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A novel cross-talk between CXCR4 and PI4KIIIα in prostate cancer cells

Abstract

Chemokine signaling regulates cell migration and tumor metastasis. CXCL12, a member of the chemokine family, and its receptor, CXCR4, a G protein coupled receptor (GPCR), are key mediators of prostate-cancer (PC) bone metastasis. In PC cells androgens activate CXCR4 gene expression and receptor signaling on lipid rafts, which induces protease expression and cancer cell invasion. To identify novel lipid-raft-associated CXCR4 regulators supporting invasion/metastasis, we performed a SILAC-based quantitative proteomic analysis of lipid-rafts derived from PC3 stable cell lines with overexpression or knockdown of CXCR4. This analysis identified the evolutionarily conserved phosphatidylinositol 4-kinase IIIα (PI4KIIIα), and SAC1 phosphatase that dephosphorylates phosphatidylinositol-4-phosphate as potential candidate CXCR4 regulators. CXCR4 interacted with PI4KIIIα membrane targeting machinery recruiting them to the plasma membrane for PI4P production. Consistent with this interaction, PI4KIIIα was found tightly linked to the CXCR4 induced PC cell invasion. Thus, ablation of PI4KIIIα in CXCR4-expressing PC3 cells reduced cellular invasion in response to a variety of chemokines. Immunofluorescence microscopy in CXCR4-expressing cells revealed localized production of PI4P on the invasive projections. Human tumor studies documented increased PI4KIIIα expression in metastatic tumors vs. the primary tumor counterparts, further supporting the PI4KIIIα role in tumor metastasis. Furthermore, we also identified an unexpected function of PI4KIIIα in GPCR signaling where CXCR4 regulates PI4KIIIα activity and mediate tumor metastasis. Altogether, our study identifies a novel cross-talk between PI4KIIIα and CXCR4 in promoting tumor metastasis and suggests that PI4KIIIα pharmacological targeting may have therapeutic benefit for advanced prostate cancer patients.

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Acknowledgements

Special thanks to Dr. Balla for providing PI4KIIIα and EFR3B expression constructs, Dr. DeCamilli for TTC7B construct and Dr. Mayinger for anti-Sac1 antibodies. We would like to thank Mr. Ussama A. Khan for technical assistance in preparing data. We also thank Astellas/Medivation for supporting clinical protocol for human tumor biopsy material collection. Supported by U.S. Department of Defense, W81XWH-09-1-0250, NIH-NCI Grant CA151557 and Fund for Cancer Research.

Author contributions

SRC conceived and coordinated the study and wrote the manuscript. AS provided expertize in preparation of data in Table 1, participated in study design and reviewed the manuscript. DS designed and performed experiments in Figs. 15 and participated in manuscript writing. LS and YL designed and performed experiments in Figs. 1, 5 and 6. BS performed experiments in Fig. 3 and supplementary figure 6. NJC and PMS designed and performed experiments in Fig. 1 and supplementary figure 2 and wrote corresponding methodology and participated in interpretation of data. MLC, SS, and UV were involved in study design of data in Fig. 6 and review of manuscript. All authors reviewed the results and approved the final version of manuscript.

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Correspondence to Sreenivasa R. Chinni.

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Sbrissa, D., Semaan, L., Govindarajan, B. et al. A novel cross-talk between CXCR4 and PI4KIIIα in prostate cancer cells. Oncogene 38, 332–344 (2019). https://doi.org/10.1038/s41388-018-0448-0

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