Endothelial progenitor cells overexpressing Grb2-associated binder 1 for in vitro-constructed tissue-engineered heart valves

Aim    Endothelial progenitor cells (EPCs) play important roles in heart valve replacement surgery. Up-regulation of Grb2‑associated binder 1 (Gab1) promotes hepatocyte growth factor (HGF) – induced endothelial progenitor cell proliferation and migration. This study aimed to investigate the effects of up-regulation of Gab1 in hepatocyte growth factor-induced EPCs in tissue-engineered heart valves (TEHV).
Material and methods    Fresh porcine aortic valves were placed in 1 % Triton X-100 and trypsin buffer for decellularization. EPCs in the control group were cultured normally, whereas those in the experimental group were both HGF stimulated and transfected with adenovirus containing the Gab1 gene. Cells in the two groups were seeded onto the decellularized valve scaffolds and cultured for 3 or 7 days. TEHV were analyzed by HE and AB-PAS staining.
Results    By day 3, the experimental group had formed confluent endothelial monolayers on top of the decellularized valves, on the basis of by HE staining and AB-PAS staining. One week later, the control group showed a imperfect endothelial layer.
Conclusion    HGF-induced EPCs overexpressing Gab1 can endothelialize the decellularized matrix and create functional TEHV, which may then be preconditioned in a bioreactor before clinical implantation.

1.

2. Dohmen PM, da Costa F, Yoshi S, Lopes SV, da Souza FP, Vilani R et al. Histological evaluation of tissue-engineered heart valves implanted in the juvenile sheep model: is there a need for in-vitro seeding? The Journal of Heart Valve Disease. 2006;15(6):823–9. PMID: 17152791

3. Cebotari S, Lichtenberg A, Tudorache I, Hilfiker A, Mertsching H, Leyh R et al. Clinical Application of Tissue Engineered Human Heart Valves Using Autologous Progenitor Cells. Circulation. 2006;114(1 Suppl): I132–7. DOI: 10.1161/CIRCULATIONAHA.105.001065

4. Sales VL, Mettler BA, Engelmayr GC, Aikawa E, Bischoff J, Martin DP et al. Endothelial Progenitor Cells as a Sole Source for Ex Vivo Seeding of Tissue-Engineered Heart Valves. Tissue Engineering Part A. 2010;16(1):257–67. DOI: 10.1089/ten.tea.2009.0424

5. Jordan JE, Williams JK, Lee S-J, Raghavan D, Atala A, Yoo JJ. Bioengineered self-seeding heart valves. The Journal of Thoracic and Cardiovascular Surgery. 2012;143(1):201–8. DOI: 10.1016/j.jtcvs.2011.10.005

6. Rippel RA, Ghanbari H, Seifalian AM. Tissue-Engineered Heart Valve: Future of Cardiac Surgery. World Journal of Surgery. 2012;36(7):1581– 91. DOI: 10.1007/s00268-012-1535-y

7. Bianconi V, Sahebkar A, Kovanen P, Bagaglia F, Ricciuti B, Calabrò P et al. Endothelial and cardiac progenitor cells for cardiovascular repair: A controversial paradigm in cell therapy. Pharmacology & Therapeutics. 2018; 181:156–68. DOI: 10.1016/j.pharmthera.2017.08.004

8. Nakaoka Y, Komuro I. Gab Docking Proteins in Cardiovascular Disease, Cancer, and Inflammation. International Journal of Inflammation. 2013; 2013:141068. DOI: 10.1155/2013/141068

9. Nakaoka Y, Nishida K, Fujio Y, Izumi M, Terai K, Oshima Y et al. Activation of gp130 Transduces Hypertrophic Signal Through Interaction of Scaffolding/Docking Protein Gab1 with Tyrosine Phosphatase SHP2 in Cardiomyocytes. Circulation Research. 2003;93(3):221–9. DOI: 10.1161/01.RES.0000085562.48906.4A

10. Shioyama W, Nakaoka Y, Higuchi K, Minami T, Taniyama Y, Nishida K et al. Docking Protein Gab1 Is an Essential Component of Postnatal Angiogenesis After Ischemia via HGF/c-Met Signaling. Circulation Research. 2011;108(6):664–75. DOI: 10.1161/CIRCRESAHA.110.232223

11. Aasrum M, Ødegård J, Sandnes D, Christoffersen T. The involvement of the docking protein Gab1 in mitogenic signalling induced by EGF and HGF in rat hepatocytes. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 2013;1833(12):3286–94. DOI: 10.1016/j.bbamcr.2013.10.004

12. Sang H, Li T, Li H, Liu J. Down-Regulation of Gab1 Inhibits Cell Proliferation and Migration in Hilar Cholangiocarcinoma. PLoS ONE. 2013;8(11): e81347. DOI: 10.1371/journal.pone.0081347

13. Zhao J, Wang W, Ha CH, Kim JY, Wong C, Redmond EM et al. Endothelial Grb2-Associated Binder 1 Is Crucial for Postnatal Angiogenesis. Arteriosclerosis, Thrombosis, and Vascular Biology. 2011;31(5):1016– 23. DOI: 10.1161/ATVBAHA.111.224493

14. Fan Q, Zhang L, Zhu W, Xue S, Song Y, Chang Q. Up-regulation of Grb2-associated binder 1 promotes hepatocyte growth factor-induced endothelial progenitor cell proliferation and migration. PeerJ. 2019;7: e6675. DOI: 10.7717/peerj.6675

15. Ravishankar P, Zeballos MA, Balachandran K. Isolation of Endothelial Progenitor Cells from Human Umbilical Cord Blood. Journal of Visualized Experiments. 2017; 127:56021. DOI: 10.3791/56021

16. Schoen FJ, Levy RJ. Calcification of Tissue Heart Valve Substitutes: Progress Toward Understanding and Prevention. The Annals of Thoracic Surgery. 2005;79(3):1072–80. DOI: 10.1016/j.athoracsur.2004.06.033

17. Siddiqui RF, Abraham JR, Butany J. Bioprosthetic heart valves: modes of failure. Histopathology. 2009;55(2):135–44. DOI: 10.1111/j.1365-2559.2008.03190.x

18. Schmidt D, Breymann C, Weber A, Guenter CI, Neuenschwander S, Zund G et al. Umbilical Cord Blood Derived Endothelial Progenitor Cells for Tissue Engineering of Vascular Grafts. The Annals of Thoracic Surgery. 2004;78(6):2094–8. DOI: 10.1016/j.athoracsur.2004.06.052

19. Schmidt D, Mol A, Neuenschwander S, Breymann C, Gössi M, Zund G et al. Living patches engineered from human umbilical cord derived fibroblasts and endothelial progenitor cells. European Journal of Cardio-Thoracic Surgery. 2005;27(5):795–800. DOI: 10.1016/j.ejcts.2005.01.064

20. Kaushal S, Amiel GE, Guleserian KJ, Shapira OM, Perry T, Sutherland FW et al. Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo. Nature Medicine. 2001;7(9):1035–40. DOI: 10.1038/nm0901-1035

21. Ackah E, Yu J, Zoellner S, Iwakiri Y, Skurk C, Shibata R et al. Akt1/protein kinase B is critical for ischemic and VEGF-mediated angiogenesis. Journal of Clinical Investigation. 2005;115(8):2119–27. DOI: 10.1172/JCI24726

22. Bonauer A, Carmona G, Iwasaki M, Mione M, Koyanagi M, Fischer A et al. MicroRNA-92a Controls Angiogenesis and Functional Recovery of Ischemic Tissues in Mice. Science. 2009;324(5935):1710–3. DOI: 10.1126/science.1174381

23. Ha CH, Wang W, Jhun BS, Wong C, Hausser A, Pfizenmaier K et al. Protein Kinase D-dependent Phosphorylation and Nuclear Export of Histone Deacetylase 5 Mediates Vascular Endothelial Growth Factor-induced Gene Expression and Angiogenesis. Journal of Biological Chemistry. 2008;283(21):14590–9. DOI: 10.1074/jbc.M800264200

24. Barrow-McGee R, Kermorgant S. Met endosomal signalling: In the right place, at the right time. The International Journal of Biochemistry & Cell Biology. 2014; 49:69–74. DOI: 10.1016/j.biocel.2014.01.009

25. Dohmen PM, Ozaki S, Nitsch R, Yperman J, Flameng W, Konertz W. A tissue engineered heart valve implanted in a juvenile sheep model. Medical Science Monitor: International Medical Journal of Experimental and Clinical Research. 2003;9(4):BR97–104. PMID: 12709659

26. Stock UA, Schenke-Layland K. Performance of decellularized xenogeneic tissue in heart valve replacement. Biomaterials. 2006;27(1):1–2. DOI: 10.1016/j.biomaterials.2005.05.100

27. Kasimir M-T, Weigel G, Sharma J, Rieder E, Seebacher G, Wolner E et al. The decellularized porcine heart valve matrix in tissue engineering: Platelet adhesion and activation. Thrombosis and Haemostasis. 2005;94(09):562–7. DOI: 10.1160/TH05-01-0025

28. Dohmen PM, Lembcke A, Hotz H, Kivelitz D, Konertz WF. Ross operation with a tissue-engineered heart valve. The Annals of Thoracic Surgery. 2002;74(5):1438–42. DOI: 10.1016/S0003-4975(02)03881-X

29. Breuer CK, Mettler BA, Anthony T, Sales VL, Schoen FJ, Mayer JE. Application of Tissue-Engineering Principles toward the Development of a Semilunar Heart Valve Substitute. Tissue Engineering. 2004;10(11– 12):1725–36. DOI: 10.1089/ten.2004.10.1725