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| Status |
Public on Apr 19, 2023 |
| Title |
Distinct microRNA and protein profiles of extracellular vesicles secreted from myotubes from morbidly obese donors with type 2 diabetes in response to electrical pulse stimulation [EPS_EXO] |
| Platform organism |
synthetic construct |
| Sample organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by array
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| Summary |
We have used a combined transcriptome-proteome approach to describe how EPS affected the cargo of extracellular vesicles derived from myotubes from morbidly obese patients with T2D, and revealed several new factors, both miRs and proteins, that might act as exercise factors.
During exercise, skeletal muscles release signaling factors that communicate with other organs and mediate beneficial effects of exercise. These factors include myokines, metabolites, and extracellular vesicles (EVs). In the present study, we have examined how electrical pulse stimulation (EPS) of myotubes, a model of exercise, affects the cargo of released EVs. Chronic low frequency EPS was applied for 24 h to human myotubes isolated and differentiated from biopsy samples from 6 morbidly obese females with T2D, and EVs, both exosomes and microvesicles (MV), were isolated from cell media 24 h thereafter.Protein content was assessed by high-resolution proteomic analysis (LC-MS/MS), non-coding RNA was quantified by Affymetrix GeneChip Multispecies miRNA-4_0 Array microarray and Flash Tag TM Biotin RNA labelling, Thermo Fisher), and selected microRNAs (miRs) validated by real time RT-qPCR. We found that human myotubes secrete both exosomes and MV. EPS treatment of the myotubes, clearly changed the protein content of both exosomes and MV, whereas the miR content was changed only in exosomes. We suggest that skeletal muscle derived EVs can contribute to circulatory EVs and mediate beneficial effects of exercise in metabolically active organs.
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| Overall design |
Number of patients: 3. Each patient was their own control by comparing the RNA and protein-cargo before and after EPS. Cellular activation by EPS was detected by IL-6 analysis. Non template controls and spike ins control probe sets were included in the microarray analysis. The Affymetrix GeneChip Command Console sodtware was used for normalization, the Robust Multichip Analysis (RMA) alogoritm was applied for signal generation, and Partek R Genomics SuiteTM software for statistical analyis. Validation of microarray results was carried out by RT-qPCR of selected miRNAs.
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| Contributor(s) |
Aas V, Øvstebø R, Brusletto BS, Aspelin T, Trøseid AM, Qureshi S, Eid DS, Olstad OK, Nyman T, Haug KF |
| Citation(s) |
37064893 |
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| Submission date |
Mar 22, 2023 |
| Last update date |
Apr 21, 2023 |
| Contact name |
Kari Bente Foss Haug |
| E-mail(s) |
[email protected]
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| Phone |
93047410
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| Organization name |
Oslo University Hospital
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| Department |
Medical Biochemistry
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| Street address |
Skogstien 28
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| City |
Blommenholm |
| ZIP/Postal code |
1365 |
| Country |
Norway |
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| Platforms (1) |
| GPL19117 |
[miRNA-4] Affymetrix Multispecies miRNA-4 Array |
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| Samples (6)
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| This SubSeries is part of SuperSeries: |
| GSE227939 |
Distinct microRNA and protein profiles of extracellular vesicles secreted from myotubes from morbidly obese donors with type 2 diabetes in response to electrical pulse stimulation |
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| Relations |
| BioProject |
PRJNA947542 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE227936_RAW.tar |
4.1 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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