INCREASED NETOSIS AND IMPAIRED DNASEI-MEDIATED NET DEGRADATION IN BLADDER CANCER PATIENTS: APPROACH FOR AN IN VITRO THERAPEUTICAL RESTORATION

You have accessJournal of UrologyCME1 Apr 2023MP14-09 INCREASED NETOSIS AND IMPAIRED DNASEI-MEDIATED NET DEGRADATION IN BLADDER CANCER PATIENTS: APPROACH FOR AN IN VITRO THERAPEUTICAL RESTORATION Raquel Herranz, Julia Oto, Marta Hueso, María Castaño, Emma Plana, Fernando Cana, Isabel Aleman, Santiago Bonamad, Javier Perez-Ardavin, César David Vera Donoso, Manuel Martinez-Sarmiento, and Pilar Medina Raquel HerranzRaquel Herranz More articles by this author , Julia OtoJulia Oto More articles by this author , Marta HuesoMarta Hueso More articles by this author , María CastañoMaría Castaño More articles by this author , Emma PlanaEmma Plana More articles by this author , Fernando CanaFernando Cana More articles by this author , Isabel AlemanIsabel Aleman More articles by this author , Santiago BonamadSantiago Bonamad More articles by this author , Javier Perez-ArdavinJavier Perez-Ardavin More articles by this author , César David Vera DonosoCésar David Vera Donoso More articles by this author , Manuel Martinez-SarmientoManuel Martinez-Sarmiento More articles by this author , and Pilar MedinaPilar Medina More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003234.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: An increased release of neutrophil extracellular traps (NETs) through NETosis, occurs in cancer. This enhances tumor growth and metastasis, which may be mediated by impaired NET degradation. However, this has never been evidenced in bladder cancer (BC). To evaluate the increase in NETosis in BC patients, to ascertain whether it is mediated by plasma DNaseI alterations and to in vitro explore therapeutic interventions. METHODS: NETs markers were measured in plasma samples of 73 BC patients and 64 controls. Plasma DNaseI activity was measured with the SRED assay. To evaluate the ability of plasma to degrade NETs, neutrophils were isolated from a healthy donor, seeded, and activated for 4 h with phorbol myristate acetate (PMA) to prompt NETosis. NETs were incubated for 6 h with 5 µl plasma from each participant and fluorescence was measured after staining with SytoxGreen. In parallel, BC plasma was supplemented with the recombinant human DNaseI Dornase Alfa (Pulmozyme, Roche), or vehicle (PBS 1X+0.1% BSA) to restore their DNaseI deficiency. We evaluated the presence of NETosis in tumor tissue by immunofluorescence (DNA, citrullinatedH3 and elastase). Statistical analysis was conducted with Graphpad (v.8.0.1). RESULTS: BC patients had significantly increased NETs markers (p<0.0001) and lower DNaseI activity (p<0.0001) in plasma compared to controls (Figure 1). In vitro NET degradation was impaired when incubated with BC plasma compared to control plasma (p<0.0001) (Figure 2). Addition of DNaseI restored NET degradation up to the level of healthy controls (p<0.0001) (Figure 2). NETosis occurs in BC tissue, more profusely in muscle-invasive BC. CONCLUSIONS: BC patients have increased NETosis in the tumor microenvironment and plasma, in part mediated by a reduced DNaseI activity. In vitro addition of Dornase Alfa, an approved treatment for cystic fibrosis, restores NET degradation in BC patients, thus becoming a potential therapeutic tool to reduce metastasis. Source of Funding: ISCIII-FEDER (PI17/00495, PI20/00075, FI21/00171), GVA (CIACIF/2021/192) and SETH © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e185 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Raquel Herranz More articles by this author Julia Oto More articles by this author Marta Hueso More articles by this author María Castaño More articles by this author Emma Plana More articles by this author Fernando Cana More articles by this author Isabel Aleman More articles by this author Santiago Bonamad More articles by this author Javier Perez-Ardavin More articles by this author César David Vera Donoso More articles by this author Manuel Martinez-Sarmiento More articles by this author Pilar Medina More articles by this author Expand All Advertisement PDF downloadLoading ...