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Transcriptional profiling of embryonic skeletal muscle stem/progenitor cells

ABSTRACT: Muscle stem cells (MuSC) change molecular and functional properties during development. Using a transgenic Tg:Pax7-nGFP mice, we FACS-isolated MuSC from embryonic (E12.5) and foetal (E17.5) stages to understand the differences and similarities amongst the myogenic stem/progenitor populations.

ORGANISM(S): Mus musculus

PROVIDER: GSE52192 | GEO | 2013/11/08

SECONDARY ACCESSION(S): PRJNA227136

REPOSITORIES: GEO

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Transcriptional profiling of embryonic skeletal muscle stem/progenitor cells

Project description:Muscle stem cells (MuSC) change molecular and functional properties during development. Using a transgenic Tg:Pax7-nGFP mice, we FACS-isolated MuSC from embryonic (E12.5) and foetal (E17.5) stages to understand the differences and similarities amongst the myogenic stem/progenitor populations. Pax7-nGFP+ cells were isolated by FACS from limb buds of E12.5 and E17.5 Tg:Pax7-nGFP embryos following enzymatic digestion. Affymetrix microarrays were performed on biological triplicates.

2013-11-08 | E-GEOD-52192 | biostudies-arrayexpress

Small RNA-seq of quiescent, activated of differentiated mouse skeletal muscle stem cells

Project description:The requirement for microRNAs in muscle homeostasis, and more precisely in quiescence/self-renewal, activation & differentiation, has been demonstrated through knock-out experiments. In this experiment, we unbiasedly probed the expression of small RNA molecules (ranging from 18-36 nt in size), during the quiescence/activation transition but also differentiation by mean of small-RNA sequencing. Quiescent cells were directly FACS-purified from resting muscle of tg:Pax7-nGFP adult mice (n=2 replicates), and a fraction of these sorted cells were cultured in vitro to obtain activated MuSC at 60h, or differentiated cells at 7 days. This dataset constitutes a key resource for further analyses of miRNAs involvement in the muscle paradigm, and more largely in the regulation of stem cells self-renewal and tissue homeostasis.

2018-03-24 | E-MTAB-5955 | biostudies-arrayexpress

Prmt5 is a crucial regulator of muscle stem cell expansion in adult mice

Project description:Skeletal muscle stem cells (MuSC), also called satellite cells, are indispensable for maintenance and regeneration of adult skeletal muscles. Yet, a comprehensive picture of the regulatory events controlling the fate of MuSC is missing. Here, we determine the proteome of MuSC to design a loss-of-function screen, and identify 120 genes important for MuSC function including the arginine methyltransferase Prmt5. MuSC-specific inactivation of Prmt5 in adult mice prevents expansion of MuSC, abolishes long-term MuSC maintenance and abrogates skeletal muscle regeneration. Interestingly, Prmt5 is dispensable for proliferation and differentiation of Pax7(+) myogenic progenitor cells during mouse embryonic development, indicating significant differences between embryonic and adult myogenesis. Mechanistic studies reveal that Prmt5 controls proliferation of adult MuSC by direct epigenetic silencing of the cell cycle inhibitor p21. We reason that Prmt5 generates a poised state that keeps MuSC in a standby mode, thus allowing rapid MuSC amplification under disease conditions.

2015-03-13 | GSE66822 | GEO

STAT3 Regulates Adult Muscle Satellite Cell Maintenance During Injury-induced Muscle Regeneration

Project description:We previously showed that STAT3 regulates myoblast differentiation in cell culture models, yet its role in adult muscle satellite cells (MuSC) in vivo was less well characterized. When Stat3 was conditionally deleted in MuSC, muscle development and adult MuSC formation were not affected. However, with repeated muscle injuries, the number of the quiescent MuSC in STAT3-null mice decreased and the regeneration was delayed, suggesting defective MuSC maintenance. Consistently, when we conditionally deleted Stat3 in MuSC of dystrophin-null mice, a mouse model for the fatal human Duchenne muscular dystrophy, the adult double knockout (dKO) mice displayed age-dependent reduction in the number of MuSC, and increase in muscle inflammation and fibrosis. Mechanistically, Stat3 ablation in dystrophin-null MuSC resulted in downregulation of several key myogenic genes including Pax7, upregulation of multiple pro-inflammatory and pro-fibrotic genes, and an increase in fibroadipogenic progenitor cells, which collectively contributed to defective MuSC maintenance and aggravated inflammation and fibrosis in dKO mice.

2016-07-13 | GSE68736 | GEO

p110α of PI3K is Indispensable for Quiescence Exit in Adult Muscle Satellite Cells

Project description:Adult muscle stem cells (MuSC) are quiescent with a localization between myofibers and basal lamina. Upon injury, MuSC exit quiescence, reenter cell cycle, expand and differentiate for muscle regeneration. By using genetic mouse model, we identified p110α/mTORC1 signaling as a indispensable pathway that permits quiescence exit and cell cycle reentry. In order to dig out the downstream effectors, we compared the transcriptome of freshly isolated MuSC from Ctrl (p110α-f/+:R26-YFP/YFP:Pax7-CreER/CreER) to MuSC-specific p110α-null (iKO, p110α-f/f:R26-YFP/YFP:Pax7-CreER/CreER) mice by RNA-sequencing, and AP1 target genes were dramatically down-regulated in iKO MuSC. Restoration of Jun could significantly rescue the cell cycle reentry defect in iKO MuSC. In summary, we provided a p110α/mTORC1/Jun axis required for quiecence exit and cell cycle reentry of MuSC.

2018-02-25 | GSE109472 | GEO

Expression data from differentiating mouse embryonic stem cells wild type and lacking functional Pax7 gene

Project description:Myogenic differentiation relies on Pax7 function. We used embryonic stem cells lacking functional Pax7 to follow its role in derivation of skeletal myoblasts. Microarray analysis allowed us to compare transcriptomes of undifferentiated and differentiating embryonic stem cells of two genotypes, i.e. Pax7+/+ and Pax7-/- at day 7 and 21 of culture. Undifferentiated mouse embryonic stem cells, embryiod bodies at day 7, and embryoid body outgrowths at day 21 of differentiation were collected for RNA extraction and hybridization on Affymetrix microarrays.

2016-02-26 | E-GEOD-66483 | biostudies-arrayexpress

Myogenic Progenitor Cell Lineage Specification by CRISPR/Cas9-based Transcriptional Activators

Project description:Engineered CRISPR/Cas9-based transcriptional activators can potently and specifically activate endogenous fate-determining genes to direct differentiation of pluripotent stem cells. Here, we demonstrate that endogenous activation of the PAX7 transcription factor results in stable epigenetic remodeling and directly reprograms human pluripotent stem cells into skeletal myoblast progenitor cells. Compared to the exogenous overexpression of PAX7 cDNA, we find that endogenous activation results in the generation of more proliferative myogenic progenitors that can maintain PAX7 expression over multiple passages in serum-free conditions while preserving the capacity for terminal myogenic differentiation. Transplantation of human myogenic progenitors derived from endogenous activation of PAX7 into immunodeficient mice resulted in a greater number of human dystrophin+ myofibers compared to exogenous PAX7 overexpression. RNA-seq analysis also revealed transcriptome-wide differences between myogenic progenitors generated via CRISPR-based endogenous activation of PAX7 and exogenous PAX7 cDNA overexpression. These studies demonstrate the utility of CRISPR/Cas9-based transcriptional activators for progenitor cell specification and their potential for regenerative medicine.

2020-04-10 | GSE145575 | GEO

Expression data from differentiating mouse embryonic stem cells wild type and lacking functional Pax7 gene

2016-02-26 | GSE66483 | GEO

Identification of PAX7-induced transcriptional changes and PAX7 genomic binding during skeletal myogenic differentiation of H9 embryonic stem cells

Project description:Skeletal myogenic commitment of human pluripotent cells can be achieved by doxycycline-inducible expression of the transcription factor PAX7. To gain further insights on PAX7 function during this process, we performed a time course whole transcriptome analysis of differentiating H9 human embryonic stem cells from doxycycline-treated and untreated cultures. In addition, we identified the genomic binding of PAX7 in one of the selected time point (referred as PAX7+ proliferating myogenic progenitors).

2017-06-30 | GSE98976 | GEO

Direct reprogramming of mouse fibroblasts into functional skeletal muscle progenitors

Project description:Skeletal muscle harbors quiescent stem cells termed satellite cells and proliferative progenitors termed myoblasts, which play pivotal roles during muscle regeneration. However, current technology does not allow permanent capture of these cell populations in vitro. Here, we show that ectopic expression of the myogenic transcription factor MyoD, combined with exposure to small molecules, reprograms mouse fibroblasts into expandable induced myogenic progenitor cells (iMPCs). iMPCs express key skeletal muscle stem and progenitor cell markers including Pax7 and Myf5 and give rise to Dystrophin-expressing myofibers upon transplantation, a subset of which maintain Pax7 expression in vivo and sustain serial regenerative responses. Similar to satellite cells, iMPCs originate from Pax7+ cells and require Pax7 itself for maintenance. Finally, we show that iMPCs can be established from muscle tissue following small molecule exposure alone. This study thus reports on a robust approach to derive expandable myogenic stem/progenitor-like cells from multiple differentiated cell types.

2018-05-01 | GSE92336 | GEO

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