Differentiation of Inflammation-Responsive Astrocytes from Glial Progenitors Generated from Human Induced Pluripotent Stem Cells

Santos, Renata; Vadodaria, Krishna C.; Jaeger, Baptiste N.; Mei, Arianna; Lefcochilos-Fogelquist, Sabrina; Mendes, Ana Cristina; Erikson, Galina; Shokhirev, Maxim N.; Randolph‐Moore, Lynne; Fredlender, Callie; Dave, Sonia; Oefner, Ruth; Fitzpatrick, Conor; Pena, Monique; Barron, Jerika J.; Ku, Manching; Denli, Ahmet M.; Kerman, Bilal E.; Charnay, Patrick; Kelsoe, John R.; Marchetto, Maria C.; Gage, Fred H.

doi:10.1016/j.stemcr.2017.05.011

Cited by 130 publications

(162 citation statements)

References 57 publications

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“…iPSCs as disease models iPSCs were initially used to model diseases with highly penetrant genetic variants (Box 1) of large phenotypic effect (Cao et al, 2016;Liu et al, 2011;Wainger et al, 2014), but more recently they have been used to study common genetic variants of modest effect size that drive complex diseases. They provide a key platform to study the impact of human cell type-specific gene regulation, as they can recapitulate the broad regulatory profile of their in vivo counterparts and also mirror tissue-specific functional genetic variation (Banovich et al, 2018;Santos et al, 2017). Moreover, large-scale iPSC-based studies have identified expression quantitative traitassociated loci (eQTLs; Box 1) that inform on the interpretation of variants identified by genome-wide association studies (GWAS; Box 1) (Carcamo-Orive et al, 2017), as well as protein quantitative trait loci that give insights into mechanisms through which diseaseassociated genetic risk modulates cell physiology (Mirauta et al, 2018 preprint).…”

Section: Introductionmentioning

confidence: 99%

Addressing variability in iPSC-derived models of human disease: guidelines to promote reproducibility

Volpato

Webber

2020

Disease Models &Amp; Mechanisms

257

169

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Induced pluripotent stem cell (iPSC) technologies have provided in vitro models of inaccessible human cell types, yielding new insights into disease mechanisms especially for neurological disorders. However, without due consideration, the thousands of new human iPSC lines generated in the past decade will inevitably affect the reproducibility of iPSC-based experiments. Differences between donor individuals, genetic stability and experimental variability contribute to iPSC model variation by impacting differentiation potency, cellular heterogeneity, morphology, and transcript and protein abundance. Such effects will confound reproducible disease modelling in the absence of appropriate strategies. In this Review, we explore the causes and effects of iPSC heterogeneity, and propose approaches to detect and account for experimental variation between studies, or even exploit it for deeper biological insight.

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Section: Introductionmentioning

confidence: 99%

Addressing variability in iPSC-derived models of human disease: guidelines to promote reproducibility

Volpato

Webber

2020

Disease Models &Amp; Mechanisms

257

169

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“…The advent of human embryonic stem cell and induced pluripotent stem cell (iPSC) technology has enabled large-scale generation of human astrocytes and other CNS cells that retain the genetic information of the patient, as powerful human in vitro models of disease (reviewed in 14 ) . Several protocols to generate astrocytes have been developed by independent groups, using either a specific gradient of patterning agents to mimic embryonic development [15][16][17][18][19][20][21] or overexpression of critical transcription factors 22,23 . Usually, iPSC differentiation into astrocytes in monolayer cultures does not require a purification step; in recent years, however, more complex 3D cultures of CNS organoids have been established that contain neural progenitor cells, neurons, oligodendrocyte lineage cells and microglia, in addition to astrocytes 22,[24][25][26][27][28][29][30][31] .…”

Section: Introductionmentioning

confidence: 99%

CD49f is a novel marker to purify functional human iPSC-derived astrocytes

Barbar

Jain

Zimmer

et al. 2019

Preprint

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Astrocytes play a central role in the central nervous system (CNS), maintaining brain homeostasis, providing metabolic support to neurons, regulating connectivity of neural circuits, and controlling blood flow as an integral part of the blood-brain barrier. They have been increasingly implicated in the mechanisms of neurodegenerative diseases, prompting a greater need for methods that enable their study. The advent of human induced pluripotent stem cell (iPSC) technology has made it possible to generate patient-specific astrocytes and CNS cells using protocols developed by our team and others as valuable disease models. Yet isolating astrocytes from primary specimens or from in vitro mixed cultures for downstream analyses has remained challenging. To address this need, we performed a screen for surface markers that allow FACS sorting of astrocytes. Here we demonstrate that CD49f is an effective marker for sorting functional human astrocytes. We sorted CD49f + cells from a protocol we previously developed that generates a complex culture of oligodendrocytes, neurons and astrocytes from iPSCs. CD49f + -purified cells express all canonical astrocyte markers and perform characteristic functions, such as neuronal support and glutamate uptake. 1Of particular relevance to neurodegenerative diseases, CD49f + astrocytes can be stimulated to take on an A1 neurotoxic phenotype, in which they secrete proinflammatory cytokines and show an impaired ability to support neuronal maturation. This study establishes a novel marker for isolating functional astrocytes from complex CNS cell populations, strengthening the use of iPSC-astrocytes for the study of their regulation and dysregulation in neurodegenerative diseases.

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“…Gene programs induced when astrocytes enter a reactive state are similar to those of the MES GBM subtype To examine connections between the MES GBM transcriptome profile and the astrocytic response during neuroinflammation, we set up an in vitro model to study the transition of astrocytes into a reactive cell state. In brief, cultured human primary astrocytes were exposed to TNFα or IL1β, which resulted in NFκB activation ( Figure 1A) and upregulation of genes coupled to proinflammatory response, CSF1, CCL2, and MX1 [20][21][22][23] ( Figure 1B). Next, we analyzed changes in the whole transcriptome during long-term treatment with IL1β (12 days).…”

Section: Resultsmentioning

confidence: 99%

Mesenchymal transition and increased therapy resistance of glioblastoma cells is related to astrocyte reactivity

Niklasson

Bergström

Jarvius

et al. 2019

The Journal of Pathology

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Grade IV astrocytoma/glioblastoma multiforme (GBM) is essentially incurable, partly due to its heterogenous nature, demonstrated even within the glioma‐initiating cell (GIC) population. Increased therapy resistance of GICs is coupled to transition into a mesenchymal (MES) cell state. The GBM MES molecular signature displays a pronounced inflammatory character and its expression vary within and between tumors. Herein, we investigate how MES transition of GBM cells relates to inflammatory responses of normal astroglia. In response to CNS insults astrocytes enter a reactive cell state and participate in directing neuroinflammation and subsequent healing processes. We found that the MES signature show strong resemblance to gene programs induced in reactive astrocytes. Likewise, astrocyte reactivity gene signatures were enriched in therapy‐resistant MES‐like GIC clones. Variable expression of astrocyte reactivity related genes also largely defined intratumoral GBM cell heterogeneity at the single‐cell level and strongly correlated with our previously defined therapy‐resistance signature (based on linked molecular and functional characterization of GIC clones). In line with this, therapy‐resistant MES‐like GIC secreted immunoregulatory and tissue repair related proteins characteristic of astrocyte reactivity. Moreover, sensitive GIC clones could be made reactive through long‐term exposure to the proinflammatory cytokine interleukin 1 beta (IL1β). IL1β induced a slow MES transition, increased therapy resistance, and a shift in DNA methylation profile towards that of resistant clones, which confirmed a slow reprogramming process. In summary, GICs enter through MES transition a reactive‐astrocyte‐like cell state, connected to therapy resistance. Thus, from a biological point of view, MES GICs would preferably be called ‘reactive GICs’. The ability of GBM cells to mimic astroglial reactivity contextualizes the immunomodulatory and microenvironment reshaping abilities of GBM cells that generate a tumor‐promoting milieu. This insight will be important to guide the development of future sensitizing therapies targeting treatment‐resistant relapse‐driving cell populations as well as enhancing the efficiency of immunotherapies in GBM. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

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Differentiation of Inflammation-Responsive Astrocytes from Glial Progenitors Generated from Human Induced Pluripotent Stem Cells

Cited by 130 publications

References 57 publications

Addressing variability in iPSC-derived models of human disease: guidelines to promote reproducibility

Addressing variability in iPSC-derived models of human disease: guidelines to promote reproducibility

CD49f is a novel marker to purify functional human iPSC-derived astrocytes

Mesenchymal transition and increased therapy resistance of glioblastoma cells is related to astrocyte reactivity

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