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| Status |
Public on Jun 20, 2024 |
| Title |
Dual inhibition of P38 MAPK and JNK pathways preserves stemness markers and alleviates premature activation of Muscle Stem Cells during isolation |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Adult skeletal muscle contains resident Muscle Stem Cells (MuSC) with high myogenic and engraftment potentials, making them suitable for cell therapy and regenerative medicine approaches. However, purification process of MuSC remains a major hurdle to their use in the clinic. Indeed, muscle tissue enzymatic dissociation triggers a massive activation of stress signaling pathways, among which P38 and JNK MAPK, associated with a premature loss of MuSC quiescence. While the role of these pathways in the myogenic progression of MuSC is well established, the extent to which their dissociation-induced activation affects the functionality of these cells remains unexplored. We assessed the effect of P38 and JNK MAPK induction on stemness marker expression and MuSC activation state during isolation by pharmacological approaches. MuSC functionality was evaluated by in vitro assays and in vivo transplantation experiments. We performed a comparative analysis of the transcriptome of human MuSC purified with pharmacological inhibitors of P38 and JNK MAPK (SB202190 and SP600125, respectively) versus available RNAseq resources. We monitored PAX7 protein levels in murine MuSC during muscle dissociation and demonstrated a two-step decline partly dependent on P38 and JNK MAPK activities. We showed that simultaneous inhibition of these pathways throughout the MuSC isolation process preserves the expression of stemness markers and limits their premature activation, leading to improved survival and amplification in vitro as well as increased engraftment in vivo. Through a comparative RNAseq analysis of freshly isolated human MuSC, we provide evidence that our findings in murine MuSC could be relevant to human MuSC. Based on these findings, we implemented a purification strategy, significantly improving the recovery yields of human MuSC. Our study highlights the pharmacological limitation of P38 and JNK MAPK activities as a suitable strategy to qualitatively and quantitatively ameliorate human MuSC purification process, which could be of great interest for cell-based therapies.
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| Overall design |
We compared the transcriptome of hMuSC purified following our protocol (referred to as hMuSC-40/PI) to that of hMuSC isolated after 120 min of dissociation without inhibitors previously published (hMuSC-120/no PI).
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| Contributor(s) |
Didier N, Ozturk T |
| Citation(s) |
38902774 |
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| Submission date |
May 28, 2024 |
| Last update date |
Jun 28, 2024 |
| Contact name |
Teoman Ozturk |
| Organization name |
INSERM IMRB U955
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| Street address |
8 rue du Général Sarrail
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| City |
Créteil |
| ZIP/Postal code |
94000 |
| Country |
France |
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| Platforms (1) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA1117437 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE268475_RAW.tar |
10.0 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
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