TCR-Based CAR Composition for Stable Expression and Co-Stimulation
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Solution Overview
Problem
Existing transgenic T cell receptors (tTCRs) and TCR-based chimeric antigen receptors (CARs) face challenges such as loss of stability for efficient expression and activity, lacking sufficient co-stimulatory signaling, and optimal cell surface expression, limiting their effectiveness in adoptive immunotherapy.
Innovation Solution
Development of novel TCR-based CAR designs incorporating varied transmembrane domains and co-stimulatory signaling elements like human 4-1BB, CD28, and CD3zeta, with specific copy numbers, to enhance T cell signaling and stability, using bicistronic or multicistronic expression vectors for improved cell surface expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If transgenic T cell receptors (tTCRs) and TCR-based CARs are used, then antigen targeting capability is improved, but stability of expression and activity deteriorates
Solution Approach 1:
The CAR receptor is divided into separate functional domains: an antigen-binding domain (TCR alpha and beta chains) and a signaling domain (CD3zeta and costimulatory molecules). This segmentation allows each domain to be independently optimized for its specific function while maintaining overall stability. The TCR chains provide stable antigen recognition, while the CD3zeta and costimulatory domains provide stable signaling, resolving the contradiction between targeting capability and expression stability.
Solution Approach 2:
The invention creates a composite CAR structure combining TCR-based antigen recognition with CD3zeta and costimulatory signaling domains. This composite design integrates the strengths of each component: the TCR chains provide MHC-restricted antigen targeting, while CD3zeta provides stable intracellular signaling, and costimulatory molecules enhance stability. The composite structure achieves both effective antigen targeting and stable expression/activity.
2Adaptability or versatility
If tTCRs and TCR-based CARs are used, then antigen recognition is improved, but co-stimulatory signaling is insufficient
Solution Approach 1:
The invention merges the TCR antigen recognition function with CD3zeta signaling and costimulatory signaling functions into a single CAR construct. By combining these functions in one receptor molecule, the system achieves both effective antigen recognition (through TCR chains) and reliable co-stimulatory signaling (through CD3zeta and costimulatory domains), resolving the contradiction between recognition and signaling capabilities.
Solution Approach 2:
The CAR construct is designed to perform multiple functions simultaneously: antigen recognition (TCR chains), signal transduction (CD3zeta), and costimulation (costimulatory molecules). This multi-functional design ensures that the receptor can effectively recognize antigens while providing sufficient co-stimulatory signaling, eliminating the limitation of separate receptor systems.
3Ease of operation
If conventional TCR-based CAR designs are used, then basic functionality is achieved, but cell surface expression is suboptimal
Solution Approach 1:
The invention optimizes parameters of the CAR construct, including the choice of transmembrane domains, cytoplasmic domain sequences, and costimulatory molecule configurations. By carefully selecting and adjusting these parameters, the CAR achieves optimal cell surface expression levels while maintaining basic functionality. The specific combination of CD3zeta and costimulatory molecules enhances surface expression stability.
Data Source
AI summary
The present invention relates to agents, compositions, and methods to confer and/or increase immune responses mediated by cellular immunotherapy.