T Cell Genomic Modification via TCR Locus Insertion
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current techniques for modifying ex vivo or intravitally gene-edited cells are limited in their ability to correct multiple mutations or introduce new synthetic genes, restricting their therapeutic applicability, especially for T cells used in adoptive cellular therapies.
Innovation Solution
The method involves inserting a nucleic acid encoding a polypeptide and a heterologous T cell receptor or synthetic antigen receptor into a specific endogenous site in the genome of human T cells, allowing for the alteration of T cell specificity and function, such as by combining a human Fas or TNFRSF12 extracellular domain with an OX40 intracellular domain via a transmembrane domain, to generate T cells with altered antigen specificity and functionality.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If current gene editing techniques are used to correct mutations or integrate synthetic genes, then therapeutic applicability is limited to single mutations or requires extensive R&D, but the ability to modify T cells with multiple functions and specificities is restricted
Solution Approach 1:
The patent divides the T cell modification process into modular components: a targeted nuclease system for precise genomic cutting, homology-directed repair templates for controlled DNA insertion, and standardized polypeptide sequences (such as Fas-OX40, TNFRSF12-OX40, LAG-3-4-1BB). Each module can be independently designed and combined to create customized T cell therapies for different diseases and applications, eliminating the need to develop entirely new synthetic genes for each therapeutic scenario.
Solution Approach 2:
The patent employs homology-directed repair (HDR) as an intermediary mechanism between the targeted nuclease and the desired genomic modification. The HDR system uses homology arms flanking the insertion site to guide precise integration of the nucleic acid construct, serving as a mediator that enables controlled genetic modification without requiring extensive de novo gene synthesis. This intermediary approach standardizes the modification process across different therapeutic applications.
2Adaptability or versatility
If nucleic acid constructs are inserted into the TCR locus to alter T cell specificity and function, then T cells gain desired antigen specificity and polypeptide function, but genomic modification complexity increases
Solution Approach 1:
The patent merges multiple functions into a single integrated nucleic acid construct that simultaneously provides: (1) the heterologous TCR or CAR for antigen recognition, (2) the polypeptide of interest (such as Fas-OX40, TNFRSF12-OX40, LAG-3-4-1BB) for therapeutic function, and (3) regulatory elements for controlled expression. This merging approach allows T cells to acquire both desired antigen specificity and polypeptide function through a single genomic insertion event, rather than requiring separate modifications for each function.
Solution Approach 2:
The patent creates universal modular components that can be applied across different T cell therapy applications. The polypeptide sequences (Fas-OX40, TNFRSF12-OX40, LAG-3-4-1BB, etc.) serve as universal building blocks that can be combined with various TCR or CAR specificities. The standardized construct design with homology arms and regulatory elements makes the system universally applicable to different disease targets and therapeutic scenarios, reducing the need for application-specific R&D.
3Reliability
If T cells are modified with heterologous polypeptides and receptors, then therapeutic functionality is enhanced, but side effects associated with T cell therapies may increase
Solution Approach 1:
The patent applies local quality control by inserting the nucleic acid construct specifically into the TCR locus, a defined genomic location that ensures proper expression regulation. The use of homology-directed repair with specific homology arms targets the modification to a precise location, ensuring that the heterologous polypeptides and receptors are expressed with appropriate tissue-specificity and temporal control. This localized approach minimizes off-target effects and ensures that therapeutic functionality is enhanced without uncontrolled side effects.
Solution Approach 2:
The patent modifies specific parameters of T cell function through controlled polypeptide expression rather than altering fundamental T cell biology. By introducing regulated polypeptides (such as Fas-OX40, TNFRSF12-OX40, LAG-3-4-1BB) with defined expression patterns and activities, the therapy enhances therapeutic functionality through parameter adjustments (polypeptide concentration, expression timing, cellular activation thresholds) rather than fundamental changes, thereby reducing unpredictable side effects.
Data Source
AI summary
Provided herein are methods and compositions for modifying the genome of human T cells.