DIRECT NEURONAL REPROGRAMMING OF OLIGODENDROCYTE PROGENITOR CELLS

Direct neuronal reprogramming is a promising avenue for cell-based therapies aimed at replacing neurons lost in neurodegenerative diseases or traumatic brain injuries with newly generated ones. Among brain-resident somatic cells, oligodendrocyte progenitor cells (OPCs) are a poorly investigated yet interesting population to target. OPCs are the only cell type that proliferate in the adult brain parenchyma under physiological and pathological conditions, and therefore their direct conversion would not cause their irreversible depletion. To investigate if, and how, OPCs can be reprogrammed; we established a protocol to isolate and culture OPCs from the cortical gray matter of post-natal mice. Subsequently, primary cultures of OPCs were transduced with retroviruses encoding for different transcription factors, such as Neurogenin2, Ascl1 and NeuroD1. We probed the differences in reprogramming rate among different transcription factors at 7 and 14 days, analysed the specific neuronal subtypes generated by these different factors and the acquisition of electrophysiological properties over time. To understand the molecular mechanisms underlying the reprogramming process, we employed single nuclei sequencing and cut&run techniques: we will present preliminary data displaying the molecular changes in OPC-neuron reprogramming and if, and how, this process differ among transcription factors. To substantiate OPCs as target population for direct neuronal reprogramming, we will also present preliminary data on the direct conversion of hIPSC-derived OPCs into neurons, using the above-mentioned reprogramming factors. Together, these data provide compelling evidence that OPCs can be reprogrammed into neurons, paving the way for investigating their direct neuronal conversion in vivo.