Global transcriptome profile of the developmental principles of in vitro iPSC-to-motor neuron differentiation

Abstract Background Human induced pluripotent stem cells (iPSC) have opened new avenues for regenerative medicine. Consequently, iPSC-derived motor neurons emerged as potentially viable therapies spinal cord injuries and neurodegenerative disorders including Amyotrophic Lateral Sclerosis. However, direct clinical application of iPSC bears in itself the risk tumorigenesis other unforeseeable genetic or epigenetic abnormalities. Results Employing RNA-seq technology, we identified characterized gene regulatory networks triggered by vitro chemical reprogramming into with molecular features (MNs) whose function vivo is to innervate effector organs. We present meta-transcriptome signatures 5 cell types: iPSCs, neural cells, neuron progenitors, early neurons, mature neurons. In strict response stimuli, along MN differentiation axis observed temporal downregulation tumor growth factor-β signaling pathway consistent activation sonic hedgehog, Wnt/β-catenin, Notch signaling. Together defining neuronal (neurogenin 2, microtubule-associated protein Pax6, neuropilin-1), steady accumulation neuron-specific genes, Islet-1 homeobox HB9. Interestingly, transcriptome profiling process showed that Ca 2+ through cAMP LPC was downregulated during conversion key activity remained inhibited until later stages formation. Pathways shaping development were well-represented including, neuroactive ligand-receptor interactions, axon guidance, cholinergic synapse A notable hallmark our maturation monoculture genes encoding G-coupled muscarinic acetylcholine receptors ionotropic nicotinic expression. formation functional spontaneous oscillations extracellular action potentials recorded on multi-electrode array chip after 20 days differentiation. Conclusions Detailed profile each developmental step from driven induction provides guidelines novel therapeutic approaches re-construction efforts muscle innervation.