Long considered primarily passive cells, astrocytes play an active role in stimulating the formation and function of new synapses to shape neural circuits during development. An emerging theme from recent research is that autism and schizophrenia are diseases of synapses, and raises the question whether functional defects in astrocytes could contribute to the pathophysiology of common devastating neurodevelopmental disorders. Here I propose the use of novel techniques for acutely purifying primary human astrocytes in addition to a protocol for generating induced pluripotent stem cell (iPSC) derived astrocytes from suspended neurospheres. The goal of these approaches is to address how astrocyte dysfunction might underlie neurodevelopmental disease. In order to demonstrate the feasibility of this new model system, I will focus on generating astrocytes from iPSCs produced from patients with 22q11 Deletion Syndrome. These individuals suffer from neurodevelopmental delay and are among the highest genetic risks of developing schizophrenia. Among the candidate genes in the chromosomal deletion for 22q11, the astrocyte-enriched enzyme proline dehydrogenase (PRODH) has already proven to be a promising player in neurological dysfunction from knockout studies in mice. My work to produce iPSC-derived astrocytes from 22q11 patients will ask whether patient-derived astrocytes contribute to defects in neuronal health, synaptic formation, and / or synaptic function through either cell autonomous or non-cell autonomous mechanisms. In addition to the findings within 22q11 patient lines, the techniques developed in this proposal offer the opportunity to investigate the role of astrocytes in numerous neurodevelopmental and neuropsychiatric disorders.
This proposal aims to study the role of astrocytes in neurodevelopmental disease by investigating primary human astrocytes as well as induced pluripotent stem cell-derived (iPSC) astrocytes from patients with 22q11 Deletion Syndrome. The proposed work involves novel methods of acute human astrocyte purification via immunopanning and a protocol for generating iPSC-derived astrocytes using suspended neurospheres.
|Sloan, Steven A; Barres, Ben A (2018) Assembling a Cellular User Manual for the Brain. J Neurosci 38:3149-3153|
|Brosius Lutz, Amanda; Chung, Won-Suk; Sloan, Steven A et al. (2017) Schwann cells use TAM receptor-mediated phagocytosis in addition to autophagy to clear myelin in a mouse model of nerve injury. Proc Natl Acad Sci U S A 114:E8072-E8080|
|Sloan, Steven A; Darmanis, Spyros; Huber, Nina et al. (2017) Human Astrocyte Maturation Captured in 3D Cerebral Cortical Spheroids Derived from Pluripotent Stem Cells. Neuron 95:779-790.e6|
|Darmanis, Spyros; Sloan, Steven A; Croote, Derek et al. (2017) Single-Cell RNA-Seq Analysis of Infiltrating Neoplastic Cells at the Migrating Front of Human Glioblastoma. Cell Rep 21:1399-1410|
|Madelaine, Romain; Sloan, Steven A; Huber, Nina et al. (2017) MicroRNA-9 Couples Brain Neurogenesis and Angiogenesis. Cell Rep 20:1533-1542|
|Zhang, Ye; Sloan, Steven A; Clarke, Laura E et al. (2016) Purification and Characterization of Progenitor and Mature Human Astrocytes Reveals Transcriptional and Functional Differences with Mouse. Neuron 89:37-53|
|Dong, Xiaomin; Chen, Kenian; Cuevas-Diaz Duran, Raquel et al. (2015) Comprehensive Identification of Long Non-coding RNAs in Purified Cell Types from the Brain Reveals Functional LncRNA in OPC Fate Determination. PLoS Genet 11:e1005669|
|Zuchero, J Bradley; Fu, Meng-Meng; Sloan, Steven A et al. (2015) CNS myelin wrapping is driven by actin disassembly. Dev Cell 34:152-67|
|Darmanis, Spyros; Sloan, Steven A; Zhang, Ye et al. (2015) A survey of human brain transcriptome diversity at the single cell level. Proc Natl Acad Sci U S A 112:7285-90|
|Pa?ca, Anca M; Sloan, Steven A; Clarke, Laura E et al. (2015) Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture. Nat Methods 12:671-8|
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