Discovering tumor-reactive T-cell receptors through single-cell sequencing of tumor-infiltrating lymphocytes

Gottschalk, E.; Aksoy, B. A.; Aksoy, P.; Swiderska-Syn, M.; Mart, C.; Macpherson, L.; Romeo, M.; Smith, A.; Knochelmann, H.; Paulos, C.; Timmers, C.; Wrangle, J.; Rubinstein, M. P.; Hammerbacher, J.

doi:10.1101/2021.11.30.470597

bioRxiv · 10.1101/2021.11.30.470597

Discovering tumor-reactive T-cell receptors through single-cell sequencing of tumor-infiltrating lymphocytes

Gottschalk, E.·Aksoy, B. A.·Aksoy, P.·Swiderska-Syn, M.·Mart, C.·Macpherson, L.·Romeo, M.·Smith, A.·Knochelmann, H.·Paulos, C.·Timmers, C.·Wrangle, J.·Rubinstein, M. P.·Hammerbacher, J.

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Abstract

We evaluated the utility of single-cell sequencing of tumor-infiltrating lymphocytes (TIL) for tumor-reactive T-cell receptor (TCR) discovery. Using the MC38 cell line as our tumor model in mice, we show that expression of exogenous TCRs via mRNA electroporation in human T cells provides an easy and quick path to validating tumor-specific candidate TCRs. We detail the identification and validation of four novel MC38-reactive mouse TCRs with varying levels of reactivity to the target cells. Validating our process, one of the MC38 TCRs is specific against a previously reported neoantigen (ASMTNMELM in the Adpgk gene). Consideration of these methodologies may aid in the development of rapid TCR-based therapies for the treatment of cancer and human disease. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=197 SRC="FIGDIR/small/470597v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@792d44org.highwire.dtl.DTLVardef@18af6aforg.highwire.dtl.DTLVardef@498c2aorg.highwire.dtl.DTLVardef@9127e8_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Gottschalk, E., Aksoy, B. A., Aksoy, P., Swiderska-Syn, M., Mart, C., Macpherson, L., Romeo, M., Smith, A., Knochelmann, H., Paulos, C., Timmers, C., Wrangle, J., Rubinstein, M. P., Hammerbacher, J.. 2021-12-01. Discovering tumor-reactive T-cell receptors through single-cell sequencing of tumor-infiltrating lymphocytes. https://doi.org/10.1101/2021.11.30.470597

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