Multi-Omics Approach to Dissect the Mechanisms of Rexinoid Signaling in Myoblast Differentiation

Stem cells represent a key resource in regenerative medicine, however, there is a critical need for pharmacological modulators to promote efficient conversion of stem cells into a myogenic lineage. We have previously shown that bexarotene, an agonist of retinoid X receptor (RXR) approved for cancer therapy, promotes the specification and differentiation of skeletal muscle progenitors. To decipher the molecular regulation of rexinoid signaling in myogenic differentiation, we have integrated RNA-seq transcription profiles with ChIP-seq of H4K8, H3K9, H3K18, H3K27 acetylation, and H3K27 methylation in addition to that of histone acetyl-transferase p300 in rexinoid-promoted myoblast differentiation. Here, we provide details regarding data collection, validation and omics integration analyses to offer strategies for future data application and replication. Our analyses also reveal molecular pathways underlying different patterns of gene expression and p300-associated histone acetylation at distinct chromatin states in rexinoid-enhanced myoblast differentiation. These datasets can be repurposed for future studies to examine the relationship between signaling molecules, chromatin modifiers and histone acetylation in myogenic regulation, providing a framework for discovery and functional characterization of muscle-specific loci.

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The effect of extracellular matrix on myogenic induction of adipose mesenchymal stem cells

10.21203/rs.3.rs-23048/v1 ◽

2020 ◽

Author(s):

Wenyong Fei ◽

bin xie ◽

Ying Liu ◽

Mingsheng Liu ◽

Xuanqi Wang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Myogenic Differentiation ◽

Subcutaneous Fat ◽

Myoblast Differentiation ◽

Differentiation Potential ◽

Culture Bottle ◽

Significant Difference ◽

Adipose Mesenchymal Stem Cells ◽

Myogenic Induction

Abstract Bankground: As one of the hot cells in the field of regenerative medicine, adipose mesenchymal stem cells(ADSCs) have been proved to have the ability of myoblast differentiation, but the disadvantage is that the efficiency of myoblast differentiation is not very high.Extracellular matrix(ECM), as a mixture of cytokines and proteins secreted by cells, is the substance secreted by cells. It has the advantages of non-toxic and harmless, and can provide the necessary material and environmental basis for cell growth and development.This study was to explore whether the myogenic differentiation potential of ADSCs cultured in ECM was improved.Method: ADSCs were extracted from subcutaneous fat of male SD rats at the age of 3 months and weight 250g. The third generation cellular of ADSCs were prepared for ECM，and the ECM was contained in the culture bottle for amplify the ADSCs. Myogenic induction was performed by 5-azacytidine using the same generation of ADSCs cultured with ECM and non-ECM.Result: ECM can significantly increase the rate of cell proliferation (P<0.05).Immunofluorescence detected the expression of β-actin, Myod and Desmin in cells ,and showed no significant difference in the expression of β-actin between groups (P>0.05). However, there were significant differences in Myod and Desmin between groups (P<0.001).RT-PCR showed that the mRNA expressions of myosin heavy chain(MHC), troponin, Myogein and Myf5 were significantly different among groups (P<0.001).Conclusion: ECM culture of ADSCs can improve its growth rate and myogenic differentiation potential.

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Faculty Opinions recommendation of Biophysical regulation of histone acetylation in mesenchymal stem cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.11973956.15692056 ◽

2012 ◽

Author(s):

David Lee ◽

Stephen Thorpe

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Histone Acetylation

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Faculty Opinions recommendation of Functional characterization of hematopoietic stem cells in the spleen.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.9509957.10159056 ◽

2011 ◽

Author(s):

Louise Purton ◽

Emma Baker ◽

Julie Quach

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Functional Characterization ◽

Hematopoietic Stem

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A Novel Strategy for Isolation, Molecular and Functional Characterization of Embryonic Mammary Stem Cells Using Molecular Genetics and Microfluidic Sorting

10.21236/ada488861 ◽

2008 ◽

Author(s):

Geoffrey M. Wahl

Keyword(s):

Stem Cells ◽

Molecular Genetics ◽

Functional Characterization ◽

Mammary Stem Cells ◽

Mammary Stem ◽

Novel Strategy ◽

Microfluidic Sorting

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The Leucine Catabolite and Dietary Supplement β-Hydroxy-β-Methyl Butyrate (HMB) as an Epigenetic Regulator in Muscle Progenitor Cells

Metabolites ◽

10.3390/metabo11080512 ◽

2021 ◽

Vol 11 (8) ◽

pp. 512

Author(s):

Virve Cavallucci ◽

Giovambattista Pani

Keyword(s):

Histone Acetylation ◽

Dietary Supplement ◽

Hdac Inhibitor ◽

Muscle Wasting ◽

C2c12 Cells ◽

Myoblast Differentiation ◽

Epigenetic Mark ◽

Epigenetic Regulator ◽

Methyl Butyrate

β-Hydroxy-β-Methyl Butyrate (HMB) is a natural catabolite of leucine deemed to play a role in amino acid signaling and the maintenance of lean muscle mass. Accordingly, HMB is used as a dietary supplement by sportsmen and has shown some clinical effectiveness in preventing muscle wasting in cancer and chronic lung disease, as well as in age-dependent sarcopenia. However, the molecular cascades underlying these beneficial effects are largely unknown. HMB bears a significant structural similarity with Butyrate and β-Hydroxybutyrate (βHB), two compounds recognized for important epigenetic and histone-marking activities in multiple cell types including muscle cells. We asked whether similar chromatin-modifying actions could be assigned to HMB as well. Exposure of murine C2C12 myoblasts to millimolar concentrations of HMB led to an increase in global histone acetylation, as monitored by anti-acetylated lysine immunoblotting, while preventing myotube differentiation. In these effects, HMB resembled, although with less potency, the histone deacetylase (HDAC) inhibitor Sodium Butyrate. However, initial studies did not confirm a direct inhibitory effect of HMB on HDACs in vitro. β-Hydroxybutyrate, a ketone body produced by the liver during starvation or intense exercise, has a modest effect on histone acetylation of C2C12 cells or in vitro HDAC inhibitor activities, and, unlike Butyrate and HMB, did not interfere with myotube formation in a myoblast differentiation assay. Instead, βHB dramatically increased lysine β-hydroxybutyrylation (Kbhb) of histone tails, an epigenetic mark associated with fasting responses and muscle catabolic states. However, when C2C12 cells were exposed to βHB in the presence of equimolar HMB this chromatin modification was drastically reduced, pointing to a role for HMB in attenuating ketosis-associated muscle wasting. In conclusion, while their mechanistic underpinnings remain to be clarified, these preliminary observations highlight novel and potentially important activities of HMB as an epigenetic regulator and βHB antagonist in muscle precursor cells, to be further explored in their biomedical implications.

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