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Published on: April 12, 2015
SDF-1 and NOTCH signaling in myogenic cell differentiation: the role of miRNA10a, 425, and 5100
Bartosz Mierzejewski1, Iwona Grabowska1, Zuzanna Michalska1
1Department of Cytology, Faculty of Biology, University of Warsaw, Miecznikowa 1 St, 02-096, Warsaw, Poland.
Background:
Skeletal muscle regeneration is a complex process regulated by many cytokines and growth factors. Among the important signaling pathways regulating the myogenic cell identity are these involving SDF-1 and NOTCH. SDF-1 participates in cell mobilization and acts as an important chemoattractant. NOTCH, on the other hand, controls cell activation and myogenic determination of satellite cells. Knowledge about the interaction between SDF-1 and NOTCH signaling is limited.
Methods:
We analyzed two populations of myogenic cells isolated from mouse skeletal muscle, that is, myoblasts derived from satellite cells (SCs) and muscle interstitial progenitor cells (MIPCs). First, microRNA level changes in response to SDF-1 treatment were analyzed with next-generation sequencing (NGS). Second, myogenic cells, i.e., SC-derived myoblasts and MIPCs were transfected with miRNA mimics, selected on the basis of NGS results, or their inhibitors. Transcriptional changes, as well as proliferation, migration, and differentiation abilities of SC-derived myoblasts and MIPCs, were analyzed in vitro. Naive myogenic potential was assessed in vivo, using subcutaneous engrafts and analysis of cell contribution to regeneration of the skeletal muscles.
Results:
SDF-1 treatment led to down-regulation of miR10a, miR151, miR425, and miR5100 in myoblasts. Interestingly, miR10a, miR425, and miR5100 regulated the expression of factors involved in the NOTCH signaling pathway, including Dll1, Jag2, and NICD. Furthermore, miR10a, miR425, and miR5100 down-regulated the expression of factors involved in cell migration: Acta1, MMP12, and FAK, myogenic differentiation: Pax7, Myf5, Myod, Mef2c, Myog, Musk, and Myh3. However, these changes did not significantly affect myogenic cell migration or fusion either in vitro or in vivo, except when miR425 was overexpressed, or miR5100 inhibitor was used. These two molecules increased the fusion of MIPCs and myoblasts, respectively. Furthermore, miR425-transfected MIPC transplantation into injured skeletal muscle resulted in more efficient regeneration, compared to control cell transplantation. However, skeletal muscles that were injected with miR10a transfected myoblasts regenerated less efficiently.
Conclusions:
SDF-1 down-regulates miR10a, miR425, and miR5100, what could affect NOTCH signaling, differentiation of myogenic cells, and their participation in skeletal muscle regeneration.
Insights
Stromal cell-derived factor 1 (SDF-1) influences skeletal muscle regeneration by down-regulating specific microRNAs (miRNAs). These miRNAs impact NOTCH signaling and myogenic cell differentiation, affecting muscle repair outcomes.
Area of Science:
- Muscle regeneration
- Cell signaling pathways
- Molecular biology
Background:
- Skeletal muscle regeneration involves complex cytokine and growth factor signaling.
- Stromal cell-derived factor 1 (SDF-1) is crucial for cell mobilization and chemoattraction.
- NOTCH signaling controls satellite cell activation and myogenic determination.
Purpose of the Study:
- To investigate the interaction between SDF-1 and NOTCH signaling in myogenic cells.
- To analyze microRNA (miRNA) expression changes in response to SDF-1.
- To assess the functional impact of specific miRNAs on myogenic cell behavior and skeletal muscle regeneration.
Main Methods:
- Analysis of miRNA expression in mouse myoblasts and muscle interstitial progenitor cells (MIPCs) using next-generation sequencing (NGS) after SDF-1 treatment.
- In vitro and in vivo functional assays of myogenic cells transfected with miRNA mimics or inhibitors.
- Assessment of transcriptional changes, proliferation, migration, differentiation, and regenerative potential in skeletal muscle.
Main Results:
- SDF-1 treatment down-regulated miR10a, miR151, miR425, and miR5100 in myoblasts.
- miR10a, miR425, and miR5100 modulated factors in NOTCH signaling, cell migration, and myogenic differentiation pathways.
- Overexpression of miR425 enhanced MIPC fusion, while miR5100 inhibition improved myoblast fusion; miR425-transfected MIPC transplantation promoted skeletal muscle regeneration.
Conclusions:
- SDF-1 down-regulates specific miRNAs (miR10a, miR425, miR5100), potentially influencing NOTCH signaling and myogenic cell differentiation.
- These miRNA changes affect skeletal muscle regeneration, with miR425 promoting regeneration via MIPC transplantation.
- The study highlights the intricate role of miRNA regulation in SDF-1-mediated skeletal muscle repair processes.
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