Single-cell transcriptomic profiling of the aging mouse brain

Nature Neuroscience, Journal Year: 2019, Volume and Issue: 22(10), P. 1696 - 1708

Published: Sept. 24, 2019

Language: Английский

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Microglia Heterogeneity in the Single-Cell Era

Cell Reports, Journal Year: 2020, Volume and Issue: 30(5), P. 1271 - 1281

Published: Feb. 1, 2020

Microglia are resident immune cells in the central nervous system (CNS) that capable of carrying out prominent and various functions during development adulthood under both homeostatic disease conditions. Although microglia traditionally thought to be heterogeneous populations, which potentially allows them achieve a wide range responses environmental changes for maintenance CNS homeostasis, lack unbiased high-throughput methods assess heterogeneity has prevented study spatially temporally distributed subsets. The recent emergence novel single-cell techniques, such as cytometry by time-of-flight mass spectrometry (CyTOF) RNA sequencing, enabled scientists overcome limitations reveal surprising context-dependent microglia. In this review, we summarize current knowledge about spatial, temporal, functional diversity development, mice humans.

Language: Английский

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GD2-CAR T cell therapy for H3K27M-mutated diffuse midline gliomas

Nature, Journal Year: 2022, Volume and Issue: 603(7903), P. 934 - 941

Published: Feb. 7, 2022

Abstract Diffuse intrinsic pontine glioma (DIPG) and other H3K27M-mutated diffuse midline gliomas (DMGs) are universally lethal paediatric tumours of the central nervous system 1 . We have previously shown that disialoganglioside GD2 is highly expressed on cells demonstrated promising preclinical efficacy GD2-directed chimeric antigen receptor (CAR) T 2 , providing rationale for a first-in-human phase I clinical trial (NCT04196413). Because CAR cell-induced brainstem inflammation can result in obstructive hydrocephalus, increased intracranial pressure dangerous tissue shifts, neurocritical care precautions were incorporated. Here we present experience from first four patients with DIPG or spinal cord DMG treated GD2-CAR at dose level (1 × 10 6 per kg administered intravenously). Patients who exhibited benefit eligible subsequent cell infusions intracerebroventricularly 3 Toxicity was largely related to location tumour reversible intensive supportive care. On-target, off-tumour toxicity not observed. Three radiographic improvement. Pro-inflammatory cytokine levels plasma cerebrospinal fluid. Transcriptomic analyses 65,598 single products fluid elucidate heterogeneity response between participants administration routes. These early results underscore promise this therapeutic approach DMG.

Language: Английский

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Microglia: Immune and non-immune functions

Immunity, Journal Year: 2021, Volume and Issue: 54(10), P. 2194 - 2208

Published: Oct. 1, 2021

Language: Английский

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Microglial subtypes: diversity within the microglial community

The EMBO Journal, Journal Year: 2019, Volume and Issue: 38(17)

Published: Aug. 2, 2019

Review2 August 2019Open Access Microglial subtypes: diversity within the microglial community Vassilis Stratoulias orcid.org/0000-0002-9724-6589 Toxicology Unit, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden Search for more papers by this author Jose Luis Venero Departamento de Bioquímica y Biología Molecular, Facultad Farmacia, Universidad Sevilla, Spain Instituto Biomedicina Sevilla-Hospital Universitario Virgen del Rocío/CSIC/Universidad Marie-Ève Tremblay Department Molecular Université Laval, Quebec, QC, Canada Axe Neurosciences, Centre Recherche du CHU Québec-Université Bertrand Joseph Corresponding Author [email protected] orcid.org/0000-0001-5655-9979 Information Stratoulias1, Venero2,3, Tremblay4,5 and *,1 1Toxicology 2Departamento 3Instituto 4Department 5Axe *Corresponding author. Tel: +46 703057405; E-mail: The EMBO Journal (2019)38:e101997https://doi.org/10.15252/embj.2019101997 PDFDownload PDF article text main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Microglia are brain-resident macrophages forming first active immune barrier in central nervous system. They fulfill multiple functions across development adulthood under disease conditions. Current understanding revolves around microglia acquiring distinct phenotypes upon exposure extrinsic cues their environment. However, emerging evidence suggests that display differences not exclusively driven milieu, rather unique properties these cells possess. This intrinsic heterogeneity has been largely overlooked, favoring prevailing view a single-cell type endowed with spectacular plasticity, allowing them acquire thereby numerous health disease. Here, we review might form which each member (or "subtype") displays performs functions. Distinctive features functional implications several subtypes considered, contexts Finally, suggest subtype categorization shall be based on function propose ways studying them. Hence, advocate plasticity (reaction states) (subtypes) should both considered when multitasking microglia. Introduction were introduced scientific literature century ago (Río-Hortega, 1919a,1919b,1919c; Fig 1). During normal physiological conditions, ramified morphology regularly distributed throughout system (CNS; Río-Hortega, 1919b). Upon pathology, transform function, leading cascade "reaction" from hypertrophic ameboid still orients research today (Flanary et al, 2007; Graeber, 2010; With recent advances genetic tools fate mapping (Ginhoux 2010), now tissue-resident CNS arise embryonic yolk sac (Alliot 1999; Schulz 2012; Kierdorf 2013; Perdiguero 2015). colonize murine day (E)9.5 (Tay 2017c) represent self-maintaining long-lived cell population persists months, if entire lifespan organism (Lawson 1992; Ajami 2007, 2011; Mildner Askew 2017; Füger Réu Tay 2017b). Beyond functioning as mediators injury, inflammation, neurodegeneration, roles healthy brain have identified at an exponential rate past decade (Cartier 2014; 2015; Figure 1. Historical overview identificationAlthough originally proposed Rio-Hortega report microglia, it was only recently idea revisited. Download figure PowerPoint exhibit widely differing depending stage life, region, context or Differences number, morphology, gene expression also reported between sexes (Schwarz Crain Lenz Pimentel-Coelho Butovsky Dorfman Hanamsagar Krasemann 2017). Adequate crucial behavioral adaptation environment (Salter Stevens, 2017a). Throughout contribute neurogenesis, neuronal circuit shaping, vascular formation remodeling, maintenance homeostasis 2017c). aging diseases, may become reactive impaired surveillance phagocytosis (Streit, 2002; Koellhoffer Spittau, contribution diseases is associated compromised (e.g., synaptic plasticity; 2017a) processes adaptive brain, yet death tissue damage pathological settings excitotoxicity, oxidative stress, inflammation; Weil 2008). reaction can triggered any kind insults disturbances CNS. Persisting reaction, often proliferation, involved conditions ranging neurodevelopmental disorders, traumatic injuries, infectious tumors, psychiatric neurodegenerative diseases. Depending stressor insult play, process shown proceed differently result sometimes contrasting outcomes (see 2A classical schematic representation, depicting gray surrounded palette colorful representing state). It recognized wide range states, tremendous shift M1/M2 classification used few years (Martinez Gordon, Ransohoff, 2016). According view, would fulfilled through toward phenotypes, molecular signature (Crain Hickman Bennett 2016; Grabert Flowers Galatro Keren-Shaul Hammond 2018; Masuda 2019). pieces indicate different pools acquired during maturation These co-exist steady state undergo further modulation phenotypic transformation response stimuli (Fig 2B). Indeed, beyond adopts stimuli, constitute members properties, perform functions, respond 2C). We distinctive putative "subtypes", structural, ultrastructural, levels, well Furthermore, categorize than signatures markers. candidates validated using methodological workflow recommend. 2. states(A) Currently, homogenous cellular (core circle gray) extremely plastic. status given developmental resulting invariably assume described literature. (B) In updated version here, heterogeneous having specializations. (C) environmental cue, stimulus, expanding and/or changing its specific phenotype. Microglia: fulfilling vast What defines subject intense debate, discussed Box 1 Accumulating indicates naïve responds identically possible assuming predetermined fact, historical perspective, notion had already 1919 his original description 1919b; He noticed some he named "satellite" found close proximity bodies. A later, satellite below, one playing cards deck 3). important acknowledge others, avant-garde scientists, paved way concept (McCluskey Lampson, 2000; Olah Hanisch, Gertig 2014). 1: How define "cell (sub)type" answer "how subtype?" probably closely related question, type?" Traditionally, defined host tissue, lineage, composition. definition term remains debate (Clevers advancement unbiased technologies transcriptome profiling, such high throughput RNAseq mass cytometry improved/related methods), revealed remarkable among traditionally homogeneous. whereas degree proteome sufficient defining subtypes, even topic (Trapnell, Okawa 2018). While allow molecularly subpopulations, approaches require complemented identification populations, order those (sub)types. Worth notice, importance confound states reaction. latter referring various stimuli. shared properties/characteristics other type. Their selective independent microenvironment. two concepts mutually exclusive, stimulus could react new phenotype, i.e., thus adding another level complexity. must steady-state unchallenged propertie(s) translate into function(s). Typically, existing foundation plan, biased respect studies aimed identifying subtype. includes work markers, most importantly staining, sorting, isolation cells. Reverse provide reliable tool studies, but they inherit technical limitations gating flow antibody unspecificity (Luo 2013). On hand, RNAseq, cytometry, electron microscopy useful tools, aware limitation terms providing static dynamics. however combined two-photon vivo imaging insights Serendipitous approach, sporadic non-systematic. All above methodologies subtypes. Considering review, need classifying evident. Deciphering whether variations instructed microenvironment prime importance, following workflow: Fate-mapping strategies visualize selectively subsets, instance non-invasive chronic imaging—could performed longitudinally development, adulthood, aging, conditions—to determine identity subsets phenotypes. remain examined longitudinally, instead another, notably determinants then studied combination protein analyses, ultrastructure, dynamic investigations. 3. Putative specializationsEmerging data support existence genomic, spatial, morphological, anticipate analyzing thoroughly, 1, similar methodology newly discovered ones, will number characteristics targeted prevention treatment. regional Although ubiquitously scattered CNS, distribution varies regions, white matter 1990). differs presence bodies, dendrites axons, myelinated blood vessels. self-renewal turnover rates lipopolysaccharide (LPS) challenge 2017b; Furube tightly transcriptional level, mouse human (Gosselin 2014, Direct variability comes isolated wild-type, adult mice, according area, determined panels pre-selected study, CD11b, CD40, CD45, CD80, CD86, F4/80, TREM2b, CX3CR1, CCR9 compared regions young mice (de Haas all markers expressed varied significantly areas. study rats, levels known showed region-specific profiles (Doorn Similar areas additionally pattern (Butovsky De Biase Additionally, RNA sequencing (RNAseq) cerebral hippocampal analyzed, 47 identified, including belonging (Zeisel findings raise intriguing possibility survival, activity, growth factor release, metabolism, myelination, blood–brain driving differentiation major contributing heterogeneity. Recently, cerebellar clearance ability, genes supporting engulfment catabolism debris (Ayata "type" reminiscent developing disease-associated (DAM) below. By contrast, striatum homeostatic epigenetic mechanisms particular, suppression striatal mediated PRC2, catalyzes repressive chromatin modification histone H3 lysine 27 trimethylation (H3K27me3). ablation PRC2 results emergence absence dying neurons, cortex. aberrant induce motor responses, decreased learning memory, together anxiety seizures characterized CNS-associated (CAM) three CAM Mrc1, Ms4at, Pf4, Stab1, Cbr2, CD163, Fcrls, compartments: leptomeninges, choroid plexus, perivascular space (Jordão Consequently, partly accounted diversity. differential expressions Differential established approach subpopulations type, GABAergic glutamatergic) observed brain. contexts, neighboring state. local cues, interactions neurons inhibitory excitatory) glial (astrocytes, oligodendrocytes, progenitors), slight signaling thresholds. Similarly, peripheral macrophage activation LPS viruses described, where subset concomitantly (Ravasi 2002). addition, directly communicate other, recruitment lead inhibition initially occupying non-overlapping territories changing, improved staining methods showing direct contacts bodies neighbor (for example, see Milior marker adjacent previous challenges considered. For instance, laser injury intact, converging site (Nimmerjahn 2005; Paris migrate cortex (Eyo 2018) cerebellum (Stowell paints layer expression, cannot excluded argue deserves investigation. below: Keratan sulfate proteoglycan (KSPG)-microglia quarter ago, rat constitutive KSPG (Bertolotto 1993), visualized situ 5D4 monoclonal located extracellular matrix surface. suggested control adhesion axonal growth. 5D4-KSPG subpopulation contrary 1993, 1998). Of note, does coincide GFAP, NG2, MAP2, relate strains inbred rats (Jander Stoll, 1996b). mammals, 5D4-KSPG-expressing spinal cord retina 1998; Jander 1996a; Jones Tuszynski, Zhang Foyez 5D4-KSPG-microglia preferential large numbers hippocampus, brainstem, olfactory bulb (OB), detected neonatal mention 5D4-KSPG-negative same (Jones KSPG-reactivity, systematic required confirm 4). 4.Toolbox. Hox8b-microglia differentiates canonical population, spatial temporal 2 ontogeny Hoxb8-microglia). Mice carrying driver Hoxb8-Cre reporter ROSA26-YFP alleles crossed trace YFP-Hoxb8 expression. YFP signal appeared YFP-positive especially OB (Chen 25–40% total YFP-negative Nagarajan Transcriptomic analyses comparing Hoxb8-positive Hoxb8-negative very state, 21 populations (De Hoxb8-microglia express genes, Tmem119, Sall1, Sall3, Gpr56, Ms4a7, hematopoietic Clel12a, Klra2, Lilra5 non-Hoxb8 (Bennett neither expresses Hoxb8 brain; instead, lineage tracer progenitors prior infiltration Selective inactivation grooming behavior, mutant strategy deletion included use Tie2 Cre affect endothelial 2010). More cell-specific prerequisite involvement behavior. (with illustration): Revisiting origin(s) An question arising relates origin(s). Do possess populating do once assumed parenchyma? convincingly derive wave hematopoiesis Hoeffel Sheng Mass 2016), follow stepwise program (Mass Matcovitch-Natan before E9.5 Based literature, differentiate inside parenchyma (a). hypothesis explain microenvironments (inhibitory, oligodendrocytes thresholds, alternative exhibiting infiltrating early investigation, tested (b c). later hypothesis, Capecchi al Hoxb8-microglia-progenitors exist E8.5 Subsequently, transit aorta-gonad-mesonephros fetal liver, expand entry E12.5 (c). lines, CSF1R−/− Erblich 2011) IL2-Tgfb1;Tgfb1−/− (Keren-Shaul 2017) transgenic expected parenchyma. E14.5 exist, Ms4a7 (Hammond great interest investigate subpopulations. zebrafish, waves (Xu Ferrero yolk-sac-equivalent structure origin populate replenished zebrafish (d). div

Language: Английский

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cGAS–STING drives ageing-related inflammation and neurodegeneration

Nature, Journal Year: 2023, Volume and Issue: 620(7973), P. 374 - 380

Published: Aug. 2, 2023

Low-grade inflammation is a hallmark of old age and central driver ageing-associated impairment disease1. Multiple factors can contribute to inflammation2; however, the molecular pathways that transduce aberrant inflammatory signalling their impact in natural ageing remain unclear. Here we show cGAS-STING pathway, which mediates immune sensing DNA3, critical chronic functional decline during ageing. Blockade STING suppresses phenotypes senescent human cells tissues, attenuates ageing-related multiple peripheral organs brain mice, leads an improvement tissue function. Focusing on brain, reveal activation triggers reactive microglial transcriptional states, neurodegeneration cognitive decline. Cytosolic DNA released from perturbed mitochondria elicits cGAS activity microglia, defining mechanism by engaged brain. Single-nucleus RNA-sequencing analysis microglia hippocampi gain-of-function mouse model demonstrates engagement sufficient direct states leading bystander cell inflammation, neurotoxicity impaired memory capacity. Our findings establish pathway as blockade potential strategy halt neurodegenerative processes age.

Language: Английский

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Microglial regional heterogeneity and its role in the brain

Molecular Psychiatry, Journal Year: 2019, Volume and Issue: 25(2), P. 351 - 367

Published: Nov. 26, 2019

Abstract Microglia have been recently shown to manifest a very interesting phenotypical heterogeneity across different regions in the mammalian central nervous system (CNS). However, underlying mechanism and functional meaning of this phenomenon are currently unclear. Baseline diversities adult microglia their cell number, cellular subcellular structures, molecular signature as well relevant functions discovered. But recent transcriptomic studies using bulk RNAseq single-cell produced conflicting results on region-specific signatures microglia. It is highly speculative whether such spatial contributes varying sensitivities individual same physiological pathological signals CNS regions, hence underlie relevance for disease development. This review aims thoroughly summarize up-to-date knowledge specific topic provide some insights potential mechanisms, starting from microgliogenesis. Understanding regional context diverse neighboring neurons other glia may an important clue future development innovative therapies neuropsychiatric disorders.

Language: Английский

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Astrocytes in chronic pain and itch

Nature reviews. Neuroscience, Journal Year: 2019, Volume and Issue: 20(11), P. 667 - 685

Published: Sept. 19, 2019

Language: Английский

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The multiplex model of the genetics of Alzheimer’s disease

Nature Neuroscience, Journal Year: 2020, Volume and Issue: 23(3), P. 311 - 322

Published: Feb. 28, 2020

Language: Английский

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Molecular architecture of the developing mouse brain

Nature, Journal Year: 2021, Volume and Issue: 596(7870), P. 92 - 96

Published: July 28, 2021

Language: Английский

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