UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Modulation of Contact Inhibition by ZO-1/ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts

Kampik, D; Basche, M; Georgiadis, A; Luhmann, UFO; Larkin, DF; Smith, AJ; Ali, RR; (2019) Modulation of Contact Inhibition by ZO-1/ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts. Investigative Ophthalmology & Visual Science , 60 (8) pp. 3170-3177. 10.1167/iovs.18-26260. Green open access

[thumbnail of Larkin_Modulation of Contact Inhibition by ZO-1 ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts_VoR.pdf]
Preview
Text
Larkin_Modulation of Contact Inhibition by ZO-1 ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts_VoR.pdf - Published Version

Download (1MB) | Preview

Abstract

PURPOSE: Endothelial cell density (ECD) is the principal factor determining the success of corneal transplants. Here we explored a strategy to increase corneal ECD in human explants via modulation of the ZO-1/ZONAB pathway. In multiple cell types, ZO-1 maintains G1 cell cycle arrest via cytoplasmic sequestration of the mitosis-inducing transcription factor ZONAB. In this study, we assessed the effects of lentiviral vector-mediated downregulation of ZO-1 or overexpression of ZONAB upon ECD and the integrity of the endothelial monolayer. METHODS: HIV-based lentiviral vectors were used to deliver either constitutively expressed ZONAB (LNT-ZONAB), or a small hairpin RNA targeting ZO-1 (LNT-shZO1). Human corneal specimens were bisected and each half was exposed to either treatment or control vector. After 1 week in ex vivo culture, effects were assessed by quantitative RT-PCR, immunohistochemistry, and ECD assessment. RESULTS: LNT-shZO1 achieved an ∼45% knockdown of ZO-1 mRNA in corneal endothelial cells cultured ex vivo, reduced ZO-1 staining, and did not affect morphologic endothelial monolayer integrity. The proliferative effect of LNT-shZO1 correlated with control ECD but not with donor age. Within a low-ECD cohort an ∼30% increase in ECD was observed. LNT-ZONAB achieved a >200-fold overexpression of ZONAB mRNA, which led to an ∼25% increase in ECD. CONCLUSIONS: ZO-1 downregulation or ZONAB upregulation increases corneal ECD via interference with contact inhibition and cell cycle control. With further development, such approaches might provide a means for improving ECD in donor corneas before transplantation.

Type: Article
Title: Modulation of Contact Inhibition by ZO-1/ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1167/iovs.18-26260
Publisher version: http://dx.doi.org/10.1167/iovs.18-26260
Language: English
Additional information: © 2019 The Authors. This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
Keywords: corneal endothelial cells, ZO-1/ZONAB, gene therapy, cell proliferation
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology
URI: https://discovery.ucl.ac.uk/id/eprint/10078955
Downloads since deposit
94Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item