarity ranges encompass a PMA concentration of 0.00625 μg/ml and an ionomycin concentration of 0.125 μg/ml (see calculations provided on Page 2 of third party observation for application number EP 2021810847.0, of record on IDS filed 10 July 2025), this therefore has a weight ratio of PMA to ionomycin of 1:20 (claims 3-4) and renders obvious the method of instant claim 2 for the same reasons as discussed in the rejection of instant claim 1. See MPEP § 2144.05(I).
Regarding claims 6-7: Following the discussion of claim 1, Crooks et al further disclose that the CD4 single positive cells are immature (claim 6) CD4+ effector T cells (claim 7) (Paragraphs [0081], [0290], [0338], [0355], [0359], [0373], [0389]). This therefore reads on the method of the instant claims.
Regarding claim 12: Following the discussion of claim 7, Crooks et al further disclose that the CD4 single positive cells are isolated from the ATO via FACS (Paragraphs [0066], [0102], [0223], [0226], [0294], [0297]). This therefore reads on the method of the instant claim.
Regarding claims 13-14: Following the discussion of claim 1, Crooks et al further disclose that the CD4 single positive cells are regulatory T cells, wherein a subset have an upregulated expression of CD25 (Paragraphs [0030], [0037], [0042], [0061], [0064], [0069], [0083], [0102], [0355]; Figure 12). Crooks et al further disclose that a subset of CD4 single positive T cells expresses CD127 (Paragraphs [0355], [0395]). Therefore, it would have been prima facie obvious to have selected CD4 single positive T regulatory cells having various expression levels of CD25 and low expression of CD127, as one of ordinary skill in the art before the effective filing date of the invention would have been motivated to accurately phenotype the resulting CD4 single positive cells. Furthermore, it would not have been outside the skillset of the ordinary artisan to sort the resulting CD4 single positive cells based on CD25 and CD127 expression levels given the flow cytometry and FACS protocols provided in Crooks et al (Paragraphs [0383], [0431]). See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al render obvious a method wherein CD4+CD25+CD127low (claim 13), CD4+CD25highCD127low, and CD4+CD25lowCD127low (claim 14) regulatory T cells are isolated from the ATO culture via FACS. This therefore renders obvious the method of the instant claims.
Regarding claims 15 and 27: Following the discussion of claim 1, Crooks et al further disclose that the iPSCs are human (claim 27) iPSCs (claim 15) (Paragraphs [0025], [0042], [0208], [0273]). This therefore reads on the method of the instant claims.
Regarding claim 32: Following the discussion of claim 1, Crooks et al further disclose that isolated CD4+ effector T cells are administered to a patient suffering from cancer (Paragraphs [0034], [0052], [0268], [0328]-[0330]). This therefore renders obvious the method of instant claim 32 for the same reasons as discussed in the rejection of instant claim 1.
Regarding claims 36 and 39: Following the discussion of claim 1, Crooks et al further disclose that isolated CD4+ regulatory T cells are administered to a patient in need of immunosuppression that is suffering from an autoimmune disease (claim 39) (Paragraphs [0034], [0061]-[0062], [0329]). This therefore renders obvious the method of instant claim 36 for the same reasons as discussed in the rejection of instant claim 1.
Regarding claim 42: Following the discussion of claim 7, Crooks et al further disclose that the CD4 single can be CD25+ or CD25- (Paragraphs [0030], [0037], [0042], [0061], [0064], [0069], [0083], [0102], [0355]; Figure 12). Therefore, it would have been prima facie obvious to have selected CD4 single positive T effector cells having various expression levels of CD25, as one of ordinary skill in the art before the effective filing date of the invention would have been motivated to accurately phenotype the resulting CD4 single positive cells. Furthermore, it would not have been outside the skillset of the ordinary artisan to sort the resulting CD4 single positive cells based on CD25 expression levels given the flow cytometry and FACS protocols provided in Crooks et al (Paragraphs [0383], [0431]). See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al render obvious a method wherein CD4+CD25low effector T cells are isolated from the ATO culture via FACS. This therefore renders obvious the method of the instant claims.
Claims 1-8, 10-15, 27, 32, 36, 39, and 42-43 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025), and further in view of Schmidt et al (PLoS One, 2016, of record).
The discussion of Crooks et al as modified by Takahama et al regarding claim 1 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Crooks et al as modified by Takahama et al render obvious claims 1-7, 12-15, 27, 32, 36, 39, and 42. Schmidt et al is considered prior art under 35 USC 102(a)(1).
Regarding claims 8 and 11: Following the discussion of claim 1 above, Crooks et al further disclose that the CD4 single positive cells are regulatory T cells (Paragraphs [0030], [0037], [0042], [0061], [0064], [0069], [0083], [0102], [0355]; Figure 12).
The combination of Crooks et al and Takahama et al fail to teach that the CD4+ T cells are further cultured in a second medium comprising TGF-β and all trans-retinoic acid (ATRA) to thereby obtain a population of Tregs, as required by instant claim 8.
Schmidt et al, however, disclose a method of inducing Tregs, wherein CD4+ T cells are cultured in the medium comprising TGF-β and ATRA for six days (Abstract; Pages 4, 20; Figure 1).
Therefore, it would have been prima facie to have modified the method of Crooks et al in view of Takahama et al such that the CD4+ T cells are cultured in medium comprising TGF-β and ATRA to obtain a population of Tregs, as detailed in Schmidt et al. One of ordinary skill in the art would have been motivated to utilize a culture medium composition that is known to generate Tregs, and would have had a reasonable expectation of success given the disclosure of Schmidt et al. See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al and Schmidt et al render obvious a method of generating Tregs, wherein the CD4+ T cells are cultured in medium comprising TGF-β and ATRA for six days (claim 11). This therefore renders obvious the method of instant claim 8.
Regarding claim 10: Following the discussion of claim 8, Schmidt et al further disclose that the medium comprising TGF-β and ATRA is serum-free X-Vivo 15 medium, which is for the culture of the T cells (Page 20). This therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of instant claim 8.
Regarding claim 43: Following the discussion of claim 8, Crooks et al further disclose that the CD4+ regulatory T cells are isolated from the ATO via FACS (Paragraphs [0066], [0102], [0223], [0226], [0294], [0297]). This therefore reads on the method of the instant claim.
Claims 1-15, 27, 32, 36, 39, and 42-43 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025) and Schmidt et al (PLoS One, 2016, of record), and further in view of Gorochov et al (WO 2013/050596 A1, of record).
The discussion of Crooks et al as modified by Takahama et al regarding claim 1 and Crooks et al as modified by Takahama et al and Schmidt et al regarding claim 8 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Crooks et al as modified by Takahama et al render obvious claims 1-7, 12-15, 27, 32, 36, 39, and 42. Crooks et al as modified by Takahama et al and Schmidt et al render obvious claims 1-8, 10-15, 27, 32, 36, 39, and 42-43. Gorochov et al is considered prior art under 35 USC 102(a)(1) and 35 USC 102(a)(2).
Regarding claim 9: As aforementioned in the discussion of claim 8 above, Crooks et al as modified by Takahama et al and Schmidt et al render obvious a method of generating Tregs, wherein the CD4+ T cells are cultured in medium comprising TGF-β and ATRA.
Schmidt et al further disclose that the medium can further comprise IL-2, an anti-CD3 antibody, and an anti-CD28 antibody, wherein the anti-CD3 and anti-28 antibodies are utilized to stimulate the CD4+ T cells (Page 20).
The combination of Crooks et al, Takahama et al, and Schmidt et al fail to teach that the medium further comprises an anti-CD2 antibody, as required by instant claim 9.
Gorochov et al, however, disclose the stimulation of Tregs with anti-CD3, anti-CD28, and anti-CD2 antibodies (Pages 5-6, 12-13).
Therefore, it would have been prima facie to have modified the method of Crooks et al in view of Takahama et al and Schmidt et al such that the culture medium further comprises an anti-CD2 antibody, as detailed in Gorochov et al. One of ordinary skill in the art would have been motivated to strengthen the stimulation of the CD4+ T cells via the addition of the anti-CD2 antibody (Gorochov et al: Pages 5-6), and would have had a reasonable expectation of success given that the disclosures of Schmidt et al and Gorochov et al are concerned with the generation and maintenance of CD4+ Tregs. See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al, Schmidt et al, and Gorochov et al render obvious a method of generating Tregs, wherein the CD4+ T cells are cultured in medium comprising TGF-β, ATRA, IL-2, an anti-CD2 antibody, an anti-CD3 antibody, and an anti-CD28 antibody. This therefore renders obvious the method of the instant claim.
Claims 1-7, 12-16, 27-28, 32, 36, 39, and 42 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025), and further in view of Kaneko et al (US 2022/0411752 A1, of record).
The discussion of Crooks et al as modified by Takahama et al regarding claims 1, 15, and 27 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Crooks et al as modified by Takahama et al render obvious claims 1-7, 12-15, 27, 32, 36, 39, and 42. Kaneko et al is considered prior art under 35 USC 102(a)(2), with an effective filing date of 01 November 2019.
Regarding claim 16: As aforementioned in the discussion of claim 15 above, Crooks et al disclose that the iPSCs from which the starting population of iPSC-derived HSPCs for the generation of the ATO are from are human iPSCs.
The combination of Crooks et al and Takahama et al fail to teach that the human iPSCs are T-cell derived iPSCs, as required by instant claim 16.
Kaneko et al, however, disclose the generation of an artificial thymic organoid (ATO) from T-iPSC-derived HSPCs, wherein the ATOs produce CD4+CD8+ T cells that are cultured in medium comprising PMA and ionomycin (Paragraphs [0003], [0005], [0013], [0064], [0066], [0078], [0080], [0133]-[0147], [0168]-[0169], [0187]-[0199]).
Therefore, it would have been prima facie obvious to have substituted the iPSCs of Crooks et al with the T-cell derived iPSCs of Kaneko et al, as doing so would have been a simple substitution of one ATO-producing iPSC for another. See MPEP § 2143(I)(B). One of ordinary skill in the art before the effective filing date of the invention would have recognized that the iPSCs and T-cell derived iPSCs are functionally comparable, as they both allow for the production of ATOs, and thereby would have been able to substitute the iPSCs with predictable results.
Consequently, Crooks et al as modified by Takahama et al and Kaneko et al render obvious a method wherein T-iPSC-derived HSPCs form an ATO that produces CD4+CD8+ T cells. This therefore renders obvious the method of the instant claim.
Regarding claim 28: Following the discussion of claim 27 above, Kaneko et al further disclose that the iPSCs are universalized iPSCs wherein an HLA gene has been knocked out. This therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of instant claim 16, as the cells of the ATO – including the CD4+CD8+ T cells – will also have an HLA gene knocked out, as evidenced by Paragraphs [0193]-[0197] of Kaneko et al.
Claims 1-7, 12-15, 17-18, 23, 25, 27, 29, 32, 36, 39, and 41-42 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025), and further in view of Gschweng et al (US 2021/0040449 A1, of record).
The discussion of Crooks et al as modified by Takahama et al regarding claims 1, 6, and 15 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Crooks et al as modified by Takahama et al render obvious claims 1-7, 12-15, 27, 32, 36, 39, and 42. Gschweng et al is considered prior art under 35 USC 102(a)(2), with an effective filing date of 16 February 2018.
Regarding claims 17 and 25: Following the discussion of claim 15 above, Crooks et al further disclose that the CD4+ T cells are genetically engineered to comprise a heterologous FOXP3 sequence, which promotes the differentiation to CD4+ regulatory T cells (Paragraphs [0061], [0063]).
The combination of Crooks et al and Takahama et al fail to teach that the iPSCs comprise a heterologous transgene sequence encoding a lineage commitment factor that promotes the differentiation of the iPSCs to CD4+ Tregs, as required by instant claim 17.
Gschweng et al, however, disclose the genetic engineering of iPSCs such that the iPSCs comprise essential cell fate regulators that promote the differentiation of the iPSCs into different T-cell lineages, including FOXP3 for the induction of Tregs (Paragraphs [0065]-[0095], [0268]-[0269], [0309]).
Therefore, it would have been prima facie obvious to have modified the method of Crooks et al in view of Takahama et al such that starting population of iPSCs comprises a FOXP3 transgene that promotes the differentiation of the iPSCs to CD4+ Tregs, as detailed in Gschweng et al. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to modify the iPSCs to ensure commitment to the Treg lineage, as that is the final desired cellular therapeutic product for the treatment of autoimmune diseases (Crooks et al: Paragraphs [0034], [0061]-[0062], [0329]; Gschweng et al: Paragraphs [0271], [0288]-[0289]), and would have had a reasonable expectation of success since both Crooks et al and Gschweng et al are concerned with the generation of CD4+ T cells from the iPSC-derived ATOs (Gschweng et al: Paragraphs [0262]-[0267]). See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al and Gschweng et al render obvious a method of generating a CD4+ Treg, wherein the starting population of iPSCs is genetically engineered to comprise a heterologous FOXP3 (claim 25) sequence in the genome. As FOXP3 is a lineage commitment factor that promotes the differentiation of the iPSCs to CD4+ Tregs, this therefore renders obvious the method of instant claim 17.
Regarding claims 18 and 23: Following the discussion of claim 1 above, Gschweng et al further disclose that exogenous protein sequences are integrated into the genome of the iPSCs at a TRAC gene locus (claim 23), wherein the exogenous protein sequence is under control of the endogenous promoter of the gene locus (Paragraphs [0030], [0032], [0057], [0106], [0180], [0206]-[0209], [0215], [0217], [0251]). Therefore, the ordinary artisan would have been motivated to insert the heterologous FOXP3 sequence into the TRAC gene locus such that the FOXP3 sequence is under control of the endogenous TRAC promoter, as doing so would have allowed for the promotion of the iPSCs towards Tregs while also preventing the host immune reactivity to the cells (Gschweng et al: Paragraph [0008]), and would have had a reasonable expectation of success given the disclosure of Gschweng et al. See MPEP § 2143(I)(G). This therefore renders obvious the method of instant claim 18.
Regarding claim 29: Following the discussion of claim 1 above, Gschweng et al further disclose that the engineered cells further comprise a suicide gene (Paragraphs [0016], [0053], [0198], [0224]-[0227]). As the engineered cells can comprise CD4+CD8+ T cells (Paragraphs [0016], [0033], [0063]-[0064], [0071], [0139]), this therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of instant claim 17.
Regarding claim 41: Following the discussion of claim 6 above, Gschweng et al further disclose that the iPSCs are engineered to comprise essential cell fate regulators that promote the differentiation of the iPSCs into different T-cell lineages, wherein the cell fate regulators can be FOXP3 and ThPOK (Paragraphs [0066], [0089], [0094], [0310]). This therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of claim 17, as the ordinary artisan would have been motivated to drive the starting population of iPSCs of Crooks et al towards the intermediate product of an immature CD4+ T cell and end product of a CD4+ regulatory T cell. See MPEP § 2143(I)(G).
Claims 1-7, 12-15, 17-27, 29, 32, 36, 39, and 41-42 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025) and Gschweng et al (US 2021/0040449 A1, of record), and further in view of Burleigh et al (WO 2019/195491 A1, of record).
The discussion of Crooks et al as modified by Takahama et al regarding claims 1, 6, and 15 and the discussion of Crooks et al as modified by Takahama et al and Gschweng et al regarding claims 17, 23, and 25 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Crooks et al as modified by Takahama et al render obvious claims 1-7, 12-15, 27, 32, 36, 39, and 42. Crooks et al as modified by Takahama et al and Gschweng et al render obvious claims 1-7, 12-15, 17-18, 23, 25, 27, 29, 32, 36, 39, and 41-42. Burleigh et al is considered prior art under 35 USC 102(a)(1) and 35 USC 102(a)(2).
Regarding claims 19-20: Following the discussion of claim 17 above, Gschweng et al further disclose that the engineered stem cells further comprise a chimeric antigen receptor (CAR) that is integrated into the TRAC gene locus (Paragraphs [0085], [0087], [0094], [0096], [0114], [0214]-[0217]).
The combination of Crooks et al, Takahama et al, and Gschweng et al fail to teach that coding sequences for the FOXP3 sequence and CAR are separated by an in-frame coding sequence for a self-cleaving peptide or by an IRES, as required by instant claim 20.
Burleigh et al, however, disclose the engineering of iPSC-derived T cells, wherein a template polynucleotide comprising one or more transgenes separated by an in-frame 2A self-cleavage peptide sequence is knocked into a TRAC gene locus of the cells, wherein the transgenes include a CAR or any polypeptide in which expression in the cell is desired (Paragraphs [0036]-[0038], [0045]-[0046], [0171], [0445], [0504]-[0505], [0510]-[0511], [0522]-[0524], [0529], [0538]-[0539]). Burleigh et al further disclose that the cell can further comprise a recombinant FOXP3 (Paragraph [0577]).
Therefore, it would have been prima facie obvious to have further modified the method of Crooks et al in view of Takahama et al and Gschweng et al such that the starting population of iPSCs is engineered to comprise a transgene comprising a FOXP3 coding sequence and CAR, wherein the FOXP3 coding sequence and CAR are separated by an in-frame 2A self-cleavage peptide sequence, as detailed in Burleigh et al. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to employ a technique that is well-known in the art to integrate multiple coding sequences of a transgene into a TRAC gene locus, and would have had a reasonable expectation given the disclosure of Burleigh et al. See MPEP § 2143(I)(A) and MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al, Gschweng et al, and Burleigh et al render obvious a method of generating CD4+ Tregs, wherein the starting population of iPSCs is genetically engineered to comprise a transgene comprising a heterologous FOXP3 sequence and CAR (claim 20) that are separated by an in-frame 2A self-cleavage peptide sequence. This therefore renders obvious the method of instant claim 19.
Regarding claim 21: Following the discussion of claim 17 above, Burleigh et al further disclose that a ribosome skipping element/self-cleavage element, such as a 2A element, is placed immediately upstream of the transgene coding sequence, such that the ribosome skipping element/self-cleavage element is placed in-frame with the endogenous gene (Paragraphs [0505]-[0508], [0551]). This therefore renders obvious the method of the instant claim for the same reasons as detailed in the rejection of instant claim 19.
Regarding claims 22 and 24: Following the discussion of claims 21 and 23 above, Burleigh et al further disclose that transgene sequence is integrated downstream of the most 5' nucleotide of exon 1 and upstream of the most 3' nucleotide of exon 1 (claim 24) of the open reading frame of the endogenous TRAC locus (Paragraphs [0013], [0058]-[0059], [0103], [0113]-[0114], [0153], [0160], [0203]-[0204], [0252], [0442], [0451], [0501]. With that, Burleigh et al further disclose that the transgene includes a recombinant TRAC that can comprise functional exonic sequences (Paragraphs [0096], [0153], [0160], [0169], [0193], [0196], [0416], [0447]-[0451], [0456], [0556]. This therefore renders obvious the method of instant claim 22 for the same reasons as detailed in the rejection of instant claim 19.
Regarding claim 26: Following the discussion of claim 25 above, Gschweng et al further disclose that the cell fate regulators can be FOXP3 and ThPOK (Paragraphs [0066], [0089], [0094], [0310]). This therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of claim 19, as the ordinary artisan would have been motivated to drive the starting population of iPSCs of Crooks et al towards the intermediate product of an immature CD4+ T cell and end product of a CD4+ Treg. See MPEP § 2143(I)(G).
Claims 1-7, 12-15, 17-21, 23-27, 32, 36, 39, and 41-42 are rejected under 35 U.S.C. 103 as being unpatentable over Crooks et al (WO 2017/075389 A1, of record on IDS filed 19 December 2025) in view of Takahama et al (J Immunol, 1996, of record on IDS filed 10 July 2025), and further in view of Conway et al (WO 2021/092581 A1, of record on IDS filed 23 July 2024).
The discussion of Crooks et al as modified by Takahama et al regarding claims 1, 6, and 15 can be observed above and is relied upon herein, the content of which is incorporated in its entirety. Crooks et al as modified by Takahama et al render obvious claims 1-7, 12-15, 27, 32, 36, 39, and 42. Conway et al is considered prior art under 35 USC 102(a)(2), with an effective filing date of 08 November 2019.
However, the applied reference of Conway et al has a common assignee and joint inventor with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2). This rejection under 35 U.S.C. 102(a)(2) might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C. 102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B) if the same invention is not being claimed; or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed in the reference and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement.
Regarding claims 17-21, 23-26, and 41: Following the discussion of claims 6 and 15 above, Crooks et al further disclose that the CD4+ T cells are genetically engineered to comprise a heterologous FOXP3 sequence, which promotes the differentiation to CD4+ regulatory T cells (Paragraphs [0061], [0063]).
The combination of Crooks et al and Takahama et al fail to teach that the iPSCs comprise a heterologous transgene sequence encoding a lineage commitment factor that promotes the differentiation of the iPSCs to CD4+ Tregs, as required by instant claim 17.
Conway et al, however, disclose genetically engineered mammalian stem and progenitor cells that have increased potential to differentiate into regulatory T cells (Abstract).
As such, Conway et al disclose a genetically engineered iPSC comprising a heterologous sequence in the genome, wherein the heterologous sequence is integrated in exon 1, 2, or 3 of the TRAC gene locus and comprises a transgene encoding sequences for FOXP3 and ThPOK, wherein the two coding sequences are in-frame and are separated by an in-frame coding sequence for a self-cleaving peptide, and an IRES immediately upstream of the transgene (Paragraphs [0007], [0010]-[0012], [0019], [0023], [0035], [0039], [0043]-[0044]; Figure 6). It is of note that FOXP3 and ThPOK are both lineage commitment factors that promote the differentiation of the iPSCs to CD4+ Tregs (Paragraphs [0033], [0039]).
Therefore, it would have been prima facie obvious to have modified the method of Crooks et al in view of Takahama et al such that the starting population of iPSCs is genetically modified to comprise a transgene encoding sequences for FOXP3 and ThPOK, as detailed in Conway et al. One of ordinary skill in the art before the effective filing date of the invention would have been motivated to modify the iPSCs to ensure commitment to the Treg lineage, as that is the final desired cellular therapeutic product for the treatment of autoimmune diseases (Crooks et al: Paragraphs [0034], [0061]-[0062], [0329]; Conway et al: Paragraphs [0091]-[0092]), and would have had a reasonable expectation of success since the disclosures of both Kaneko et al and Conway et al are concerned with the differentiation of iPSCs into CD4+ Tregs. See MPEP § 2143(I)(G).
Consequently, Crooks et al as modified by Takahama et al and Conway et al render obvious a method of generating CD4+ Tregs, wherein the starting population of iPSCs is genetically engineered to comprise a heterologous sequence in the genome, wherein the heterologous sequence is integrated in exon 1, 2, or 3 of the TRAC gene locus (claims 18, 23-24) and comprises a transgene encoding sequences for FOXP3 and ThPOK, wherein the two coding sequences are in-frame and are separated by an in-frame coding sequence for a self-cleaving peptide (claims 19-20, 25-26), and an IRES immediately upstream of the transgene (claim 21). As FOXP3 and ThPOK are both lineage commitment factors that promote the differentiation of the iPSCs to immature CD4+ T cells (claim 41) to CD4+ Tregs, this therefore renders obvious the method of instant claim 17.
Regarding claim 22: Following the discussion of claim 21, Conway et al further disclose that the heterologous sequence further comprises, immediately upstream of the IRES, a nucleotide sequence comprising all the exonic sequences of the T cell specific gene locus that are downstream of the integration site, such that the T cell specific gene locus remains able to express an intact T cell specific gene product (Paragraphs [0010], [0019]). This therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of instant claim 17.
Regarding claim 29: Following the discussion of claim 1, Conway et al further disclose that the engineered cells further comprise a suicide gene (Paragraphs [0015], [0049]-[0050], [0075]-[0077]). As the engineered cells can comprise CD4+CD8+ T cells (Paragraphs [0025], [0028], [0087], [00108]), this therefore renders obvious the method of the instant claim for the same reasons as discussed in the rejection of instant claim 17.
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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/ALYSSA G WESTON/Examiner, Art Unit 1633
/CHRISTOPHER M BABIC/Supervisory Patent Examiner, Art Unit 1633
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