PDCD1 Knockout CAR-T Cells for Lower Allogeneic Immunogenicity
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Solution Overview
Problem
Existing CAR-T cell therapies face challenges in immunogenicity and reactivity, leading to harmful reactions such as rejection by the host or graft-versus-host-disease, limiting the accessibility of allogeneic adoptive transfer strategies for universal CAR cells.
Innovation Solution
Inactivation of the PDCD1 gene in cells using a CRISPR nuclease and RNA molecule with a guide sequence targeting specific regions of the PDCD1 gene, such as Exons 1-5, to modify cells for improved activity, retention, and expansion in allogeneic adoptive cancer immunotherapy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If CAR-T cell therapy is developed using allogeneic adoptive transfer strategy, then accessibility and universality are improved, but immunogenicity and reactivity increase causing harmful reactions
Solution Approach 1:
The patent extracts and removes the harmful PDCD1 gene (which encodes PD-1 protein) from the CAR-T cells using CRISPR-Cas9 gene editing. By taking out this specific gene responsible for immunogenicity and reactivity, the therapy maintains its anti-tumor activity while eliminating the harmful effects such as graft-versus-host disease and host rejection, thus enabling universal allogeneic application
Solution Approach 2:
The patent changes the genetic parameter of the CAR-T cells by modifying the PDCD1 gene status from present to knocked out. This parameter change fundamentally alters the immune properties of the cells, transforming them from a state that causes harmful reactions to one that is suitable for universal allogeneic transfer while maintaining therapeutic efficacy
2Reliability
If PDCD1 gene is knocked out in CAR-T cells, then activity and retention are improved, but manufacturing complexity increases
Solution Approach 1:
The patent replaces complex traditional gene editing methods with the CRISPR-Cas9 system, which uses a simpler mechanism involving guide RNA and Cas9 nuclease. This substitution reduces the manufacturing complexity compared to earlier gene editing approaches while achieving reliable and consistent knockout of the PDCD1 gene, thereby improving cell activity and retention
Solution Approach 2:
The patent uses CRISPR-Cas9 as an intermediary tool to achieve the desired genetic modification. This intermediary system simplifies the overall manufacturing process by providing a standardized, efficient, and controllable method for gene knockout, making the complexity manageable while achieving the therapeutic goals of improved activity and retention
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
Enhances the performance of modified cells by improving activity, retention, and expansion, reducing immunogenicity and reactivity, thereby facilitating safer and more effective allogeneic adoptive transfer therapies.
Implementation Method 1
introducing to the cell a composition comprising: a CRISPR nuclease, or a polynucleotide molecule encoding the CRISPR nuclease; and an RNA molecule comprising a guide sequence portion having 17-50 nucleotides, or a polynucleotide molecule encoding the RNA molecule, wherein a complex of the CRISPR nuclease and the RNA molecule affects a double strand break in the allele of the PDCD1 gene
Data Source
AI summary
Compositions comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-6220 and methods and uses thereof.