MP17-09 CXCL12-CXCR4 AXIS ACTIVATION PROMOTES COP II VESICLE-MEDIATED SECRETION OF COLLAGEN BY PROSTATE MYOFIBROBLASTS

Abstract

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2017MP17-09 CXCL12-CXCR4 AXIS ACTIVATION PROMOTES COP II VESICLE-MEDIATED SECRETION OF COLLAGEN BY PROSTATE MYOFIBROBLASTS Susan Patalano-Salsman, Jose A. Rodriguez-Nieves, Diego Almanza, Amy Avery, Andrew Judell-Halfpenney, Todd Riley, and Jill Macoska Susan Patalano-SalsmanSusan Patalano-Salsman More articles by this author , Jose A. Rodriguez-NievesJose A. Rodriguez-Nieves More articles by this author , Diego AlmanzaDiego Almanza More articles by this author , Amy AveryAmy Avery More articles by this author , Andrew Judell-HalfpenneyAndrew Judell-Halfpenney More articles by this author , Todd RileyTodd Riley More articles by this author , and Jill MacoskaJill Macoska More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.598AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Factors that promote lower urinary tract voiding dysfunction (LUTD) in aging men include excessive prostatic proliferation and muscle contraction. Recent studies suggest that tissue fibrosis, e.g., peri-urethral collagen accumulation, may also contribute to LUTD. Our group recently reported that activation of the CXCL12/CXCR4 axis and downstream MEK/ERK activation, as well as canonical TGFβ/TGFβR axis activation and downstream Smad3 activation, promote prostate fibroblast to myofibroblast phenoconversion and collagen production. Based on this finding, we hypothesized that downstream MEK/ERK and Smad signaling would converge at the transcriptional level to promote the activation of genes encoding pro-fibrotic proteins. To test this, RNA sequencing analysis was performed on prostate fibroblasts treated with CXCL12 or TGFβ. METHODS Prostate fibroblasts were treated with 4ng/ml TGFβ or 100pM CXCL12 for 12 hrs (RNA) or 24, 48 and 72 hrs (protein). Total RNA was purified and subjected to qRT-PCR or RNASeq. Sequence data was pipelined through the Bowtie/TopHat/Cufflinks/CummeRbund (Tuxedo) packages. Pathview v1.10.1 and Cytoscape were used to perform Kegg pathway and network analysis, respectively. Immunoblot analysis was performed using whole protein lysates or concentrated conditioned media. RESULTS RNASeq analysis showed that prostate fibroblasts treated with CXCL12 up-regulated the transcript levels of genes that encoded members of the Cullin-RING 3 (CRL3) ubiquitin ligase family of proteins. These proteins form COPII vesicles that transport procollagen from the endoplasmic reticulum to the cell membrane and extracellular space. Western blot analysis showed that CXCL12-treated cells up-regulated CRL3 proteins and secreted higher levels of procollagen compared to vehicle and TGFβ-treated cells. Procollagen secretion was ablated upon treatment with AMD3100, a CXCR4 antagonist, demonstrating that the observed increased collagen secretion was specifically coupled to CXCL12/CXCR4 axis activation. CONCLUSIONS The results of these studies are consistent with our hypothesis and present a major new discovery: Activation of the CXCL12/CXCR4 axis promotes fibrosis by increasing both the expression and secretion of collagen. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e214-e215 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Susan Patalano-Salsman More articles by this author Jose A. Rodriguez-Nieves More articles by this author Diego Almanza More articles by this author Amy Avery More articles by this author Andrew Judell-Halfpenney More articles by this author Todd Riley More articles by this author Jill Macoska More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...