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Rapid Identification and Evaluation of Neoantigen-reactive T-Cell Receptors From Single Cells
Biman C Paria1, Noam Levin, Frank J Lowery
1Surgery Branch, National Cancer Institute, Bethesda, MD.
Abstract:
Engineered T cells expressing tumor-specific T-cell receptors (TCRs) are emerging as a mode of personalized cancer immunotherapy that requires identification of TCRs against the products of known driver mutations and novel mutations in a timely fashion. We present a nonviral and non-next-generation sequencing platform for rapid, and efficient neoantigen-specific TCR identification and evaluation that does not require the use of recombinant cloning techniques. The platform includes an innovative method of TCRα detection using Sanger sequencing, TCR pairings and the use of TCRα/β gene fragments for putative TCR evaluation. Using patients' samples, we validated and compared our new methods head-to-head with conventional approaches used for TCR discovery. Development of a unique demultiplexing method for identification of TCRα, adaptation of synthetic TCRs for gene transfer, and a reliable reporter system significantly shortens TCR discovery time over conventional methods and increases throughput to facilitate testing prospective personalized TCRs for adoptive cell therapy.
Insights
This study introduces a novel platform for quickly identifying tumor-specific T-cell receptors (TCRs) for personalized cancer immunotherapy. The method bypasses complex sequencing and cloning, accelerating the discovery of effective TCRs for adoptive cell therapy.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Personalized cancer immunotherapy relies on identifying tumor-specific T-cell receptors (TCRs).
- Current methods for TCR discovery are often time-consuming and complex, hindering rapid therapeutic development.
Purpose of the Study:
- To present a novel, nonviral, and non-next-generation sequencing platform for rapid and efficient neoantigen-specific TCR identification and evaluation.
- To validate and compare this new platform against conventional TCR discovery approaches using patient samples.
Main Methods:
- Utilized a unique Sanger sequencing method for TCRα detection.
- Employed TCR pairing and TCRα/β gene fragments for putative TCR evaluation.
- Developed a demultiplexing method for TCRα identification and adapted synthetic TCRs for gene transfer with a reporter system.
Main Results:
- The developed platform significantly shortens TCR discovery time compared to conventional methods.
- The platform demonstrated increased throughput, facilitating the testing of personalized TCRs.
- Validation using patient samples confirmed the efficacy and efficiency of the new methods.
Conclusions:
- The nonviral, non-NGS platform offers a rapid and efficient approach for neoantigen-specific TCR discovery.
- This technology accelerates the development of personalized TCRs for adoptive cell therapy.
- The innovative methods enhance the potential for timely and effective cancer immunotherapy.
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