Abstract
Rapalogs (everolimus and temsirolimus) are allosteric mTORC1 inhibitors and approved agents for advanced clear cell renal cell carcinoma (ccRCC), although only a subset of patients derive clinical benefit. Progress in genomic characterization has made it possible to generate comprehensive profiles of genetic alterations in ccRCC; however, the correlations between recurrent somatic mutations and rapalog efficacy remain unclear. Here, we demonstrate by using multiple patient-derived ccRCC cell lines that compared to PTEN-proficient cells, PTEN-deficient cells exhibit hypersensitivity to rapalogs. Rapalogs inhibit cell proliferation by inducing G0/G1 arrest without inducing apoptosis in PTEN-deficient ccRCC cell lines. Using isogenic cell lines generated by CRISPR/Cas9, we validate the correlation between PTEN loss and rapalog hypersensitivity. In contrast, deletion of VHL or chromatin-modifying genes (PBRM1, SETD2, BAP1, or KDM5C) fails to influence the cellular response to rapalogs. Our mechanistic study shows that ectopic expression of an activating mTOR mutant (C1483F) antagonizes PTEN-induced cell growth inhibition, while introduction of a resistant mTOR mutant (A2034V) enables PTEN-deficient ccRCC cells to escape the growth inhibitory effect of rapalogs, suggesting that PTEN loss generates vulnerability to mTOR inhibition. PTEN-deficient ccRCC cells are more sensitive to the inhibitory effects of temsirolimus on cell migration and tumor growth in zebrafish and xenograft mice, respectively. Of note, PTEN protein loss as detected by immunohistochemistry is much more frequent than mutations in the PTEN gene in ccRCC patients. Our study suggests that PTEN loss correlates with rapalog sensitivity and could be used as a marker for ccRCC patient selection for rapalog therapy.
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Data availability
The RNA-seq data have been deposited to the National Center for Biotechnology Information (NCBI) via the Sequence Read Archive (SRA) database repository under accession code PRJNA777372. All other relevant data supporting the findings of this study are available upon reasonable request.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (81973357), Guangdong Basic and Applied Basic Research Foundation (2020A1515010027), Guangzhou Basic and Applied Basic Research Foundation (4001616289), Wu Jieping Medical Foundation (3206750.2020-10-120), Bethune Charitable Foundation Research Programme (B-19-H-20200622) and HUI LAN Public Welfare (2021-HLJJ0328). We thank Dr. Kun-Liang Guan (University of California, San Diego) for critically discussing and revising the manuscript.
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XLL and GMZ substantially contributed to designing the study, performing experiments, and analyzing the data. WHS, LXY, and SSH performed experiments and analyzed the data. ZLR analyzed the data. JJZ and SWL provided experimental resources. LY and YLL conceived and designed the study and experiments, assembled and interpreted the data, wrote and revised the manuscript, and approved the manuscript. All authors read and approved the final manuscript.
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Liu, Xl., Zhang, Gm., Huang, Ss. et al. PTEN loss confers sensitivity to rapalogs in clear cell renal cell carcinoma. Acta Pharmacol Sin 43, 2397–2409 (2022). https://doi.org/10.1038/s41401-022-00862-1
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DOI: https://doi.org/10.1038/s41401-022-00862-1
