Single-Cell TCR Sequencing for Rapid T Cell Isolation
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Solution Overview
Problem
Current methods for isolating cancer-reactive T cells and T cell receptors from blood samples are time-consuming, invasive, and require prior knowledge of HLA composition and tumor mutations, limiting their effectiveness in treating advanced cancer patients.
Innovation Solution
A method involving the isolation of T cells from blood samples based on specific gene expression profiles, followed by sorting and sequencing of T cell receptor complementarity determining regions to identify and isolate T cells reactive against cancer-specific or viral antigens, allowing for the construction of TCRs without the need for invasive tumor resection or extensive screening.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current methods are used to produce cancer-reactive T cells, then T cells can be obtained for adoptive cell therapy, but the process requires significant time and may not readily identify the desired T cell receptors
Solution Approach 1:
The patent applies preliminary action by performing single-cell RNA sequencing and gene expression profiling on T cells isolated from peripheral blood before functional screening. This allows identification of T cell clones with high likelihood of cancer reactivity based on their transcriptional signatures, thereby reducing the time and resources needed for subsequent functional validation and T cell receptor identification
Solution Approach 2:
The patent replaces traditional mechanical/functional screening methods with molecular biology-based identification. Instead of relying on time-consuming functional assays to identify T cell receptors, the method uses gene expression profiling and single-cell sequencing to directly identify T cell clones and their receptors based on molecular signatures, significantly accelerating the identification process
2Reliability
If invasive tumor resection is performed to obtain tumor-infiltrating T cells, then T cells reactive to tumor antigens can be isolated, but the procedure is invasive and time-consuming
Solution Approach 1:
The patent extracts the essential function of obtaining tumor-reactive T cells without requiring actual tumor tissue. By isolating T cells from peripheral blood and identifying those with gene expression profiles characteristic of tumor-reactive clones, the method extracts the desired T cell population through a minimally invasive blood draw rather than invasive tumor resection
Solution Approach 2:
The patent introduces gene expression profiling as an intermediary between blood T cell isolation and tumor reactivity identification. This intermediary step uses molecular signatures to predict tumor reactivity, serving as a surrogate marker that eliminates the need for direct tumor tissue analysis and invasive procedures
3Measurement precision
If extensive screening is performed to identify desired T cell receptors, then accurate identification can be achieved, but the process becomes time-consuming and complex
Solution Approach 1:
The patent segments the T cell population into individual single cells for analysis, allowing parallel processing and identification of multiple T cell clones simultaneously. This segmentation enables high-throughput single-cell RNA sequencing and T cell receptor sequencing, achieving comprehensive screening without requiring sequential functional assays for each clone, thereby reducing overall process complexity
Data Source
AI summary
Provided are methods of preparing an enriched population of T cells having antigenic specificity for a target antigen. The method may comprise isolating T cells from a blood sample of a patient; selecting the isolated T cells which have a gene expression profile; and separating the selected T cells from the unselected cells. The separated selected T cells provide an enriched population of T cells having antigenic specificity for the target antigen. Methods of isolating a TCR, preparing a population of cells that express a TCR, isolated TCRs, isolated populations of cells, pharmaceutical compositions, and methods of treating or preventing a condition in a mammal are also provided.