Abstract: SA-PO251
Characterization of Extracellular Matrix Composition in Wilms Tumor and Its Impact on Cellular Behavior
Session Information
- Onconephrology: Immunological Cross-Talk
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Onconephrology
- 1700 Onconephrology
Authors
- Perin, Laura, University of Southern California Keck School of Medicine, Los Angeles, California, United States
- Koos, David S., Children's Hospital Los Angeles, Los Angeles, California, United States
- Thornton, Matthew Edward, University of Southern California Keck School of Medicine, Los Angeles, California, United States
- Grubbs, Brendan, University of Southern California Keck School of Medicine, Los Angeles, California, United States
- Da Sacco, Stefano, University of Southern California Keck School of Medicine, Los Angeles, California, United States
- Petrosyan, Astgik, University of Southern California Keck School of Medicine, Los Angeles, California, United States
Background
Wilms Tumor (WT) is a 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
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 Proteome Profiler Human XL Oncology Array. Changes in gene expression of seeded SIX2+CITED1+ WT and human fetal kidney (hFK) progenitor cells on different dECM scaffolds were examined by immunofluorescence and bulk RNA-seq.
Results
Our study found differences in ECM fiber expression in WT dECM, with the outer layer comprising dense and elongated fibers, followed by pocketed and mesh-like structures. WT dECM exhibited elevated levels of oncoproteins ERBB2/3, SERPINE1, and MMP2 compared to normal kidney dECM. The dECM supported the long-term survival, migration, and proliferation of WT and hFK cells. When seeded on different dECM scaffolds, WT and hFK cells showed altered transcriptomic profiles and changes in cellular behavior. WT and hFK cells on WT dECM exhibited variations in ECM binding proteins (integrins) and increased expression of EMT and cancer stem cell markers like vimentin and SOX9 compared to normal kidney dECM.
Conclusion
This study provides valuable insights into the role of the ECM in regulating cancer cell behavior. The findings have significant implications for developing physiologically relevant in vitro tumor models and identifying novel therapeutic targets and mechanisms.
Funding
- Private Foundation Support