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A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states

Microglia are emerging as key drivers of neurological diseases. However, we lack a systematic understanding of the underlying mechanisms. Here, we present a screening platform to systematically elucidate functional consequences of genetic perturbations in human induced pluripotent stem cell-derived...

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Autores principales: Dräger, Nina M., Sattler, Sydney M., Huang, Cindy Tzu-Ling, Teter, Olivia M., Leng, Kun, Hashemi, Sayed Hadi, Hong, Jason, Aviles, Giovanni, Clelland, Claire D., Zhan, Lihong, Udeochu, Joe C., Kodama, Lay, Singleton, Andrew B., Nalls, Mike A., Ichida, Justin, Ward, Michael E., Faghri, Faraz, Gan, Li, Kampmann, Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group US 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9448678/
https://www.ncbi.nlm.nih.gov/pubmed/35953545
http://dx.doi.org/10.1038/s41593-022-01131-4
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author Dräger, Nina M.
Sattler, Sydney M.
Huang, Cindy Tzu-Ling
Teter, Olivia M.
Leng, Kun
Hashemi, Sayed Hadi
Hong, Jason
Aviles, Giovanni
Clelland, Claire D.
Zhan, Lihong
Udeochu, Joe C.
Kodama, Lay
Singleton, Andrew B.
Nalls, Mike A.
Ichida, Justin
Ward, Michael E.
Faghri, Faraz
Gan, Li
Kampmann, Martin
author_facet Dräger, Nina M.
Sattler, Sydney M.
Huang, Cindy Tzu-Ling
Teter, Olivia M.
Leng, Kun
Hashemi, Sayed Hadi
Hong, Jason
Aviles, Giovanni
Clelland, Claire D.
Zhan, Lihong
Udeochu, Joe C.
Kodama, Lay
Singleton, Andrew B.
Nalls, Mike A.
Ichida, Justin
Ward, Michael E.
Faghri, Faraz
Gan, Li
Kampmann, Martin
author_sort Dräger, Nina M.
collection PubMed
description Microglia are emerging as key drivers of neurological diseases. However, we lack a systematic understanding of the underlying mechanisms. Here, we present a screening platform to systematically elucidate functional consequences of genetic perturbations in human induced pluripotent stem cell-derived microglia. We developed an efficient 8-day protocol for the generation of microglia-like cells based on the inducible expression of six transcription factors. We established inducible CRISPR interference and activation in this system and conducted three screens targeting the ‘druggable genome’. These screens uncovered genes controlling microglia survival, activation and phagocytosis, including neurodegeneration-associated genes. A screen with single-cell RNA sequencing as the readout revealed that these microglia adopt a spectrum of states mirroring those observed in human brains and identified regulators of these states. A disease-associated state characterized by osteopontin (SPP1) expression was selectively depleted by colony-stimulating factor-1 (CSF1R) inhibition. Thus, our platform can systematically uncover regulators of microglial states, enabling their functional characterization and therapeutic targeting.
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spelling pubmed-94486782022-09-08 A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states Dräger, Nina M. Sattler, Sydney M. Huang, Cindy Tzu-Ling Teter, Olivia M. Leng, Kun Hashemi, Sayed Hadi Hong, Jason Aviles, Giovanni Clelland, Claire D. Zhan, Lihong Udeochu, Joe C. Kodama, Lay Singleton, Andrew B. Nalls, Mike A. Ichida, Justin Ward, Michael E. Faghri, Faraz Gan, Li Kampmann, Martin Nat Neurosci Article Microglia are emerging as key drivers of neurological diseases. However, we lack a systematic understanding of the underlying mechanisms. Here, we present a screening platform to systematically elucidate functional consequences of genetic perturbations in human induced pluripotent stem cell-derived microglia. We developed an efficient 8-day protocol for the generation of microglia-like cells based on the inducible expression of six transcription factors. We established inducible CRISPR interference and activation in this system and conducted three screens targeting the ‘druggable genome’. These screens uncovered genes controlling microglia survival, activation and phagocytosis, including neurodegeneration-associated genes. A screen with single-cell RNA sequencing as the readout revealed that these microglia adopt a spectrum of states mirroring those observed in human brains and identified regulators of these states. A disease-associated state characterized by osteopontin (SPP1) expression was selectively depleted by colony-stimulating factor-1 (CSF1R) inhibition. Thus, our platform can systematically uncover regulators of microglial states, enabling their functional characterization and therapeutic targeting. Nature Publishing Group US 2022-08-11 2022 /pmc/articles/PMC9448678/ /pubmed/35953545 http://dx.doi.org/10.1038/s41593-022-01131-4 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Dräger, Nina M.
Sattler, Sydney M.
Huang, Cindy Tzu-Ling
Teter, Olivia M.
Leng, Kun
Hashemi, Sayed Hadi
Hong, Jason
Aviles, Giovanni
Clelland, Claire D.
Zhan, Lihong
Udeochu, Joe C.
Kodama, Lay
Singleton, Andrew B.
Nalls, Mike A.
Ichida, Justin
Ward, Michael E.
Faghri, Faraz
Gan, Li
Kampmann, Martin
A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
title A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
title_full A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
title_fullStr A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
title_full_unstemmed A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
title_short A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
title_sort crispri/a platform in human ipsc-derived microglia uncovers regulators of disease states
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9448678/
https://www.ncbi.nlm.nih.gov/pubmed/35953545
http://dx.doi.org/10.1038/s41593-022-01131-4
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