Proteomic Profiling of Human Prostate Cancer-associated Fibroblasts (CAF) Reveals LOXL2-dependent Regulation of the Tumor Microenvironment

Nguyen, Elizabeth V. and Pereira, Brooke A. and Lawrence, Mitchell G. and Ma, Xiuquan and Rebello, Richard J. and Chan, Howard and Niranjan, Birunthi and Wu, Yunjian and Ellem, Stuart and Guan, Xiaoqing and Wu, Jianmin and Skhinas, Joanna N. and Cox, Thomas R. and Risbridger, Gail P. and Taylor, Renea A. and Lister, Natalie L. and Daly, Roger J. (2019) Proteomic Profiling of Human Prostate Cancer-associated Fibroblasts (CAF) Reveals LOXL2-dependent Regulation of the Tumor Microenvironment. Molecular and Cellular Proteomics, 18 (7). pp. 1410-1427. ISSN 1535-9476

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Abstract

In prostate cancer, cancer-associated fibroblasts (CAF) exhibit contrasting biological properties to non-malignant prostate fibroblasts (NPF) and promote tumorigenesis. Resolving intercellular signaling pathways between CAF and prostate tumor epithelium may offer novel opportunities for research translation. To this end, the proteome and phosphoproteome of four pairs of patient-matched CAF and NPF were characterized to identify discriminating proteomic signatures. Samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with a hyper reaction monitoring data-independent acquisition (HRM-DIA) workflow. Proteins that exhibited a significant increase in CAF versus NPF were enriched for the functional categories “cell adhesion” and the “extracellular matrix.” The CAF phosphoproteome exhibited enhanced phosphorylation of proteins associated with the “spliceosome” and “actin binding.” STRING analysis of the CAF proteome revealed a prominent interaction hub associated with collagen synthesis, modification, and signaling. It contained multiple collagens, including the fibrillar types COL1A1/2 and COL5A1; the receptor tyrosine kinase discoidin domain-containing receptor 2 (DDR2), a receptor for fibrillar collagens; and lysyl oxidase-like 2 (LOXL2), an enzyme that promotes collagen crosslinking. Increased activity and/or expression of LOXL2 and DDR2 in CAF were confirmed by enzymatic assays and Western blotting analyses. Pharmacological inhibition of CAF-derived LOXL2 perturbed extracellular matrix (ECM) organization and decreased CAF migration in a wound healing assay. Further, it significantly impaired the motility of co-cultured RWPE-2 prostate tumor epithelial cells. These results indicate that CAF-derived LOXL2 is an important mediator of intercellular communication within the prostate tumor microenvironment and is a potential therapeutic target.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Faculty/School / Institute/Centre: Current - Faculty of Health, Engineering and Sciences - School of Health and Wellbeing (1 Jan 2015 -)
Faculty/School / Institute/Centre: Current - Faculty of Health, Engineering and Sciences - School of Health and Wellbeing (1 Jan 2015 -)
Date Deposited: 19 Jan 2020 23:22
Last Modified: 29 Jan 2020 02:10
Uncontrolled Keywords: Phosphoproteome; Prostate cancer; Prostate cancer biomarkers; Tumor microenvironment; Cancer biomarker(s); Cancer-associated fibroblasts; Fibroblasts; LOXL2; Non-malignant prostate fibroblasts
Fields of Research (2008): 11 Medical and Health Sciences > 1112 Oncology and Carcinogenesis > 111201 Cancer Cell Biology
11 Medical and Health Sciences > 1103 Clinical Sciences > 110312 Nephrology and Urology
11 Medical and Health Sciences > 1112 Oncology and Carcinogenesis > 111299 Oncology and Carcinogenesis not elsewhere classified
Socio-Economic Objectives (2008): C Society > 92 Health > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920119 Urogenital System and Disorders
C Society > 92 Health > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920114 Reproductive System and Disorders
Identification Number or DOI: 10.1074/mcp.RA119.001496
URI: http://eprints.usq.edu.au/id/eprint/36891

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