Abstract 4700: Spatial analysis of the immune microenvironment and tumor cell transition in sarcomatoid renal cell carcinoma
Authors: May Allison , Williams Claire , The Stephanie , Kim Sean , Schurman Nathan , Shelley Greg , Robinson Tyler , Salami Simpa , Mehra Rohit , Udager Aaron , Keller Evan 
Published: April 2023

Background: Sarcomatoid renal cell carcinoma (sRCC) is thought to arise by epithelial to mesenchymal transition (EMT) of the parental tumor in diverse RCC including clear cell RCC (ccRCC). sRCC is also known to be highly immunogenic and also express high levels of inflammatory pathway genes. Cells undergoing EMT and immune cells can have reciprocal feedback on each other; however, the role of such crosstalk in sRCC is unknown. Here, we use single cell spatial transcriptomics to evaluate the heterogeneity of EMT within sRCC in correlation with the immune microenvironment and in-vitro studies to explore mechanisms of crosstalk.

Methods: A sRCC nephrectomy specimen was subjected to single cell spatial transcriptomics of 1000 mRNA targets (NanoString CosMX). Semi-supervised clustering was performed and cell types were assigned by reference to single cell RNAseq data and then mapped spatially. Differential gene expression and cell to cell distance analysis was performed. In-vitro assays included treatment of 786-O and Caki-1 ccRCC lines with recombinant SPP1 and protein and mRNA analysis.

Results: Histopathologic evaluation revealed a ccRCC zone, a transition zone (tz) between the ccRCC and sRCC in which the ccRCC cells gained an increased mesenchymal morphology, and a well-developed sRCC zone. Transcriptomic data revealed four distinct tumor cell populations: a ccRCC, a tzRCC, and two sRCC populations. Importantly, a subset of the histologically-categorized ccRCC had a tzRCC transcriptional signature, demonstrating this signature is identifiable prior to development of morphologic features of sRCC. FZD4, a factor that may function in preventing EMT, was uniquely expressed on ccRCC and lost in tzRCC and sRCC cells. CCL20, a promoter of macrophage recruitment, was highly expressed in tzRCC. Distance analysis showed macrophages were highly spatially correlated with tzRCC and sRCC cells. Macrophages near tzRCC differentially expressed SPP1. In-vitro treatment of ccRCC cells with recombinant SPP1 led to induction of EMT and further upregulation of CCL20 expression.

Conclusion: We report a unique tumor cell population characterized by ccRCC morphology but harboring a transcriptome suggestive of progression towards sRCC. This provides rationale for the development of a novel molecular signature to subclassify ccRCC. We identified FZD4, CCL20 and SPP1 as key genes in the transition of ccRCC to sRCC which may mediate crosstalk between RCC cells and macrophages, with potential for therapeutic targets.

Citation Format: Allison May, Claire Williams, Stephanie The, Sean Kim, Nathan Schurman, Greg Shelley, Tyler Robinson, Simpa Salami, Rohit Mehra, Aaron Udager, Evan Keller. Spatial analysis of the immune microenvironment and tumor cell transition in sarcomatoid renal cell carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4700.