Identification of highly conserved alternative splicing and RNA binding proteins regulators contributing to immune microenvironment regulation in patients with clear cell renal cell carcinoma

Background Clear cell renal cell carcinoma (ccRCC) is a common renal malignant tumor, and its abnormal immune microenvironment is closely related to tumor progression, but the regulatory mechanism of alternative splicing and RNA binding protein (RBP) in this process is not clear. The purpose of this study was to investigate the effects of abnormal alternative splicing events on the immune microenvironment of ccRCC, and to identify the RBPs that mediate these events. Methods The RNA sequencing data in TCGA database were used to identify the differentially regulated alternative splicing events (RASE) between ccRCC and adjacent tissues by SUVA process. CIBERPORT algorithm was used to evaluate the degree of immune cell infiltration, screen the differentially expressed RBP, and construct the RBP-RASE-immune cell co regulatory network. Finally, immunohistochemistry was used to verify the expression of key RBP in tumor tissues. Results a total of 744 highly conserved rases in ccrcc were identified, of which 22 were significantly associated with immune cell infiltration, involving T cells, natural killer cells, macrophages and other immune groups. By overlapping the differentially expressed genes with the known RBP list, 225 differentially expressed RBPs were screened out, and the regulatory network was constructed. Four key RBPs (DDX47, RNASE2, RPL36, RPL36A) were identified, and their high expression was associated with poor prognosis. Immunohistochemistry showed that the expressions of RNASE2, RPL36 and RPL36A in ccRCC tumor tissues were significantly higher than those in adjacent tissues. Conclusion In this study, the RBP-RNASE-immune cell regulatory network was constructed systematically in ccRCC for the first time, revealing the potential mechanism of abnormal alternative splicing affecting tumor immune microenvironment through RBP, and providing a new molecular target for immunotherapy of ccRCC.