Proteomic profiling of human prostate cancer-associated fibroblasts (CAF) reveals LOXL2-dependent regulation of the tumor microenvironment

Elizabeth V. Nguyen, Brooke A. Pereira, Mitchell G. Lawrence, Xiuquan Ma, Richard J. Rebello, Howard Chan, Birunthi Niranjan, Yunjian Wu, Stuart Ellem, Xiaoqing Guan, Jianmin Wu, Joanna N. Skhinas, Thomas R. Cox, Gail P. Risbridger, Renea A. Taylor, Natalie L. Lister, Roger J. Daly

Research output: Contribution to journalArticleResearchpeer-review

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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.

Original languageEnglish
Pages (from-to)1410-1427
Number of pages18
JournalMolecular and Cellular Proteomics
Volume18
Issue number7
DOIs
Publication statusPublished - 1 Jul 2019

Cite this

Nguyen, Elizabeth V. ; Pereira, Brooke A. ; Lawrence, Mitchell G. ; Ma, Xiuquan ; Rebello, Richard J. ; Chan, Howard ; Niranjan, Birunthi ; Wu, Yunjian ; Ellem, Stuart ; Guan, Xiaoqing ; Wu, Jianmin ; Skhinas, Joanna N. ; Cox, Thomas R. ; Risbridger, Gail P. ; Taylor, Renea A. ; Lister, Natalie L. ; Daly, Roger J. / Proteomic profiling of human prostate cancer-associated fibroblasts (CAF) reveals LOXL2-dependent regulation of the tumor microenvironment. In: Molecular and Cellular Proteomics. 2019 ; Vol. 18, No. 7. pp. 1410-1427.
@article{27385060d7514a32b22462a6e1962d17,
title = "Proteomic profiling of human prostate cancer-associated fibroblasts (CAF) reveals LOXL2-dependent regulation of the tumor microenvironment",
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.",
author = "Nguyen, {Elizabeth V.} and Pereira, {Brooke A.} and Lawrence, {Mitchell G.} and Xiuquan Ma and Rebello, {Richard J.} and Howard Chan and Birunthi Niranjan and Yunjian Wu and Stuart Ellem and Xiaoqing Guan and Jianmin Wu and Skhinas, {Joanna N.} and Cox, {Thomas R.} and Risbridger, {Gail P.} and Taylor, {Renea A.} and Lister, {Natalie L.} and Daly, {Roger J.}",
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Proteomic profiling of human prostate cancer-associated fibroblasts (CAF) reveals LOXL2-dependent regulation of the tumor microenvironment. / Nguyen, Elizabeth V.; Pereira, Brooke A.; Lawrence, Mitchell G.; Ma, Xiuquan; Rebello, Richard J.; Chan, Howard; Niranjan, Birunthi; Wu, Yunjian; Ellem, Stuart; Guan, Xiaoqing; Wu, Jianmin; Skhinas, Joanna N.; Cox, Thomas R.; Risbridger, Gail P.; Taylor, Renea A.; Lister, Natalie L.; Daly, Roger J.

In: Molecular and Cellular Proteomics, Vol. 18, No. 7, 01.07.2019, p. 1410-1427.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Proteomic profiling of human prostate cancer-associated fibroblasts (CAF) reveals LOXL2-dependent regulation of the tumor microenvironment

AU - Nguyen, Elizabeth V.

AU - Pereira, Brooke A.

AU - Lawrence, Mitchell G.

AU - Ma, Xiuquan

AU - Rebello, Richard J.

AU - Chan, Howard

AU - Niranjan, Birunthi

AU - Wu, Yunjian

AU - Ellem, Stuart

AU - Guan, Xiaoqing

AU - Wu, Jianmin

AU - Skhinas, Joanna N.

AU - Cox, Thomas R.

AU - Risbridger, Gail P.

AU - Taylor, Renea A.

AU - Lister, Natalie L.

AU - Daly, Roger J.

PY - 2019/7/1

Y1 - 2019/7/1

N2 - 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.

AB - 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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U2 - 10.1074/mcp.RA119.001496

DO - 10.1074/mcp.RA119.001496

M3 - Article

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EP - 1427

JO - Molecular & Cellular Proteomics

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SN - 1535-9476

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