The ASPIRE Path in Molecular Medicine

Mitochondrial fragmentation is a key property of the pluripotent state and an essential feature of reprogramming. In the Gama Laboratory, we have demonstrated that the anti-apoptotic mitochondrial protein Myeloid Cell Leukemia-1 (MCL-1), a member of the B-cell lymphoma-2 (BCL-2) family, is a fundamental regulator of mitochondrial dynamics in human induced pluripotent stem cells (hiPSCs). The main goal of my project is to examine whether the MCL-1-mediated mitochondrial morphology and function are maintained after hiPSCs are differentiated into human neural progenitor cells (hNPCs). We have found that knocking down MCL-1 in hNPCs using RNAi does not lead to cell death of hNPCs. Importantly, MCL-1 knockdown leads to the downregulation of key neuronal identity markers (e.g., PAX6 and Nestin). The mechanism in which MCL-1 is required to maintain neural progenitor identity has not been elucidated. I hypothesize that MCL-1 downregulation affects mitochondria morphology, and consequently, the respiratory and metabolic capacity of neural progenitor cells. To test this, we have engineered an inducible hiPSC line in which we can modulate the expression of MCL-1 using CRISPRoff. We plan to elucidate the effect of MCL-1 during cell transitions and better understand the function of MCL-1 in early neurogenesis.