Engineered TCR Residue Substitution for Surface Expression
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Solution Overview
Problem
There is a need to increase the cell surface expression of weakly expressed T cell receptors (TCRs) to enhance the efficacy of TCR therapy, as high levels of TCR expression on the cell surface result in greater avidity and better therapeutic outcomes, but existing TCRs differ significantly in their ability to be expressed and often get outcompeted by endogenous TCRs.
Innovation Solution
Identifying and substituting specific amino acid residues at key positions in the TCR framework regions, such as L96 of the α chain, R9 of the β chain, Y10 of the β chain, and T24 of the α chain, with residues like leucine, arginine, tyrosine, and threonine, to enhance dimerization and reduce mispairing, thereby increasing cell surface expression without affecting peptide/MHC recognition or TCR structure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If TCR is expressed as an exogenous TCR, then antigen-specific T cell function is achieved, but cell surface expression level is reduced and mispairing with endogenous TCR occurs
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid residues at specific positions within the TCR framework regions. By changing the chemical properties, charge, or size of residues at key positions (such as CDR-HL3, CDR-HL2, CDR-LL3, CDR-LL2, FR3, FR4, or transmembrane domains), the patent optimizes dimerization efficiency and cell surface expression while maintaining antigen-specific function.
Solution Approach 2:
The patent applies local quality by making targeted modifications at specific localized regions of the TCR structure rather than global changes. By focusing mutations on particular framework regions or CDRs that influence dimerization and surface expression, the patent achieves improved cell surface expression and reduced mispairing while preserving the overall TCR structure and antigen recognition capability.
2Quantity of substance
If amino acid substitutions are made to enhance TCR expression, then cell surface expression is improved, but TCR structure and folding may be affected
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid residues at specific positions within the TCR framework regions. By changing the chemical properties, charge, or size of residues at key positions (such as CDR-HL3, CDR-HL2, CDR-LL3, CDR-LL2, FR3, FR4, or transmembrane domains), the patent optimizes dimerization efficiency and cell surface expression while maintaining antigen-specific function.
Solution Approach 2:
The patent applies preliminary action by performing computational predictions and in silico modeling of amino acid substitutions before actual experimentation. By predicting which residue changes are most likely to improve expression without disrupting structure, and by testing combinations systematically, the patent reduces the risk of adverse structural effects.
3Quantity of substance
If multiple amino acid residues are substituted to maximize expression, then TCR expression is significantly improved, but complexity of the engineering process increases
Solution Approach 1:
The patent applies segmentation by dividing the TCR structure into distinct functional regions (CDRs and framework regions) and addressing each region separately with targeted mutations. By segmenting the engineering approach into modular steps—selecting positions, predicting substitutions, testing combinations—the patent makes the complex process more manageable and systematic.
Solution Approach 2:
The patent applies universality by developing a systematic framework that can be applied to different TCR specificities and types. The methods for identifying and modifying key framework region residues represent a universal approach that can enhance expression of various exogenous TCRs, reducing the need for de novo engineering for each new TCR.
Data Source
Figure 1A~1B
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AI summary
TCR The present invention relates to an engineered T cell receptor (TCR). In particular, the present invention relates to methods for expression of a TCR when expressed as an exogenous TCR, to methods for selecting a TCR with high cell surface expression when expressed as an exogenous TCR and to methods for identifying residues which contribute to the cell surface expression level of a TCR. The present invention also relates to an engineered TCR which has a high level of cell surface expression when expressed as an exogenous TCR compared to the corresponding germline TCR sequence.