KLF4 as a rheostat of osteolysis and osteogenesis in prostate tumors in the bone

Abstract

We previously showed that KLF4, a gene highly expressed in murine prostate stem cells, blocks the progression of indolent intraepithelial prostatic lesions into aggressive and rapidly growing tumors. Here, we show that the anti-tumorigenic effect of KLF4 extends to PC3 human prostate cancer cells growing in the bone. We compared KLF4 null cells with cells transduced with a DOX-inducible KLF4 expression system, and find KLF4 function inhibits PC3 growth in monolayer and soft agar cultures. Furthermore, KLF4 null cells proliferate rapidly, forming large, invasive, and osteolytic tumors when injected into mouse femurs, whereas KLF4 re-expression immediately after their intra-femoral inoculation blocks tumor development and preserves a normal bone architecture. KLF4 re-expression in established KLF4 null bone tumors inhibits their osteolytic effects, preventing bone fractures and inducing an osteogenic response with new bone formation. In addition to these profound biological changes, KLF4 also induces a transcriptional shift from an osteolytic program in KLF4 null cells to an osteogenic program. Importantly, bioinformatic analysis shows that genes regulated by KLF4 overlap significantly with those expressed in metastatic prostate cancer patients and in three individual cohorts with bone metastases, strengthening the clinical relevance of the findings in our xenograft model.

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Acknowledgements

We thank Dr. E. Hernando-Monge (NYU School of Medicine) for the GFP-luciferase plasmid (BLIV513PA-1) and Drs. S. Logan and M. Garabedian (NYU School of Medicine) for helpful discussions. This study was supported by the NIH (R01CA132641 to E.L.W., R01AG056169 and K08AR069099 to P.L., R01CA181111 to M.S., T32CA009161 and T32AR064184 to A.S-P.), the American Cancer Society (RSG-16-033-01-DDC to M.S.), the Department of Urology and the Kimmel Center for Stem Cell Biology. Core funding (Applied Bioinformatics Laboratories, Genome Technology Center, High Throughput Biology Laboratory, Experimental Pathology Research Laboratory, Micro-CT) is partially supported by the Perlmutter Cancer Center (P30CA016087), NYSTEM (contract C026719), and NIH (S10 OD010751).

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E.T., P.L., M.S., and E.L.W. conceived and designed the study. E.T., V.B-C., X.X., and A.M.J. performed the experiments and analyzed the data. M.S., A.S-P., and A.K-J. performed the bioinformatics analysis. J.M. and P.L. interpreted histological data. L.B. provided the BFP sequence of the KLF4-expressing vector and helped with the cloning strategy. D.J.K. acquired the images of soft agar cultures and analyzed the data. E.T., L.O., M.S., and E.L.W. wrote the manuscript. All authors revised and approved the manuscript.

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Correspondence to Markus Schober or Elaine L. Wilson.

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Tassone, E., Bradaschia-Correa, V., Xiong, X. et al. KLF4 as a rheostat of osteolysis and osteogenesis in prostate tumors in the bone. Oncogene 38, 5766–5777 (2019). https://doi.org/10.1038/s41388-019-0841-3

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