biostudies-literatureLeng KNew York Stem Cell FoundationNEI NIH HHSU.S. Department of Health & Human Services | NIH | National Eye InstituteU.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and StrokeNational Multiple Sclerosis SocietyNIA NIH HHSU.S. Department of Health & Human Services | NIH | National Institute on AgingNINDS NIH HHSCalifornia Institute for Regenerative MedicineChan Zuckerberg Initiative Ben Barres Early Career Acceleration Award1528-1542https://www.ebi.ac.uk/biostudies/studies/S-EPMC9633461biostudies-literatureUnknown25(11)Astrocytes become reactive in response to insults to the central nervous system by adopting context-specific cellular signatures and outputs, but a systematic understanding of the underlying molecular mechanisms is lacking. In this study, we developed CRISPR interference screening in human induced pluripotent stem cell-derived astrocytes coupled to single-cell transcriptomics to systematically interrogate cytokine-induced inflammatory astrocyte reactivity. We found that autocrine-paracrine IL-6 and interferon signaling downstream of canonical NF-κB activation drove two distinct inflammatory reactive signatures, one promoted by STAT3 and the other inhibited by STAT3. These signatures overlapped with those observed in other experimental contexts, including mouse models, and their markers were upregulated in human brains in Alzheimer's disease and hypoxic-ischemic encephalopathy. Furthermore, we validated that markers of these signatures were regulated by STAT3 in vivo using a mouse model of neuroinflammation. These results and the platform that we established have the potential to guide the development of therapeutics to selectively modulate different aspects of inflammatory astrocyte reactivity.Nature neuroscienceCRISPRi screens in human iPSC-derived astrocytes elucidate regulators of distinct inflammatory reactive states.PMC9633461Druckenmiller FellowshipP30 EY002162R21 NS119954P30 AG066514P30 EY02162-39R03 AG063157R01 NS084030Harry Weaver Neuroscience ScholarshipR01 NS09755T32 NS115706EDUC4-12812K01 AG062683P01 NS083513F30 AG066418R01 NS097551Kim HLi EKrawczyk MUllian EMFancy SPJRooney BKoontz MKampmann MLeng KXia WSofroniew MVRomero-Fernandez WWang SZhang YAo YGoate ALippmann ESTcw JSchrag MSRose IVLfalseCRISPRi screens in human iPSC-derived astrocytes elucidate regulators of distinct inflammatory reactive states.Astrocytes become reactive in response to insults to the central nervous system by adopting context-specific cellular signatures and outputs, but a systematic understanding of the underlying molecular mechanisms is lacking. In this study, we developed CRISPR interference screening in human induced pluripotent stem cell-derived astrocytes coupled to single-cell transcriptomics to systematically interrogate cytokine-induced inflammatory astrocyte reactivity. We found that autocrine-paracrine IL-6 and interferon signaling downstream of canonical NF-κB activation drove two distinct inflammatory reactive signatures, one promoted by STAT3 and the other inhibited by STAT3. These signatures overlapped with those observed in other experimental contexts, including mouse models, and their markers were upregulated in human brains in Alzheimer's disease and hypoxic-ischemic encephalopathy. Furthermore, we validated that markers of these signatures were regulated by STAT3 in vivo using a mouse model of neuroinflammation. These results and the platform that we established have the potential to guide the development of therapeutics to selectively modulate different aspects of inflammatory astrocyte reactivity.2022-01-01T00:00:00Z2022 Nov2026-05-27T10:52:04.381Z2025-04-04T11:26:27.216ZS-EPMC96334613630306910.1038/s41593-022-01180-9