Pro-Fibrotic Phenotype in a Patient with Segmental Stiff Skin Syndrome via TGF-β Signaling Overactivation
Abstract
:1. Introduction
2. Results
2.1. Clinical and Histological Findings
2.2. Germline and Somatic FBN1 Variants Analysis
2.3. Altered Smad- and Non-Smad-Dependent TGF-β Signaling Pathways
2.4. Increased Expression Profile of Key Pro-Fibrotic Molecules
2.5. Reduced Metalloproteinase 2 Activity
2.6. Increased Cell Proliferation and Migration
2.7. Losartan Reduces the Activation Status of TGF-β Signaling Pathways in SSS Cells
3. Discussion
4. Materials and Methods
4.1. Patient and Samples
4.2. Cell Cultures
4.3. DNA Extraction and Next-Generation Sequencing
4.4. CytoScan XON
4.5. Immunohistochemistry Analysis
4.6. RNA Extraction and Reverse Transcription
4.7. Quantitative Real-Time PCR
4.8. Western Blotting and Quantification of Phosphorylated Proteins
4.9. Confocal Study
4.10. Zymography
4.11. PrestoBlue Assay
4.12. Transwell Migration Assay
4.13. Losartan Treatment
4.14. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
CNVs | copy number variations |
COLL | collagens |
Ctrl | control |
D-MEM | Dulbecco’s Modified Eagle Medium |
ECM | extracellular matrix |
ERK | extracellular signal-regulated kinase |
FBN1 | fibrillin-1 |
LTBP | Latent transforming growth factor beta binding protein |
MMP2 | Matrix metalloproteinase 2 |
MRI | Magnetic Resonance Imaging |
NGS | next generation sequencing |
NF-κB | Nuclear factor κ-B |
PBS | Phosphate-buffered saline |
Pt | patient |
qPCR | quantitative polymerase chain reaction |
ROI | region of interest |
SDS-PAGE | sulfate: polyacrylamide gel electrophoresis |
SSS | stiff skin syndrome |
TGF-β | Transforming growth factor β |
Tsk | Tight skin mice |
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Fusco, C.; Nardella, G.; Augello, B.; Boccafoschi, F.; Palumbo, O.; Fusaro, L.; Notarangelo, A.; Barbano, R.; Parrella, P.; Annicchiarico, G.; et al. Pro-Fibrotic Phenotype in a Patient with Segmental Stiff Skin Syndrome via TGF-β Signaling Overactivation. Int. J. Mol. Sci. 2020, 21, 5141. https://doi.org/10.3390/ijms21145141
Fusco C, Nardella G, Augello B, Boccafoschi F, Palumbo O, Fusaro L, Notarangelo A, Barbano R, Parrella P, Annicchiarico G, et al. Pro-Fibrotic Phenotype in a Patient with Segmental Stiff Skin Syndrome via TGF-β Signaling Overactivation. International Journal of Molecular Sciences. 2020; 21(14):5141. https://doi.org/10.3390/ijms21145141
Chicago/Turabian StyleFusco, Carmela, Grazia Nardella, Bartolomeo Augello, Francesca Boccafoschi, Orazio Palumbo, Luca Fusaro, Angelantonio Notarangelo, Raffaela Barbano, Paola Parrella, Giuseppina Annicchiarico, and et al. 2020. "Pro-Fibrotic Phenotype in a Patient with Segmental Stiff Skin Syndrome via TGF-β Signaling Overactivation" International Journal of Molecular Sciences 21, no. 14: 5141. https://doi.org/10.3390/ijms21145141