Investigating Extracellular Matrix Composition and Role in Wilms Tumor Development and Progression
David S. Koos, PhD1, Matthew E. Thornton, MS2, Brendan H. Grubbs, MD2, Roger De Filippo, MD3, Stefano Da Sacco, PhD3, Laura Perin, PhD3, Astgik Petrosyan, PhD3.
1Children's Hospital Los Angeles, Los Angeles, CA, USA, 2University of Southern California, Los Angeles, CA, USA, 3University of Southern California/Children's Hospital Los Angeles, Los Angeles, CA, US
BACKGROUND: Wilms Tumor (WT) is a common pediatric renal cancer that arises from abnormal kidney development. The extracellular matrix (ECM) in the tumor microenvironment plays a critical role in supporting normal and cancerous cells and is essential for tumor growth and development. This study aims to understand the differences in ECM composition between normal kidneys and WT samples and how these variations affect cellular behavior. METHODS: Using an optimized decellularization technique developed in our laboratory, we decellularized human fetal kidneys and biopsies obtained from healthy pediatric kidneys and WT patients. Second-harmonic generation (SHG) microscopy and two-photon excited fluorescence (TPEF), combined with immunohistochemistry, were used to characterize the decellularized matrices (dECM ) and reveal differences in ECM composition and distribution. Expression of various cancer-related proteins in WT dECM samples was compared to normal kidney dECM samples using Proteome Profiler Human XL Oncology Array. Finally, changes in cellular behavior and transcriptomics of seeded cells on different dECM scaffolds were examined.
RESULTS: We found differences in ECM fiber expression in the WT, with the outer layer comprising dense and elongated fibers, followed by pocketed and mesh-like structures within the ECM. WT dECM samples showed elevated expression of oncoproteins ERBB2/3, SERPINE1, and MMP2 compared to normal human kidney dECM samples. Additionally, changes in transcriptomic profiles and cellular behavior of WT and hFK cells were observed when seeded on different dECM scaffolds.
CONCLUSIONS: The results of this study offer valuable insights into how the ECM regulates cancer cell behavior. These findings have significant implications for developing in vitro tumor models with physiologically relevant properties and identifying novel therapeutic targets and mechanisms.
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