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Microarrays analysis of SREBP-1a and SREBP-1c target genes identify new regulatory pathways.

ABSTRACT: In this study we have identified the target genes of SREBP1a and SREBP1c in primary cultures of human skeletal muscle cells using adenoviral vectors expressing the mature nuclear form of human SREBP1a or SREBP1c combined with oligonucleotide microarrays. Keywords: comparison of human myotubes infected either by SREBP1a or 1C Human myotubes were prepared from 3 different skeletal muscle biopsies. After 7 days of differentiation, myotubes were infected for 48 hours with recombinant adenovirus expressing either Renilla, nuclear SREBP1a or nuclear SREBP1c. Each SREBP1a or SREBP1c infected myotubes culture was compared to Renilla infected myotubes culture. Renilla infected myotubes were considered as the control. Three biological replicates were processed.

ORGANISM(S): Homo sapiens

SUBMITTER: Jennifer Rieusset

PROVIDER: E-GEOD-10918 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Microarray analyses of SREBP-1a and SREBP-1c target genes identify new regulatory pathways in muscle.

Rome Sophie S Lecomte Virginie V Meugnier Emmanuelle E Rieusset Jennifer J Debard Cyrille C Euthine Vanessa V Vidal Hubert H Lefai Etienne E

Physiological genomics 20080617 3

In this study we have identified the target genes of sterol regulatory element binding protein (SREBP)-1a and SREBP-1c in primary cultures of human skeletal muscle cells, using adenoviral vectors expressing the mature nuclear form of human SREBP-1a or SREBP-1c combined with oligonucleotide microarrays. Overexpression of SREBP-1a led to significant changes in the expression of 1,315 genes (655 upregulated and 660 downregulated), whereas overexpression of SREBP-1c modified the mRNA level of 514 ge ...[more]

PMID: 18559965

Similar Datasets

Microarrays analysis of SREBP-1a and SREBP-1c target genes identify new regulatory pathways.

Project description:In this study we have identified the target genes of SREBP1a and SREBP1c in primary cultures of human skeletal muscle cells using adenoviral vectors expressing the mature nuclear form of human SREBP1a or SREBP1c combined with oligonucleotide microarrays. Keywords: comparison of human myotubes infected either by SREBP1a or 1C

2008-06-19 | GSE10918 | GEO

FTO: gene expression and function in skeletal muscle

Project description:We identified the target genes of FTO ("fat mass and obesity associated") in primary cultures of human skeletal muscle cells using adenoviral vectors expressing FTO or GFP and oligonucleotide microarrays. Human myotubes were prepared from 4 different skeletal muscle biopsies. After differentiation, myotubes were infected for 48 hours with recombinant adenovirus expressing either GFP or FTO. Each FTO-infected myotubes culture was compared to GFP-infected myotubes culture. GFP-infected myotubes were regarded as the control. Four biological replicates were processed.

2011-07-05 | E-GEOD-22857 | biostudies-arrayexpress

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Project description:In this study we have identified the target genes of BHLHB2 and BHLHB3 in primary cultures of human skeletal muscle cells using adenoviral vectors expressing either BHLHB2 or BHLHB3, and oligonucleotide microarrays. Human myotubes were prepared from 4 different skeletal muscle biopsies. After 7 days of differentiation, myotubes were infected for 48 hours with recombinant adenovirus expressing either GFP, BHLHB2 or BHLHB3. Each BHLHB2- or BHLHB3-infected myotubes culture was compared to GFP-infected myotubes culture. GFP-infected myotubes were considered the control. Four biological replicates were processed.

2010-04-11 | E-GEOD-12947 | biostudies-arrayexpress

Effect of ATF4 overexpression on mRNA levels in differentiated C2C12 myotubes.

Project description:ATF4 is a bZIP transcription factor that that promotes skeletal muscle atrophy. The goal of these studies was to determine the effects of ATF4 overexpression on mRNA levels in differentiated C2C12 myotubes. For additional details see Ebert et al, Stress-Induced Skeletal Muscle Gadd45a Expression Reprograms Myonuclei and Causes Muscle Atrophy. JBC epub. June 12,2012 C2C12 myotubes were infected with adenovirus co-expressing eGFP and ATF4-FLAG. Control myotubes were infected with adenovirus co-expressing eGFP and a transcriptionally inactive ATF4 construct (ATF4∆bZIP).

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Deep muscle proteome

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Expression data from C2C12 myotubes infected with adenovirus expressing Gli1

Project description:We sought to determine the effects of over-expression of Gli1 on gene expression in C2C12 myotube cultures. C2C12 myoblasts were induced to differentiate for 4 days. At that time, when >80% of nuclei were incorporated into multi-nucleated syncitial myotubes, we infected the cultures with recombinant adenovirus expressing GFP alone or GFP and a full length human Gli1. Media was changed 12 hours later. Cultures were lysed 60 hours after the initial infection. Gli1 over-expression induces de-differentiation of myotubes and proliferation of myoblasts. Results provide insight into the molecular basis of SHH signaling on skeletal muscle cells.

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Large-scale expansion of human iPSC-derived skeletal muscle cells for disease modeling and cell-based therapeutic strategies

Project description:Although skeletal muscle cells can be generated from human iPSCs, transgene-free protocols include only limited options for their purification and expansion. In this study we found that FACS-purified myogenic progenitors generated from healthy controls and Pompe disease iPSCs can be robustly expanded as much as 5 x 1011 fold. At all steps during expansion, cells could be cryopreserved or differentiated into myotubes with a high fusion index. In vitro, cells were amenable to maturation into striated and contractile myofibers. Insertion of the acid alpha glucosidase cDNA into the AAVS1 locus in iPSCs using CRISPR/cas9 prevented glycogen accumulation in myotubes generated from a patient with classic infantile Pompe disease. In vivo, the expression of human-specific nuclear and sarcolemmar antigens indicated that myogenic progenitors engraft into murine muscle to form human myofibers. This protocol is useful for modeling of skeletal muscle disorders and for using patient-derived, gene-corrected cells to develop cell-based strategies.

2018-05-03 | GSE111163 | GEO

miR-19b-3p Overexpression in Human Skeletal Muscle Cells

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E4BP4 is an insulin-induced stabilizer of nuclear SREBP-1c and promotes SREBP-1c-mediated lipogenesis.

Project description:Upon food intake, insulin stimulates de novo lipogenesis (DNL) in hepatocytes via the AKT-mTORC1-sterol regulatory element-binding protein (SREBP)-1c pathway. How insulin maintains the maximal SREBP-1c activities during the entire feeding state remains elusive. We previously reported that insulin induced b-ZIP transcription factor, E4-binding protein 4 (E4BP4), in hepatocytes. In the current study, we show that insulin injection increases hepatic E4bp4 expression by activating the AKT-mTORC1-SREBP-1c pathway in hepatocytes. E4bp4-deficient hepatocytes not only fail to maintain robust DNL but also become resistant to SREBP-1c-induced lipogenesis. In vivo, acute depletion of E4bp4 in the liver by adenoviral shRNA reduces the expression of lipogenic enzymes and results in reduced levels of serum triglycerides and cholesterol during the postprandial phase. In hepatocytes, E4BP4 interacts with nuclear SREBP-1c to preserve its acetylation, and subsequently protects it from ubiquitination-dependent degradation. In conclusion, the current studies uncover a novel positive feedback pathway mediated by E4BP4 to augment SREBP-1c-mediated DNL in the liver during the fed state.

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A self-renewing biomimetic skeletal muscle construct engineered using induced myogenic progenitor cells

Project description:Skeletal muscle is a highly organized and regenerative tissue that maintains its homeostasis primarily by activation and differentiation of muscle stem cells. Mimicking an in vitro skeletal muscle differentiation program that contains self-renewing adult muscle stem cells and aligned myotubes has been challenging. Here, we set out to engineer a biomimetic skeletal muscle construct that can self-regenerate and produce aligned myotubes using induced myogenic progenitor cells (iMPCs), a heterogeneous culture consisting of skeletal muscle stem, progenitor and differentiated cells. Utilizing electrospinning, we fabricated polycaprolactone (PCL) substrates that enabled iMPC-differentiation into aligned myotubes by controlling PCL fiber orientation. Newly-conceived constructs contained highly organized multinucleated myotubes in conjunction with self-renewing muscle stem cells, whose differentiation capacity was augmented by Matrigel supplementation. Additionally, we demonstrate using single cell RNA sequencing (scRNA-seq) that iMPC-derived constructs faithfully recapitulate a step-wise myogenic differentiation program. Notably, when the constructs were subjected to a damaging myonecrotic agent, self-renewing muscle stem cells rapidly differentiated into aligned myotubes, akin to skeletal muscle repair in vivo. Taken together, we report on a novel in vitro system that mirrors myogenic regeneration and muscle fiber alignment, and can serve as a platform to study myogenesis, model muscular dystrophies or perform drug screens.

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