Pd20-09 Three-Dimensional Imaging and Spatial Analysis of Intact Tissue Improve Diagnostic Yield and Accuracy in Upper Tract Urothelial Carcinoma Diagnosis

You have accessJournal of UrologyBladder Cancer: Upper Tract Transitional Cell Carcinoma I (PD20)1 May 2024PD20-09 THREE-DIMENSIONAL IMAGING AND SPATIAL ANALYSIS OF INTACT TISSUE IMPROVE DIAGNOSTIC YIELD AND ACCURACY IN UPPER TRACT UROTHELIAL CARCINOMA DIAGNOSIS Keishiro Fukumoto, Yota Yasumizu, Nobuyuki Tanaka, Toshikazu Takeda, Kazuhiro Matsumoto, Shinya Morita, Takeo Kosaka, Ryuichi Mizuno, Hiroshi Asanuma, Per Uhlén, and Mototsugu Oya Keishiro FukumotoKeishiro Fukumoto , Yota YasumizuYota Yasumizu , Nobuyuki TanakaNobuyuki Tanaka , Toshikazu TakedaToshikazu Takeda , Kazuhiro MatsumotoKazuhiro Matsumoto , Shinya MoritaShinya Morita , Takeo KosakaTakeo Kosaka , Ryuichi MizunoRyuichi Mizuno , Hiroshi AsanumaHiroshi Asanuma , Per UhlénPer Uhlén , and Mototsugu OyaMototsugu Oya View All Author Informationhttps://doi.org/10.1097/01.JU.0001009556.54476.eb.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Biopsies of Upper Tract Urothelial Carcinoma (UTUC) are often minute and can be challenging to diagnose using standard pathology procedures and conventional microscopy. To address this, we developed an innovative gelatin embedding technique and a 3D tissue-clearing imaging pipeline. This approach allows for the exploration of the inner landscape of clinical UTUC biopsies without sectioning. Our findings reveal that 3D analysis of phosphorylated ribosomal protein S6 (pS6) could enhance prediction and prognosis accuracy. METHODS: Twenty-one clinical UTUC samples were collected from nineteen patients between 2017 and 2020 using biopsy forceps. The samples underwent a series of processes including formalin fixation, bleaching, immunolabeling, gelatin embedding, and clearing (Figure 1A), followed by capture using light-sheet microscopy. The primary antibodies used were anti-CD34, pS6, AE1/AE3 and histone. Utilizing 3D imaging enabled spatial distribution analysis, cluster analysis, and tumor vasculature analysis (Figure 1B). RESULTS: Spatial distribution analysis of these 21 samples revealed cell counts ranging from 24,978 to 247,270, with pS6 positive cells varying from 78 to 68,536. The percentage of pS6-positive cells was significantly higher in high-grade UTUC than in low-grade UTUC (27% vs 11%, p=0.002). Detailed 3D image analysis showed that high-grade UTUC had more clusters of pS6 positive cells compared to low-grade UTUC. The ratio of clustered pS6-positive cells was higher in high-grade tumors (p=0.048) and was associated with shorter relapse-free survival (p=0.022). Visualization of tumor vasculature revealed that pS6-positive cells were significantly more localized near blood vessels in high-grade tumors (p=0.01), and a high density of pS6-positive cells around blood vessels was significantly associated with relapse-free survival (p=0.003). CONCLUSIONS: Our study demonstrates that 3D volume imaging of the structural niches of pS6-cells deep inside UTUC samples can significantly improve diagnostic yield and prognosis prediction. Download PPT Source of Funding: None © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e450 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Keishiro Fukumoto More articles by this author Yota Yasumizu More articles by this author Nobuyuki Tanaka More articles by this author Toshikazu Takeda More articles by this author Kazuhiro Matsumoto More articles by this author Shinya Morita More articles by this author Takeo Kosaka More articles by this author Ryuichi Mizuno More articles by this author Hiroshi Asanuma More articles by this author Per Uhlén More articles by this author Mototsugu Oya More articles by this author Expand All Advertisement PDF downloadLoading ...