CRISPRi-based screens in iAssembloids to elucidate neuron-glia interactions.

Autor:	Li E; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA.; Biomedical Sciences Graduate Program, University of California, San Francisco, San Francisco, CA, USA., Benitez C; TETRAD Graduate Program, University of California, San Francisco, San Francisco, CA, USA., Boggess SC; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA., Koontz M; Department of Ophthalmology, School of Medicine, University of California, San Francisco, San Francisco, CA, USA., Rose IVL; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA.; Neuroscience Graduate Program, University of California, San Francisco, San Francisco, CA, USA., Martinez D; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA., Draeger N; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA., Teter OM; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA.; UC Berkeley-UCSF Graduate Program in Bioengineering, University of California, San Francisco, San Francisco, CA, USA., Samelson AJ; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA., Pierce N; FirstGen Internship, Emerson Collective, USA; University of California, Berkeley, Berkeley, CA, USA., Ullian EM; Department of Ophthalmology, School of Medicine, University of California, San Francisco, San Francisco, CA, USA., Kampmann M; Institute for Neurodegenerative Diseases, University of California, San Francisco, San Francisco, CA, USA.; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Nov 24. Date of Electronic Publication: 2024 Nov 24.
DOI:	10.1101/2023.04.26.538498
Abstrakt:	The sheer complexity of the brain has complicated our ability to understand the cellular and molecular mechanisms underlying its function in health and disease. Genome-wide association studies have uncovered genetic variants associated with specific neurological phenotypes and diseases. In addition, single-cell transcriptomics have provided molecular descriptions of specific brain cell types and the changes they undergo during disease. Although these approaches provide a giant leap forward towards understanding how genetic variation can lead to functional changes in the brain, they do not establish molecular mechanisms. To address this need, we developed a 3D co-culture system termed iAssembloids (induced multi-lineage assembloids) that enables the rapid generation of homogenous neuron-glia spheroids. We characterize these iAssembloids with immunohistochemistry and single-cell transcriptomics and combine them with large-scale CRISPRi-based screens. In our first application, we ask how glial and neuronal cells interact to control neuronal death and survival. Our CRISPRi-based screens identified that GSK3β inhibits the protective NRF2-mediated oxidative stress response in the presence of reactive oxygen species elicited by high neuronal activity, which was not previously found in 2D monoculture neuron screens. We also apply the platform to investigate the role of APOE- ε 4, a risk variant for Alzheimer's Disease, in its effect on neuronal survival. We find that APOE- ε 4-expressing astrocytes may promote more neuronal activity as compared to APOE- ε 3-expressing astrocytes. This platform expands the toolbox for the unbiased identification of mechanisms of cell-cell interactions in brain health and disease.
Databáze:	MEDLINE
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