"When Good Matrix Goes Bad" by Dorothee Weihrauch, DVM, PhD
•Download as PPSX, PDF•
1 like•1,237 views
This document summarizes research on how a defective extracellular matrix in scleroderma impacts heart function. Key findings include: 1) Endothelial cells grown on the defective matrix of scleroderma mouse models have impaired proliferation and increased apoptosis, expressing proteins linked to fibrosis. 2) The defective matrix binds higher levels of proteins like TGFβ and PDGF linked to fibrosis. It also contains more oxidative damage products like advanced glycation end products. 3) Human cardiomyocytes on the defective matrix show increased expression of genes associated with fibrosis and stress responses. 4) The results suggest the defective matrix in scleroderma contributes to cardiac dysfunction by promoting fibrosis and oxidative stress in heart cells.
Recommended
More Related Content
What's hot
What's hot (20)
Similar to "When Good Matrix Goes Bad" by Dorothee Weihrauch, DVM, PhD
Similar to "When Good Matrix Goes Bad" by Dorothee Weihrauch, DVM, PhD (20)
Recently uploaded
Recently uploaded (20)
"When Good Matrix Goes Bad" by Dorothee Weihrauch, DVM, PhD
- 1. What happens when the matrix goes bad? Dorothée Weihrauch DVM, PhD Department of Anesthesiology Medical College of Wisconsin
- 2. Acknowledgements • John G. Krolikowski • Hao Xu • Omoshalewa Bamkole • Kirkwood A. Pritchard • Ulrich Broeckel
- 3. The Disease
- 4. Scleroderma • Autoimmune, connective tissue disorder • Marked increases in fibrosis of the skin and internal organs • Increases myocardial inflammation • Chronic states of oxidative stress and inflammation impair heart function in other well-known diseases thedoctorwillseeyounow.com
- 5. Anti T15 idiotype Endostatin Negative Control Angiostatin Endostatin/ Collagen 18H+E Angiostatin Negative Control CD45 CD64 Anti T15 idiotype Endostatin Negative Control Immunofluorescence studies of Hearts of SSc Patients
- 6. Animal Model
- 7. The Tight Skin Mouse • Tight skin (Tsk) mutation is a genomic duplication within the fibrillin-1 (Fbn 1) gene • Synthesize and secrete normal fibrillin (~305 kD), mutant oversized Tsk fibrillin-1 (~405 kD) • Increases in fibrosis of the skin and internal organ • Myocardial inflammation and fibrosis impair heart function • Tsk fibrillin-1 is stably incorporated into cell layers. • Defects in microtubule assembly and stability • Experience chronic increases in oxidative stress and inflammation • Chronic vascular disease
- 8. C57/Bl6 Tsk-/+ Tsk-/+ +D4F T15- oxidized Phosphorylcholine
- 10. Collagen XVIII/ Endostatin C57/Bl6 Tsk-/+ Tsk-/+ +D4F
- 11. In Vitro
- 13. #ofcells/well 30000 20000 15000 10000 5000 0 25000 A Absorbence(450nm) B 0.8 0.6 0.4 0.2 0.0 1.0 MTS assayEC Proliferation Tsk-/+ Tsk + D-4F -/+ C57 Bl/6J Tsk-/+ Tsk + D-4F -/+ C57 Bl/6J # # Endothelial Cells seeded on defective matrix have impaired proliferation
- 14. RelativeFluorescenceUnits %ofTUNELpositivecells/well 500 400 300 200 100 0 Caspase Activity by FLICAApoptosis by TUNEL Tsk-/+ Tsk + D-4F -/+ C57 Bl/6J Tsk-/+ Tsk + D-4F -/+ C57 Bl/6J Viable Cells Tunnel Positive Cells # 80 60 40 20 0 100 # A B Endothelial Cells seeded on defective matrix undergo more often apoptosis
- 15. Endothelial Cells seeded on defective matrix express FSP-1 VE- Cadherin FSP-1 twist Tsk-/+ +D-4F Tsk-/+ C57Bl/6J
- 16. Endothelial Cells seeded on defective matrix express Collagen-I related proteins Tsk-/+ +D-4F Tsk-/+ C57Bl/6J Hsp47 DDR-2 Coll-1 b- Actin
- 17. IL-6 (Endothelial Cells on matrix)
- 18. 720 740 760 780 800 820 840 860 880 Gelatin C57 Tsk/+ MeanGreenFluorescence c-fos expression in human cardiomyocytes on different matrices
- 19. 600 700 800 900 C57 Tsk/+ MeanGreenFluorescence BNP expression in human cardiomyocytes on different matrices
- 20. Characterization of the Matrix
- 21. Tsk/+ C57 PDGF TGFβ and PDGF are bound to ECM isolated from Tsk/+ Tsk/+ C57 TGFb
- 22. Advanced Oxidized Protein Products (matrix) 0.38 0.39 0.4 0.41 0.42 0.43 0.44 0.45 0.46 0.47 0.48 0.49 C57 Tsk Absorption * *
- 23. 4 - Hydroxynonenal (matrix)
- 24. Density 0 200 400 600 800 1000 1200 C57 Tsk C57 Tsk Advanced Glycated End Products (matrix) *
- 25. 3 – Nitrotyrosine (matrix) 75 kDa 37 kDa 25 kDa 100 kDa C57 Tsk 0 40000 80000 120000 160000 C57 Tsk 3-Nitrotyrosine *
- 26. Oxidized adducts on Matrix • What else to look for • How to remove them Oxidative Stress • How to better measure it Other Suggestions What comes to your mind
Editor's Notes
- Looking at IL6 (gene target for IRF5), there was increased expression in the Ecs on Tsk matrix as compared to controls
- AOPP assay involves a mixture of reagents, an incubation period, and then absorption units are read in a plate reader As we expected, we saw more AOPP in the Tsk matrix as compared to controls
- And again with 4HNE , we see more of its expression in tsk matrix as compared to controls
- Expect to see more Nitrotyrosine in the matrix of the Tsk than controls