The transcription factor PU.1 is critical for viability and function of human brain microglia

Smith, Amy M.; Gibbons, Hannah M.; Oldfield, Robyn; Bergin, Peter M.; Mee, Edward; Faull, Richard L.M.; Dragunow, Mike

doi:10.1002/glia.22486

Cited by 100 publications

(109 citation statements)

References 57 publications

(78 reference statements)

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“…We hypothesize that the pericyte cells originate from the microvasculature and meninges and, in fact, we see a consistency in cell specific markers and response to inflammatory cues in our dissociated and leptomeningeal explant cultures. Using a number of markers in this and previous studies (CD45, PU.1, GFAP), we are confident with the identification of microglia and astrocytes in our cultures [17,26,41]. …”

Section: Discussionsupporting

confidence: 76%

“…In this initial culture, there are approximately 80% of cells that stain positively for both αSMA and PDGFRβ (Figure 1H). After subsequent passaging, CD45 and GFAP positive cells were lost and only the proliferating fibroblast-like and pericyte-like cells remained [26]. All cells in the later cultures are positive for fibronectin, and approximately 90% of cells are positive for both αSMA and PDGFRβ (Figure 1H).…”

Section: Resultsmentioning

confidence: 99%

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A role for human brain pericytes in neuroinflammation

Jansson

Rustenhoven

Feng

et al. 2014

J Neuroinflammation

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138

144

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BackgroundBrain inflammation plays a key role in neurological disease. Although much research has been conducted investigating inflammatory events in animal models, potential differences in human brain versus rodent models makes it imperative that we also study these phenomena in human cells and tissue.MethodsPrimary human brain cell cultures were generated from biopsy tissue of patients undergoing surgery for drug-resistant epilepsy. Cells were treated with pro-inflammatory compounds IFNγ, TNFα, IL-1β, and LPS, and chemokines IP-10 and MCP-1 were measured by immunocytochemistry, western blot, and qRT-PCR. Microarray analysis was also performed on late passage cultures treated with vehicle or IFNγ and IL-1β.ResultsEarly passage human brain cell cultures were a mixture of microglia, astrocytes, fibroblasts and pericytes. Later passage cultures contained proliferating fibroblasts and pericytes only. Under basal culture conditions all cell types showed cytoplasmic NFκB indicating that they were in a non-activated state. Expression of IP-10 and MCP-1 were significantly increased in response to pro-inflammatory stimuli. The two chemokines were expressed in mixed cultures as well as cultures of fibroblasts and pericytes only. The expression of IP-10 and MCP-1 were regulated at the mRNA and protein level, and both were secreted into cell culture media. NFκB nuclear translocation was also detected in response to pro-inflammatory cues (except IFNγ) in all cell types. Microarray analysis of brain pericytes also revealed widespread changes in gene expression in response to the combination of IFNγ and IL-1β treatment including interleukins, chemokines, cellular adhesion molecules and much more.ConclusionsAdult human brain cells are sensitive to cytokine challenge. As expected ‘classical’ brain immune cells, such as microglia and astrocytes, responded to cytokine challenge but of even more interest, brain pericytes also responded to such challenge with a rich repertoire of gene expression. Immune activation of brain pericytes may play an important role in communicating inflammatory signals to and within the brain interior and may also be involved in blood brain barrier (BBB) disruption . Targeting brain pericytes, as well as microglia and astrocytes, may provide novel opportunities for reducing brain inflammation and maintaining BBB function and brain homeostasis in human brain disease.

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Section: Discussionsupporting

confidence: 76%

Section: Resultsmentioning

confidence: 99%

A role for human brain pericytes in neuroinflammation

Jansson

Rustenhoven

Feng

et al. 2014

J Neuroinflammation

Self Cite

138

144

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“…Large domains of the YS-EBs were positive for the master myeloid transcription factor PU.1 (Fig. 1d), which is necessary for microglial differentiation and maintenance 21,22 and can induce macrophages from fibroblasts 23 . Under phase-contrast microscopy, clusters of cells were seen to delaminate towards the lumen of the cysts (Fig.…”

Section: Resultsmentioning

confidence: 99%

Efficient derivation of microglia-like cells from human pluripotent stem cells

Muffat

Yuan

et al. 2016

Nat Med

512

556

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Microglia, the only lifelong resident immune cells of the central nervous system (CNS), are highly specialized macrophages which have been recognized to play a crucial role in neurodegenerative diseases such as Alzheimer’s, Parkinson’s and Adrenoleukodystrophy (ALD). However, in contrast to other cell types of the human CNS, bona fide microglia have not yet been derived from cultured human pluripotent stem cells. Here we establish a robust and efficient protocol for the rapid production of microglia-like cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells that uses defined serum-free culture conditions. These in vitro pluripotent stem cell-derived microglia-like cells (termed pMGLs) faithfully recapitulate the expected ontogeny and characteristics of their in vivo counterparts and resemble primary fetal human and mouse microglia. We generated these cells from multiple disease-specific cell lines, and find that pMGLs derived from MeCP2 mutant hES cells are smaller than their isogenic controls. We further describe a culture platform to study integration and live behavior of pMGLs in organotypic 3D-cultures. This modular differentiation system allows the study of microglia in highly defined conditions, as they mature in response to developmentally relevant cues, and provides a framework to study the long-term interactions of microglia residing in a tissue-like environment.

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“…8E). Given that the transcription factor PU.1 plays a critical role in the development and function of microglia (Yeamans et al, 2007; Feng et al, 2008; Kierdorf et al, 2013; Smith et al, 2013), we used q-PCR to test the effects of age and BDS on mRNA levels of PU.1. Consistent with our promoter analysis, we found a significant effect of age (F(1,31) = 56.9, p < 0.0005, Fig.…”

Section: Resultsmentioning

confidence: 99%

Early life stress perturbs the maturation of microglia in the developing hippocampus

Delpech

Lan

Jin

et al. 2016

Brain, Behavior, and Immunity

157

152

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Children exposed to abuse or neglect show abnormal hippocampal development and similar findings have been reported in rodent models. Using brief daily separation (BDS), a mouse model of early life stress, we previously showed that exposure to BDS impairs hippocampal function in adulthood and perturbs synaptic maturation, synaptic pruning, axonal growth and myelination in the developing hippocampus. Given that microglia are involved in these developmental processes, we tested whether BDS impairs microglial activity in the hippocampus of 14 (during BDS) and 28-day old mice (one week after BDS). We found that BDS increased the density and altered the morphology of microglia in the hippocampus of 14-day old pups, effects that were no longer present on postnatal day (PND) 28. Despite the normal cell number and morphology seen at PND28, the molecular signature of hippocampal microglia, assessed using the NanoString immune panel, was altered at both ages. We showed that during normal hippocampal development, microglia undergo significant changes between PND14 and PND28, including reduced cell density, decreased ex vivo phagocytic activity, and an increase in the expression of genes involved in inflammation and cell migration. However, microglia harvested from the hippocampus of 28-day old BDS mice showed an increase in phagocytic activity and reduced expression of genes that normally increase across development. Promoter analysis indicated that alteration in the transcriptional activity of PU.1, Creb1, Sp1, and RelA accounted for most of the transcriptional changes seen during normal microglia development and for most of the BDS-induced changes at PND14 and PND28. These findings are the first to demonstrate that early life stress dysregulates microglial function in the developing hippocampus and to identify key transcription factors that are likely to mediate these changes.

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The transcription factor PU.1 is critical for viability and function of human brain microglia

Cited by 100 publications

References 57 publications

A role for human brain pericytes in neuroinflammation

A role for human brain pericytes in neuroinflammation

Efficient derivation of microglia-like cells from human pluripotent stem cells

Early life stress perturbs the maturation of microglia in the developing hippocampus

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