TY - JOUR T1 - Tumor and immune remodeling following radiotherapy in human renal cell carcinoma JF - Journal for ImmunoTherapy of Cancer JO - J Immunother Cancer DO - 10.1136/jitc-2022-006392 VL - 11 IS - 4 SP - e006392 AU - Jacky Chow AU - Adil Khan AU - Madeline Gaudieri AU - Brianna J Wasik AU - Alexis Conway AU - Kah Teong Soh AU - Elizabeth A Repasky AU - Thomas Schwaab AU - Paul K Wallace AU - Scott I Abrams AU - Anurag K Singh AU - Jason B Muhitch Y1 - 2023/04/01 UR - http://jitc.bmj.com/content/11/4/e006392.abstract N2 - Background Studies evaluating peripheral patient samples show radiation can modulate immune responses, yet the biological changes in human tumors particularly at the cellular level remain largely unknown. Here, we address how radiation treatment shapes the immune compartment and interactions with cancer cells within renal cell carcinoma (RCC) patient tumors.Methods To identify how radiation shaped the immune compartment and potential immune interactions with tumor cells we evaluated RCC tumors from patients treated only with nephrectomy or with radiation followed by nephrectomy. Spectral flow cytometry using a 35-marker panel was performed on cell suspensions to evaluate protein expression within immune subsets. To reveal how radiation alters programming of immune populations and interactions with tumor cells, we examined transcriptional changes by single-cell RNA sequencing (scRNAseq).Results Spectral flow cytometry analysis revealed increased levels of early-activated as well as effector programmed cell death protein-1 (PD-1)+ CD8 T-cell subsets within irradiated tumors. Following quality control, scRNAseq of tumor samples from nephrectomy-only or radiation followed by nephrectomy-treated patients generated an atlas containing 34,626 total cells. Transcriptional analysis revealed increased transition from stem-like T-cell populations to effector T cells in irradiated tumors. Interferon (IFN) pathways, that are central to radiation-induced immunogenicity, were enriched in irradiated lymphoid, myeloid, and cancer cell populations. Focused cancer cell analysis showed enhanced antigen presentation and increased predicted TRAIL-mediated and IFN-mediated interactions between tumor cells and the same effector T-cell subsets increased by radiation. TRAIL and IFN pathways enriched in irradiated tumors were associated with survival in patients treated with immunotherapy.Conclusions These findings identify the source of IFN enrichment within irradiated RCC and reveal heightened levels of PD-1+ CD8+ T-cell subsets and increased probability of interactions with tumor cells following standalone radiation treatment. This study provides a window into the irradiated tumor-immune microenvironment of patients and rationale for treatment combinations.Data are available in a public, open access repository. All data relevant to the study are included in the article or uploaded as supplementary information. Single-cell RNA-sequencing data sets generated are available on GEO database (GSE202374). Patient response data were from the European Genome-Phenome Archive (EGA) under accession number EGAS00001002928. Annotated single-cell RNA data sets and TCGA barcodes of all KIRC samples are deposited in GitHub (github.com/JackyLChow/Chow_et_al_JITC_2023). Spectral flow cytometry data are available in ImmPort under access code SDY1998. ER -