CRISPRi screens in human iPSC-derived astrocytes elucidate regulators of distinct inflammatory reactive states

Leng, Kun; Rose, Indigo V.L.; Kim, Hyosung; Xia, Wenlong; Romero-Fernández, Wilber; Rooney, Brendan; Koontz, Mark; Li, Emmy; Ao, Yan; Wang, Shinong; Krawczyk, Mitchell C.; Tcw, Julia; Goate, Alison M.; Zhang, Ye; Ullian, Erik M.; Sofroniew, Michael V.; Fancy, Stephen P.J.; Schrag, Matthew; Lippmann, Ethan S.; Kampmann, Martin

doi:10.1038/s41593-022-01180-9

Cited by 43 publications

(41 citation statements)

References 115 publications

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“…Thus, our ability to generate mixed GRN +/+ neurons + GRN −/− astrocytes showing nearly as strong crSTMN2 and TDP-43 phenotypes of full GRN −/− neuron and astrocyte cultures indicate that GRN LoF in human astrocytes can lead to TDP-43 LoF in neurons. This finding is mechanistically important and adds to a growing body of literature suggesting that disease-associated astrocytes and more generally glia can drive cell death and neuronal dysfunction ( Huang et al., 2022 ; Leng et al, 2022 ; Liddelow et al., 2017 ; Taha et al., 2022 ; Zhang et al., 2020 ).…”

Section: Discussionsupporting

confidence: 53%

Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

et al. 2023

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Section: Discussionsupporting

confidence: 53%

Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

et al. 2023

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“…Reactive astrocytes play an important role in the pathogenesis of many neurodegenerative diseases (Escartin et al, 2021). To investigate this, several studies have modeled inflammation-stimulated reactivity in iPSC-derived astrocytes (Barbar et al, 2020; Leng et al, 2022; Perriot et al, 2018; Roybon et al, 2013; Santos et al, 2017; Tchieu et al, 2019; Tcw et al, 2017). We also characterized the transcriptomic profile of our astrocytes stimulated with pro-inflammatory cytokines TNF-α, IL-1α, and C1q that drive an A1-like reactive state (Liddelow et al, 2017) using qPCR.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, we investigated the responses of astrocytes to pro-inflammatory stimuli by treating the cells with TNF-α, IL-1α, and C1q that drive a reactive phenotype (Escartin et al, 2021; Liddelow et al, 2017). This assay has been widely used to study neuroinflammation in vitro and in vivo (Barbar et al, 2020; Leng et al, 2022; Liddelow et al, 2017; Zhou et al, 2019). Moreover, the morphology and transcriptomic profile of stimulated astrocytes have been well characterized.…”

Section: Discussionmentioning

confidence: 99%

deCLUTTER²⁺pipeline to analyze calcium traces in a novel stem cell model for ventral midbrain patterned astrocytes

Grochowska

Ferraro

Mascaro

et al. 2022

Preprint

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Astrocytes are the most populous cell type of the human central nervous system and are essential for physiological brain function. Increasing evidence suggests multiple roles for astrocytes in Parkinson's disease (PD), nudging a shift in the research focus, which historically pivoted around the ventral midbrain dopaminergic neurons (vmDANs). Studying human astrocytes and other cell types in vivo remains technically and ethically challenging. However, in vitro reprogrammed human stem cell-based models provide a promising alternative. Here, we describe a novel protocol for astrocyte differentiation from human stem cell-derived vmDANs-generating progenitors. This protocol simulates the regionalization, gliogenic switch, radial migration, and final differentiation that occur in the developing human brain. We have characterized the morphological, molecular, and functional features of these ventral midbrain astrocytes with a broad palette of techniques. In addition, we have developed a new pipeline for calcium imaging data analysis called deCLUTTER2+(deconvolution of Ca2+fluorescent patterns) that can be used to discover spontaneous or cue-dependent patterns of Ca2+transients. Altogether, our protocol enables the characterization of the functional properties of human ventral midbrain astrocytes under physiological conditions and in PD.

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“…Furthermore, we investigated whether human iPSC-derived reactive astrocytes displayed similar inflammatory responses to those in stab wound-injured mice. Therefore, we mapped the genes characterizing the two inflammatory clusters of iPSC-derived reactive astrocytes from Leng et al 48 (IRAS 1 and IRAS2) on our integrated single-cell dataset (Fig. 4b, E.D.…”

Section: Injury Induces Common Transcriptomic Changes In Glial Cellsmentioning

confidence: 99%

Shared inflammatory glial cell signature after brain injury, revealed by spatial, temporal and cell-type-specific profiling of the murine cerebral cortex

Koupourtidou¹,

Schwarz²,

Aliee

et al. 2023

Preprint

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Traumatic brain injury leads to a highly orchestrated immune- and glial cell response partially responsible for long-lasting disability and the development of secondary neurodegenerative diseases. A holistic understanding of the mechanisms controlling the responses of specific cell types and their crosstalk is required to develop an efficient strategy for better regeneration. Here, we combined spatial and single-cell transcriptomics to chart the transcriptomic signature of the injured murine cerebral cortex, and identified specific states of astrocytes, microglia, and oligodendrocyte precursor cells contributing to this signature. Interestingly, these cellular populations share a large fraction of injury-regulated genes, including inflammatory programs downstream of the innate immune-associated pathways Cxcr3 and Tlr1/2. Systemic manipulation of these pathways decreased the reactivity state of glial cells associated with poor regeneration. The functional relevance of the newly discovered shared signature of glial cells highlights the importance of our resource enabling comprehensive analysis of early events after brain injury.

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CRISPRi screens in human iPSC-derived astrocytes elucidate regulators of distinct inflammatory reactive states

Cited by 43 publications

References 115 publications

Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

deCLUTTER²⁺pipeline to analyze calcium traces in a novel stem cell model for ventral midbrain patterned astrocytes

Shared inflammatory glial cell signature after brain injury, revealed by spatial, temporal and cell-type-specific profiling of the murine cerebral cortex

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CRISPRi screens in human iPSC-derived astrocytes elucidate regulators of distinct inflammatory reactive states

Cited by 43 publications

References 115 publications

Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

deCLUTTER2+pipeline to analyze calcium traces in a novel stem cell model for ventral midbrain patterned astrocytes

Shared inflammatory glial cell signature after brain injury, revealed by spatial, temporal and cell-type-specific profiling of the murine cerebral cortex

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Product

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About

deCLUTTER²⁺pipeline to analyze calcium traces in a novel stem cell model for ventral midbrain patterned astrocytes