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Modulation of apoptotic signaling pathways to promote survival of endothelial cells by gene therapy
Author(s) -
FUCHSLUGER T,
JURKUNAS U,
KAZLAUSKAS A,
DANA R
Publication year - 2010
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2010.3135.x
Subject(s) - apoptosis , transplantation , programmed cell death , economic shortage , corneal transplantation , flow cytometry , microbiology and biotechnology , genetic enhancement , cell , biology , cancer research , medicine , immunology , gene , biochemistry , linguistics , philosophy , government (linguistics)
Purpose Corneal transplantation is the most common transplantation worldwide. Surgeons and eye banks face major problems: (1) shortage of tissue in aging populations, (2) loss of high‐quality tissue due to cell loss during storage, (3) graft failure. It has been demonstrated that EC loss is mediated by the cells’ intrinsic death machinery resulting in apoptosis. Identification of survival strategies could raise the availability of tissue, with a significant impact on transplantation by lowering graft rejection rate. The purpose of this study different apoptotic pathways and to determine the protective effect of the anti‐apoptotic proteins bcl‐xL and p35. Methods Intrinsic (mitochondria‐mediated) and extrinsic (ligand‐mediated) apoptotic pathways were selectively activated to provoke apoptosis of murine and human corneal endothelial cells suspensions and corneas. Gene transfer of bcl‐xL or p35 was accomplished, survival of EC was determined by flow cytometry and laser scanning microscopy. Results Interestingly, we were able to determine distinct differences in cell survival enhancement depending on the type of overexpressed protein. Whereas uninfected controls showed significant EC death, gene‐therapeutically treated EC demonstrated significantly increased cell survival. We will present data on the efficacy of certain anti‐apoptotic proteins in select pathways. Conclusion Exploring inhibitory strategies of EC death can lead to clinically relevant survival strategies with significant impact on corneal grafting.