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Quaking orchestrates a post-transcriptional regulatory network of endothelial cell cycle progression critical to angiogenesis and metastasis

Abstract

Angiogenesis is critical to cancer development and metastasis. However, anti-angiogenic agents have only had modest therapeutic success, partly due to an incomplete understanding of tumor endothelial cell (EC) biology. We previously reported that the microRNA (miR)-200 family inhibits metastasis through regulation of tumor angiogenesis, but the underlying molecular mechanisms are poorly characterized. Here, using integrated bioinformatics approaches, we identified the RNA-binding protein (RBP) quaking (QKI) as a leading miR-200b endothelial target with previously unappreciated roles in the tumor microenvironment in lung cancer. In lung cancer samples, both miR-200b suppression and QKI overexpression corresponded with tumor ECs relative to normal ECs, and QKI silencing phenocopied miR-200b-mediated inhibition of sprouting. Additionally, both cancer cell and endothelial QKI expression in patient samples significantly corresponded with poor survival and correlated with angiogenic indices. QKI supported EC function by stabilizing cyclin D1 (CCND1) mRNA to promote EC G1/S cell cycle transition and proliferation. Both nanoparticle-mediated RNA interference of endothelial QKI expression and palbociclib blockade of CCND1 function potently inhibited metastasis in concert with significant effects on tumor vasculature. Altogether, this work demonstrates the clinical relevance and therapeutic potential of a novel, actionable miR/RBP axis in tumor angiogenesis and metastasis.

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Acknowledgements

The authors would like to especially thank members of the Victoria Bautch laboratory for their advice on performing the endothelial sprouting assay, the UNC Animal Histopathology Core, the UNC Biomedical Research Imaging Center, Janet Dow from the UNC Flow Cytometry Core Facility, and the Tissue Pathology Lab Core. The authors would also like to thank Adolfo Alfonso for providing the ZsGreen fluorescent labeled 10T1/2 (pericyte) cells used in the pericyte-coated sprouting assay. C.V.P. was supported in part by the National Institutes of Health R01-CA215075, R01-CA042978 and U54-CA198999, a Mentored Research Scholar Grants in Applied and Clinical Research (MRSG-14-222-01-RMC) from the American Cancer Society, the Jimmy V Foundation Scholar award, the UCRF Innovator Award, the Stuart Scott V Foundation/Lung Cancer Initiative Award for Clinical Research, the University Cancer Research Fund, the Lung Cancer Research Foundation, the Free to Breathe Metastasis Research Award and the Susan G. Komen Career Catalyst Award. S.H.A. was supported in part by a grant from the National Institute of General Medical Sciences under award 5T32 GM007092. E.B.H. was supported in part by a grant from the National Cancer Institute of the National Institutes of Health under award number T32CA196589. AKS was supported in part by The American Cancer Society Research Professor Award and grants from the National Cancer Institute (P50 CA217685 and R35 CA209904). The UNC Flow Cytometry Core Facility and Lineberger Comprehensive Cancer Center Animal Histopathology and Animal Studies Cores are all supported in part by a National Cancer Institute Center Core Support Grant (CA016086) to the UNC Lineberger Comprehensive Cancer Center. The UNC Flow Cytometry Core Facility is also supported in part by the North Carolina Biotech Center Institutional Support Grant 2012-IDG-1006.

Author information

Conception: CVP; Experimental design and methodology: SHA, AP, EBH, PLL, TAW, CVP; Acquisition of data: SHA, AP, EBH, PLL, TAW, AB, SM, XL, CVP; Analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): AP, SHA, CVP; Writing, review, and/or revision of the manuscript: All authors; Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): CVP, AP; Study supervision: CVP.

Correspondence to Chad V. Pecot.

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