Heterogenous expression of endoglin marks advanced renal cancer with distinct tumor microenvironment fitness

Intratumoral heterogeneity, including in clear cell renal cell carcinoma, is a potential cause of drug resistance and metastatic cancer progression. We specified the heterogeneous population marked by endoglin (also known as CD105) in a preclinical model of clear cell renal cell carcinoma progression. Highly malignant derivatives of human clear cell renal cell carcinoma OS-RC-2 cells were established as OS5Ks by serial orthotopic inoculation in our previous study. Expression of both ENG (encoding endoglin) mRNA and protein were heterogeneously upregulated in OS5Ks, and the endoglin-positive (ENG⁺ ) population exhibited growth dependency on endoglin in anchorage-independent cultures. Despite the function of endoglin as a type III receptor, transforming growth factor β and bone morphogenetic protein-9 signaling were unlikely to contribute to the proliferative phenotype. Although endoglin has been proposed as a marker for renal cancer-initiating cells, the OS5K-3 ENG⁺ population did not enrich other reported cancer-initiating cell markers or differentiate into the ENG^- population. Mouse tumor inoculation models revealed that the tumor-forming capabilities of OS5K-3 ENG⁺ and ENG^- cells in vivo were highly dependent on the microenvironment, with the renal microenvironment most preferable to ENG⁺ cells. In conclusion, the renal microenvironment, rather than the hypothesized ENG⁺ cell-centered hierarchy, maintains cellular heterogeneity in clear cell renal cell carcinoma. Therefore, the effect of the microenvironment should be considered when evaluating the proliferative capability of renal cancer cells in the experimental settings.