April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Anti-Apoptotic Gene Therapy to Promote Survival of Allogeneic Corneal Epithelial Grafts
Author Affiliations & Notes
  • Thomas A. Fuchsluger
    Center of Ophthalmology/Inst of Anatomy, Essen University Hospital, Essen, Germany
    Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts
  • Daniel R. Saban
    Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts
  • Andre Okanobo
    Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts
  • Reza Dana
    Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  Thomas A. Fuchsluger, None; Daniel R. Saban, None; Andre Okanobo, None; Reza Dana, None
  • Footnotes
    Support  DFG Fu 726/1-1 (TF), R01EY012963 (RD), K24EY019098 (RD), EBAA Richard Lindstrom Research Grant (TF)
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 2057. doi:
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    • Get Citation

      Thomas A. Fuchsluger, Daniel R. Saban, Andre Okanobo, Reza Dana; Anti-Apoptotic Gene Therapy to Promote Survival of Allogeneic Corneal Epithelial Grafts. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2057.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To study whether anti-apoptotic gene therapy to allogeneic corneal epithelial sheets in mice promotes graft survival. Engraftment of cultured allogeneic epithelium currently is a viable modality for patients bilaterally blind due to bilateral limbal stem cell deficiencies, e.g. after chemical burn. Despite full corticosteroid intervention, graft failure rate entailing apoptosis of donor cells is >50%. Gene transfer to cultured epithelial cell sheets in vitro before transplantation could improve the outcome in this most demanding ocular surface pathology .

Methods: : Epithelial denuded syngeneic Balb/C corneal buttons were reconstituted with allo- or syngeneic epithelium, transplanted and compared to epithelial grafts transduced with the anti-apoptotic gene p35 (total n=40). Feasibility of gene transfer was determined by studying kinetics of IZsGreen expression. Epithelial integrity and graft survival were assessed by fluoresceine staining and corneal opacity. Allo-response was evaluated by delayed type hypersensitivity (DTH).

Results: : Transfer of IZsGreen in vitro was successfully achieved obtaining high expression rates. Denuded corneas or those reconstituted with allogeneic epithelium showed a significantly higher opacity rate and considerably prolonged periods for epithelial regeneration relative to syngeneic controls. Surprisingly, gene-therapeutically modulated allogeneic epithelial grafts showed a significantly lower opacity rate and significantly expedited restoration of the epithelial barrier function compared to allo-untreated epithelial grafts (p<0.01, respectively). In addition, DTH analysis consistently showed significantly reduced T-cell response in the treated group (p<0.01).

Conclusions: : We herein established a novel model of allogeneic epithelial graft rejection. We were able to demonstrate that gene therapeutic modification of grafted cell sheets may be a viable option to increase allogeneneic epithelial graft survival. This sets the stage to further study gene therapeutic interventions of epithelial allo-grafts, and its impact on graft survival and immune response.

Keywords: gene transfer/gene therapy • transplantation • cornea: basic science 
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