High-content synaptic phenotyping in human cellular models reveals a role for BET proteins in synapse assembly
Martin H Berryer,
Gizem Rizki,
Anna Nathanson,
Jenny A Klein,
Darina Trendafilova,
Sara G Susco,
Daisy Lam,
Angelica Messana,
Kristina M Holton,
Kyle W Karhohs,
Beth A Cimini,
Kathleen Pfaff,
Anne E Carpenter,
Lee L Rubin,
Lindy E Barrett
Affiliations
Martin H Berryer
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Gizem Rizki
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Anna Nathanson
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Jenny A Klein
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Darina Trendafilova
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Sara G Susco
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Daisy Lam
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States
Angelica Messana
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States
Kristina M Holton
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Kyle W Karhohs
Imaging Platform, Broad Institute of MIT and Harvard, Cambridge, United States
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States
Resolving fundamental molecular and functional processes underlying human synaptic development is crucial for understanding normal brain function as well as dysfunction in disease. Based upon increasing evidence of species-divergent features of brain cell types, coupled with emerging studies of complex human disease genetics, we developed the first automated and quantitative high-content synaptic phenotyping platform using human neurons and astrocytes. To establish the robustness of our platform, we screened the effects of 376 small molecules on presynaptic density, neurite outgrowth, and cell viability, validating six small molecules that specifically enhanced human presynaptic density in vitro. Astrocytes were essential for mediating the effects of all six small molecules, underscoring the relevance of non-cell-autonomous factors in synapse assembly and their importance in synaptic screening applications. Bromodomain and extraterminal (BET) inhibitors emerged as the most prominent hit class and global transcriptional analyses using multiple BET inhibitors confirmed upregulation of synaptic gene expression. Through these analyses, we demonstrate the robustness of our automated screening platform for identifying potent synaptic modulators, which can be further leveraged for scaled analyses of human synaptic mechanisms and drug discovery efforts.
