Abstract
Corneal transplantation is the most common form of grafting performed worldwide. Corneal endothelial cells (EC) form a monolayer in the posterior portion of the cornea and are essential for corneal transparency. EC loss during storage before transplantation is a principal reason for rendering donor tissue unsuitable for transplantation, and apoptosis has been shown to be the major contributor to EC loss during storage and after transplantation. Therefore, the potential use of anti-apoptotic gene therapy to promote both graft storage and graft survival is of major interest. The goal of this study was to transduce human donor corneas in vitro to enhance EC survival during storage conditions used in eye banking. We utilized a lentiviral vector to perform gene transfer of baculoviral p35 or mammalian Bcl-xL to corneal endothelium in different storage conditions utilizing a lentiviral vector. Our results show significantly enhanced survival and prolonged retention of physiological EC morphology in cells expressing either p35 or Bcl-xL. The clinical application of this technology could lead to a higher availability of donor tissue for transplantation, extend storage periods and reduce graft failure after transplantation.
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References
Whitcher JP, Srinivasan M, Upadhyay MP . Corneal blindness: a global perspective. Bull World Health Organ 2001; 79: 214–221.
Garg P, Krishna PV, Stratis AK, Gopinathan U . The value of corneal transplantation in reducing blindness. Eye (London, England) 2005; 19: 1106–1114.
Resnikoff S, Pascolini D, Etya’ale D, Kocur I, Pararajasegaram R, Pokharel GP et al. Global data on visual impairment in the year 2002. Bull World Health Organ 2004; 82: 844–851.
Jones GL, Ponzin D, Pels E, Maas H, Tullo AB, Claerhout I . European eye bank association. Dev Ophthalmol 2009; 43: 15–21.
Joyce NC, Harris DL, Mello DM . Mechanisms of mitotic inhibition in corneal endothelium: contact inhibition and TGF-beta2. Invest Ophthalmol Vis Sci 2002; 43: 2152–2159.
Armitage WJ, Easty DL . Factors influencing the suitability of organ-cultured corneas for transplantation. Invest Ophthalmol Vis Sci 1997; 38: 16–24.
Vasara K, Setala K, Ruusuvaara P . Follow-up study of human corneal endothelial cells, photographed in vivo before enucleation and 20 years later in grafts. Acta Ophthalmol Scand 1999; 77: 273–276.
Ruusuvaara P, Setala K . Long-term follow-up of cryopreserved corneal endothelium. A specular microscopic study. Acta Ophthalmol (Copenh) 1988; 66: 687–691.
Williams KA, Lowe M, Bartlett C, Kelly TL, Coster DJ . Risk factors for human corneal graft failure within the Australian corneal graft registry. Transplantation 2008; 86: 1720–1724.
Komuro A, Hodge DO, Gores GJ, Bourne WM . Cell death during corneal storage at 4 degrees C. Invest Ophthalmol Vis Sci 1999; 40: 2827–2832.
Means TL, Geroski DH, Hadley A, Lynn MJ, Edelhauser HF . Viability of human corneal endothelium following Optisol-GS storage. Arch Ophthalmol 1995; 113: 805–809.
Albon J, Tullo AB, Aktar S, Boulton ME . Apoptosis in the endothelium of human corneas for transplantation. Invest Ophthalmol Vis Sci 2000; 41: 2887–2893.
Barcia RN, Dana MR, Kazlauskas A . Corneal graft rejection is accompanied by apoptosis of the endothelium and is prevented by gene therapy with bcl-xL. Am J Transplant 2007; 7: 2082–2089.
Okumura N, Ueno M, Koizumi N, Sakamoto Y, Hirata K, Hamuro J et al. Enhancement on primate corneal endothelial cell survival in vitro by a ROCK inhibitor. Invest Ophthalmol Vis Sci 2009; 50: 3680–3687.
Bourges JL, Torriglia A, Valamanesh F, Benezra D, Renard G, Behar-Cohen FF . Nitrosative stress and corneal transplant endothelial cell death during acute graft rejection. Transplantation 2007; 84: 415–423.
Gong N, Ecke I, Mergler S, Yang J, Metzner S, Schu S et al. Gene transfer of cyto-protective molecules in corneal endothelial cells and cultured corneas: analysis of protective effects in vitro and in vivo. Biochem Biophys Res Commun 2007; 357: 302–307.
Hori J, Wang M, Miyashita M, Tanemoto K, Takahashi H, Takemori T et al. B7-H1-induced apoptosis as a mechanism of immune privilege of corneal allografts. J Immunol 2006; 177: 5928–5935.
Hengartner MO . The biochemistry of apoptosis. Nature 2000; 407: 770–776.
Adams JM, Cory S . The Bcl-2 protein family: arbiters of cell survival. Science (New York, NY) 1998; 281: 1322–1326.
Xue D, Horvitz HR . Caenorhabditis elegans CED-9 protein is a bifunctional cell-death inhibitor. Nature 1997; 390: 305–308.
Clem RJ . Baculoviruses and apoptosis: the good, the bad, and the ugly. Cell Death Differ 2001; 8: 137–143.
Clem RJ . Baculoviruses and apoptosis: a diversity of genes and responses. Curr Drug Targets 2007; 8: 1069–1074.
Toniolo M, Camposampiero D, Griffoni C, Jones GL . Quality management in European eye banks. Dev Ophthalmol 2009; 43: 70–86.
Pels E, Rijneveld WJ . Organ culture preservation for corneal tissue. Technical and quality aspects. Dev Ophthalmol 2009; 43: 31–46.
Parker DG, Kaufmann C, Brereton HM, Anson DS, Francis-Staite L, Jessup CF et al. Lentivirus-mediated gene transfer to the rat, ovine and human cornea. Gene Therapy 2007; 14: 760–767.
Beutelspacher SC, Ardjomand N, Tan PH, Patton GS, Larkin DF, George AJ et al. Comparison of HIV-1 and EIAV-based lentiviral vectors in corneal transduction. Exp Eye Res 2005; 80: 787–794.
Wang X, Appukuttan B, Ott S, Patel R, Irvine J, Song J et al. Efficient and sustained transgene expression in human corneal cells mediated by a lentiviral vector. Gene Therapy 2000; 7: 196–200.
McMahon JM, Conroy S, Lyons M, Greiser U, O’Shea C, Strappe P et al. Gene transfer into rat mesenchymal stem cells: a comparative study of viral and nonviral vectors. Stem Cells Dev 2006; 15: 87–96.
Zhang XY, La Russa VF, Reiser J . Transduction of bone-marrow-derived mesenchymal stem cells by using lentivirus vectors pseudotyped with modified RD114 envelope glycoproteins. J Virol 2004; 78: 1219–1229.
Puthalakath H, Strasser A . Keeping killers on a tight leash: transcriptional and post-translational control of the pro-apoptotic activity of BH3-only proteins. Cell Death Differ 2002; 9: 505–512.
Sattler M, Liang H, Nettesheim D, Meadows RP, Harlan JE, Eberstadt M et al. Structure of Bcl-xL-Bak peptide complex: recognition between regulators of apoptosis. Science (New York, NY) 1997; 275: 983–986.
Zhou Q, Krebs JF, Snipas SJ, Price A, Alnemri ES, Tomaselli KJ et al. Interaction of the baculovirus anti-apoptotic protein p35 with caspases. Specificity, kinetics, and characterization of the caspase/p35 complex. Biochemistry 1998; 37: 10757–10765.
Xu G, Cirilli M, Huang Y, Rich RL, Myszka DG, Wu H . Covalent inhibition revealed by the crystal structure of the caspase-8/p35 complex. Nature 2001; 410: 494–497.
Gain P, Thuret G, Chiquet C, Dumollard JM, Mosnier JF, Burillon C et al. Value of two mortality assessment techniques for organ cultured corneal endothelium: trypan blue versus TUNEL technique. Br J Ophthalmol 2002; 86: 306–310.
Crewe JM, Armitage WJ . Integrity of epithelium and endothelium in organ-cultured human corneas. Invest Ophthalmol Vis Sci 2001; 42: 1757–1761.
Barinaga M . Stroke-damaged neurons may commit cellular suicide. Science (New York, NY) 1998; 281: 1302–1303.
Nakamura A, Akahane M, Shigematsu H, Tadokoro M, Ohgushi H, Dohi Y et al. Cell sheet transplantation of cultured mesenchymal stem cells enhances bone formation in a rat nonunion model. Bone 2009; 46: 418–424.
Hayashi R, Yamato M, Takayanagi H, Oie Y, Kubota A, Hori Y et al. Validation system of tissue engineered epithelial cell sheets for corneal regenerative medicine. Tissue Eng 2009; 16: 553–560.
Lombardo M, Lombardo G, Friend DJ, Serrao S, Terry MA . Long-term anterior and posterior topographic analysis of the cornea after deep lamellar endothelial keratoplasty. Cornea 2009; 28: 408–415.
Gardlik R, Palffy R, Hodosy J, Lukacs J, Turna J, Celec P . Vectors and delivery systems in gene therapy. Med Sci Monit 2005; 11: RA110–RA121.
Pels E, Beele H, Claerhout I . Eye bank issues: II. Preservation techniques: warm versus cold storage. Int Ophthalmol 2008; 28: 155–163.
Goren MB . The Eye Bank Association of America. Compr Ophthalmol Update 2006; 7: 261–262.
Acknowledgements
This work was supported by NIH R01EY012963 (RD), K24EY019098 (RD), Research to Prevent Blindness Lew R. Wasserman Merit Award (RD), the German Research Foundation (DFG/FU 726/1-1, TF) and the Eye Bank Association of America (TF). We thank Tissue Banks International and the Lions Eye Institute for Transplant and Research (Tampa, Florida) for providing corneas for research purposes. We thank Kim Fechtel, PhD, for the critical review of the manuscript.
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The Schepens Eye Research Institute and the University of Duisburg-Essen filed a patent cooperative treaty to protect rights emanating from this work. Dr Kazlauskas received compensation in 2009 from Celegene. Dr Fuchsluger, Dr Jurkunas and Dr Dana have nothing to disclose.
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Fuchsluger, T., Jurkunas, U., Kazlauskas, A. et al. Anti-apoptotic gene therapy prolongs survival of corneal endothelial cells during storage. Gene Ther 18, 778–787 (2011). https://doi.org/10.1038/gt.2011.20
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DOI: https://doi.org/10.1038/gt.2011.20
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