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Mouse Fetal Cortical-derived Neurosphere cultures transfected with null control or miRNA-153 mimetic

ABSTRACT: The expression of the mciroRNA, miR-153 is significantly altered in neural stem cells (NSCs) following exposure to the teratogens, alcohol and nicotine (PMIDs:22458409, 17687032). To better understand how miR-153 may mediate teratogenesis, we first needed to identify miR153 targets in fetal NSCs. Gestational day 12.5 fetal mouse cortical neural epithelium was isolated and grown as neurospheres in defined medium for studies. We overexpressed miR-153 in fetal NSCs for 24 hours in a transient transfection assay, and profiled mRNA expression using a microarray platform (codelink array) to identify potential target genes. Over expression of pre-miR-153 resulted in a ~32-fold induction of miR-153 compared to the transfection control. Transcript profiles were assessed from RNA samples (6 independent replicates in each condition) hybridized to codelink microarrays according to manufacturer's protocols. Data analysis showed that miR-153 overexpression resulted in statistically significant down-regulation of 102 genes (at a false discovery rate α=0.1, of a total of 860 transcripts down-regulated by ≥1.3-fold). These data suggested the role of this miRNA in moderately controlling expression hundreds of gene transcripts. These regulated genes may be important in controlling neural stem cell differentiation and downstream signaling pathways.

ORGANISM(S): Mus musculus

PROVIDER: GSE49684 | GEO | 2014/08/01

SECONDARY ACCESSION(S): PRJNA214708

REPOSITORIES: GEO

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Mouse Fetal Cortical-derived Neurosphere cultures transfected with null control or miRNA-153 mimetic

Project description:The expression of the mciroRNA, miR-153 is significantly altered in neural stem cells (NSCs) following exposure to the teratogens, alcohol and nicotine (PMIDs:22458409, 17687032). To better understand how miR-153 may mediate teratogenesis, we first needed to identify miR153 targets in fetal NSCs. Gestational day 12.5 fetal mouse cortical neural epithelium was isolated and grown as neurospheres in defined medium for studies. We overexpressed miR-153 in fetal NSCs for 24 hours in a transient transfection assay, and profiled mRNA expression using a microarray platform (codelink array) to identify potential target genes. Over expression of pre-miR-153 resulted in a ~32-fold induction of miR-153 compared to the transfection control. Transcript profiles were assessed from RNA samples (6 independent replicates in each condition) hybridized to codelink microarrays according to manufacturer's protocols. Data analysis showed that miR-153 overexpression resulted in statistically significant down-regulation of 102 genes (at a false discovery rate α=0.1, of a total of 860 transcripts down-regulated by ≥1.3-fold). These data suggested the role of this miRNA in moderately controlling expression hundreds of gene transcripts. These regulated genes may be important in controlling neural stem cell differentiation and downstream signaling pathways. Six independent replicates were performed for control and treatment groups, and twelve independent single channel intensity values were uploaded after normalization for analysis independent single channel intensity values were uploaded after normalization

2014-08-01 | E-GEOD-49684 | biostudies-arrayexpress

miR-153 overexpressing NSPCs derived from mouse ES cells

Project description:Mammalian neural stem/progenitor cells (NSPCs) sequentially generate neurons and glia during central nervous system (CNS) development. Several transcription factors and microRNAs (miRNAs) are involved in the temporal regulation of NSPC differentiation. miRNA-153 (miR-153) as a modulator of NSPC specification. Overexpression (OE) of miR-153 delayed the onset of astrogliogenesis and maintained NSPCs in an undifferentiated state in vitro. miR-153-OE and control NSPCs (tertiary neurospheres (TNs) derived from mouse ES Cells via embryoid body formation) subjected to the gene expression microarray analysis.

2015-08-01 | E-GEOD-70129 | biostudies-arrayexpress

miR-153 overexpressing NSPCs derived from mouse ES cells

2015-08-01 | GSE70129 | GEO

Neurosphere cultures transfected with nonsense & miRNA-335 knockout reagents

Project description:To understand the mechanism by which miR-335 acts anti proliferative factor, we profiled mRNA expression using microarray platform ( codelink array). Embryonic day 12.5 neural cortical epithelium was isolated and grown as neurospheres in defined medium. Cultures were transfected with 2'-O-methyl (2'OMe)-base modified miRNA-335 antisense (2'OMe-335) and control (2' O Me-EGFP). To verify the suppression of miR-335 after the 2'-OMe treatment, we used real time RT-PCR which confirmed the down regulation of miR-335 up to ~4.5 cycles (~24 fold). For the microarray experiment, mRNA isolated from these cells was hybridized to codelink microarrays. Three independent replicate were done for treatment and control, six independent single channel intensity values were uploaded after normalization for analysis. Our array results indicated that there is general up-regulation in gene expression due to miR-335 Knock Out ( KO). We identified 116 genes statistically significant genes (adjusted p value < 0.05), out of these 95.7% were up-regulated genes. The highest fold difference we observed was between 2 and 3 fold, indicating the role of this miRNA in moderately controlling expression hundreds of genes. Of the several genes the highly expressed are important in maturation of neural stem cells and downstream signaling pathway genes

2019-07-31 | GSE14624 | GEO

Transcription profiling of mouse C57BL/6 Mouse Embryo Culture Model treated with ethanol

Project description:Mouse model for Fetal Alcohol Syndrome. Embryos exposed to alcohol in controlled environment to assess teratogenic effects. Fetal Alcohol Syndrome (FAS) is a leading developmental disorder. To date, a holistic view of molecular gene changes is largely unexplored. Using microarray analysis of whole embryo mouse culture with strict-control over alcohol-level, we found, directly related alcohol-metabolism, a reduction of retinol binding protein 1(Rbp1), and a, de novo expression of aldehyde dehydrogenase 1B1 (ALDH1B1). Remarkably, four key hemopoiesis genes (glycophorin A, adducin 2, beta-2 microglobulin, and ceruloplasmin) became absent, and many histone variants genes were reduced. Hypothesis-driven informatics analysis and intersection analysis of two independent experiments indicated that the altered genes are involved in cell growth, hemopoiesis, histone modification, eye and heart development, and a collective reduction in expression of growth factor genes (Igf1, Efemp1, Tieg, and Edil3) and neural specification genes (neurogenin, Sox 5, bHLHb5). Down-regulated neural specification phenotypes further supported the above findings. Further more, the gene expression profile indicated distinct subgroups which overlapped with the teratogenesis of the open- and the closed-neural tubes known in FAS. In summary, our data reveal genes alteration with causal potential for dysmorpology (e.g. retinoic acid, neuronal specification, and neurotrophic factors, and epigenetics related histone genes) and those downstream responsive genes related to alcohol metabolism, and developmental teratogenesis. Experiment Overall Design: Comparison of whole embryo gene expression after exposure to ethanol for 46 hours. Note 2 independent experiments completed.

2009-09-05 | E-GEOD-9545 | biostudies-arrayexpress

Impact of miR-199-5p overexpression and CAV1 silencing on human lung fibroblasts

Project description:To identify putative novel specific targets of mir-199-5p, we overexpressed miR-199a-5p as well as miR-21 and a siRNA targeted against CAV1 in human HFL1 pulmonary fibroblasts (CCL-153) by transfecting them with synthetic pre-miRNAs or a synthetic “negative” pre-miRNA as control (miR-Neg). RNA samples were harvested at 48 hours post-transfection and 2 independent experiments were carried out. Additional samples correspond to HFL1 cells treated or not with 10ng/ml TGFbeta for 48 hours in the absence of serum (2 independent experiments).

2013-02-01 | GSE34815 | GEO

Gene therapy using genome-edited human iPS cell-derived neural stem cells overcomes refractory course of invasive glioma stem cells

Project description:Spatiotemporal analyses using brain slice culture and brain clearing demonstrated that human induced pluripotent stem cell-derived neural stem cells (iPSC-NSCs) possess higher tumor-trophic migratory capacity than fetal NSCs, adipose tissue and bone marrow derived mesenchymal stem cells (MSCs). NSCs expressing prodrug converting enzyme fusion gene exhibited strong anti-tumor effect for glioma stem cell in vivo models. The present research concepts may become a platform of cell-based gene therapy for glioma.

2022-09-16 | GSE150470 | GEO

Efficient Generation of Integration-free and Engraftable induced Neural Stem Cells (iNSC) from Epithelial-like Cells in Human Urine

Project description:Human neural stem cells (NSCs) hold great promise in reparative therapy for neural diseases and injuries. However, the isolation of NSCs from either fetal or adult tissue and their application remain difficult due to ethical and immune-rejection concerns. One promising solution is to generate patient specific induced pluripotent stem cells (iPSCs) and subsequently differentiate them into NSCs. Alternatively, induced neurons (iN) can be generated directly from fibroblasts by retroviral delivery of neural specific transcription factors or miroRNAs. The later two approaches remain inefficient and with safety concerns. Here, we describe a process to generate, from epithelial-like cells present in human urine, integration free and engraftable NSCs (UiNSC) efficiently. Using an episomal system to deliver reprogramming factors and microRNAs into human urine cells and subsequently culture them in a chemically defined medium supplemented with a cocktail of small molecules, we generated UiNSCs capable of proliferation and without the transgenes upon prolonged expansion. These transgene free UiNSCs express typical neural stem cell markers and are able to differentiate into all neuronal subtypes and glial cells in vitro and more importantly could engraft and differentiate in vivo upon transplantation into the brain of new born rat. Thus, our work provides an efficient and highly practical platform to generate patient specific NSCs for regenerative medicine. This is a general expression microarray design (NimbleGen platform). It includes 6 samples.

2012-12-01 | E-GEOD-36395 | biostudies-arrayexpress

Identity and Nature of Neural Stem Cells in the Adult Human Subventricular Zone

Project description:The existence of neural stem cells (NSCs) in adult human brain neurogenic regions remains unresolved. To address this, we created a cell atlas of the adult human subventricular zone (SVZ) derived from fresh neurosurgical samples using single-cell transcriptomics. We discovered 2 adult radial glia (RG)-like populations, aRG1 and aRG2. aRG1 shared features with fetal early RG (eRG) and aRG2 were transcriptomically similar to fetal outer RG (oRG). We also captured early neuronal and oligodendrocytic NSC states. We found that the biological programs driven by their transcriptomes support their roles as early-lineage NSCs. Finally, we show that these NSCs have the potential to transition between states and along lineage trajectories. These data reveal that multipotent NSCs reside in the adult human SVZ.

2023-12-27 | GSE248995 | GEO

Gene Expression Profiles Associated with Ethanol-Induced Teratogenic Effects in a C57BL/6 Mouse Embryo Culture Model

Project description:Mouse model for Fetal Alcohol Syndrome. Embryos exposed to alcohol in controlled environment to assess teratogenic effects. Fetal Alcohol Syndrome (FAS) is a leading developmental disorder. To date, a holistic view of molecular gene changes is largely unexplored. Using microarray analysis of whole embryo mouse culture with strict-control over alcohol-level, we found, directly related alcohol-metabolism, a reduction of retinol binding protein 1(Rbp1), and a, de novo expression of aldehyde dehydrogenase 1B1 (ALDH1B1). Remarkably, four key hemopoiesis genes (glycophorin A, adducin 2, beta-2 microglobulin, and ceruloplasmin) became absent, and many histone variants genes were reduced. Hypothesis-driven informatics analysis and intersection analysis of two independent experiments indicated that the altered genes are involved in cell growth, hemopoiesis, histone modification, eye and heart development, and a collective reduction in expression of growth factor genes (Igf1, Efemp1, Tieg, and Edil3) and neural specification genes (neurogenin, Sox 5, bHLHb5). Down-regulated neural specification phenotypes further supported the above findings. Further more, the gene expression profile indicated distinct subgroups which overlapped with the teratogenesis of the open- and the closed-neural tubes known in FAS. In summary, our data reveal genes alteration with causal potential for dysmorpology (e.g. retinoic acid, neuronal specification, and neurotrophic factors, and epigenetics related histone genes) and those downstream responsive genes related to alcohol metabolism, and developmental teratogenesis. Keywords: comparison of gene expression profiles for treated vs. control

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