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ScRNA-seq time course of cerebellar Nestin-expressing progenitors from neonatal mice with and without injury at birth

ABSTRACT: scRNA-seq time course of cerebellar Nestin-expressing progenitors from neonatal mice with and without injury at birth

PROVIDER: PRJEB66207 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR12057800_1.fastq.gz	Fastqsanger.gz
	ERR12057800_2.fastq.gz	Fastqsanger.gz
	ERR12057801_1.fastq.gz	Fastqsanger.gz
	ERR12057801_2.fastq.gz	Fastqsanger.gz
	ERR12057802_1.fastq.gz	Fastqsanger.gz

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Childhood Cerebellar Tumors Mirror Conserved Fetal Transcriptional Programs

Project description:The study of the origin and development of cerebellar tumours has been hampered by the complexity and heterogeneity of cerebellar cells that change over the course of development. We used single-cell transcriptomics to study >60,000 cells from the developing murine cerebellum, and show that different molecular subgroups of childhood cerebellar tumours mirror the transcription of cells from distinct, temporally restricted cerebellar lineages. Sonic Hedgehog medulloblastoma transcriptionally mirrors the granule cell hierarchy as expected, whereas Grp3-MB resemble Nestin+ve stem cells, Group 4 medulloblastomas resemble unipolar brush cells, and PFA/PFB ependymoma and cerebellar pilocytic astrocytoma resemble the pre-natal gliogenic progenitor cells. Furthermore, single-cell transcriptomics of human childhood cerebellar tumours demonstrates that many bulk tumours contain a mixed population of cells with divergent differentiation. Our data highlight cerebellar tumours as a disorder of early brain development, and provide a proximate explanation for the peak incidence of cerebellar tumours in early childhood.

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scRNA-seq time course of cerebellar Nestin-expressing progenitors from neonatal mice with and without injury at birth

Project description:To investigate the molecular changes that NEP subpopulations endure upon injury and to determine whether specific populations of NEPs have a transient increase in the signaling pathways associated with injury induced cellular stress and ROS, we performed multiplexed scRNA-seq of NEPs (CFP+) isolated by fluorescence activated cell sorting (FACS) from Nes-CFP/+ transgenic neonates with and without irradiation at timepoints P1, P2, P3 and P5.

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scRNA-seq of primary mouse and human cerebellar nestin-expressing progenitor neurosphere cultures

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Data in support of Prenatal intra-amniotic lipopolysaccharide induces neonatal renal immune activation after preterm birth

Project description:Preterm birth is often predisposed by chorioamnionitis (CA) and CA affects the fetal gut and lungs via intra-amniotic (IA) inflammation, thus accentuating the proinflammatory effects of preterm birth. It is not known if IA inflammation also affects other perfusion-sensitive organs (e.g., kidneys) before and after preterm birth. Using preterm pigs as model for preterm infants, we hypothesized that CA induces fetal and neonatal renal dysfunctions that can intially be detected via plasma proteome, partly explaining the frequent renal dysfunction in preterm infants. Fetal pigs (88% gestation) were given an IA dose of lipopolysaccharide (LPS, 1 mg/kg, n=28), delivered preterm by cesarean section three days later, and compared with controls (CON, n=26) at birth and postnatal day five. Plasma proteome and protein markers of inflammatory pathways were evaluated.

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Comparison between Nestin+ and Nestin- Ptch1 deficient GNPs

Project description:The intermediate filament protein Nestin serves as a biomarker for stem cells and has been used to identify subsets of cancer stem-like cells. However, the mechanistic contributions of Nestin to cancer pathogenesis are not understood. Here we report that Nestin binds the hedgehog pathway transcription factor Gli3 to mediate the development of medulloblastomas of the hedgehog subtype. In a mouse model system, Nestin levels increased progressively during medulloblastoma formation resulting in enhanced tumor growth. Conversely, loss of Nestin dramatically inhibited proliferation and promoted differentiation. Mechanistic investigations revealed that the tumor-promoting effects of Nestin were mediated by binding to Gli3, a zinc finger transcription factor that negatively regulates hedgehog signaling. Nestin binding to Gli3 blocked Gli3 phosphorylation and its subsequent proteolytic processing, thereby abrogating its ability to negatively regulate the hedgehog pathway. Our findings show how Nestin drives hedgehog pathway-driven cancers and uncover in Gli3 a therapeutic target to treat these malignancies.

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Prenatal LPS infection impacting on neonatal mice lung at birth

Project description:Prenatal LPS infection impacting on neonatal mice lung at birth

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Cerebellar granule cell replenishment postinjury by adaptive reprogramming of Nestin+ progenitors

Project description:We report the application of RNA-sequencing for high-throughput profiling of gene expression in Nestin expressing cells of P5 mouse cerebellum Non-Irradiated or Irradiated at P1. By using the Nes-CFP reporter mouse line, we isolated Nes-CFP positive cells of P5 cerebellum to compare the transcriptomes between Nes-CFP positive cells from Non-irradiated cerebellum and cerebellum irradiated at P1.

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An increase in ROS is required for repair of the neonatal cerebellum

Project description:The neonatal mouse cerebellum shows remarkable regenerative potential upon injury at birth. Nestin-expressing progenitors (NEPs) undergo adaptive reprogramming to replenish the lost granule cell progenitors. Here, we investigate how the injury microenvironment affects NEPs’ regenerative potential and adaptive reprogramming. Single cell transcriptomic and bulk chromatin accessibility analyses of the NEPs from control and injured neonatal cerebella show a temporary increase in mechanisms involved in the response to reactive oxygen species (ROS), a known damage-associated molecular pattern. Analysis of ROS levels also confirms a temporary increase in ROS levels upon injury at postanal day 1, 1-2 days after injury, overlapping with the induction in cell death in the cerebellum. Using a transgenic mouse model that overexpresses mitochondrial catalase (mCAT) and reduces ROS levels globally, we showed that the regenerative potential of NEPs decreases upon injury to the granule cell progenitors at birth and the repair is impaired. Finally, we demonstrated that microglia are involved in adaptive reprogramming by regulating NEP migration to the external granule layer, the site of injury. Collectively, our results highlight that the changes in the tissue microenvironment such as an increase in the ROS levels are required for the adaptative reprogramming of NEPs upon injury to the granule cell progenitors at birth, highlighting the instructive roles of microenvironmental signals during regeneration in the neonatal brain.

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scRNA-seq of primary mouse and human cerebellar nestin-expressing progenitor neurosphere cultures

Project description:Nestin-expressing progenitors (NEPs) of the developing cerebellum give rise to Bergmann glia, astrocytes and molecular layer interneurons via molecularly distinct subtypes. NEPs can be cultured in vitro as neurosphere in presence of growth factors. In order to characterise the primary neurosphere cultures generated from mouse and human developing cerebella, we performed single cell RNA-sequencing of primary NEP cultures established from 9-17 post-conception week human fetal cerebella and postnatal day 1 old mouse cerebella from two different mouse strains.

2025-12-04 | GSE301654 | GEO

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