Pd16-06 Extracellular Matrix in the Tumor Microenvironment and Its Influence on Cancer

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology I (PD16)1 May 2024PD16-06 EXTRACELLULAR MATRIX IN THE TUMOR MICROENVIRONMENT AND ITS INFLUENCE ON CANCER David S. Koos, Xiaogang Hou, Esteban Fernandez, Matthew E. Thornton, Brendan H. Grubbs, Roger E. De Filippo, Stefano Da Sacco, Laura Perin, and Astgik Petrosyan David S. KoosDavid S. Koos , Xiaogang HouXiaogang Hou , Esteban FernandezEsteban Fernandez , Matthew E. ThorntonMatthew E. Thornton , Brendan H. GrubbsBrendan H. Grubbs , Roger E. De FilippoRoger E. De Filippo , Stefano Da SaccoStefano Da Sacco , Laura PerinLaura Perin , and Astgik PetrosyanAstgik Petrosyan View All Author Informationhttps://doi.org/10.1097/01.JU.0001009560.23593.56.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The tumor microenvironment's extracellular matrix (ECM) is critical in supporting normal and cancerous cell growth and development. In Wilms Tumor (WT), a pediatric renal cancer that arises from abnormal kidney development, we investigate the influence of the ECM on cellular behavior. We use a combination of decellularization techniques with two-photon excited fluorescence (TPEF) microscopy and transcriptomics to identify how cancer ECM vs. normal kidney ECM influences normal and cancer cells. METHODS: We decellularized normal kidneys and WT patient samples using an optimized decellularization technique. Second-harmonic generation (SHG) microscopy and two-photon excited fluorescence (TPEF), combined with immunohistochemistry, were used to characterize the decellularized matrices (dECM). Expression of various cancer-related proteins in WT dECM samples was compared to normal kidney dECM samples using the Proteome Profiler Human XL Oncology Array. Changes in gene expression of seeded WT and human fetal kidney (hFK) nephron progenitor cells on different dECM scaffolds were examined by immunofluorescence and bulk RNA-seq. RESULTS: Tumor ECM imaged at 700 um in-depth shows a difference in fiber area and angle, with the outer 250 um layer comprising dense and elongated fibers, followed by pocketed and mesh-like structures when imaged at 300 um in-depth. Analysis of tumor ECM vs. normal kidney ECM showed high expression of several oncoproteins, such as ERB2/3, PAI-1, and reduced CCL2 and BCL-X expression. Cellular behavior and transcriptomic profiles were altered when cultured on normal vs. tumor ECM for 21 days. RNA-Seq data of tumor cells seeded on tumor ECM vs. normal kidney ECM expressed Gene Ontology (GO) pathways favoring proliferation and reduction of pathways favoring differentiation. Normal cells seeded on tumor ECM vs. normal kidney ECM showed predicted activation of cancer development pathways and inhibition of apoptotic pathways. CONCLUSIONS: This study provides valuable insights into the role of the ECM in regulating cancer cell behavior. These findings have significant future implications for developing physiologically relevant in vitro tumor models and identifying novel therapeutic targets and mechanisms against tumor development and progression targeting the ECM. Source of Funding: Children's Hospital Los Angeles Intramural RCDA 2023 The GOFARR Research Fund © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e367 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information David S. Koos More articles by this author Xiaogang Hou More articles by this author Esteban Fernandez More articles by this author Matthew E. Thornton More articles by this author Brendan H. Grubbs More articles by this author Roger E. De Filippo More articles by this author Stefano Da Sacco More articles by this author Laura Perin More articles by this author Astgik Petrosyan More articles by this author Expand All Advertisement PDF downloadLoading ...