title: Functional genomic screen of human stem cell differentiation reveals pathways involved in neurodevelopment and neurodegeneration.
abstract: Human embryonic stem cells (hESCs) can be induced and differentiated to form a relatively homogeneous population of neuronal precursors in vitro. We have used this system to screen for genes necessary for neural lineage development by using a pooled human short hairpin RNA (shRNA) library screen and massively parallel sequencing. We confirmed known genes and identified several unpredicted genes with interrelated functions that were specifically required for the formation or survival of neuronal progenitor cells without interfering with the self-renewal capacity of undifferentiated hESCs. Among these are several genes that have been implicated in various neurodevelopmental disorders (i.e., brain malformations, mental retardation, and autism). Unexpectedly, a set of genes mutated in late-onset neurodegenerative disorders and with roles in the formation of RNA granules were also found to interfere with neuronal progenitor cell formation, suggesting their functional relevance in early neurogenesis. This study advances the feasibility and utility of using pooled shRNA libraries in combination with next-generation sequencing for a high-throughput, unbiased functional genomic screen. Our approach can also be used with patient-specific human-induced pluripotent stem cell-derived neural models to obtain unparalleled insights into developmental and degenerative processes in neurological or neuropsychiatric disorders with monogenic or complex inheritance.
authors.affiliation: Program in Neurodevelopment and Regeneration, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Vincent P
authors.initials: VP
authors.lastname: Schulz
authors.forename: Brian D
authors.initials: BD
authors.lastname: Reed
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Xinghua
authors.initials: X
authors.lastname: Pan
authors.forename: Jessica
authors.initials: J
authors.lastname: Mariani
authors.forename: Ghia
authors.initials: G
authors.lastname: Euskirchen
authors.forename: Michael P
authors.initials: MP
authors.lastname: Snyder
authors.forename: Flora M
authors.initials: FM
authors.lastname: Vaccarino
authors.forename: Natalia
authors.initials: N
authors.lastname: Ivanova
authors.forename: Sherman M
authors.initials: SM
authors.lastname: Weissman
authors.forename: Anna M
authors.initials: AM
authors.lastname: Szekely
mesh_terms: D001321:Autistic Disorder / Q000235:genetics; D002454:Cell Differentiation; D020868:Gene Silencing; D018390:Gene Targeting; D015894:Genome, Human; D006801:Humans; D008607:Intellectual Disability / Q000235:genetics; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D012313:RNA
keywords: Mendelian disorders of the nervous system; RNA-binding proteins; high-throughput sequencing; neural differentiation; pooled RNAi screening