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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 04/09/2026 has been entered.
1. Claims 1, 5, 13, 18 – 19, 22, 26, 31, 33, 36, 38, 49, 53 – 55, 60, 68, 70, and 76 are pending. Claims 1, 5, 13, 18 – 19, 22, 26, 31, 33, 36, 38, 49, 53 – 55, and 76 are under consideration.
Withdrawn Claim Rejections
2. The rejection of claims 1, 5, 13, 18, 19, 22, 26, 31, 33, 36, 38, 49, 53, 55, and 76 under 35 U.S.C. 112(b) is withdrawn in view of Applicant’s amendment to claim 1 to clarify “resultant population”.
3. The rejection of claim 8 under 35 U.S.C. 112(b) is rendered moot in view of Applicant’s cancellation of the claim.
4. The rejection of claim 8 under 35 U.S.C. 112(d) is rendered moot in view of Applicant’s cancellation of the claim.
5. The rejection of claim 8 under 35 U.S.C. 103 is rendered moot in view of Applicant’s cancellation of the claim.
6. The rejection of claims 1, 5, 13, 18, 26, 33, 36, 38, 49, 55, and 76 under 35 U.S.C. 103 is withdrawn in view of Applicant’s arguments that Beck teaches culturing CD34+ hematopoietic progenitor cells (HPCs) with Delta4 fused with the Fc region of human IgG1 that was immobilized onto Dyna1 beads to produce natural killer (NK) cells as opposed to CD3+ T cells as recited in claim 1.
7. The rejection of claims 19 and 20 under 35 U.S.C. 103 is withdrawn in view of Applicant’s arguments that Beck teaches culturing CD34+ hematopoietic progenitor cells (HPCs) with Delta4 fused with the Fc region of human IgG1 that was immobilized onto Dyna1 beads to produce natural killer (NK) cells as opposed to CD3+ T cells as recited in claim 1.
8. The rejection of claim 31 under 35 U.S.C. 103 is withdrawn in view of Applicant’s arguments that Beck teaches culturing CD34+ hematopoietic progenitor cells (HPCs) with Delta4 fused with the Fc region of human IgG1 that was immobilized onto Dyna1 beads to produce natural killer (NK) cells as opposed to CD3+ T cells as recited in claim 1.
9. The rejection of claim 53 under 35 U.S.C. 103 is withdrawn in view of Applicant’s arguments that Dallas teaches differentiating HSCs while claim 53 recites hemogenic endothelium.
10. The rejection of claim 54 under 35 U.S.C. 103 is withdrawn in view of Applicant’s arguments that Dallas teaches differentiating HSCs while claim 54 recites hemogenic endothelium.
11. The provisional rejection of claim 8 on the ground of nonstatutory double patenting are rendered moot in view of Applicant’s cancellation of the claim.
12. The provisional rejection of claims 1, 5, 13, 18, 26, 31, 33, 36, 38, and 49 on the ground of nonstatutory double patenting is withdrawn in view of the amendment to the claims of application 18279336 to recite “producing a population of CD56+ NK cells”.
13. The provisional rejection of claim 53 on the ground of nonstatutory double patenting is withdrawn in view of the amendment to the claims of application 18279336 to recite “producing a population of CD56+ NK cells”.
14. The provisional rejection of claim 54 on the ground of nonstatutory double patenting is withdrawn in view of the amendment to the claims of application 18279336 to recite “producing a population of CD56+ NK cells”.
Claim Interpretation
15. It is noted that claims 1, 5, 13, 19, 22, 26, 31, 33, 36, 38, and 49 contain optional limitations. The MPEP 2111.04 states that “a claim scope is not limited by claim language that suggests or makes optional but does not require steps to be performed, or by claim language that does not limit a claim to a particular structure”. Therefore, in this case for the purpose of compact prosecution, the limitations following the phrase “optionally” in these claims will be interpreted as the optional components and therefore is/are non-limiting or not required.
16. For the purpose of applying prior art, “aggregation medium” is being interpreted as any medium in which pluripotent stem cells are cultured in to produce hemogenic endothelium.
17. Claims 13, 53, and 54 recite “FLT3”. For the purpose of applying prior art “FLT3” is being interpreted as the ligand of FLT3 (i.e., the cytokine and not the protein FLT3).
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.
18. Claim(s) 1, 5, 13, 18, 19, 22, 26, 33, 36, 38, 49, 55, and 76 is/are rejected under 35 U.S.C. 103 as being unpatentable over Daley (WO2018048828; Filed 09/06/2017; Published 03/15/2018; previously cited), hereinafter Daley which is cited on the IDS filed 01/02/2025 in view of Vodyanyk (US-12163155-B2; Filed 04/18/2018; Published 12/10/2024), hereinafter Vodyanyk.
Regarding step a of claim 1 and claim 26, Daley teaches culturing human iPS cells (“iPS” of claim 26) in a medium to form CD34+ hemogenic endothelium (page 20, para. 0130; page 82, 0456 – 0458; page 89, para. 0487). Daley teaches is now widely accepted that HSCs originate from hemogenic endothelium (page 38, para. 0259).
Regarding step b of claim 1, claim 36 and claim 38, Daley teaches inhibiting a histone methyltransferase by knocking down EZH1 (“inhibiting a histone methyltransferase” and “wherein the histone methyltransferase catalyzes the addition of methyl group to histone 3 lysine residue 27” of claim 1 and “genetically modifying the resultant population of CD34+ hemogenic endothelium” of claim 36 and “EZH1” of claim 38) with shRNA in the CD34+ cells (page 83, para. 0460; page 87 – 88, 0482 – 0483; page 10, 0061 – 0063; page 27, 0215; page 62, 0374; Figure 1A). Daley teaches EZH1 mediates methylation on histone lysine 27 (H3K27) (page 88, 0485).
Regarding step c of claim 1 and claim 5, Daley teaches co-culturing CD34+ EB with EZH1 inhibited by shRNA with Notch ligand delta like 1-expressing OP9 cells in culture media (“simultaneously to step b)” of claim 1) and T cells were produced (page 83, 0462; page 85, 0470; Figure 1A; page 87, 0482). Daley also teaches that the method can comprise activating the Notch signaling pathway instead of co-culture with OP9-DL1/4 cells to produce T cells where the Notch ligand is Delta-like-1, Deltal-like-4, or immobilized Delta1ext-IgG (claim 5) (page 3, 0013; page 5, 0023 – 0024; page 6, 0026; page10, 0058; page 26, 0213; page 27, 0214; page 32, 0238; page 62, 0373 – 0376; page 63, 0381 – 0383). Daley does not teach “a Notch ligand attached to a solid substrate” and “the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand” of claim 1.
Regarding claim 13, Daley teaches the media comprises Flt3 and IL7 (page 83, 0462).
Regarding claim 18, Daley teaches after day 20 co-culture, the differentiated cells are CD4/CD8 double positive (page 84, 0467).
Regarding claims 19 and 22, Daley teaches sorting the CD4+CD8+ cells and stimulating with plate-bound anti-CD3 antibody (“T cell activator” of claim 19) in the medium in the presence of IL-2 and anti-CD28 antibody (“T cell activator” of claim 19) (page 84, para. 0468). Daley does not teach IL-15 of claim 22.
Regarding claim 33, Daley teaches CD34+ cells were isolated by cell sorting by CD34+CD45+ surface markers from bulk EB culture (page 18, para. 0109; page 34, 0243; page 82, 0458; Figure 1A). Daley teaches the CD34+ population of cells are cells that are also CD38 negative or low (page 35, para. 0245).
Regarding claim 49, Daley teaches the histone methyltransferase inhibitor inhibits the G9a/GLP heteromeric complex and a histone methyltransferase inhibitor includes AMI-1, A-366, BIX-01294, BIX01338, BRD4770, chaetocin, UNC0224, UNC0631, UNC0638, UNC0642, UNC0646, EPZ5676, EPZ005687, GSK343, EPZ-6438, 3-deazaneplanocin A (DZNeP) HCl ,UNC1999, MM-102, SGC 0946, Entacapone, EPZ015666, UNC0379, Ell, MI-2 (Menin-MLL Inhibitor), MI-3 (Menin-MLL Inhibitor), PFI-2, GSK126, EPZ004777, BRD4770, and EPZ-6438 (page 10, 0062 and 0068; page 51, 0320).
Daley does not teach “a Notch ligand attached to a solid substrate” and “the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand” of claim 1 or “a population of CD4+ cells and a population of CD8+ cells” of claim 19 or “the single-positive-T-Cell-differentiation medium comprises 10 ng/mL IL-15” of claim 22 or the descriptive limitations of the CD3+ T cells of claims 55 and 76. However, Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, therapy for a host of diseases, disorders and conditions, and for the in vitro studies of disease modeling, drug screening, and hematological diseases (page 1, 0004). Daley teaches there is a lack of supply of functional HLA-matched immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions, and for the in vitro studies of disease modeling, drug screening, and hematological diseases (page 2, 0009). Daley teaches the method is useful as a cell preparation method in immunotherapy (page 2, 0009). Daley teaches knockdown of histone methyltransferases enhanced CD4+CD8+ T cell potential from CD34+ cells (page 87 – 88, para. 0482 – 0483).
Regarding “a Notch ligand attached to a solid substrate” and “the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand” of claim 1, Vodyanyk teaches a method of differentiating iPS cells expressing a chimeric antigen receptor (CAR) to CD3+ T cells comprising differentiating iPS cells to CD34+ hematopoietic progenitors through aggregate suspension culture, followed by enriching for CD34+ HPC cells by MACS using CD34 microbeads, followed by differentiating the enriched CD34+ cells to CD3+ T cell by plating the CD34+ cells on plastic plates coated with Notch ligand hDLL4-Fc chimeric protein in a T cell differentiation medium (col. 48, lines 10 – 48; col. 49, lines 51 – 67; col. 50; Figure 1A - 1D). Vodyanyk teaches the T cell differentiation medium does not contain serum, and comprises FLT3L and IL7 (col. 50, lines 45 – 55). Vodyanyk teaches the method produces CD4+ and CD8+ single and double positive cells in Figure 1D (col. 51, lines 41 – 45). Vodyanyk teaches the CD3+ T cells acquired CD8 expression (col. 52, lines 1 – 5; Figure 1G).
Regarding claims 19 and 22, Vodyanyk teaches T cell expansion with anti-CD3 mAb (“T cell activator”) and 10 ng/mL IL-15 (“IL-15”) and the method produces CD4+ and CD8+ single positive cells (col. 51, lines 5 – 21 and 41 – 45).
Vodyanyk teaches the CAR-expressing T cells produced by the method displayed cytotoxicity against the CD19+ targets, produced IFNg, TNFa, IL2, IL3, and GM-CSF, suppressed tumors in vivo, and prolonged survival (col. 48, lines 66 – 67; col. 49, lines 1 and 14 – 50; Figure 5A and 5B). Vodyanyk teaches despite technological advancements in the diagnosis and treatment options available to patients diagnosed with cancer, the prognosis still often remains poor and many patients cannot be cured (col. 1, lines 22 – 25). Vodyanyk teaches adoptive T cell therapy has many drawbacks including lack of patient and tumor-specific T cells and therefore, there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy (col. 1, lines 25 – 50).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Daley regarding a method of generating T cells from pluripotent stem cells by forming CD34+ hemogenic endothelium and culturing these cells with an inhibitor of a histone methyltransferase and a Notch ligand with the teachings of Vodyanyk regarding differentiating pluripotent stem cells to CD3+ T cells by forming CD34+ cells and culturing these cells on a plate coated with a Notch ligand in differentiation media to arrive at the claimed method for differentiating CD3+ T cells, the method comprising: a) differentiating a population of pluripotent stem cells in an aggregation medium for a sufficient time to promote differentiation into a population of CD34+ hemogenic endothelium; b) inhibiting a hi stone methyltransferase in the resultant population of CD34+ hemogenic endothelium from step a); wherein the histone methyltransferase catalyzes the addition of methyl group to hi stone 3 lysine residue 9 (H3K9) and/or histone 3 lysine residue 27 (H3K27); and c) simultaneously to step b), differentiating the resultant population of CD34+ hemogenic endothelium from step a) in a CD3+-T-cell differentiation medium in the presence of a Notch ligand attached to a solid substrate for a sufficient time to promote differentiation into a population of CD3+ T cells; wherein differentiating the hemogenic endothelium in the presence of the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand. One would have been motivated to combine the teachings of Daley and Vodyanyk in a method to produce CD3+ T cells for immunotherapy as Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions and Vodyanyk teaches there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy. One would have a reasonable expectation of success in combining the teachings as Daley teaches the method is useful as a cell preparation method in immunotherapy and Vodyanyk teaches the method produces CD3+ CAR-T cells that suppressed tumors in vivo and prolonged survival.
Regarding claims 55 and 76, as Daley in view of Vodynayk make obvious the method of differentiation of iPS cells to hemogenic endothelium, followed by further differentiation to CD3+ T cells in a stromal-free method in which Notch ligand is immobilized to a solid surface, Daley in view of Vodyanyk make obvious CD3+ T cells that meet the limitations of claim 55 and 76.
19. Claim(s) 31 is/are rejected under 35 U.S.C. 103 as being unpatentable over Daley (WO2018048828; Filed 09/06/2017; Published 03/15/2018; previously cited), hereinafter Daley which is cited on the IDS filed 01/02/2025 in view of Vodyanyk (US-12163155-B2; Filed 04/18/2018; Published 12/10/2024), hereinafter Vodyanyk as applied to claims 1, 5, 13, 18, 19, 22, 26, 33, 36, 38, 49, 55, and 76 above, and further in view of Sturgeon (Sturgeon, Christopher M., et al. Nature biotechnology 32.6 (2014): 554-561.), hereinafter Sturgeon which is cited on the IDS filed 01/02/2025.
Daley in view of Vodyanyk make obvious the limitations of claim 1 as set forth above.
Daley teaches the medium for EB differentiation comprises BMP4, SCF, and IL-6 (page 82, 0457). Daley does not teach SB-431542, CHIR99021, bFGF, VEGF, IL-11, IGF-1 or EPO.
Vodyanyk teaches the media for differentiating pluripotent stem cells to CD34+ cells comprises CHIR99021, IGF1, VEGF, FGF2 (bFGF), BMP4, SCF, FLT3L, IL-11 (col. 49, lines 60 – 67; col. 50, lines 1 – 31).
Daley and Vodyanyk do not teach SB431542 and EPO.
Sturgeon teaches a method of culturing pluripotent stem cell aggregates in media containing BMP4, bFGF, SB431542, CHIR99021, VEGF, IL-6, IGF-1, IL-11, SCF, and EPO resulted in the formation of CD34+ hemogenic endothelium (page 562, left col. para. 1; Figure 1a; page 555, right col. para. 2; Figure 2; page 557, left col. last para. and right col. para. 1; Figure 5). Sturgeon teaches isolating the CD34+ cells by FACS and culturing with OP9-DL4 to produce T cells (page 557, left col. para. 2; Figure 2e; page 559, right col. last para.). Sturgeon teaches culturing with SB431542 and CHIR99021 specifies the definitive program that leads to the formation of T cells (Figure 4e; page 558, right col. para. 1; Figure 5; page 559, left col.; Figure 3i and 3j; Figure 4d). Sturgeon teaches the formation of mesoderm can proceed either through the primitive or definitive lineages via hemogenic endothelium that are distinguished based on T-lymphoid potential where the primitive lineage does not produce T cells (page 560, left col. para. 2 and right col. para. 2; Figure 5, blue pathway; Figure 3j).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Daley regarding a method of generating T cells from pluripotent stem cells by forming CD34+ hemogenic endothelium and culturing these cells with an inhibitor of a histone methyltransferase and a Notch ligand where the aggregate media comprises BMP4, SCF, and IL-6 with the teachings of Vodyanyk regarding differentiating pluripotent stem cells to CD3+ T cells by forming CD34+ cells and culturing these cells on a plate coated with a Notch ligand in differentiation media where the aggregate media comprises CHIR99021, IGF1, VEGF, FGF2 (bFGF), BMP4, SCF, FLT3L, IL-11 with the teachings of Sturgeon regarding differentiating pluripotent stem cells to T cells where the aggregate media comprises BMP4, bFGF, SB431542, CHIR99021, VEGF, IL-6, IGF-1, IL-11, SCF, and EPO to arrive at the claimed method wherein the aggregation medium comprises BMP4, SB-431542, CHIR99021, bFGF, VEGF, IL-6, IL-11, IGF-1, SCF, and EPO. One would have been motivated to combine the teachings of Daley, Vodyanyk, and Sturgeon in a method to produce CD3+ T cells for immunotherapy from the definitive and not primitive lineage as Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions and Vodyanyk teaches there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy and Sturgeon teaches the primitive lineage does not produce T cells. One would have a reasonable expectation of success in combining the teachings as Daley teaches the method is useful as a cell preparation method in immunotherapy and Vodyanyk teaches the method produces CD3+ CAR-T cells that suppressed tumors in vivo and prolonged survival and Sturgeon teaches the method restricts the definitive lineage that when cultured with a Notch ligand forms T cells.
20. Claim(s) 53 is/are rejected under 35 U.S.C. 103 as being unpatentable over Daley (WO2018048828; Filed 09/06/2017; Published 03/15/2018; previously cited), hereinafter Daley which is cited on the IDS filed 01/02/2025 in view of Kaneko (US20180298337-A1; Filed 10/14/2016; Published 10/18/2018), hereinafter Kaneko which is cited on the IDS filed 01/02/2025 in view of Vodyanyk (US-12163155-B2; Filed 04/18/2018; Published 12/10/2024), hereinafter Vodyanyk.
Regarding step a, Daley teaches culturing human iPS cells in a medium to form CD34+ hemogenic endothelium (page 20, para. 0130; page 82, 0456 – 0458; page 89, para. 0487). Daley teaches is now widely accepted that HSCs originate from hemogenic endothelium (page 38, para. 0259).
Regarding step b, Daley teaches inhibiting a histone methyltransferase by knocking down EZH1 (“inhibiting a histone methyltransferase” and “wherein the histone methyltransferase catalyzes the addition of methyl group to histone 3 lysine residue 27”) with shRNA in the CD34+ cells (page 83, para. 0460; page 87 – 88, 0482; page 10, 0061 – 0063; page 27, 0215; page 62, 0374; Figure 1A). Daley teaches EZH1 mediates methylation on histone lysine 27 (H3K27) (page 88, 0485).
Regarding step c, Daley teaches co-culturing CD34+ EB with EZH1 inhibited by shRNA with Notch ligand delta like 1-expressing OP9 cells in culture media (“simultaneously to step b)”) and T cells were produced (page 83, 0462; page 85, 0470; Figure 1A; page 87, 0482). Daley also teaches that the method can comprise activating the Notch signaling pathway instead of co-culture with OP9-DL1/4 cells to produce T cells where the Notch ligand is Delta-like-1, Deltal-like-4, or immobilized Delta1ext-IgG (“the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand”) (page 3, 0013; page 5, 0023 – 0024; page 6, 0026; page10, 0058; page 26, 0213; page 27, 0214; page 32, 0238; page 62, 0373 – 0376; page 63, 0381 – 0383). Daley teaches the media comprises Flt3 ligand and IL-7 at 5 ng/ml each (page 83, para. 0462) but does not teach 15 ng/mL FLT3 or 25 ng/mL IL7. Daley teaches co-culture for 20 days in the media with OP9-DL1 (page 84, para. 0467) and after 5 weeks of co-culture (“at least 4 weeks”) of CD34+ cells with shRNA and OP9-DL1, T cell potential was analyzed where knockdown of methyl transferases enhanced CD4+CD8+ T cell potential from CD34+ cells (page 87 – 88, para. 0482 – 0483) but does not teach “in the presence of 10 µg/mL Notch ligand attached to a solid substrate”. Daley teaches 30 ng/mL of SCF in the media (page 83, para. 0462) but does not teach TPO.
Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, therapy for a host of diseases, disorders and conditions, and for the in vitro studies of disease modeling, drug screening, and hematological diseases (page 1, 0004). Daley teaches there is a lack of supply of functional HLA-matched immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions, and for the in vitro studies of disease modeling, drug screening, and hematological diseases (page 2, 0009). Daley teaches the method is useful as a cell preparation method in immunotherapy (page 2, 0009).
Regarding 15 ng/mL FLT3 or 25 ng/mL IL7, and “5 ng/mL TPO”, Kaneko teaches a method of differentiating iPS cells to CD34+ hemogenic endothelium in iCD34-A and iCD34-B media, followed by stromal-free differentiation of the CD34+ hemogenic endothelium in iT-A2 media comprising 0.2 – 50 ng/mL Flt3L, 0.2 – 50 ng/mL IL7 and 1 – 250 ng/mL SCF with soluble DLL4 protein (15 ng/mL FLT3 or 25 ng/mL IL7 ) (page 47, para. 0324 – 0328; page 48, para. 0329 – 0331; page 50, para. 0343 and 0347; page 27, Table 14; Figure 12, 13). Kaneko teaches the stromal free method differentiated iPSC-derived hemogenic endothelium towards T cells (Figure 32; page 10, para. 0086). Kaneko teaches CD34+ cells were differentiated towards T cells in iTC-A1 media containing 5 – 1500 ng/mL soluble DLL1 and DLL4, 0.2 – 50 ng/mL Flt3L, 0.2 – 50 ng/mL IL7 and 1 – 250 ng/mL SCF (15 ng/mL FLT3 or 25 ng/mL IL7 ) (page 46 – 47, para. 0321; page 19, Table 4). Kaneko teaches after approximately 10 days, the culture setting was switched to iTC-B1 containing 5 – 1500 ng/mL soluble DLL1 and DLL4, 0.2 – 50 ng/mL Flt3L, 0.04 – 10 ng/mL IL7 and 0.2 – 50 ng/mL SCF to complete T cell maturation (page 47, para. 0321; page 20, Table 5). Kaneko teaches after approximately 30 – 40 days, the cell population was assessed for the composition of T cells including surface expression of CD3, CD7, TCRαβ, CD4, and CD8, and Figure 10 shows the resulting T cells (page 47, para. 0321). Kaneko teaches a method of differentiating hemogenic endothelium to multipotent progenitors in iMPP-A medium comprising 0.5 – 150 ng/mL TPO (“5 ng/mL TPO”) that can subsequently be differentiated to hematopoietic lineages that is dependent on Notch signaling (page 27, Table 13; Figure 12; page 48, para. 0332 – 0333).
Kaneko does not teach “10 µg/mL Notch ligand attached to a solid substrate”. However, Kaneko teaches to advance the promise of iPSC and genomically engineered iPSC technology as an allogeneic source of hematopoietic cellular therapeutics, it is essential to be able to efficiently and reproducibly generate immune effector populations including T cells (page 1, para. 0003). Kaneko teaches there is a need for methods and compositions of differentiating stem cell to definitive hematopoiesis without relying on co-culturing or serum-containing media, and without requiring the formation of embryoid body aggregates as intermediates (page 1, para. 0006). One would have been motivated to combine the teachings of Daley and Kaneko as both teach methods of differentiating iPS cells to hemogenic endothelium and then to T cells for the use of T cells in immunotherapy.
Regarding “10 µg/mL Notch ligand attached to a solid substrate”, Vodyanyk teaches a method of differentiating iPS cells expressing a chimeric antigen receptor (CAR) to CD3+ T cells comprising differentiating iPS cells to CD34+ hematopoietic progenitors, followed by differentiating the CD34+ cells to CD3+ T cell by plating the CD34+ cells on plastic plates coated with 0.5 µg/cm2 Notch ligand hDLL4-Fc chimeric protein in a T cell differentiation medium (col. 48, lines 10 – 48; col. 49, lines 51 – 67; col. 50; Figure 1A - 1D). Vodyanyk teaches 0.25 mL/cm2 medium was added to the plate and 5000 cells/cm2 were plated (col. 50, lines 50 – 56). A surface concentration of 0.5 µg/cm2 Notch ligand hDLL4-Fc chimeric protein on a plate with 0.25 mL/cm2 medium is the same as 2 µg/mL [0.5 µg/cm2 = X µg/mL x 0.25 mL/cm2 = 2 µg/mL]. It would have been obvious to adjust the concentration of Notch ligand to arrive at 10 µg/mL as it is a result-effective variable dependent on the surface area of the solid substrate and number of cells plated. Vodyanyk teaches the T cell differentiation medium comprises SCF, TPO, FLT3L and IL7 each at 50 ng/mL for two weeks (col. 50, lines 45 – 55). Vodanyk teaches the differentiation process can be continued and improve the yield of T cells by re-plating harvested cells to freshly-prepared DLL4 plates at 10,000 cells/cm2 until the desired yield of T cells is achieved (col. 50, lines 62 – 67). Vodyanyk teaches the CAR-expressing T cells produced by the method displayed cytotoxicity against the CD19+ targets, produced IFNg, TNFa, IL2, IL3, and GM-CSF, suppressed tumors in vivo, and prolonged survival (col. 48, lines 66 – 67; col. 49, lines 1 and 14 – 50; Figure 5A and 5B). Vodyanyk teaches despite technological advancements in the diagnosis and treatment options available to patients diagnosed with cancer, the prognosis still often remains poor and many patients cannot be cured (col. 1, lines 22 – 25). Vodyanyk teaches adoptive T cell therapy has many drawbacks including lack of patient and tumor-specific T cells and therefore, there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy (col. 1, lines 25 – 50). One would have been motivated to combine the teachings of Daley and Vodyanyk because both teach method of forming T cells for immunotherapy.
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Daley regarding a method of generating T cells from pluripotent stem cells by forming CD34+ hemogenic endothelium and culturing these cells with an inhibitor of a histone methyltransferase and a Notch ligand in media comprising Flt3 ligand and IL-7 with the teachings of Kaneko regarding a method of differentiating iPS cells to CD34+ hemogenic endothelium, followed by stromal-free differentiation of the CD34+ hemogenic endothelium towards T cells in media comprising 0.2 – 50 ng/mL Flt3L, 0.2 – 50 ng/mL IL7 and 1 – 250 ng/mL SCF with soluble DLL4 protein with the teachings of Vodyanyk regarding differentiating pluripotent stem cells to CD3+ T cells by forming CD34+ cells and culturing these cells on a plate coated with 0.5 µg/cm2 Notch ligand hDLL4-Fc chimeric protein in a T cell differentiation medium comprising SCF, TPO, FLT3L and IL7 to arrive at the claimed method for differentiating CD3+ T cells, the method comprising: a) differentiating a population of pluripotent stem cells in an aggregation medium for a sufficient time to promote differentiation into a population of CD34+ hemogenic endothelium; and b) inhibiting a histone methyltransferase in the resultant population of CD34+ hemogenic endothelium from step a): wherein the histone methyltransferase catalyzes the addition of methyl group to hi stone 3 lysine residue 9 (H3K9) and/or histone 3 lysine residue 27 (H3K27); and c) simultaneously to step b ), differentiating the resultant population of CD34+ hemogenic endothelium from step a) in a CD3+-T-cell-differentiation medium comprising 15 ng/ml FLT3 and 25 ng/ml IL 7 in the presence of 10 μg/mL Notch ligand attached to a solid substrate for at least 4 weeks to promote differentiation into a population of CD3+ T cells; wherein differentiating the hemogenic endothelium in the presence of the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand; and wherein the CD3+-T-cell-differentiation medium further comprises 5 ng/mL TPO and 30 ng/ml SCF for at least the first two weeks. One would have been motivated to combine the teachings of Daley, Kaneko, and Vodyanyk in a method to produce CD3+ T cells for immunotherapy as Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions and Kaneko teaches to advance the promise of iPSC and genomically engineered iPSC technology as an allogeneic source of hematopoietic cellular therapeutics, it is essential to be able to efficiently and reproducibly generate immune effector populations including T cells and Vodyanyk teaches there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy. One would have a reasonable expectation of success in combining the teachings as Daley teaches the method is useful as a cell preparation method in immunotherapy and Daley, Kaneko and Vodyanyk teach stromal free methods that form CD3, CD4, and CD8 T cells from iPS cells, and Vodyanyk teaches the method produces CD3+ CAR-T cells that suppressed tumors in vivo and prolonged survival.
21. Claim(s) 54 is/are rejected under 35 U.S.C. 103 as being unpatentable over Daley (WO2018048828; Filed 09/06/2017; Published 03/15/2018; previously cited), hereinafter Daley which is cited on the IDS filed 01/02/2025 in view of Kaneko (US20180298337-A1; Filed 10/14/2016; Published 10/18/2018), hereinafter Kaneko which is cited on the IDS filed 01/02/2025 in view of Vodyanyk (US-12163155-B2; Filed 04/18/2018; Published 12/10/2024), hereinafter Vodyanyk.
Regarding step a, Daley teaches culturing human iPS cells in a medium to form CD34+ hemogenic endothelium (page 20, para. 0130; page 82, 0456 – 0458; page 89, para. 0487). Daley teaches is now widely accepted that HSCs originate from hemogenic endothelium (page 38, para. 0259).
Regarding step b, Daley teaches co-culturing CD34+ EB with a histone methyltransferase inhibitor with Notch ligand delta like 1-expressing OP9 cells in culture media and T cells were produced (page 83, 0462; page 85, 0470; Figure 1A; page 87, 0482). Daley teaches the media comprises Flt3 ligand and IL-7 at 5 ng/ml each (page 83, para. 0462) but does not teach 15 ng/mL FLT3 or 25 ng/mL IL7. Daley teaches co-culture for 20 days in the media with OP9-DL1 (page 84, para. 0467). Daley teaches the method can comprise activating the Notch signaling pathway instead of co-culture with OP9-DL1/4 cells to produce T cells where the Notch ligand is Delta-like-1, Deltal-like-4, or immobilized Delta1ext-IgG (“the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand”) (page 3, 0013; page 5, 0023 – 0024; page 6, 0026; page10, 0058; page 26, 0213; page 27, 0214; page 32, 0238; page 62, 0373 – 0376; page 63, 0381 – 0383). Daley teaches 30 ng/mL of SCF in the media (page 83, para. 0462) but does not teach TPO.
Regarding G9a/GLP inhibitor and “for at least the first two weeks”, Daley teaches the method comprises a histone methyltransferase inhibitor that inhibits G9a/GLP (page 10, 0062; Figure 7A). Daley teaches inhibition of G9a/GLP by nucleic acids or small molecules (page 15, 0093). Daley teaches treating MHPCs with a histone methyltransferase inhibitor for directing the differentiation of these cells to immune cells by activating the Notch signaling pathway in these cells (page 3, 0013). Daley teaches knockdown of histone H3 lysine 9 methyltransferases enhanced T cell potential and G9a catalyzes H3K9 methylation (page 49, 0307; page 88, 0482). Daley teaches co-culture for 20 days in the media with OP9-DL1 (page 84, para. 0467) and after 5 weeks of co-culture of CD34+ cells with shRNA and OP9-DL1, T cell potential was analyzed where knockdown of methyl transferases enhanced CD4+CD8+ T cell potential from CD34+ cells (page 87 – 88, para. 0482 – 0483).
Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, therapy for a host of diseases, disorders and conditions, and for the in vitro studies of disease modeling, drug screening, and hematological diseases (page 1, 0004). Daley teaches there is a lack of supply of functional HLA-matched immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions, and for the in vitro studies of disease modeling, drug screening, and hematological diseases (page 2, 0009). Daley teaches the method is useful as a cell preparation method in immunotherapy (page 2, 0009).
Regarding 15 ng/mL FLT3 or 25 ng/mL IL7, and “5 ng/mL TPO”, Kaneko teaches a method of differentiating iPS cells to CD34+ hemogenic endothelium in iCD34-A and iCD34-B media, followed by stromal-free differentiation of the CD34+ hemogenic endothelium in iT-A2 media comprising 0.2 – 50 ng/mL Flt3L, 0.2 – 50 ng/mL IL7 and 1 – 250 ng/mL SCF with soluble DLL4 protein (15 ng/mL FLT3 or 25 ng/mL IL7 and “the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand”) (page 47, para. 0324 – 0328; page 48, para. 0329 – 0331; page 50, para. 0343 and 0347; page 27, Table 14; Figure 12, 13). Kaneko teaches the stromal free method differentiated iPSC-derived hemogenic endothelium towards T cells (Figure 32; page 10, para. 0086). Kaneko teaches CD34+ cells were differentiated towards T cells in iTC-A1 media containing 5 – 1500 ng/mL soluble DLL1 and DLL4, 0.2 – 50 ng/mL Flt3L, 0.2 – 50 ng/mL IL7 and 1 – 250 ng/mL SCF (15 ng/mL FLT3 or 25 ng/mL IL7 and “the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand”) (page 46 – 47, para. 0321; page 19, Table 4). Kaneko teaches after approximately 10 days, the culture setting was switched to iTC-B1 containing 5 – 1500 ng/mL soluble DLL1 and DLL4, 0.2 – 50 ng/mL Flt3L, 0.04 – 10 ng/mL IL7 and 0.2 – 50 ng/mL SCF to complete T cell maturation (page 47, para. 0321; page 20, Table 5). Kaneko teaches after approximately 30 – 40 days, the cell population was assessed for the composition of T cells including surface expression of CD3, CD7, TCRαβ, CD4, and CD8, and Figure 10 shows the resulting T cells (page 47, para. 0321). Kaneko teaches a method of differentiating hemogenic endothelium to multipotent progenitors in iMPP-A medium comprising 0.5 – 150 ng/mL TPO (“5 ng/mL TPO”) that can subsequently be differentiated to hematopoietic lineages that is dependent on Notch signaling (page 27, Table 13; Figure 12; page 48, para. 0332 – 0333).
Kaneko teaches to advance the promise of iPSC and genomically engineered iPSC technology as an allogeneic source of hematopoietic cellular therapeutics, it is essential to be able to efficiently and reproducibly generate immune effector populations including T cells (page 1, para. 0003). Kaneko teaches there is a need for methods and compositions of differentiating stem cell to definitive hematopoiesis without relying on co-culturing or serum-containing media, and without requiring the formation of embryoid body aggregates as intermediates (page 1, para. 0006). One would have been motivated to combine the teachings of Daley and Kaneko as both teach methods of differentiating iPS cells to hemogenic endothelium and then to T cells for the use of T cells in immunotherapy.
Vodyanyk teaches a method of differentiating iPS cells expressing a chimeric antigen receptor (CAR) to CD3+ T cells comprising differentiating iPS cells to CD34+ hematopoietic progenitors, followed by differentiating the CD34+ cells to CD3+ T cell by plating the CD34+ cells on plastic plates coated with 0.5 µg/cm2 Notch ligand hDLL4-Fc chimeric protein in a T cell differentiation medium (col. 48, lines 10 – 48; col. 49, lines 51 – 67; col. 50; Figure 1A - 1D). Vodyanyk teaches 0.25 mL/cm2 medium was added to the plate and 5000 cells/cm2 were plated (col. 50, lines 50 – 56). Vodyanyk teaches the T cell differentiation medium comprises SCF, TPO, FLT3L and IL7 each at 50 ng/mL for two weeks (col. 50, lines 45 – 55). Vodanyk teaches the differentiation process can be continued and improve the yield of T cells by re-plating harvested cells to freshly-prepared DLL4 plates at 10,000 cells/cm2 until the desired yield of T cells is achieved (col. 50, lines 62 – 67). Vodyanyk teaches the CAR-expressing T cells produced by the method displayed cytotoxicity against the CD19+ targets, produced IFNg, TNFa, IL2, IL3, and GM-CSF, suppressed tumors in vivo, and prolonged survival (col. 48, lines 66 – 67; col. 49, lines 1 and 14 – 50; Figure 5A and 5B). Vodyanyk teaches despite technological advancements in the diagnosis and treatment options available to patients diagnosed with cancer, the prognosis still often remains poor and many patients cannot be cured (col. 1, lines 22 – 25). Vodyanyk teaches adoptive T cell therapy has many drawbacks including lack of patient and tumor-specific T cells and therefore, there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy (col. 1, lines 25 – 50). One would have been motivated to combine the teachings of Daley and Vodyanyk because both teach method of forming T cells for immunotherapy.
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Daley regarding a method of generating T cells from pluripotent stem cells by forming CD34+ hemogenic endothelium and culturing these cells with an inhibitor of a histone methyltransferase that is a G9a/GLP inhibitor and a Notch ligand in media comprising Flt3 ligand and IL-7 with the teachings of Kaneko regarding a method of differentiating iPS cells to CD34+ hemogenic endothelium, followed by stromal-free differentiation of the CD34+ hemogenic endothelium towards T cells in media comprising 0.2 – 50 ng/mL Flt3L, 0.2 – 50 ng/mL IL7 and 1 – 250 ng/mL SCF with soluble DLL4 protein with the teachings of Vodyanyk regarding differentiating pluripotent stem cells to CD3+ T cells by forming CD34+ cells and culturing these cells on a plate coated with Notch ligand hDLL4-Fc chimeric protein in a T cell differentiation medium comprising SCF, TPO, FLT3L and IL7 to arrive at the claimed method for differentiating CD3+ T cells, the method comprising: a) differentiating a population of pluripotent stem cells in an aggregation medium for a sufficient time to promote differentiation into a population of CD34+ hemogenic endothelium; and b) differentiating the resultant population of CD34+ hemogenic endothelium in a CD3+-T-cell-differentiation medium comprising 15 ng/ml FLT3 and 25 ng/ml IL7 in the presence of 10 μg/mL Notch ligand attached to a solid substrate for at least 4 weeks to promote differentiation into a population of CD3+ T cells; wherein differentiating the hemogenic endothelium in the presence of the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand; and wherein the CD3+-T-cell-differentiation medium further comprises 5 ng/mL TPO, 30 ng/ml SCF, and a G9a/GLP inhibitor for at least the first two weeks. One would have been motivated to combine the teachings of Daley, Kaneko, and Vodyanyk in a method to produce CD3+ T cells for immunotherapy as Daley teaches there is a lack of supply of functional immune cells for the in vivo cellular replacement therapy, the treatment of diseases, disorders and medical conditions and Daley teaches knockdown of histone H3 lysine 9 methyltransferases enhanced T cell potential and G9a catalyzes H3K9 methylation and Kaneko teaches to advance the promise of iPSC and genomically engineered iPSC technology as an allogeneic source of hematopoietic cellular therapeutics, it is essential to be able to efficiently and reproducibly generate immune effector populations including T cells and Vodyanyk teaches there is an unmet need for therapeutically sufficient and functional antigen-specific immune cells for effective use in immunotherapy. One would have a reasonable expectation of success in combining the teachings as Daley teaches the method is useful as a cell preparation method in immunotherapy and Daley, Kaneko and Vodyanyk teach stromal free methods that form CD3, CD4, and CD8 T cells from iPS cells, and Vodyanyk teaches the method produces CD3+ CAR-T cells that suppressed tumors in vivo and prolonged survival.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
22. Claims 1, 5, 13, 18, 19, 22, 26, 31, 33, 36, 38, 49, 53, and 76 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1, 2, 5, 8, 10, 11, 12, 15, 18, 21, 26, 29, 30, 33, 41, 43, 45, and 52 of copending Application No. 18994861 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because the instant and pending claims read on a method of generating CD3+ T cells from CD34+ hemogenic endothelium.
Instant claim 1 recites a method for differentiating CD3+ T cells, the method comprising: a) differentiating a population of pluripotent stem cells in an aggregation medium for a sufficient time to promote differentiation into a population of CD34+ hemogenic endothelium; b) inhibiting a hi stone methyltransferase in the resultant population of CD34+ hemogenic endothelium from step a); wherein the histone methyltransferase catalyzes the addition of methyl group to hi stone 3 lysine residue 9 (H3K9) and/or histone 3 lysine residue 27 (H3K27); and c) simultaneously to step b), differentiating the resultant population of CD34+ hemogenic endothelium from step a) in a CD3+-T-cell differentiation medium in the presence of a Notch ligand attached to a solid substrate for a sufficient time to promote differentiation into a population of CD3+ T cells; wherein differentiating the hemogenic endothelium in the presence of the Notch ligand does not comprise co-culturing with a stromal cell expressing the Notch ligand.
Instant claim 1 recites a method for generating CD3+ T cells, the method comprising: (a) contacting a CD34+ hemogenic endothelial (HE) cells with a CD5+CD7+ differentiation medium comprising interleukin-3 (IL-3) under conditions and for a sufficient time to generate CD5+ CD7+ T cell progenitor cells, (b) contacting the CD5+CD7+ proT cells with a CD3+ T cell differentiation medium under conditions and for a sufficient time to generate CD3+ T cells.
Instant claim 1 aligns to reference claim 1, 30, and 52 as all claims are drawn to differentiating CD34+ hemogenic endothelium to generate CD3+ T cells. Instant claim 1 broadly recites “CD3+-T cell-differentiation medium”, and therefore encompasses the media of reference claim 1. Step a of instant claim 1 and instant claim 26 align to reference claim 33 at “generating CD34+ hemogenic endothelium from a population of pluripotent stem cells, optionally induced pluripotent stem cells (iPSCs)”, and “wherein the population of pluripotent stem cells is differentiated into a population of CD34+ hemogenic endothelium by way of embryoid bodies or 2D adherent cultures”. Step b of instant claim 1 and instant claim 38 align to reference claim 45 at “wherein the method further comprises inhibition of EZH1 activity and/or expression in the CD34+ HE cells”. Step b of instant claim 1 broadly recites “inhibiting a histone methyl transferase”, and EZH1 is a histone methyltransferase as taught by the reference specification at para. 00124 and instant claim 38. Step c of instant claim 1 and instant claim 5 aligns to reference claim 15 at “wherein the CD34+ HE cells are cultured in the presence of a Notch ligand; wherein the Notch ligand is attached to a solid surface; and/or the Notch ligand is attached to a cell culture dish” and reference claim 18 at “wherein the Notch ligand is not expressed by a stromal cell; wherein the method does not comprise co-culturing with a stromal cell expressing a Notch ligand”.
Instant claim 5 aligns to reference claim 15.
Instant claim 13 aligns to reference claims 5, 8, 30 (FLT3 and IL-7 and concentrations)reference claim 29 (“serum-free”).
Instant claim 31 aligns to reference claim 33 at “BMP4, SB-431542, CHIR99021, bFGF, VEGF, IL-6, IL-11, IGF-1, SCF, and EPO”.
Instant claim 33 aligns to reference claim 33 at “further comprising selecting or isolating the resultant population of CD34+ hemogenic endothelium using expression of surface markers on the population of CD34+ hemogenic endothelium”.
Instant claim 36 aligns to reference claim 41 at “further comprising a step of genetically modifying the resultant CD34+ hemogenic endothelial cells or the resultant CD3+ T cells”.
Instant claims 55 and 76 align to reference claim 43 at “wherein the CD3+ T cells are CD3+ TCRαβ+ T cells; and/or wherein the CD3+ T cells comprise a diverse T cell receptor (TCR) repertoire”.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
23. Claim 53 is provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 2, 5, 8, 12, 15, 18, 21, 26, 29, 30, 33, and 45 of copending Application No. 18994861 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because the instant and pending claims read on a method of generating CD3+ T cells from CD34+ hemogenic endothelium.
Instant claim 53 aligns to reference claim 1, 30, and 52 as all claims are drawn to differentiating CD34+ hemogenic endothelium to generate CD3+ T cells. Instant claim 53 broadly recites “CD3+-T cell-differentiation medium”, and therefore encompasses the media of reference claim 1. Step a of instant claim 53 aligns to reference claim 33 at “generating CD34+ hemogenic endothelium from a population of pluripotent stem cells, optionally induced pluripotent stem cells (iPSCs)”, and “wherein the population of pluripotent stem cells is differentiated into a population of CD34+ hemogenic endothelium by way of embryoid bodies or 2D adherent cultures”. Step b of instant claim 53 aligns to reference claim 45 at “wherein the method further comprises inhibition of EZH1 activity and/or expression in the CD34+ HE cells”. Step b of instant claim 53 broadly recites “inhibiting a histone methyl transferase”, and EZH1 is a histone methyltransferase as taught by the reference specification at para. 00124 and instant claim 38. Step c of instant claim 53 aligns to reference claim 5 and 8 at “the concentration of SCF is 5 – 50 ng/mL” and “the concentration of FLT-3 is 5 – 30 ng/mL” and “the concentration is 10 – 50 ng/mL” and “the concentration of TPO is 1 – 10 ng/mL”. Step c of instant claim 53 aligns to reference claim 15 at “wherein the CD34+ HE cells are cultured in the presence of a Notch ligand; wherein the Notch ligand is attached to a solid surface; and/or the Notch ligand is attached to a cell culture dish” and reference claim 18 at “wherein the Notch ligand is not expressed by a stromal cell; wherein the method does not comprise co-culturing with a stromal cell expressing a Notch ligand”. Step c of instant claim 53 aligns to reference claim 12 at “wherein step (b) is performed for at least 1 week”. Step c of instant claim 53 aligns to reference claim 30 at “stem cell factor (SCF), FLT-3, IL-7 and thrombopoietin (TPO)”.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
24. Claim 54 is provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 2, 5, 8, 12, 15, 18, 21, 26, 29, 30, 33, and 45 of copending Application No. 18994861 in view of Daley (WO2018048828; Filed 09/06/2017; Published 03/15/2018; previously cited), hereinafter Daley which is cited on the IDS filed 01/02/2025.
Instant claim 54 aligns to reference claim 1, 30, and 52 as all claims are drawn to differentiating CD34+ hemogenic endothelium to generate CD3+ T cells. Instant claim 53 broadly recites “CD3+-T cell-differentiation medium”, and therefore encompasses the media of reference claim 1. Step a of instant claim 53 aligns to reference claim 33 at “generating CD34+ hemogenic endothelium from a population of pluripotent stem cells, optionally induced pluripotent stem cells (iPSCs)”, and “wherein the population of pluripotent stem cells is differentiated into a population of CD34+ hemogenic endothelium by way of embryoid bodies or 2D adherent cultures”. Step b of instant claim 53 aligns to reference claim 5 and 8 at “the concentration of SCF is 5 – 50 ng/mL” and “the concentration of FLT-3 is 5 – 30 ng/mL” and “the concentration is 10 – 50 ng/mL” and “the concentration of TPO is 1 – 10 ng/mL”. Step c of instant claim 54 aligns to reference claim 15 at “wherein the CD34+ HE cells are cultured in the presence of a Notch ligand; wherein the Notch ligand is attached to a solid surface; and/or the Notch ligand is attached to a cell culture dish” and reference claim 18 at “wherein the Notch ligand is not expressed by a stromal cell; wherein the method does not comprise co-culturing with a stromal cell expressing a Notch ligand”. Step b of instant claim 54 aligns to reference claim 12 at “wherein step (b) is performed for at least 1 week”. Step c of instant claim 54 aligns to reference claim 30 at “stem cell factor (SCF), FLT-3, IL-7 and thrombopoietin (TPO)”.
Reference claims 1, 30, and 52 do not recite “a G9a/GLP inhibitor” of instant claim 54. Daley teaches a method of differentiating CD34+ hemogenic endothelium comprising a histone methyltransferase inhibitor where G9a/GLP is a histone methyltransferase, and that inhibition promotes lymphoid potential of hematopoietic progenitors derived from pluripotent stem cells (page 15, para. 0093; page 49, para. 0307; Figure 1A).
It would have been obvious prior to the effective filing date of the invention as claimed to have modified the method of reference claims 1, 30, and 52 to include a G9a/GLP inhibitor in order to promote lymphoid potential of the CD34+ hemogenic endothelium to produce CD3+ T cells.
This is a provisional nonstatutory double patenting rejection.
25. Claims 1, 5, 13, 18, 19, 21, 26, 31, 33, 36, 38, 55, and 76 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 8, and 10 – 19 of U.S. Patent No. 11525119. Although the claims at issue are not identical, they are not patentably distinct from each other because the instant claims and Patent claims are drawn to a method of producing T cells by culturing CD34+ hemogenic endothelium generated from iPS cells with a histone methyltransferase inhibitor and a Notch ligand.
Step a and b of instant claim 1 aligns to step a of Patent claim 1 and step c of instant claim 1 aligns to step b of Patent claim 1. Instant claim 5 aligns to Patent claim 8 as both recite the notch ligand is selected from selected from the group consisting of Delta-like-I (DLL1), Delta-like-4 (DLL4), immobilized Deltal ext-IgG and immobilized Delta4ext-IgG, wherein optionally the immobilized Delta1ext-IgG consists of an extracellular domain of human Delta-like-I fused to the Fe domain of human IgG1.
Step b of instant claim 1 and instant claim 38 aligns with Patent claim 10 – 19 because instant claim 1 broadly recites “inhibiting a histone methyltransferase” and instant claim 38 recites EZH1 is a histone methyltransferase that catalyzes the addition of methyl group to H3K27 and the inhibitor is a small molecule inhibitor or a nucleic acid inhibitor and the small molecule inhibitor is a heterorganic compound or an organometallic compound; or selected from the group consisting of UNC0224, 3-deazaneplanocin A (DZNep), and UNC1999 (as recited in Patent claim 13) and others; and optionally the nucleic acid inhibitor is a nucleic acid targeting the expression of the histone methyltransferase (Patent claims 14 – 19).
26. Claims 53 is rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 8, and 10 – 19 of U.S. Patent No. 11525119. Although the claims at issue are not identical, they are not patentably distinct from each other because the instant claim and Patent claims are drawn to a method of producing T cells by culturing CD34+ hemogenic endothelium generated from iPS cells with a histone methyltransferase inhibitor and a Notch ligand.
Steps a – c of instant claim 53 aligns to Patent claim 1 as both recite differentiating pluripotent stem cells to CD34+ hemogenic endothelium, inhibiting a histone methyltransferase (Patent claims 10 – 19), and further differentiating to T cells with a notch ligand. The limitations of the Notch ligand of instant claim 53 map to Patent claim 8 as both are drawn to an immobilized Notch ligand.
27. Claims 54 is rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 8, and 10 – 13 of U.S. Patent No. 11525119. Although the claims at issue are not identical, they are not patentably distinct from each other because the instant claim and Patent claims are drawn to a method of producing T cells by culturing CD34+ hemogenic endothelium generated from iPS cells with a histone methyltransferase inhibitor and a Notch ligand.
Steps a and b of instant claim 54 align to Patent claim 1 as both recite differentiating pluripotent stem cells to CD34+ hemogenic endothelium, inhibiting a histone methyltransferase (Patent claims 10 – 13), and further differentiating to T cells with a notch ligand. The limitations of the Notch ligand of instant claim 54 map to Patent claim 8 as both are drawn to an immobilized Notch ligand. Instant claim 54 recites “a G9a/GLP inhibitor” and Patent claim 13 recites UNC0224, 3-deazaneplanocin A (DZNep) and UNC1999. The instant specification teaches the G9a/GLP inhibitor is UNC0224 (page 6, para. 0056). Therefore, “a G9a/GLP inhibitor” of instant claim 54 aligns to Patent claim 13 at recitation of UNC0224
Applicant’s Arguments/ Response to Arguments
28. Applicant Argues: Applicant asserts that Beck teaches a method of NK cell differentiation while claim 1 is drawn to differentiating CD3+ T cells.
Response to Argument: Applicant’s arguments have been fully considered and are persuasive, and the previous rejections of the claims citing the teachings of Beck have been withdrawn. In the new rejection, Vodyanyk teaches a stromal-free method of producing CD3+ T cells with immobilized Notch ligand hDLL4-Fc chimeric protein.
Applicant Argues: Applicant argues that the EZ_T cells are highly similar to PBMC alpha beta T cells and the Office does not demonstrate how Daley renders obvious the claimed methods that produce CD3+ T cells according to claim 76.
Response to Argument: The EZ_T cells are stroma-free ipsc-generated T cells with EZH1 knockdown (Applicant’s specification at para. 0085). Because Daley teaches that the method of producing iPSC-derived T cells with EZH1 knockdown can be performed in the absence of stroma with immobilized Notch ligand, and because Vodynayk teaches that CD3+ T cells are produced by a stroma-free method with immobilized Notch ligand, Daley in view of Vodynayk make obvious the method of claim 1 and thus make obvious the cells of claim 76.
Applicant Argues: Applicant disagrees with the cited teachings of Dallas in the rejections of claims 53 and 54 because the Office does not explain how murine bone marrow cells of Dallas are similar to a population of CD34+ hemogenic endothelium.
Response to Argument: Applicant’s arguments have been fully considered and are persuasive, and the previous rejections of the claims citing the teachings of Dallas have been withdrawn. In the new rejection, Kaneko teaches a stromal-free method of culturing CD34+ hemogenic endothelium with soluble Notch ligand to produce T cells and Vodyanyk teaches a stromal-free method of producing CD3+ T cells with immobilized Notch ligand hDLL4-Fc chimeric protein from CD34+ hemogenic endothelium.
Applicant Argues: Applicant asserts that the double patenting rejections should be withdrawn because the present application is the earlier filed application and the MPEP states that if a provisional nonstatutory rejection is the only rejection remaining it should be withdrawn.
Response to Argument: The previous provisional double patenting rejections have been withdrawn because of the amendments to the claims of reference application 18279336.
Conclusion
No claims allowed.
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/ZANNA MARIA BEHARRY/Examiner, Art Unit 1632
Prosecution Insights