IP Library › Granted Patent US 12,241,089
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)
Assignees: Kyoto University; Thyas Co. Ltd.
C12N5/0696A61K35/14A61K39/4611A61K39/4632A61K39/464453A61K39/464838A61P35/00C12N5/0636C12N5/0638C12N5/10C12N2501/515C12N2506/45C12N2510/00
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Quick Facts
Patent No.
US 12,241,089
App. No.
15/326,977
Granted
Mar 4, 2025
Kind
B2
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
Related Publication 20170369850A1 · Dec 28, 2017
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