PT - JOURNAL ARTICLE AU - Pizzolla, Angela AU - Keam, Simon Paul AU - Vergara, Ismael A AU - Caramia, Franco AU - Thio, Niko AU - Wang, Minyu AU - Kocovski, Nikolce AU - Tantalo, Daniela AU - Jabbari, Jafar AU - Au-Yeung, George AU - Sandhu, Shahneen AU - Gyorki, David E AU - Weppler, Alison AU - Perdicchio, Maurizio AU - McArthur, Grant A AU - Papenfuss, Anthony T AU - Neeson, Paul Joseph TI - Tissue-resident memory T cells from a metastatic vaginal melanoma patient are tumor-responsive T cells and increase after anti-PD-1 treatment AID - 10.1136/jitc-2022-004574 DP - 2022 May 01 TA - Journal for ImmunoTherapy of Cancer PG - e004574 VI - 10 IP - 5 4099 - http://jitc.bmj.com/content/10/5/e004574.short 4100 - http://jitc.bmj.com/content/10/5/e004574.full SO - J Immunother Cancer2022 May 01; 10 AB - Background Vaginal melanoma (VM) is a rare cancer and has a poor response to immune checkpoint blockade (ICB). CD8+Tissue Resident Memory (TRM) T cells proliferate in response to ICB and correlate with longer survival in metastatic cutaneous melanoma. However, their capacity to respond to VM and their neoantigens is not known.Methods Using longitudinal samples, we explored the evolution of VM mutations by whole-exome sequencing and RNAseq, we also defined the immune context using multiplex immunohistochemistry and nanostring pan cancer immune profile. Then using fresh single cell suspensions of the metastatic samples, we explored VM T cells via mass cytometry and single cell RNAseq and T cell receptor sequencing (TCRseq). Finally, we investigated TRM, pre-TRM and exhausted T cell function against melanoma neo-antigens and melanoma differentiation antigens in vitro.Results Primary VM was non-inflamed and devoid of CD8+ TRM cells. In contrast, both metastases showed proliferating CD8+ TRM were clustered at the tumor margin, with increased numbers in the second ICB-refractory metastasis. The first metastasis showed dense infiltration of CD8+ T cells, the second showed immune exclusion with loss of melanoma cell Major histocompatibility complex (MHC)-I expression associated with downregulation of antigen presentation pathway gene expression. CD8+ TRM from both metastases responded to autologous melanoma cells more robustly than all other CD8+ T cell subsets. In addition, CD8+ TRM shared TCR clones across metastases, suggesting a response to common antigens, which was supported by recognition of the same neoantigen by expanded tumor infiltrating lymphocytes.Conclusions In this study, we identified TRM clusters in VM metastases from a patient, but not primary disease. We showed TRM location at the tumor margin, and their superior functional response to autologous tumor cells, predicted neoantigens and melanoma differentiation antigens. These CD8+ TRM exhibited the highest tumor-responsive potential and shared their TCR with tumor-infiltrating effector memory T cells. This suggests VM metastases from this patient retain strong antitumor T cell functional responses; however, this response is suppressed in vivo. The loss of VG MHC-I expression is a common immune escape mechanism which was not addressed by anti-PD-1 monotherapy; rather an additional targeted approach to upregulate MHC-I expression is required.Data are available on reasonable request. We are in the process to upload the data in dbGAP. It will be available on request as per our institute policy.