(T1340) THE EFFECT OF GRAPHENE OXIDE SCAFFOLDS ON YAP1-MEDIATED CYTOSKELETAL REMODELING, MITOCHONDRIAL DYNAMICS, AND ADHESION IN INDUCED PLURIPOTENT STEM CELL-DERIVED CORNEAL ENDOTHELIAL CELLS
Abstract: Graphene oxide (GO) scaffolds are promising for supporting cell adhesion, survival, and growth. This study investigates GO's effect on iPSC-derived corneal endothelial cells (CECs), which are difficult to proliferate in vitro, focusing on the YAP mechanotransduction pathway and its role in mitochondrial function and actin cytoskeletal remodeling. Two types of CECs—an immortalized cell line and iPSC-derived CECs—were cultured on 20 nm thick GO-coated surfaces for up to 5 days. Western blotting, immunocytochemistry (ICC), RT-PCR, and RNA interference (RNAi) assays were used to explore YAP signaling and mitochondrial interaction. Antibodies specific to YAP1, TAZ, F-actin, and mitochondrial marker ATP5A were used to analyze protein expression, while qPCR assessed gene changes. GO scaffolds were non-cytotoxic and promoted CEC proliferation, as demonstrated by increased expression of Ki-67, phospho-histone H3, and phosphorylated ERK. GO enhanced cell adhesion through upregulation of N-cadherin and Focal Adhesion Kinase while preserving the intrinsic characteristics of CECs. YAP signaling was significantly activated in the GO group. YAP1-RNAi analysis confirmed that GO scaffolds regulate mitochondrial dynamics, cytoskeletal remodeling, and adhesion via YAP1 signaling, emphasizing their mechanotransduction effects on iPSC-derived CECs. Furthermore, a reduction in ATP5A expression suggests that GO-YAP1 interactions influence mitochondrial functionality, linking YAP signaling to cellular energy dynamics and highlighting the role of GO scaffolds in supporting iPSC-derived CECs growth through mechanotransduction pathways. This study confirms that GO scaffolds improve iPSC-derived CECs adhesion and proliferation by activating YAP signaling in response to mechanical cues, influencing mechanotransduction, cytoskeletal remodeling, and mitochondrial dynamics to regulate iPSC-derived CECs growth.
Funding Source: This work was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (RS-2024-00438366)
