Utilization of A 3D Culture System of Collagen-Mimic Peptide Gfoger-Based Hydrogel to Model Osteosarcoma from Engineered Ipsc

Abstract

Modeling the early stages of human osteosarcoma development remains a significant challenge. Most existing human models are derived from patient tumor tissue which is used to establish tumor cell lines or xenograft models in immunodeficient mice. These models are largely derived from late or end stage disease and do not allow the study of the early events of transformation. Further, 2D cell lines are largely homogenous and do not replicate the heterogeneity of primary tumors. Xenografted models more closely replicate the primary tumor but can have low engraftment rates and are logistically challenging to maintain. The immunocompromised nature of xenografted mice limits the potential for immunotherapy studies. The work presented here establishes a 3D culture system to model early-stage osteosarcoma development from engineered human iPSC. When cultured as aggregates in a GFOGER (integrin-specific glycine-phenylalanine-hydroxyproline-glycine-glutamate-arginine) based hydrogel known to promote osteoblastic differentiation, osteoblasts engineered with osteosarcoma-associated mutations readily form 3D organoids. Histological analysis supports that 3D culture of iPSC-derived osteoblasts promotes a more tissue-like phenotype with increased mineralization and ECM development within the tissue construct. In addition, preliminary functional studies suggest that 3D culture promotes transformative properties and an osteosarcoma phenotype. This novel approach has potential for future applications in disease modeling, in vivo studies, and drug discovery.