RNA-Seq-Based TCR Profiling Reveals Persistently Increased Intratumoral Clonality in Responders to Anti-PD-1 Therapy

Abstract

Substantial effort is being invested in the search for peripheral or intratumoral T cell receptor (TCR) repertoire features that could predict the response to immunotherapy. Here we demonstrate the utility of MiXCR software for TCR and immunoglobulin repertoire extraction from RNA-Seq data obtained from sorted tumor-infiltrating T and B cells. We use this approach to extract TCR repertoires from RNA-Seq data obtained from sorted tumor-infiltrating CD4⁺ and CD8⁺ T cells in an HKP1 (Kras^G12Dp53^-/-) syngeneic mouse model of lung cancer after anti-PD-1 treatment. For both subsets, we demonstrate decreased TCR diversity in response to therapy. At a later time point, repertoire diversity is restored in progressing disease but remains decreased in responders to therapy in both CD4⁺ and CD8⁺ subsets. These observations complement previous studies and suggest that stably increased intratumoral CD4⁺ and CD8⁺ T cell clonality after anti-PD-1/PD-L1 therapy could serve as a predictor of long-term response.

Keywords: MiXCR; RNA-Seq; T cell clonality; TCR repertoire; anti-PD-1; tumor-infiltrating lymphocytes.