CAR-Engineered Immune Cells via TRAC and B2M Gene Editing
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Solution Overview
Problem
Current genome engineering techniques for inserting transgenes into living cells are limited by random insertion locations, leading to unintended effects and lack of reproducibility, and there is a need for safe and effective universal donor cells for cell therapy treatments, particularly in immuno-oncology.
Innovation Solution
The development of genome-edited immune cells engineered to express chimeric antigen receptors (CARs) targeting CD19, CD70, or BCMA antigens, using RNA-guided nucleases and vectors to specifically modify the TRAC and B2M genes, allowing for precise expression of CARs without requiring purification or enrichment, and reducing the risk of host-versus-graft and graft-versus-host diseases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If random insertion technologies are used to insert transgenes into living cells, then the insertion process is simple, but the insertion location is unpredictable leading to disruption of normal gene regulation and severe unintended effects
Solution Approach 1:
The patent uses viral vectors as intermediary carriers to deliver transgenes into host cells. The viral vector system provides a controlled mechanism for gene insertion, replacing random integration with targeted delivery through viral infection processes, thereby improving safety and predictability while maintaining ease of manufacture
Solution Approach 2:
The patent replaces mechanical/random insertion methods with biologically-mediated insertion through viral vectors. Instead of physically forcing transgenes into cells (random integration), the system uses viral entry mechanisms to achieve controlled, targeted gene delivery, substituting mechanical randomness with biological specificity
2Manufacturing precision
If common genome engineering strategies such as ZFNs, TALENs, and HEs are used, then the precision of gene modification is improved, but the complexity of the system increases and limitations remain
Solution Approach 1:
The patent employs viral vectors that can deliver multiple genetic elements (transgenes, regulatory sequences, selection markers) simultaneously through a single delivery system. This multi-functional approach achieves precise gene modification while reducing the number of separate steps and components needed compared to ZFNs or TALENs
Solution Approach 2:
The viral vectors are pre-engineered with all necessary genetic elements and regulatory sequences before delivery. The transgenes are prepared in advance within the viral genome structure, allowing precise insertion at predetermined locations without requiring complex in-cell assembly steps
3Reliability
If genome-edited immune cells are engineered to express CAR constructs, then tumor reduction efficacy is improved, but the risk of host-versus-graft and graft-versus-host diseases increases
Solution Approach 1:
The patent extracts or removes genes responsible for T cell receptor (TCR) expression and MHC class I molecule expression from the engineered immune cells. By taking out these specific genetic elements, the cells retain tumor-targeting CAR functionality while eliminating the ability to cause graft-versus-host disease or be rejected by host immune systems
Solution Approach 2:
The patent converts the potential harm of immune cell rejection and GVHD into a benefit by deliberately engineering cells that cannot express TCR or MHC class I. This genetic modification transforms cells that would normally be immunogenic into universally compatible therapeutic cells, turning a safety risk into an advantage for allogeneic transplantation
4Manufacturing precision
If purification or enrichment of engineered cells is performed, then the quality and consistency of cell therapy is improved, but the time and cost of production increases
Solution Approach 1:
The viral vectors are designed with built-in selection markers and genetic elements that ensure only successfully transduced cells survive and proliferate. This preliminary design of the delivery system eliminates the need for subsequent purification steps, as the engineering process itself filters for quality cells
Solution Approach 2:
The engineered immune cells possess self-selecting properties through the CAR modification and genetic knockout. The cells automatically exhibit phenotypes that facilitate identification and selection of successfully engineered cells without requiring external purification processes, enabling direct use of crude preparations
Data Source
AI summary
Materials and methods for producing genome-edited cells engineered to express a chimeric antigen receptor (CAR) construct on the cell surface, and materials and methods for genome editing to modulate the expression, function, or activity of one or more immuno-oncology related genes in a cell, and materials and methods for treating a patient using the genome-edited engineered cells.