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Production of T cells from RAG-inactivated iPSC

A technology of cells and cell groups, applied in the field of progenitor cells, which can solve problems such as lack and limitation

Pending Publication Date: 2022-04-01
ADAPTIMMUNE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, current adoptive T cell therapy is limited by the lack of suitable patient- and tumor-specific T cells, and sufficient therapeutic and functional antigen-specific T cells are required for effective use in immunotherapy

Method used

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  • Production of T cells from RAG-inactivated iPSC
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  • Production of T cells from RAG-inactivated iPSC

Examples

Experimental program
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Embodiment

[0208] method

[0209] hiPSC culture

[0210] iPSCs are routinely cultured in mTeSR1(SCT) on Matrigel (BD brand, Corning "Corning") using tissue culture plasticware in 5% CO 2 , 5% O 2 and 37°C. HiPSCs were obtained according to the instructions using the EasyPassage tool (Invitrogen), and seeded in a medium with 10 μm Y27632 (Andy Biotechnology "R&D Systems") at a ratio of 1:6 or 1:12 in the first 48 hours of culture. For differentiation, hiPSCs were passaged in low-density culture onto matrigel or vitronectin using a split ratio of 1:48 or 1:98. 24 hours after inoculation, the inoculation density was approximately 1 colony per field of view when observed under a microscope with a magnification of ×4. hiPSCs were cultured in mTeSR2 or E8flex (SCT) for approximately 4-5 days, depending on the cell culture substrate used, until colonies became dense and distinct cells were no longer visible.

[0211] T cell differentiation from pluripotent stem cells

[0212] The hi...

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Abstract

The present invention relates to the differentiation of progenitor cells inactivating a recombinant activation gene (RAG) into T cells by expression of an exogenous T cell receptor (TCR). A population of T cells can be produced by (i) differentiating a population of RAG-inactivated induced pluripotent stem cells (iPSC) into mesoderm cells, (ii) differentiating the mesoderm cells (MC) to produce a population of hematopoietic endothelial cells (HECs), (iii) differentiating the HECs into a population of hematopoietic progenitor cells (HPC), (iv) differentiating the population of HPCs into T cell progenitor cells, and (v) differentiating the population of T cell progenitor cells. And (v) maturing the T cell progenitor cells to produce a double positive CD4 + CD8 + T cell population. The method may also include introducing a heterologous nucleic acid encoding an antigen receptor (e.g., a T cell receptor (TCR) or a chimeric antigen receptor (CAR)) into one of the following groups: RAG inactivated (a) iPSC, (b) MC, (c) HEC, (d) HPC, or (e) progenitor T cells. This may contribute, for example, to the production of T cells for immunotherapy.

Description

technical field [0001] The present invention relates to a method for generating T cells from RAG-inactivated progenitor cells, such as induced pluripotent stem cells (iPSCs). Background technique [0002] Immunotherapy promises to transform the field of cancer treatment with the promise of long-term survival (McDermott et al. Cancer Treat Rev. 2014 Oct;40(9):1056-64). There is a clear unmet medical need for new immunomodulatory agents across a broad spectrum of patient populations and tumor types. In addition, new agents are needed to increase the magnitude and duration of antitumor responses. The development of these reagents has been made possible by an improved understanding of the fundamentals governing T cell immunity over the past two decades (Sharma and Allison, Cell. 2015 Apr 9;161(2):205-14 ). This typically requires recognition of tumor-associated peptide antigens presented by MHC molecules by tumor-specific CD4+ and CD8+ T cells. Different vaccination strategi...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/0789C12N5/10C12N15/864C12N15/867A61K35/17A61P35/00
CPCC12N5/0638C12N5/0647C12N2506/45C12N2506/11C12N2501/16C12N2501/155C12N2501/165C12N2501/115C12N2501/125C12N2501/727C12N5/10C12N2740/16043A61P35/00A61K39/464486A61K39/4611A61K39/464481A61K39/4632A61K39/464488C12N15/86C07K14/7051
Inventor L·J·巴克L·卡朋特A·西德韦
Owner ADAPTIMMUNE
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