MP66-15 PATIENT-CENTRIC 3 DIMENSIONAL-BIOSYSTEMS FOR DEVELOPING PERSONALIZED IMMUNOTHERAPY FOR PATIENTS WITH UPPER TRACT UROTHELIAL CARCINOMA

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP66)1 Sep 2021MP66-15 PATIENT-CENTRIC 3 DIMENSIONAL-BIOSYSTEMS FOR DEVELOPING PERSONALIZED IMMUNOTHERAPY FOR PATIENTS WITH UPPER TRACT UROTHELIAL CARCINOMA Sanghee Lee, Michelle Muldong, Jonathan Hsu, Fady Ghali, Sung Gu Kang, Andrew Karim Kader, Amirali Salmasi, and Christina Jamieson Sanghee LeeSanghee Lee More articles by this author , Michelle MuldongMichelle Muldong More articles by this author , Jonathan HsuJonathan Hsu More articles by this author , Fady GhaliFady Ghali More articles by this author , Sung Gu KangSung Gu Kang More articles by this author , Andrew Karim KaderAndrew Karim Kader More articles by this author , Amirali SalmasiAmirali Salmasi More articles by this author , and Christina JamiesonChristina Jamieson More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002106.15AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Upper urinary tract urothelial carcinoma (UTUC) affects both men and women and accounts for approximately 5% of urothelial malignancies and 5 to 10% of renal cancers. Although surgery remains the mainstay of UTUC therapy, there is an increasing need for multi-modality approaches. Recent studies reviewed the genetic landscape of UTUC and revealed significant recurrent mutations. These observations emphasize the great potential of personalized immunotherapy which requires a thorough understanding of the adoptive and innate immune response to tumor development and progression. Therefore, we aimed to establish 3D-Biosystems in which a patient’s tumor infiltrating lymphocytes (TILs) were combined with their own ureter-derived organoids. METHODS: Ureter tissue samples from a patient with UTUC were collected immediately after surgical removal. 3D-organoids were established as described in Mullenders et al (2019). In parallel, TILs were isolated and expanded as described in Dudley et al (2003) and Tran et al (2007). 3D-Biosystems were established by adding 48 x 104 TILS to the 3D-organoids. Time course imaging of organoids was performed before and after the addition of TILs (1, 4, 28, 56, 84 and 91 hr) utilizing an EVOS microscope. Organoids and TILs in 3D-Biosystems were processed for histology as described in Lee et al (2020) and stained using Immuno-fluorescent cytochemistry (IFC) for markers: Cytokeratin 5 (CK5, basal urothelial cells), Uroplakin III (UPK III, intermediate and superficial urothelial cells) and P63 (basal and intermediate urothelial cells). Gene expression profiles were obtained by RNA-Seq analysis. RESULTS: In 3D-Biosystems, TILs migrated to 3D-organoids by 1 hr after co-culture. Sub-populations of 3D-organoids had extruded compartments by 91 hr after interacting with TILs. The patient’s ureter-derived organoids were CK5 and P63 positive but UPK III negative as was previously seen in urothelial organoids by Clevers group. Transcriptomic data revealed TILs expression profiles. CONCLUSIONS: Our 3D-Biosystems demonstrated functional responsiveness of TILs to tumor-derived organoids from the same patient. The TILs recognized and interacted with 3D-organoids that retain their original characteristics. Therefore, this personalized 3D-Biosystem has great potential as a screening platform to determine immunotherapy responses in combination with existing therapies and as a research platform to determine mechanisms of action. Source of Funding: This study is supported by the JM Foundation. © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1135-e1135 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Sanghee Lee More articles by this author Michelle Muldong More articles by this author Jonathan Hsu More articles by this author Fady Ghali More articles by this author Sung Gu Kang More articles by this author Andrew Karim Kader More articles by this author Amirali Salmasi More articles by this author Christina Jamieson More articles by this author Expand All Advertisement Loading ...