P2.08. THE OVEREXPRESSION OF PGC1Α SUPPORTS NEURONAL MATURATION IN DORSAL FOREBRAIN ORGANOID MODEL
Zuzanna Kuczyńska, Konrad Wasilewski, Paweł Leszczyński, Michał Liput, Erkan Metin, Leonora Bużańska
Mossakowski Medical Research Institute, Department of Stem Cell Bioengineering, Pawinskiego 5, Warsaw, Poland
INTRODUCTION: The PGC1α protein, encoded by the PPARGC1A gene, is a transcriptional coactivator regulating energy metabolism. Recently, several studies suggested the importance of the PGC1α in the development of neural system. Due to its role in promoting mitochondrial biogenesis, this protein may have therapeutic potential in neuronal disorders characterized by mitochondrial dysfunction.
AIM(S): The aim of this study was to investigate the effects of PGC1α overexpression on the neuronal differentiation of human dorsal forebrain organoids.
METHOD(S): Human induced pluripotent stem cells (iPSCs) were genetically modified using lentiviral vectors carrying either the CMV-GFP-puro control plasmid or the CMV-PPARGC1A-puro plasmid to achieve overexpression of PGC1α. After puromycin selection of the iPSCs, the expression of pluripotency markers was assessed by qRT-PCR and immunofluorescence staining. Subsequently, dorsal forebrain organoids were generated from the modified iPSC lines and collected on day 100 of development. Neuronal marker expression was analyzed by qRT-PCR and immunohistochemistry.
RESULTS: Analysis of PPARGC1A expression confirmed the successful generation of an iPSC line with increased levels of PGC-1α. The expression of pluripotency markers POU5F1 and NANOG remained unchanged, although SOX2 expression decreased in no significant manner. Dorsal forebrain organoids derived from engineered line revealed elevated expression of PPARGC1A as compared to controls. The expression of neural progenitor markers—NESTIN, CDH2, and SOX2 was not significantly altered. However, the expression of mature neuronal markers, FABP7, NEUROD1 and NEFL was increased in organoids with PPARGC1A overexpression.
CONCLUSIONS: The increased expression of PPARGC1A does not affect the expression of early neural markers, but it promotes neuronal differentiation, suggesting that PGC-1α may support neuronal maturation in dorsal forebrain organoids.
FINANCIAL SUPPORT: This work was supported by the National Science Centre Grants No. 2022/45/N/NZ1/02754 and MMRI PAS.