The recent identification of transcription factors (TFs) that can induce conversion of fibroblasts into pluripotent stem (iPS) cells makes it potentially possible to generate patient-specific neurons from fibroblasts. However, the neurons thus produced are difficult to obtain. The present project builds on preliminary results in which we demonstrate direct conversion of adult fibroblasts cells into neurons, referred to as 'induced neuronal cells'(iN cells), without an iPS intermediate. The resulting iN cells have the functional properties of neurons, including the ability to form functional synapses as assayed by electrophysiology. Thus, the iN cell technology provides a novel, more facile approach to generating and studying human neurons, and opens up a new avenue to investigating human disease processes. However, at this point the iN cell technology has only been developed for mouse fibroblasts, fundamental questions regarding the conversion process and the molecular identity of iN cells were not determined, the generation of iN cells from human fibroblasts has not yet been established, and most importantly, the feasibility of the iN cell technology to study diseases affecting neuronal function has not been demonstrated. In this project, we propose to address these important challenges in an interdisciplinary approach capitalizing from the combined expertises of the Wernig and S|dhof laboratories. We propose experiments that will systematically investigate the cellular and molecular identity of iN cells, and develop protocols to induce specific neuronal subpopulations from fibroblasts. These protocols will then be employed to model genetic diseases in mouse iN cells. Furthermore, we will extend our findings to human fibroblasts, with the long term goal to establish a cell model for neuropsychiatric diseases. Our goals will be pursued by a combination of tissue culture experiments with cells cultured from mice and humans, cell biology, molecular biology, and electrophysiology. We believe our proposed experiments have the potential to fundamentally change existing paradigms of cellular differentiation and epigenetic gene regulation, and could provide a novel platform to study human neurons from patients suffering from a variety of brain diseases.
This application will develop methods to generate neurons directly from non-neuronal cells, allowing the production of neurons from skin fibroblasts of human patients. These methods will then be used to test the effects of mutations associated with neuropsychiatric disorders on neuronal biology, with the long-term goal of establishing a better understanding of the pathomechanism of these diseases in human neurons.
|Zhong, Lei R; Chen, Xin; Park, Esther et al. (2018) Retinoic Acid Receptor RAR?-Dependent Synaptic Signaling Mediates Homeostatic Synaptic Plasticity at the Inhibitory Synapses of Mouse Visual Cortex. J Neurosci 38:10454-10466|
|Zhang, Zhenjie; Marro, Samuele G; Zhang, Yingsha et al. (2018) The fragile X mutation impairs homeostatic plasticity in human neurons by blocking synaptic retinoic acid signaling. Sci Transl Med 10:|
|Südhof, Thomas C (2018) Towards an Understanding of Synapse Formation. Neuron 100:276-293|
|Tanabe, Koji; Ang, Cheen Euong; Chanda, Soham et al. (2018) Transdifferentiation of human adult peripheral blood T cells into neurons. Proc Natl Acad Sci U S A 115:6470-6475|
|Chanda, Soham; Hale, W Dylan; Zhang, Bo et al. (2017) Unique versus Redundant Functions of Neuroligin Genes in Shaping Excitatory and Inhibitory Synapse Properties. J Neurosci 37:6816-6836|
|Seigneur, Erica; Südhof, Thomas C (2017) Cerebellins are differentially expressed in selective subsets of neurons throughout the brain. J Comp Neurol 525:3286-3311|
|Yang, Nan; Chanda, Soham; Marro, Samuele et al. (2017) Generation of pure GABAergic neurons by transcription factor programming. Nat Methods 14:621-628|
|Patzke, Christopher; Acuna, Claudio; Giam, Louise R et al. (2016) Conditional deletion of L1CAM in human neurons impairs both axonal and dendritic arborization and action potential generation. J Exp Med 213:499-515|
|Yi, Fei; Danko, Tamas; Botelho, Salome Calado et al. (2016) Autism-associated SHANK3 haploinsufficiency causes Ih channelopathy in human neurons. Science 352:aaf2669|
|Chanda, S; Aoto, J; Lee, S-J et al. (2016) Pathogenic mechanism of an autism-associated neuroligin mutation involves altered AMPA-receptor trafficking. Mol Psychiatry 21:169-77|
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