MP77-04 H-IPSE, A PARASITE-DERIVED CANDIDATE DRUG FOR BLADDER PAIN, IS INTERNALIZED THROUGH CLATHRIN- AND CAVEOLIN-MEDIATED UPTAKE AND LOCALIZES WITHIN NON-UROTHELIAL CELLS

Abstract

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Kidney & Bladder II (MP77)1 Apr 2020MP77-04 H-IPSE, A PARASITE-DERIVED CANDIDATE DRUG FOR BLADDER PAIN, IS INTERNALIZED THROUGH CLATHRIN- AND CAVEOLIN-MEDIATED UPTAKE AND LOCALIZES WITHIN NON-UROTHELIAL CELLS Olivia Lamanna*, Evaristus Mbanefo, Kenji Ishida, Franco Falcone, Theodore Jardetzky, Luke Pennington, and Michael Hsieh Olivia Lamanna*Olivia Lamanna* More articles by this author , Evaristus MbanefoEvaristus Mbanefo More articles by this author , Kenji IshidaKenji Ishida More articles by this author , Franco FalconeFranco Falcone More articles by this author , Theodore JardetzkyTheodore Jardetzky More articles by this author , Luke PenningtonLuke Pennington More articles by this author , and Michael HsiehMichael Hsieh More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000963.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: We have demonstrated that H-IPSE, the Schistosoma haematobium ortholog of the IL-4 Inducing Principle from Schistosoma mansoni Eggs, is therapeutic in ifosfamide-induced hemorrhagic cystitis through the downregulation of pro-inflammatory pathways. We also have data suggesting H-IPSE alleviates resiniferatoxin (capsaicin receptor)-induced bladder pain. As an infiltrin, H-IPSE translocates into host cell nuclei to alter host transcription. We have shown nuclear translocation-deficient mutants of H-IPSE lose their therapeutic effects, suggesting that intracellular localization of H-IPSE to facilitate nuclear activities is particularly important for this protein’s actions. Our objective was to determine IPSE’s mechanism of entry into urothelial cells and compare the magnitude of IPSE intracellular localization in non-urothelial cell types within the bladder (endothelial, dendritic, and neuronal cells) to further identify and exploit IPSE’s therapeutic functions in inflammation-driven bladder pain. METHODS: H03, an H-IPSE ortholog, was conjugated to Alexa-488 fluorophore. Urothelial, endothelial, hepatocyte, immature dendritic, and neuronal cell lines were incubated with H03, and analyzed by flow cytometry before and after extracellular signal quenching. Urothelial cells were treated with chlorpromazine and/or filipin (to inhibit clathrin- and caveoli-mediated endocytosis), then incubated with H03, quenched, and analyzed by flow cytometry. RESULTS: All cell lines demonstrated IPSE internalization; however, urothelial cells internalized IPSE most efficiently at 83%, while internalization by neurons was significantly less, at 6.7%. Internalization by urothelial cells decreased by 7.0% when caveolin-mediated endocytosis was inhibited (filipin) and by 11.8% when clathrin-mediated endocytosis was inhibited (chlorpromazine). Upon treatment with both inhibitors, internalization decreased by 23.1%, suggesting IPSE utilizes both clathrin- and caveolin-mediated endocytosis to enter urothelial cells. CONCLUSIONS: H-IPSE can be internalized by various non-urothelial cell types, suggesting that IPSE’s therapeutic function may not be isolated to the urothelium. We found that IPSE interacts with neurons minimally, indicating IPSE is not inducing direct therapeutic effects on capsaicin receptor-expressing nociceptive neurons as hypothesized, and could be using an alternative pathway. We have shown that IPSE is internalized by urothelial cells using both clathrin- and caveolin-mediated endocytosis. Additional research on IPSE’s endosomal trafficking into the nucleus and nuclear targets is needed to further understand how IPSE downregulates pro-inflammatory pathways in the bladder and thereby increase its therapeutic potential. Source of Funding: NIH R01-DK113504 © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e1164-e1164 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Olivia Lamanna* More articles by this author Evaristus Mbanefo More articles by this author Kenji Ishida More articles by this author Franco Falcone More articles by this author Theodore Jardetzky More articles by this author Luke Pennington More articles by this author Michael Hsieh More articles by this author Expand All Advertisement PDF downloadLoading ...