Pd27-12 antagonism of the p75ntr receptor increases the release of nerve growth factor in urothelial cells by inhibiting matrix metalloproteinase-9

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology (PD27)1 Sep 2021PD27-12 ANTAGONISM OF THE P75NTR RECEPTOR INCREASES THE RELEASE OF NERVE GROWTH FACTOR IN UROTHELIAL CELLS BY INHIBITING MATRIX METALLOPROTEINASE-9 Aalya Hamouda, Philippe Cammisotto, Stephanie Sirmakesyan, Uri Saragovi, and Lysanne Campeau Aalya HamoudaAalya Hamouda More articles by this author , Philippe CammisottoPhilippe Cammisotto More articles by this author , Stephanie SirmakesyanStephanie Sirmakesyan More articles by this author , Uri SaragoviUri Saragovi More articles by this author , and Lysanne CampeauLysanne Campeau More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002020.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Decreased levels of nerve growth factor (NGF) have been observed in the urine of patients with overactive bladder syndrome (OAB) and of diabetic animal models. This unbalance appeared to result from an increased activity of the proteolytic enzyme metalloproteinase-9 (MMP-9). On the other hand, THX-B, an inhibitor of p75NTR, restored normal levels of NGF in mice with type 1 diabetes. We here examine in vitro the effect of THX-B on the activity of MMP-9 in bladder cells. METHODS: Primary culture of urothelial and smooth muscle cells (SMCs) were grown from mouse bladder and exposed to p75NTR antagonist THX. The expression of NGF and MMP-9 were assessed by RT-qPCR, by immunohistochemistry and by immunoblotting. Levels of microRNAs were measured by RT-qPCR. NGF and proNGF secretion were measured by ELISA kits and MMP-9 activity by enzymatic assays. RESULTS: The mRNAs for NGF and MMP-9 were detected and found expressed in both cell types at similar level. Microscopy confirmed the localization of both proteins in cells. Urothelial cells and SMCs were both major source of NGF and proNGF. On the other hand, MMP-9 protein content was 7 times higher in SMCs than in urothelial cells, which was confirmed by high levels of miR-491-5p in the latter. However, secretion of active MMP-9 in the medium was 40 times higher in urothelial cells medium. Incubation with THX-B (5 µg/mL) for 24 hours abolished the synthesis and secretion of MMP-9 and doubled the concentration of NGF in the medium of urothelial cells. ProNGF secretion levels were not affected. THX-B had little effects on SMCs both at the level of NGF and MMP-9. CONCLUSIONS: Urothelial cells appear to be the primary target for THX-B to modulate secretion of NGF and MMP-9. These results are in accordance with our previous publications on OAB patients and diabetes type 1 in rodents, and suggest that THX-B could be a therapeutic tool to improve OAB by targeting primarily the urothelium. Source of Funding: Canadian Urology AssociationQuebec Network for Research on Aging © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e448-e448 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Aalya Hamouda More articles by this author Philippe Cammisotto More articles by this author Stephanie Sirmakesyan More articles by this author Uri Saragovi More articles by this author Lysanne Campeau More articles by this author Expand All Advertisement PDF downloadLoading ...