Large-scale analysis of cell-cell communication reveals angiogenin-dependent tumor progression in clear cell renal cell carcinoma

Cellular crosstalk in the tumor microenvironment (TME) is still largely uncharacterized, while it plays an essential role in shaping immunosuppression or anti-tumor response. Large-scale analyses are needed to better decipher cell-cell communication in cancer. In this work, we used original and publicly available single-cell RNA sequencing (scRNAseq) data to characterize in-depth the communication networks in human clear cell renal cell carcinoma (ccRCC). We identified 50 putative communication channels specifically used by cancer cells to interact with other cells, including two novel angiogenin-mediated interactions. Expression of angiogenin and its receptors was validated at the protein level in primary ccRCC. Mechanistically, angiogenin enhanced ccRCC cell line proliferation and down-regulated secretion of IL-6, IL-8, and MCP-1 proinflammatory molecules. This study provides novel biological insights into molecular mechanisms of ccRCC, and suggests angiogenin and its receptors as potential therapeutic targets in clear cell renal cancer. Fresh samples of ccRCC human tumors and adjacent healthy tissue were obtained from the pathology departments of Saint-Louis and Bichat hospitals (Paris, France), from 3 patients that underwent partial or total nephrectomy for localized clear cell renal cell carcinoma. After tissue dissociation and single-cell suspension generation, cell sorting was done by FACS to enrich for rare immune and stromal cell types. Single-cell libraries were generated using the Chromium 10X Next GEM Single-cell 3’ Kit (v3.1) according to the manufacturer instructions (10X Genomics). Quality of the libraries was assessed by High Sensitivity D1000 ScreenTape assay and next-generation sequencing was performed on an Illumina NovaSeq 6000, with a sequencing depth of 200 000 reads/cell.