RESUMEN
Fibrotic scar tissue formation occurs in humans and mice. The fibrotic scar impairs tissue regeneration and functional recovery. However, the origin of scar-forming fibroblasts is unclear. Here, we show that stromal fibroblasts forming the fibrotic scar derive from two populations of perivascular cells after spinal cord injury (SCI) in adult mice of both sexes. We anatomically and transcriptionally identify the two cell populations as pericytes and perivascular fibroblasts. Fibroblasts and pericytes are enriched in the white and gray matter regions of the spinal cord, respectively. Both cell populations are recruited in response to SCI and inflammation. However, their contribution to fibrotic scar tissue depends on the location of the lesion. Upon injury, pericytes and perivascular fibroblasts become activated and transcriptionally converge on the generation of stromal myofibroblasts. Our results show that pericytes and perivascular fibroblasts contribute to the fibrotic scar in a region-dependent manner.
Asunto(s)
Cicatriz , Fibroblastos , Fibrosis , Pericitos , Traumatismos de la Médula Espinal , Animales , Fibroblastos/patología , Fibroblastos/metabolismo , Fibrosis/patología , Traumatismos de la Médula Espinal/patología , Ratones , Pericitos/patología , Pericitos/metabolismo , Masculino , Femenino , Cicatriz/patología , Ratones Endogámicos C57BL , Células del Estroma/patologíaRESUMEN
Current methods for epigenomic profiling are limited in their ability to obtain genome-wide information with spatial resolution. We introduce spatial ATAC, a method that integrates transposase-accessible chromatin profiling in tissue sections with barcoded solid-phase capture to perform spatially resolved epigenomics. We show that spatial ATAC enables the discovery of the regulatory programs underlying spatial gene expression during mouse organogenesis, lineage differentiation and in human pathology.
Asunto(s)
Cromatina , Transposasas , Animales , Humanos , Ratones , Cromatina/genética , Transposasas/genética , Transposasas/metabolismo , Epigenómica/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Análisis de Secuencia de ADN/métodosRESUMEN
The adult spinal cord stem cell potential resides within the ependymal cell population and declines with age. Ependymal cells are, however, heterogeneous, and the biological diversity this represents and how it changes with age remain unknown. Here, we present a single-cell transcriptomic census of spinal cord ependymal cells from adult and aged mice, identifying not only all known ependymal cell subtypes but also immature as well as mature cell states. By comparing transcriptomes of spinal cord and brain ependymal cells, which lack stem cell abilities, we identify immature cells as potential spinal cord stem cells. Following spinal cord injury, these cells re-enter the cell cycle, which is accompanied by a short-lived reversal of ependymal cell maturation. We further analyze ependymal cells in the human spinal cord and identify widespread cell maturation and altered cell identities. This in-depth characterization of spinal cord ependymal cells provides insight into their biology and informs strategies for spinal cord repair.
Asunto(s)
Neuroglía , Traumatismos de la Médula Espinal , Adulto , Animales , Humanos , Ratones , Diferenciación Celular , Neuroglía/metabolismo , Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/metabolismoRESUMEN
Spinal cord ependymal cells display neural stem cell properties in vitro and generate scar-forming astrocytes and remyelinating oligodendrocytes after injury. We report that ependymal cells are functionally heterogeneous and identify a small subpopulation (8% of ependymal cells and 0.1% of all cells in a spinal cord segment), which we denote ependymal A (EpA) cells, that accounts for the in vitro stem cell potential in the adult spinal cord. After spinal cord injury, EpA cells undergo self-renewing cell division as they give rise to differentiated progeny. Single-cell transcriptome analysis revealed a loss of ependymal cell gene expression programs as EpA cells gained signaling entropy and dedifferentiated to a stem-cell-like transcriptional state after an injury. We conclude that EpA cells are highly differentiated cells that can revert to a stem cell state and constitute a therapeutic target for spinal cord repair.
Asunto(s)
Células-Madre Neurales , Traumatismos de la Médula Espinal , Diferenciación Celular/fisiología , Humanos , Células-Madre Neurales/metabolismo , Neuroglía , Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/metabolismoRESUMEN
Mature oligodendrocytes (MOLs) show transcriptional heterogeneity, the functional consequences of which are unclear. MOL heterogeneity might correlate with the local environment or their interactions with different neuron types. Here, we show that distinct MOL populations have spatial preference in the mammalian central nervous system (CNS). We found that MOL type 2 (MOL2) is enriched in the spinal cord when compared to the brain, while MOL types 5 and 6 (MOL5/6) increase their contribution to the OL lineage with age in all analyzed regions. MOL2 and MOL5/6 also have distinct spatial preference in the spinal cord regions where motor and sensory tracts run. OL progenitor cells (OPCs) are not specified into distinct MOL populations during development, excluding a major contribution of OPC intrinsic mechanisms determining MOL heterogeneity. In disease, MOL2 and MOL5/6 present different susceptibility during the chronic phase following traumatic spinal cord injury. Our results demonstrate that the distinct MOL populations have different spatial preference and different responses to disease.
Asunto(s)
Oligodendroglía/citología , Oligodendroglía/patología , Traumatismos de la Médula Espinal/fisiopatología , Animales , Axones/patología , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Biomarcadores/metabolismo , Linaje de la Célula , Cuerpo Calloso/citología , Encefalomielitis Autoinmune Experimental/patología , Femenino , Perfilación de la Expresión Génica , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Noqueados , Ratones Transgénicos , Oligodendroglía/fisiología , Análisis de la Célula Individual , Médula Espinal/citologíaRESUMEN
Injuries to the central nervous system (CNS) are inefficiently repaired. Resident neural stem cells manifest a limited contribution to cell replacement. We have uncovered a latent potential in neural stem cells to replace large numbers of lost oligodendrocytes in the injured mouse spinal cord. Integrating multimodal single-cell analysis, we found that neural stem cells are in a permissive chromatin state that enables the unfolding of a normally latent gene expression program for oligodendrogenesis after injury. Ectopic expression of the transcription factor OLIG2 unveiled abundant stem cell-derived oligodendrogenesis, which followed the natural progression of oligodendrocyte differentiation, contributed to axon remyelination, and stimulated functional recovery of axon conduction. Recruitment of resident stem cells may thus serve as an alternative to cell transplantation after CNS injury.
Asunto(s)
Células-Madre Neurales/fisiología , Neurogénesis/fisiología , Oligodendroglía/fisiología , Regeneración de la Medula Espinal/fisiología , Animales , Astrocitos/fisiología , Axones/fisiología , Linaje de la Célula , Epéndimo/citología , Epéndimo/metabolismo , Ratones , Ratones Endogámicos C57BL , Neurogénesis/genética , Factor de Transcripción 2 de los Oligodendrocitos/metabolismo , Oligodendroglía/citología , Recuperación de la Función/genética , Recuperación de la Función/fisiología , Remielinización/genética , Remielinización/fisiología , Análisis de la Célula Individual , Traumatismos de la Médula Espinal/fisiopatología , Regeneración de la Medula Espinal/genéticaRESUMEN
Parenchymal astrocytes have emerged as a potential reservoir for new neurons in non-neurogenic brain regions. It is currently unclear how astrocyte neurogenesis is controlled molecularly. Here we show that Notch signaling-deficient astrocytes can generate new neurons after injury. Using single-cell RNA sequencing, we found that, when Notch signaling is blocked, astrocytes transition to a neural stem cell-like state. However, only after injury do a few of these primed astrocytes unfold a neurogenic program, including a self-amplifying progenitor-like state. Further, reconstruction of the trajectories of individual cells allowed us to uncouple astrocyte neurogenesis from reactive gliosis, which occur along independent branches. Finally, we show that cortical neurogenesis molecularly recapitulates canonical subventricular zone neurogenesis with remarkable fidelity. Our study supports a widespread potential of parenchymal astrocytes to function as dormant neural stem cells.
Asunto(s)
Neocórtex , Células-Madre Neurales , Astrocitos , Neurogénesis , NeuronasRESUMEN
Regulation of adult neural stem cell (NSC) number is critical for lifelong neurogenesis. Here, we identified a post-transcriptional control mechanism, centered around the microRNA 204 (miR-204), to control the maintenance of quiescent (q)NSCs. miR-204 regulates a spectrum of transcripts involved in cell cycle regulation, neuronal migration, and differentiation in qNSCs. Importantly, inhibition of miR-204 function reduced the number of qNSCs in the subependymal zone (SEZ) by inducing pre-mature activation and differentiation of NSCs without changing their neurogenic potential. Strikingly, we identified the choroid plexus of the mouse lateral ventricle as the major source of miR-204 that is released into the cerebrospinal fluid to control number of NSCs within the SEZ. Taken together, our results describe a novel mechanism to maintain adult somatic stem cells by a niche-specific miRNA repressing activation and differentiation of stem cells.
Asunto(s)
Plexo Coroideo/química , MicroARNs/genética , Células-Madre Neurales/citología , Adulto , Animales , Ciclo Celular , Diferenciación Celular , Movimiento Celular , Femenino , Regulación de la Expresión Génica , Humanos , Masculino , Ratones , MicroARNs/líquido cefalorraquídeo , Persona de Mediana Edad , Células-Madre Neurales/química , Nicho de Células MadreRESUMEN
The function of somatic stem cells declines with age. Understanding the molecular underpinnings of this decline is key to counteract age-related disease. Here, we report a dramatic drop in the neural stem cells (NSCs) number in the aging murine brain. We find that this smaller stem cell reservoir is protected from full depletion by an increase in quiescence that makes old NSCs more resistant to regenerate the injured brain. Once activated, however, young and old NSCs show similar proliferation and differentiation capacity. Single-cell transcriptomics of NSCs indicate that aging changes NSCs minimally. In the aging brain, niche-derived inflammatory signals and the Wnt antagonist sFRP5 induce quiescence. Indeed, intervention to neutralize them increases activation of old NSCs during homeostasis and following injury. Our study identifies quiescence as a key feature of old NSCs imposed by the niche and uncovers ways to activate NSCs to repair the aging brain.
Asunto(s)
Encéfalo/fisiología , Factores de Edad , Animales , Encéfalo/citología , Diferenciación Celular/fisiología , División Celular/fisiología , Proliferación Celular/fisiología , Senescencia Celular/fisiología , Homeostasis , Masculino , Ratones , Ratones Endogámicos C57BL , Regeneración Nerviosa , Células-Madre Neurales/citología , Células-Madre Neurales/fisiología , Neurogénesis , Nicho de Células MadreRESUMEN
Adult somatic stem cells are generally defined as cells with the ability to differentiate into multiple different lineages and to self-renew during long periods of time. These features were long presumed to be represented in one single tissue-specific stem cell. Recent development of single-cell technologies reveals the existence of diversity in fate and activation state of somatic stem cells within the blood, skin and intestinal compartments [1] but also in the adult brain. Here we review how recent advances have expanded our view of neural stem cells (NSCs) as a diverse pool of cells and how the specialized microenvironment in which they reside acts to maintain this diversity. In addition, we discuss the plasticity of the system in the injured brain.
Asunto(s)
Células Madre Adultas/citología , Células Madre Adultas/fisiología , Células-Madre Neurales/citología , Células-Madre Neurales/fisiología , Adulto , Encéfalo/citología , Lesiones Encefálicas/fisiopatología , Microambiente Celular/fisiología , HumanosRESUMEN
Whether new neurons are added in the postnatal cerebral cortex is still debated. Here, we report that the meninges of perinatal mice contain a population of neurogenic progenitors formed during embryonic development that migrate to the caudal cortex and differentiate into Satb2+ neurons in cortical layers II-IV. The resulting neurons are electrically functional and integrated into local microcircuits. Single-cell RNA sequencing identified meningeal cells with distinct transcriptome signatures characteristic of (1) neurogenic radial glia-like cells (resembling neural stem cells in the SVZ), (2) neuronal cells, and (3) a cell type with an intermediate phenotype, possibly representing radial glia-like meningeal cells differentiating to neuronal cells. Thus, we have identified a pool of embryonically derived radial glia-like cells present in the meninges that migrate and differentiate into functional neurons in the neonatal cerebral cortex.
Asunto(s)
Diferenciación Celular , Movimiento Celular , Corteza Cerebral/citología , Meninges/citología , Neurogénesis , Neuroglía/citología , Neuronas/citología , Animales , Animales Recién Nacidos , Linaje de la Célula , Embrión de Mamíferos/citología , Transportador 1 de Aminoácidos Excitadores/metabolismo , Perfilación de la Expresión Génica , Células HEK293 , Humanos , Ratones Endogámicos C57BL , Nestina/metabolismo , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Reproducibilidad de los Resultados , Análisis de la Célula Individual , Esferoides Celulares/citología , Coloración y Etiquetado , Transcriptoma/genéticaRESUMEN
Heterogeneous pools of adult neural stem cells (NSCs) contribute to brain maintenance and regeneration after injury. The balance of NSC activation and quiescence, as well as the induction of lineage-specific transcription factors, may contribute to diversity of neuronal and glial fates. To identify molecular hallmarks governing these characteristics, we performed single-cell sequencing of an unbiased pool of adult subventricular zone NSCs. This analysis identified a discrete, dormant NSC subpopulation that already expresses distinct combinations of lineage-specific transcription factors during homeostasis. Dormant NSCs enter a primed-quiescent state before activation, which is accompanied by downregulation of glycolytic metabolism, Notch, and BMP signaling and a concomitant upregulation of lineage-specific transcription factors and protein synthesis. In response to brain ischemia, interferon gamma signaling induces dormant NSC subpopulations to enter the primed-quiescent state. This study unveils general principles underlying NSC activation and lineage priming and opens potential avenues for regenerative medicine in the brain.
Asunto(s)
Lesiones Encefálicas/patología , Perfilación de la Expresión Génica/métodos , Células-Madre Neurales/metabolismo , Células-Madre Neurales/patología , Análisis de la Célula Individual/métodos , Animales , Isquemia Encefálica/patología , Diferenciación Celular , Linaje de la Célula , Interferón gamma/metabolismo , Masculino , Ratones Endogámicos C57BL , Transcripción GenéticaRESUMEN
Neuroinflammation is increasingly recognized as a hallmark of neurodegeneration. Activated central nervous system-resident microglia and infiltrating immune cells contribute to the degeneration of dopaminergic neurons (DNs). However, how the inflammatory process leads to neuron loss and whether blocking this response would be beneficial to disease progression remains largely unknown. CD95 is a mediator of inflammation that has also been proposed as an apoptosis inducer in DNs, but previous studies using ubiquitous deletion of CD95 or CD95L in mouse models of neurodegeneration have generated conflicting results. Here we examine the role of CD95 in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP)-induced neurodegeneration using tissue-specific deletion of CD95 or CD95L. We show that DN death is not mediated by CD95-induced apoptosis because deletion of CD95 in DNs does not influence MPTP-induced neurodegeneration. In contrast, deletion of CD95L in peripheral myeloid cells significantly protects against MPTP neurotoxicity and preserves striatal dopamine levels. Systemic pharmacological inhibition of CD95L dampens the peripheral innate response, reduces the accumulation of infiltrating myeloid cells, and efficiently prevents MPTP-induced DN death. Altogether, this study emphasizes the role of the peripheral innate immune response in neurodegeneration and identifies CD95 as potential pharmacological target for neurodegenerative disease.
Asunto(s)
Apoptosis/inmunología , Neuronas Dopaminérgicas/inmunología , Proteína Ligando Fas/inmunología , Inmunidad Innata , Células Mieloides/inmunología , Trastornos Parkinsonianos/inmunología , Animales , Apoptosis/genética , Cuerpo Estriado/inmunología , Cuerpo Estriado/patología , Dopamina/genética , Dopamina/inmunología , Neuronas Dopaminérgicas/patología , Proteína Ligando Fas/antagonistas & inhibidores , Proteína Ligando Fas/genética , Inflamación , Ratones , Ratones Noqueados , Células Mieloides/patología , Trastornos Parkinsonianos/genética , Trastornos Parkinsonianos/patología , Receptor fas/inmunologíaRESUMEN
The role of CD95 (Fas/Apo1) in cancer has been a matter of debate for over 30 years. First discovered as an apoptosis-inducing molecule, CD95 soon emerged as a potential anticancer therapy. Yet accumulating evidence indicates a profound role for CD95 in alternative nonapoptotic signaling pathways that increase tumorigenesis. This fact challenges the initial clinical idea of using CD95 as a 'tumor killer' while setting the stage for clinical studies targeting the nonapoptotic signaling branch of CD95. This review summarizes the findings surrounding manipulation of the CD95 pathway for cancer therapy, considering how one receptor can both promote and prevent cell growth.