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| Status |
Public on May 28, 2020 |
| Title |
RNA-seq from human hepatic stellate cells treated with GIPC shRNA reveals contribution of GIPC to hepatic fibrosis |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Background and Aims: Transforming growth factor (TGF-β) induced activation of quiescent hepatic stellate cells (HSC) and their transformation to myofibroblasts is a key event in liver fibrosis and portal hypertension. GIPC (also referred to as synectin) is a downstream signal activation molecule of TGF-β and other receptors. In this study, we sought to identify novel genes targeted by TGF-β and GIPC and elucidate if and how they may contribute to liver fibrosis.
Methods and Results: We performed sequential mRNA sequencing analysis on TGF-β stimulated HSC and then on TGF-β-stimulated HSC in presence and absence of GIPC knockdown. IGFBP-3, an insulin growth factor transport protein, emerged as a top activation target of both TGF-β and GIPC, which was confirmed by qPCR, ELISA and Western blot (WB) analysis. Targeted chromatin immunoprecipitation (ChIP) revealed that GIPC increases the histone 3 lysine 27 (H3K27) acetylation activating mark and concurrently decreases the H3K27 inhibitory trimethylation (H3K27m3) mark providing an epigenetic correlate to the gene regulation changes. In vivo, global knockout of IGFBP-3 mice resulted in attenuation of HSC activation markers and attenuation of portal pressure in response to chronic liver injury models. Analysis of serum levels from cirrhotic patients also showed IGFBP-3 increase of more than 2-fold compared to healthy controls. Finally, in vitro mechanism studies revealed that IGFBP-3 promotes HSC migration through integrin dependent phosphorylation of AKT.
Conclusion: TGF-β upregulates IGFBP-3 through GIPC leading to increased HSC migration in vitro and promotes portal hypertension in vivo. These studies support the role of IGFBP-3 as a potential pathophysiologic target or biomarker in chronic liver disease.
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| Overall design |
next generation RNAseq data from human hepatic stellate cells with GIPC knockdown (GIPC shRNA) compared to controls shows that HSC activation/fibrosis pathway is differentially regulated by GIPC.
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| Contributor(s) |
Shah VH, Yaqoob U |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
May 27, 2020 |
| Last update date |
Jun 04, 2020 |
| Contact name |
Aditya Bhagwate |
| E-mail(s) |
bhagwate.adityavijay@mayo.edu
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| Organization name |
Mayo Clinic
|
| Street address |
200 1st Street SW
|
| City |
Rochester |
| State/province |
MN |
| ZIP/Postal code |
55905 |
| Country |
USA |
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| Platforms (1) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
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| Samples (6)
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| This SubSeries is part of SuperSeries: |
| GSE151771 |
The role of IGFBP-3 as a potential pathophysiologic target or biomarker in chronic liver disease |
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| Relations |
| BioProject |
PRJNA635292 |
| SRA |
SRP264915 |
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