Granted Patent
B2
US 12,241,089 · App. 15/326,977 · Granted Mar 4, 2025
Method for inducing t cells for cell-based immunotherapy from pluripotent stem cells
Inventors:
Shin Kaneko (Kyoto, JP); Hiroshi Kawamoto (Kyoto, JP); Kyoko Masuda (Kyoto, JP); Atsutaka Minagawa (Kyoto, JP); Akitsu Hotta (Kyoto, JP); Yutaka Shimazu (Kyoto, JP); Hiroshi Ichise (Kyoto, JP)
C12N5/0696A61K35/14A61K39/4611A61K39/4632A61K39/464453A61K39/464838A61P35/00C12N5/0636C12N5/0638C12N5/10C12N2501/515C12N2506/45C12N2510/00
View Patent ↗
Loading inventors, assignments & file history…
Abstract
Provided is a method for inducing T cells for a cell-based immunotherapy, comprising the steps of: (1) providing Rag 1 and/or Rag 2 gene knockout human pluripotent stem cells bearing genes encoding a T cell receptor specific for a desired antigen, and (2) inducing T cells from the pluripotent stem cells of step (1). Further provided are a cell-based immunotherapy method that uses the T cells for the cell-based immunotherapy and an iPS cell bank for the cell-based immunotherapy.
Claims (12)
1. An in vitro method for inducing T cells for a cell-based immunotherapy, where the T cells are CD8 single positive T cells derived from human pluripotent stem cells, comprising the steps of:
(i) providing human pluripotent stem cells having both alleles of Rag-1 or Rag-2 gene knocked out and comprising genes encoding an alpha chain and a beta chain of a T cell receptor specific for a desired antigen,
wherein the human pluripotent stem cells of step (i) are prepared by a method comprising the steps of:
(a) introducing genes encoding the alpha chain and beta chain of the antigen specific T cell receptor for the desired antigen into the pluripotent stem cells; and
(b) knocking out both alleles of the Rag-1 or Rag-2 gene in the human pluripotent stem cells provided in step (a) by genome editing;
(ii) inducing the pluripotent stem cells of step (i) into a population of CD3 positive T cells, wherein the population of CD3 positive T cells comprise CD4CD8 double positive T cells, and CD4CD8 double negative T cells;
(iii) isolating only the CD4CD8 double positive T cells from the population of T cells in step (ii) using flow cytometry; and
(iv) inducing CD8 single positive T cells, which are differentiated from the isolated CD4CD8 double positive T cells of step (iii), and
wherein the CD8 single positive T cells have no rearrangement of the alpha chain of the T cell receptor specific for the desired antigen.
2. The method according to claim 1 , wherein the human pluripotent stem cells are human iPS cells.
3. The method according to claim 1 , wherein the Rag-1 or Rag-2 gene is knocked out by CRISPR-Cas9.
4. The method of claim 2 , wherein the iPS cells have a homozygous HLA haplotype that matches at least one of HLA haplotypes of a subject to be treated.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 13, 2017
From: KANEKO, SHIN; KAWAMOTO, HIROSHI; MASUDA, KYOKO; MINAGAWA, ATSUTAKA; HOTTA, AKITSU; SHIMAZU, YUTAKA; ICHISE, HIROSHI
To: KYOTO UNIVERSITY; THYAS CO. LTD.
Reel/Frame 042688/0766 →
Continuity (3)
Provisional Application
62026341
· Jul 18, 2014
Provisional Application
62026332
· Jul 18, 2014
References Cited (120)
US 8048999B2
· Yamanaka et al.
· 2011
[cited by applicant]
US 8278104B2
· Yamanaka et al.
· 2012
[cited by applicant]
US 8877493B2
· Sekiguchi et al.
· 2014
[cited by applicant]
US 9127256B2
· Fusaki et al.
· 2015
[cited by applicant]
US 9546384B2
· Frendewey
· 2017
[cited by examiner]
US 9683232B2
· Yamanaka et al.
· 2017
[cited by applicant]
US 20110117645A1
· Yasuda
· 2011
[cited by applicant]
EP 2853590A1
· 2015
[cited by applicant]
WO WO2006132524A1
· 2006
[cited by applicant]
WO WO2007069666A1
· 2007
[cited by applicant]
WO WO2008118820A2
· 2008
[cited by applicant]
WO WO2009007852A2
· 2009
[cited by applicant]
WO WO2009032194A1
· 2009
[cited by applicant]
WO WO2009057831A1
· 2009
[cited by applicant]
WO WO2009058413A1
· 2009
[cited by applicant]
WO WO2009075119A1
· 2009
[cited by applicant]
WO WO2009079007A1
· 2009
[cited by applicant]
WO WO2009091659A2
· 2009
[cited by applicant]
WO WO2009101084A1
· 2009
[cited by applicant]
WO WO2009101407A2
· 2009
[cited by applicant]
WO WO2009102983A2
· 2009
[cited by applicant]
WO WO2009114949A1
· 2009
[cited by applicant]
WO WO2009117439A2
· 2009
[cited by applicant]
WO WO2009123349A1
· 2009
[cited by applicant]
WO WO2009126250A2
· 2009
[cited by applicant]
WO WO2009126251A2
· 2009
[cited by applicant]
WO WO2009126655A2
· 2009
[cited by applicant]
WO WO2009157593A1
· 2009
[cited by applicant]
WO WO2010008054A1
· 2010
[cited by applicant]
WO WO2010009015A2
· 2010
[cited by applicant]
WO WO2010013845A1
· 2010
[cited by applicant]
WO WO2010033906A2
· 2010
[cited by applicant]
WO WO2010033920A2
· 2010
[cited by applicant]
WO WO2010042800A1
· 2010
[cited by applicant]
WO WO2010050626A1
· 2010
[cited by applicant]
WO WO2010056831A2
· 2010
[cited by applicant]
WO WO2010068955A2
· 2010
[cited by applicant]
WO WO2010098419A1
· 2010
[cited by applicant]
WO WO2010102267A2
· 2010
[cited by applicant]
WO WO2010111409A2
· 2010
[cited by applicant]
WO WO2010111422A2
· 2010
[cited by applicant]
WO WO2010115050A2
· 2010
[cited by applicant]
WO WO2010124290A2
· 2010
[cited by applicant]
WO WO2010137746A1
· 2010
[cited by applicant]
WO WO2010147395A2
· 2010
[cited by applicant]
WO WO2010147612A1
· 2010
[cited by applicant]
WO WO2011043405A1
· 2011
[cited by applicant]
WO WO2011096482A1
· 2011
[cited by applicant]
WO 2013176197A1
· 2013
[cited by applicant]
WO WO2014165707A2
· 2014
[cited by examiner]
WO 2015099134A1
· 2015
[cited by applicant]
Delmonte et al., Journal of Clinical Immunology (2018) 38:646-655. (Year: 2018).
[cited by examiner]
Nakatsuji et al., Nature Biotechnology, 26(7): 739-740, Jul. 2008). (Year: 2008).
[cited by examiner]
Vizcardo et al., Cell Stem Cell, 12:31-36, Jan. 3, 2013, supplemental materials, pp. 1-9.
[cited by examiner]
Fong et al., Molecular Immunology 37: 391-402, 2000.
[cited by examiner]
Eljaafari J Immunol ; 190:184-194 (Year: 2013).
[cited by examiner]
Yu et al The Journal of Experimental Medicine ⋅ vol. 197, No. 4, 475-487 (Year: 2003).
[cited by examiner]
Eminli et al. (2008) “Reprogramming of Neural Progenitor Cells into Induced Pluripotent Stem Cells in the Absence of Exogenous Sox2 Expression,” Stem Cells, 26, pp. 2467-2474.
[cited by applicant]
Feng et al. (2008) “Reprogramming of fibroblasts into induced pluripotent stem cells with orphan nuclear receptor Esrrb,” Nat Cell Biol., 11, pp. 197-203.
[cited by applicant]
Han et al. (2010) “Tbx3 improves the germ-line competency of induced pluripotent stem cells,” Nature, 463, pp. 1096-1100.
[cited by applicant]
Heng et al. (2009) “The Nuclear Receptor Nr5a2 Can Replace Oct4 in the Reprogramming of Murine Somatic Cells to Pluripotent Cells,” Cell Stem Cell, 6, pp. 167-174.
[cited by applicant]
Huangfu et al. (2008) “Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2,” Nat. Biotechnol., 26, pp. 1269-1275.
[cited by applicant]
Ichida et al. (2009) “A Small-Molecule Inhibitor of Tgf-b Signaling Replaces Sox2 in Reprogramming by Inducing Nanog” Cell Stem Cell, 5, pp. 491-503.
[cited by applicant]
Yu et al. (2007) “Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells,” Science, 318, pp. 1917-1920.
[cited by applicant]
Takahashi et al. (2007) “Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors,” Cell, 131, pp. 861-872.
[cited by applicant]
Kim et al. (2009) “Direct reprogramming of human neural stem cells by OCT4,” Nature, 461, pp. 649-643.
[cited by applicant]
Lyssiotis et al. (2009) “Reprogramming of murine fibroblasts to induced pluripotent stem cells with chemical complementation of Klf4,” Proc. Natl. Acad. Sci., 106, pp. 8912-8917.
[cited by applicant]
Mali et al. (2010) “Butyrate Greatly Enhances Derivation of Human Induced Pluripotent Stem Cells by Promoting Epigenetic Remodeling and the Expression of Pluripotency-Associated Genes,” Stem Cells., 28, pp. 713-720.
[cited by applicant]
Marson et al. (2008) “Wnt signaling promotes reprogramming of somatic cells to pluripotency,” Cell Stem Cell., 3, pp. 132-135.
[cited by applicant]
Nakagawa et al. (2008) “Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts,” Nat. Biotechnol., 26, pp. 101-106.
[cited by applicant]
Judson et al. (2009) “Embryonic stem cell specific microRNAs promote induced pluripotency,” Nat. Biotech., 27, pp. 459-461.
[cited by applicant]
Shi et al. (2008) “A Combined Chemical and Genetic Approach for the Generation of Induced Pluripotent Stem Cells,” Cell Stem Cell., 2, pp. 525-5283.
[cited by applicant]
Shi et al. (2008) “A Combined Chemical and Genetic Approach for the Generation of Induced Pluripotent Stem Cells,” Cell Stem Cell., 3, pp. 568-574.
[cited by applicant]
Zhao et al. (2008) “Two Supporting Factors Greatly Improve the Efficiency of Human iPSC Generation,” Cell Stem Cell., 3, pp. 475-479.
[cited by applicant]
Bendle et al. (2010) “Two Supporting Factors Greatly Improve the Efficiency of Human iPSC Generation,” Nat. Med., 16(5), pp. 565-570.
[cited by applicant]
Takahashi et al. (2006) “Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors,” Cell, 126, pp. 663-676.
[cited by applicant]
Cyranoski et al. (2012) “Stem-cell pioneer banks on future therapies,” Nature, 488, 139, 2 pp.
[cited by applicant]
Huangfu et al. (2008) “Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds,” Nat. Biotechnol., 26, pp. 795-797.
[cited by applicant]
Ito et al. (1988) “Change of HLA phenotype in postoperative erythroderma,” Lancet, 331, p. 413.
[cited by applicant]
Cong et al. (2013) “Multiplex Genome Engineering Using CRISPR/Cas Systems,” Science, 39; 819, 9 pp.
[cited by applicant]
Maekawa et al. (2011) “Direct reprogramming of somatic cells is promoted by maternal transcription factor Glis1,” Nature, 474, 7 pp.
[cited by applicant]
Morgan et al. (2006) “Cancer Regression in Patients After Transfer of Genetically Engineered Lymphocytes,” Science, 314, 5 pp.
[cited by applicant]
Nishimura et al. (2013) “Generation of Rejuvenated Antigen-Specific T Cells by Reprogramming to Pluripotency and Redifferentiation,” Cell Stem Cell, pp. 114-126.
[cited by applicant]
Mali et al. (2013) “RNA-Guided Human Genome Engineering via Cas9,” Science, 39: 8 pp.
[cited by applicant]
Okita et al. (2008) “Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors,” Science, 322, pp. 949-953.
[cited by applicant]
Kaneko (2014), The Medical Frontline, 69, pp. 724-733.
[cited by applicant]
Sun et al. (2009) “Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells,” Proc. Natl. Acad. Sci., 106, pp. 15720-15725.
[cited by applicant]
Takahashi et al. (2009) “Human Induced Pluripotent Stem Cells on Autologous Feeders,” PLoS One, 4, 6 pp.
[cited by applicant]
Timmermans et al. (2009) “Generation of T Cells from Human Embryonic Stem Cell-Derived Hematopoietic Zones,” Journal of Immunology, 182, pp. 6879-6888.
[cited by applicant]
Vizcardo et al. (2013) “Regeneration of Human Tumor Antigen-Specific T Cells from iPSCs Derived from Mature CD8+ T Cells,” Cell Stem Cell, 12, pp. 31-36.
[cited by applicant]
Warren et al. (2010) “Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells with Synthetic Modified mRNA,” Cell Stem Cell., 7, pp. 618-630.
[cited by applicant]
Yoshida et al. (2009) “Hypoxia Enhances the Generation of Induced Pluripotent Stem Cells,” Cell Stem Cell., 5, pp. 237-241.
[cited by applicant]
Nakajima (2013), Hematology Frontier, vol. 23, No. 8, pp. 1105-1110.
[cited by applicant]
Gattinoni et al. (2006) “Adoptive immunotherapy for cancer: building on success,” The Journal of Immunology, vol. 6, No. 5, pp. 383-393.
[cited by applicant]
Lei et al. (Jul. 2011) “In Vivo Programming of Tumor Antigen-Specific T Lymphocytes from Pluripotent Stem Cells to Promote Cancer Immunosurveillance,” Cancer Research, vol. 71, No. 14, pp. 4742-4747.
[cited by applicant]
Duda et al. (2014) “High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPRjCas9 nickase pairs,” Nucleic Acids Research, vol. 42, No. 10, 16 pp.
[cited by applicant]
Nishimura et al. (2013) “Generation of Rejuvenated Antigen-Specific T Cells by Reprogramming to Pluripotency and Redifferentiation,” Cell Stem Cell, vol. 12, No. 1, pp. 114-126.
[cited by applicant]
English translation of International Preliminary Report on Patentability for PCT/JP2015/070608 mailed on Jan. 24, 2017.
[cited by applicant]
Extended European Search Report issued in the corresponding EP Application No. 15821273.8 mailed on Feb. 2, 2018, 7 pp.
[cited by applicant]
International Search Report and Written Opinion of PCT/JP2015/070608 mailed Oct. 20, 2015, 10 pp.
[cited by applicant]
Hale et al. (2010) “Cutting Edge: Rag Deletion in Peripheral T Cells Blocks TCR Revision,” J. Immunol. 184:5964-5968.
[cited by applicant]
Mombaerts et al. (1992) “RAG-1-deficient mice have no mature B and T lymphocytes,” Cell. 68:869-877.
[cited by applicant]
Shinkai et al. (1992) “RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement,” Cell. 68:855-867.
[cited by applicant]
Turka et al. (1991) “Thymocyte expression of RAG-1 and RAG-2: termination by T cell receptor cross-linking,” Science. 253:778-781.
[cited by applicant]
Watanabe et al. (Feb. 2013) “Establishment of a stable T lymphoma cell line transduced with HLA-A*24:02-restricted WT1-specific TCR genes and its application to antigen-specific immunomonitoring,” Biomed. Res. 34(1):41-…
[cited by applicant]
International Search Report with Written Opinion corresponding to International Patent Application No. PCT/JP2015/070608, mailed Oct. 28, 2015.
[cited by applicant]
Minagawa et al., “Enhancing T Cell Receptor Stability in Rejuvenated Ipsc-Derived T Cells Improves Their Use in Cancer Immunotherapy”, Cell Stem Cell vol. 23, pp. 850-858, Dec. 6, 2018.
[cited by applicant]
Riolobos et al., “HLA Engineering of Human Pluripotent Stem Cells”, Molecular Therapy, vol. 21, No. 6, pp. 1232-1241, Jun. 2013.
[cited by applicant]
Themeli et al., “Generation of tumor-targeted human T lymphocytes from induce pluripotent stem cells for cancer therapy”, Nature Biotechnology, vol. 31, No. 10, pp. 928-933 doi: 10.1038/nbt.2678.
[cited by applicant]
Maeda et al., “Regeneration of CD8aβ T Cells from T-cell-Derived iPSC Imparts Potent Tumor Antigen-Specific Cytotoxicity”, Cancer Research, Dec. 1, 2016, vol. 76, No. 23, pp. 6839-6850.
[cited by applicant]
Shlyahtenko et al., “Molecular Mechanism Underlying RAG1/RAG2 Synaptic Complex Formation.” The Journal of Biological Chemistry 284(31):20956-20964 (2009).
[cited by applicant]
Cheroutre et al., “Doubting the TCR Coreceptor Function of CD8aa.” Immunity 28:149-159 (2008).
[cited by applicant]
Parel et al., “CD4+ CD8+ double positive (DP) T cells in health and disease.” Autoimmunity Reviews 3:215-220 (2004).
[cited by applicant]
McMahan et al., “RAG Reexpression and DNA Recombination at T Cell Receptor Loci in Peripheral CD4+ T Cells,” Immunity, 9:637-647, 1998.
[cited by applicant]
Michie et al., “Allelic exclusion and differentiation by protein kinase C-mediated signals in immature thymocytes,” Proc. Natl. Acad. Sci., 98(2):609-614, 2001.
[cited by applicant]
Minagawa et al., “Enhancing T Cell Receptor Stability in Rejuvenated iPSC-Derived T Cells Improves Their Use in Cancer Immunotherapy,” Cell Stem Cell, 23:850-858, 2018.
[cited by applicant]
Shinkai et al., “RAG-2-Deficient Mice Lack Mature Lymphocytes Owing to Inability to Initiate V(D)J Rearrangement,” Cell, 68:855-867, 1992.
[cited by applicant]
Villa et al., “V(D)J recombination defects in lymphocytes due to RAG mutations: severe immunodeficiency with a spectrum of clinical presentations,” Blood, 97(1):81-88, 2001.
[cited by applicant]