PD05-09 FLUORESCENT-DIHYDROTESTOSTERONE BASED INHIBITOR IMPAIRED PROSTATE CANCER CELL LINE GROWTH AND ANDROGEN RECEPTOR SIGNALING

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I (PD05)1 Apr 2019PD05-09 FLUORESCENT-DIHYDROTESTOSTERONE BASED INHIBITOR IMPAIRED PROSTATE CANCER CELL LINE GROWTH AND ANDROGEN RECEPTOR SIGNALING Michael Fiandalo*, Vitaliy Sviripa, John Stocking, Danny Holbert, John Wilton, Krystin Mantione, Kristopher Attwood, Hans Minderman, Yue Wu, David Watt, and James Mohler Michael Fiandalo*Michael Fiandalo* More articles by this author , Vitaliy SviripaVitaliy Sviripa More articles by this author , John StockingJohn Stocking More articles by this author , Danny HolbertDanny Holbert More articles by this author , John WiltonJohn Wilton More articles by this author , Krystin MantioneKrystin Mantione More articles by this author , Kristopher AttwoodKristopher Attwood More articles by this author , Hans MindermanHans Minderman More articles by this author , Yue WuYue Wu More articles by this author , David WattDavid Watt More articles by this author , and James MohlerJames Mohler More articles by this author View All Author Informationhttps://doi.org/10.1097/01.JU.0000555069.86414.52AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVES: Men with advanced prostate cancer (CaP) receive androgen deprivation therapy (ADT), which lowers circulating testosterone (T) levels, impairs androgen receptor (AR) activation and results in CaP regression. ADT is palliative and CaP recurs as lethal castration-recurrent/resistant CaP CRPC). The small molecule anti-androgen, enzalutamide, is used to treat CRPC. However, expression of the AR-V7 splice variant impairs enzalutamide activity. A fluorescent-DHT surrogate, DHT-coumarin-1 (DHT-C-1), was one of many fluorescent-androgen surrogates synthesized to track androgen metabolism in CaP models. DHT-C-1, unexpectedly, inhibited growth of AR positive and AR-V7 positive CaP cell lines. The objective of these studies was to characterize DHT-C-1 activity in AR and AR-V7 positive and AR negative CaP cell lines. METHODS: Androgen sensitive and AR-positive LAPC-4, VCaP and LNCaP, castration-recurrent AR and AR-V7 positive CWR-R1 and 22rv1, and androgen-independent and AR negative PC-3 and DU145 cell lines were treated with serum-free complete media (to simulate androgen deprivation), coumarin alone, to show the fluorophore did not impair CaP cell growth or AR signaling, DHT or DHT-C-1. PC-3 cells that stably expressed AR-green fluorescent protein (AR-GFP) were used for ImageStream analysis. ImageStream, a hybrid technique of flow cytometry and fluorescent-microscopy, was used to assess CaP cell uptake of DHT-C-1. MTT was used to assess cell growth and quantitative real-time polymerase chain reaction (qRT-PCR) was used to assess kallikrein-related peptidase 2 (KLK2) and prostate-specific antigen (PSA) transcript abundance levels. RESULTS: ImageStream revealed DHT-C-1 was taken up by all 7 CaP cell lines. DHT-C-1 demonstrated nuclear co-localization with AR-GFP and nuclear marker DRAQ5 occurred only in AR positive CaP cell lines. ImageStream revealed that DHT-C-1 co-localized with the AR-ligand binding domain. DHT-C-1 treatment impaired growth of all 5 of the AR positive and AR-V7 positive CaP cell lines during androgen deprivation. VCaP cell growth inhibition required 1000 times lower concentration of DHT-C-1 than enzalutamide. qRT-PCR revealed that DHT-C-1 treatment impaired induction of PSA and KLK2 transcription, which suggested DHT-C-1 interfered with AR-regulated signaling pathways. CONCLUSIONS: DHT-C-1 demonstrated activity against CaP cells that express AR or AR-V7. DHT-C-1 may be effective alone or in combination to improve enzalutamide response. DHT-C-1 in vivo testing is in progress using CaP xenografts. Source of Funding: Acknowledgement. P01-CA77739, P20-RR020171, DoD Prostate Cancer Research Program Award No. W81XWH-16-1-0635 and Post-doctoral Training Award W81XWH-15-1-0409 and, in part, by the NCI Cancer Center Support Grant to Roswell Park (P30-CA016056) for the Bioanalytics, Metabolomics and Pharmacokinetics, Pathology Network and Biostatistics and Bioinformatics.DSW was supported by NIH R21 CA205108 (J. Mohler, PI) from the National Cancer Institute. DSW was also supported by NIH P30 GM110787 from the National Institute of General Medical Sciences (L. Hersh, PI) and DoD Prostate Cancer Research Program under Award No. W81XWH-16-1-0635 Buffalo, NY; Lexington, KY; Buffalo, NY; Morgantown, WV; Buffalo, NY; Lexington, KY; Buffalo, NY© 2019 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 201Issue Supplement 4April 2019Page: e83-e83 Advertisement Copyright & Permissions© 2019 by American Urological Association Education and Research, Inc.MetricsAuthor Information Michael Fiandalo* More articles by this author Vitaliy Sviripa More articles by this author John Stocking More articles by this author Danny Holbert More articles by this author John Wilton More articles by this author Krystin Mantione More articles by this author Kristopher Attwood More articles by this author Hans Minderman More articles by this author Yue Wu More articles by this author David Watt More articles by this author James Mohler More articles by this author Expand All Advertisement PDF downloadLoading ...