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Cited by 258Neuroinflammation and Neurodegeneration MechanismsPluripotent Stem Cells ResearchMultiple Sclerosis Research Studies

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

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Dräger, Nina M., Sattler, Sydney M., Huang, Cindy Tzu-Ling, Teter, Olivia M., Leng, Kun, Hashemi, Sayed Hadi, Hong, Jason, Aviles, Giovanni +11 more·2022·Nature Neuroscience
DOI 10.1038/s41593-022-01131-4PMC PMC9448678
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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.

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Authors
Dräger, N. M., Sattler, S. M., Huang, C. T.-L., Teter, O. M., Leng, K., Hashemi, S. H., Hong, J., Aviles, G., Clelland, C. D., Zhan, L., Udeochu, J. C., Kodama, L., Singleton, A. B., Nalls, M. A., Ichida, J., Ward, M. E., Faghri, F., Gan, L., & Kampmann, M.
Affiliations
  • University of California, San Francisco (US)
  • Gladstone Institutes (US)
  • University of Illinois Urbana-Champaign (US)
  • National Institutes of Health (US)
  • University of Southern California (US)
Corresponding author
  • Gan, Li
  • Kampmann, Martin
ORCID
Venue
Nature Neuroscience
Publisher
Springer Science and Business Media LLC
Published
2022-08-11
Volume
25
Issue
9
Pages
1149-1162
ISSN
  • 1097-6256 (ISSN-L)
  • 1546-1726
DOI
10.1038/s41593-022-01131-4
PMC
PMC9448678
OpenAlex
W4292229868
Cited by
258
References
25
Citations by year
2026: 71 · 2025: 70 · 2024: 54 · 2023: 42 · 2022: 20 · 2021: 1
FWCI
17.06
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Neuroscience (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • National Science Foundation (Graduate Research Fellowship)
  • U.S. Department of Health & Human Services | NIH | National Institute on Aging (F30 AG066418, F30 AG062043, R01 AG051390)
  • U.S. Department of Health & Human Services | National Institutes of Health (ZO1 AG000534-02)
  • U.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and Stroke (U54 NS100717)
  • U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (DP2 GM119139)
  • U.S. Department of Health & Human Services | NIH | National Institute of Mental Health (U01 MH115747)
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Multiple Sclerosis Research Studies
Keywords
microglia, induced pluripotent stem cell-derived microglia, CRISPR interference, CRISPR activation, microglial states, microglial survival, microglial activation, phagocytosis, neurodegeneration, osteopontin, CSF1R inhibition, single-cell RNA sequencing, disease-associated microglia, transcription factors
MeSH terms
Brain, Humans, Phagocytosis, Microglia, Induced Pluripotent Stem Cells, Clustered Regularly Interspaced Short Palindromic Repeats

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