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.

Public Health Relevance

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.

Agency
National Institute of Health (NIH)
Type
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
Project #
1F30MH106261-01
Application #
8833635
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Rosemond, Erica K
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Stanford University
Department
Neurology
Type
Schools of Medicine
DUNS #
City
Stanford
State
CA
Country
United States
Zip Code
94304
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
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
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
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
PaÅŸca, 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
Sloan, Steven A; Barres, Ben A (2014) Mechanisms of astrocyte development and their contributions to neurodevelopmental disorders. Curr Opin Neurobiol 27:75-81
Sloan, Steven A; Barres, Ben A (2014) The detrimental role of glial acidification during ischemia. Neuron 81:221-3
Sloan, Steven A; Barres, Ben A (2014) Looks can be deceiving: reconsidering the evidence for gliotransmission. Neuron 84:1112-5