Using single cell RNAseq to unveil characteristics of CD133+ neural stem cells during development
使用单细胞 RNAseq 揭示 CD133 神经干细胞发育过程中的特征
基本信息
- 批准号:9751415
- 负责人:
- 金额:$ 19.5万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2018
- 资助国家:美国
- 起止时间:2018-08-01 至 2021-07-31
- 项目状态:已结题
- 来源:
- 关键词:AdultAnatomyAntibodiesAtlasesBiological AssayBiotechnologyBrainCell Differentiation processCell LineageCell surfaceCellsCharacteristicsComplexDataData AnalysesDatabasesDevelopmentEmbryoEmotionsEndotheliumEpendymal CellForebrain DevelopmentFutureGene Expression RegulationGenesGenetic TranscriptionGenomicsGlial Fibrillary Acidic ProteinGoalsGrantHematopoieticHeterogeneityImmunohistochemistryIn Situ HybridizationIndividualKnowledgeLabelLightLocationMapsMesenchymalMessenger RNAMicrospheresMitoticMolecularMolecular ProfilingMovementMusNervous system structureNeural Tube ClosureNeuraxisNeuroepithelialNeuronsPathway AnalysisPopulationPropertyProsencephalonProteinsPublishingRegulator GenesReporterReportingResolutionStem cellsStructureSurfaceSynapsesTechniquesTechnologyTimeTissuesTomatoesTubeValidationVascular Endothelial Growth FactorsVentricularWorkbasebrain cellcell cortexcell typeclinical applicationcostexperimental studygenetic signatureinjuredloss of functionmultipotent cellnerve stem cellnervous system developmentnestin proteinneural circuitneurodevelopmentneurogenesisnovelprogenitorprogramspromininranpirnaserelating to nervous systemrepairedself-renewalsingle cell sequencingsingle-cell RNA sequencingstem cell nichestem cell populationtranscriptometranscriptome sequencing
项目摘要
Using single cell RNAseq to unveil characteristics of CD133+ neural stem cells during development
Project Summary
Neural stem cells (NSCs), by definition, are self-renewing, multipotent cells that generate major cell types of
the nervous system. Our current knowledge about NSC is still quite limited, insufficient to fulfill the goal of
achieving good understanding of neurodevelopment or to engage endogenous NSCs to repair degenerated or
injured central nervous system (CNS). Like other tissue specific stem cells, NSCs reside in highly complex
cellular microenvironment and are in close contact with both stem cell niche and their downstream progeny cell
types, which had made the characterization of the cellular and molecular signatures of NSCs extremely
challenging. Recently developed single cell RNAseq technology pave the way to uncover molecular
characteristics of any cell types including stem cells with precision. Particularly, the novel droplet-based
technique allowed thousands of single cells to be sequenced in one tube yet still produce transcriptomes with
single cell resolution. In this application, we propose to apply this scalable and low cost single cell RNAseq
analysis developed by 10X Genomics to characterize CD133 positive ependymal cells lining the ventricular
surface of the brain. The transciptome data of thousands of single CD133+ cells from cortex at different time
points during development will be subjected to comprehensive data analyses, including Weighted Gene
Coexpression Network Analysis (WGCNA), Pseudo-time analysis, principle component analysis (PCA) / t-
Distributed Stochastic Neighbor Embedding (tSNE). We aim to characterize the molecular features unique to
CD133+ cell subtypes in a temporally and spatially specific manner, through which we can obtain a better
understanding of NSCs activities during development, as well as their potential interactions with the
vasculature niche. Immunohistochemistry and/or in situ hybridization analyses will be performed to validate the
sequencing result and provide physical locations of each of the specific cell types. Through lineage tracing
studies, we will be able to map out lineage relationships of heterogeneous NSC populations. We believe this
study will provide valuable information for building the atlas or roadmap of the origins of NSCs and how they
evolve during development. Moreover, we aim to bridge this project into a RO1 grant where the function of
some of the interesting core regulatory genes identified by this study will be further studied by gain- and loss-of
function analyses and lineage tracing studies.
利用单细胞RNAseq揭示CD133+神经干细胞发育过程中的特征
项目成果
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