학술논문
A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states
Document Type
article
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
Source
Nature Neuroscience. 25(9)
Subject
Language
Abstract
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.