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| Status |
Public on Nov 30, 2016 |
| Title |
Intronic polyadenylation of PDGFRα in resident stem cells attenuates muscle fibrosis |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
The platelet-derived growth factor receptor alpha (PDGFRα) exhibits divergent effects in skeletal muscle. At physiological levels, signaling through this receptor promotes muscle development in growing embryos and proper angiogenesis in regenerating adult muscle. However, either increased PDGF ligands or enhanced PDGFRα pathway activity causes pathological fibrosis. This excessive collagen deposition, which is seen in aged and diseased muscle, interferes with proper muscle function and limits the effectiveness of gene- and cell-based therapies for muscle disorders. Although compelling evidence exists for the role of PDGFRα in fibrosis, little is known about the cells through which this pathway acts. Here we show that PDGFRα signaling regulates a population of muscle-resident fibro/adipogenic progenitors (FAPs) that play a supportive role in muscle regeneration but may also cause fibrosis when aberrantly regulated. We found that FAPs produce multiple transcriptional variants of PDGFRα with different polyadenylation sites, including an intronic variant that codes for a protein isoform containing a truncated kinase domain. This variant, upregulated during regeneration, acts as a decoy to inhibit PDGF signaling and to prevent FAP over-activation. Moreover, increasing expression of this isoform limits fibrosis in vivo, suggesting both biological relevance and therapeutic potential of modulating polyadenylation patterns in stem cell populations.
We used microarrays to explore the biological effects of altering intronic polyadenylation of PDGFRα in FAPs.
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| Overall design |
FAPs were isolated from the uninjured hindlimbs of C57Bl/6 mice. Cells were plated at 1 x 10^6 cells per well in 12-well plates. Cells were grown for 2.5 days in DMEM supplemented with 10% FBS. The media was switched to Ham’s F10 supplemented with 10% horse serum and transfected with 10 µM control or experimental AMO as indicated for 48 hours. The media was then replaced with Opti-Mem and cells re-transfected with 10 µM AMO with 50uM PDGF-AA or control. After 48 hours, the cells were lysed and RNA prepared with the RNeasy Mini Kit as per the manufacturer’s instructions (Qiagen). The microarray data was obtained using Affymetrix Mouse 1.0 ST. Each biological replicate represents cells pooled from 2 mice.
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| Contributor(s) |
Mueller AA, Rando TA |
| Citation(s) |
27894125 |
| |
| Submission date |
Aug 05, 2014 |
| Last update date |
Mar 04, 2019 |
| Contact name |
Thomas A Rando |
| E-mail(s) |
rando@stanford.edu
|
| Organization name |
Stanford University School of Medicine
|
| Department |
Neurology and Neurological Sciences
|
| Street address |
3801 Miranda Ave
|
| City |
Palo Alto |
| State/province |
CA |
| ZIP/Postal code |
94304 |
| Country |
USA |
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| Platforms (1) |
| GPL6246 |
[MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version] |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA257474 |
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