Rag1 Rag2 Knockout Pluripotent Stem Cells for T Cell Immunotherapy
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Solution Overview
Problem
Current methods for inducing T cells for cell-based immunotherapy face challenges such as the loss of antigen specificity due to additional TCRα chain rearrangement during differentiation from pluripotent stem cells, and the risk of graft-versus-host reactions in allogenic transplantation.
Innovation Solution
The method involves providing Rag1 and/or Rag2 gene knockout human pluripotent stem cells bearing genes encoding a T cell receptor (TCR) specific for a desired antigen, and inducing T cells from these stem cells to create a cell bank for cell-based immunotherapy. This approach prevents TCR rearrangement and reduces the risk of graft-versus-host reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If T cells are induced from pluripotent stem cells through conventional differentiation, then T cells can be generated for immunotherapy, but additional TCRα chain rearrangement occurs causing loss of antigen specificity and risk of autoreactive T cells
Solution Approach 1:
The patent extracts and removes the problematic TCRα chain rearrangement step from the differentiation process by using pre-assembled TCRα and TCRβ chains. Instead of allowing endogenous rearrangement during differentiation, the designed TCR chains are introduced exogenously to replace the natural rearrangement process, thereby eliminating loss of antigen specificity while maintaining the ability to generate functional T cells from pluripotent stem cells
Solution Approach 2:
The patent applies preliminary action by pre-assembling the TCRα and TCRβ chains with desired antigen specificity before introducing them to the pluripotent stem cells. This pre-preparation ensures that the T cells will express the correct TCR configuration from the start of differentiation, preventing subsequent rearrangement issues and ensuring consistent antigen specificity throughout the process
2Productivity
If allogenic transplantation is performed with iPS cells from donors with homozygous HLA haplotype to recipients with heterozygous HLA haplotypes, then a cell bank can be created, but graft-versus-host reactions may occur
Solution Approach 1:
The patent applies local quality by differentiating iPS cells into specific T cell subsets (such as regulatory T cells or specific cytotoxic T cell phenotypes) that have reduced alloreactivity. By controlling the differentiation outcome to produce T cells with specific functional properties rather than bulk T cells, the patent reduces the harmful graft-versus-host reaction risk while maintaining the productivity benefits of using homozygous HLA donor iPS cells for cell bank creation
3Duration of action of stationary object
If T cells are immortalized through fusion with cancer cells or long term culture with stimulating TCR and cytokines, then infinite proliferation is achieved, but cell function to kill cancer cells is lost and safety concerns arise
Solution Approach 1:
The patent applies dynamics by creating a system where T cells can be continuously generated from pluripotent stem cells on demand, rather than attempting to immortalize the T cells themselves. This dynamic approach allows fresh, functional T cells to be produced continuously through controlled differentiation, maintaining both infinite supply capability and preserved cancer cell killing function without the need for T cell immortalization
Data Source
AI summary
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.