MP16-18 THE ROLE OF CANCER-ASSOCIATED FIBROBLASTS AND THEIR EXTRACELLULAR VESICLES IN PROSTATE CANCER PROGRESSION

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I (MP16)1 Apr 2020MP16-18 THE ROLE OF CANCER-ASSOCIATED FIBROBLASTS AND THEIR EXTRACELLULAR VESICLES IN PROSTATE CANCER PROGRESSION Johannes Linxweiler*, Angela Zaccagnino, Dirk Himbert, Turkan Hajili, Philip Zeuschner, Christina Körbel, Michael Stöckle, Michael D. Menger, Matthias Saar, and Kerstin Junker Johannes Linxweiler*Johannes Linxweiler* More articles by this author , Angela ZaccagninoAngela Zaccagnino More articles by this author , Dirk HimbertDirk Himbert More articles by this author , Turkan HajiliTurkan Hajili More articles by this author , Philip ZeuschnerPhilip Zeuschner More articles by this author , Christina KörbelChristina Körbel More articles by this author , Michael StöckleMichael Stöckle More articles by this author , Michael D. MengerMichael D. Menger More articles by this author , Matthias SaarMatthias Saar More articles by this author , and Kerstin JunkerKerstin Junker More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000841.018AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The interaction between cancer cells und cancer-associated fibroblasts (CAFs) plays an important role in the biology of prostate cancer (PCa) and seems to be also mediated by the mutual exchange of extracellular vesicles (EVs). In this project, we analyzed the functional effect of CAFs and their EVs on PCa cells in vitro and in vivo. METHODS: Primary cultures of CAFs, not-cancer-associated (NCAFs) and benign prostate hyperplasia-associated (BPHFs) fibroblasts were established from human samples and characterized by immunofluorescence staining (IF; Pan-CK, Vimentin, αSMA). Their effect on the proliferation of PCa cells (LuCaP136 spheroids, LNCaP cells) was analyzed in vitro via a MTS Assay. In an orthotopic xenograft model, we performed a coinjection of CAFs/NCAFs/BPHFs with PCa cells (LuCaP136 spheroids, LNCaP cells) and subsequently monitored tumor burden for 10 weeks via serum PSA measurements and small animal imaging. Fibroblast-secreted EVs were isolated via ultracentrifugation and characterized by Western Blot, Nanoparticle Tracking Analysis and transmission electron microscopy. The uptake of fibroblast-secreted EVs by PCa cells was analyzed in vitro by fluorescence labeling experiments. RESULTS: CAF, NCAF and BPHF primary cultures could be established successfully and were characterized by IF (all Pan-CK- and Vimentin+; only CAFs αSMA+). The viability of LuCaP136 spheroids and LNCaP cells was significantly stimulated by cocultivation with fibroblasts in vitro (LuCaP136 spheroids by CAFs, LNCaP cells by all fibroblast types). In the xenograft experiments, an intraprostatic (orthotopic) coinjection of cancer cells together with CAFs resulted in significantly larger primary tumors, more lymph node and lung metastases compared to the coinjection of cancer cells together with NCAFs or BPHFs. EVs secreted by primary prostate fibroblasts could be successfully isolated by ultracentrifugation from conditioned media as confirmed by the expression of EV markers (CD9, Syntenin), typical round shape with lipid bilayer and a median size of about 100nm. EVs from CAFs were successfully taken up by PCa cells in fluorescence labeling experiments. CONCLUSIONS: The isolation and characterization of prostate fibroblast primary cultures and their EVs was successfully established. CAFs stimulated the growth of PCa cells in vitro and in vivo and their EV´s were taken up by PCa cells. The molecular mechanisms being involved in this interaction, especially the role of EVs, will be elucidated in future work. Source of Funding: Braun-Stiftung, Else Kröner-Fresenius-Stiftung © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e222-e222 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Johannes Linxweiler* More articles by this author Angela Zaccagnino More articles by this author Dirk Himbert More articles by this author Turkan Hajili More articles by this author Philip Zeuschner More articles by this author Christina Körbel More articles by this author Michael Stöckle More articles by this author Michael D. Menger More articles by this author Matthias Saar More articles by this author Kerstin Junker More articles by this author Expand All Advertisement PDF downloadLoading ...