Universal CAR-T Cell Preparation via HLA-A/B Knockout
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Solution Overview
Problem
Current CAR-T cell therapy is constrained by the need for individualized patient-specific treatment, high costs, and the risk of graft-versus-host disease, limiting the scalability and uniformity of universal CAR-T cell production.
Innovation Solution
A method for preparing universal chimeric antigen receptor T cells using CRISPR-Cas9 gene editing to knock out HLA-A and HLA-B genes, inhibiting their binding to TCR, while maintaining normal β2m expression, allowing for the expression of a chimeric antigen receptor targeting tumor cells and avoiding immune recognition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional CAR-T cell therapy is used with individualized patient-specific treatment, then treatment efficacy is improved, but production scalability and uniformity deteriorate
Solution Approach 1:
The patent creates universal CAR-T cells that can be used for multiple patients through gene editing to eliminate HLA-A and HLA-B expression. The CAR-T cells are designed with a universal structure that includes a specific CAR molecule (e.g., anti-CD19) combined with gene modifications (CRISPR/Cas9 editing of HLA-A and HLA-B loci) that allow them to function across different patients without individual customization, thus achieving both efficacy and scalability
2Ease of manufacture
If universal CAR-T cells are produced without gene editing, then production cost and time are reduced, but graft-versus-host disease risk increases
Solution Approach 1:
The patent extracts and removes the harmful HLA-A and HLA-B molecules from the CAR-T cell surface through gene editing. By specifically targeting and eliminating these HLA loci using CRISPR/Cas9 technology, the method removes the source of graft-versus-host disease while maintaining the therapeutic function of the CAR-T cells, thus achieving safety without significantly complicating the production process
3Reliability
If HLA-A and HLA-B expression is maintained in CAR-T cells, then immune recognition and survival are improved, but graft-versus-host disease and host-versus-graft responses occur
Solution Approach 1:
The patent applies local quality modification by specifically targeting only the HLA-A and HLA-B loci for gene editing while preserving other important immune recognition molecules. The CRISPR/Cas9 system is designed to edit only specific HLA genes, leaving HLA-C and other protective molecules intact, thus maintaining necessary immune function while eliminating the specific harmful HLA-A and HLA-B molecules that cause GVHD and HVG responses
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The approach enables the production of universal CAR-T cells that can be used for allogeneic tumor treatment without inducing GVHD or HVG responses, improving their survival and anti-tumor efficacy, and can be scaled up for uniform quality across patients.
Implementation Method 1
modifying the T cell to express the CAR, and inhibiting the binding of HLA-A and HLA-B of the CAR-T cell to TCR, thereby obtaining the CAR-T cell; wherein in step (B), HLA-A and HLA-B genes are knocked out using a gene editing system, and the gene editing system comprises CRISPR-Cas9 system
Implementation Method 2
the gene editing system comprises CRISPR-Cas9 system, wherein said CRISPR-Cas9 system comprises gRNA and Cas9 protein; said gRNA comprises a gRNA targeting HLA-A and a gRNA targeting HLA-B
Data Source
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AI summary
Provided are a universal chimeric antigen receptor (CAR) T cell and preparation method and application thereof. The binding of the cell HLA-A/HLA-B to the TCR is inhibited; the TCR gene expression is silenced; the invention man be used for allogeneic tumor treatment and does not cause GVHD and HVG reaction during allogeneic infusion, and thus improves the survival and antineoplastic effect of allogeneic CAR-T cells in the receptor.