دورية أكاديمية
Modulation of Contact Inhibition by ZO-1/ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts.
| العنوان: | Modulation of Contact Inhibition by ZO-1/ZONAB Gene Transfer-A New Strategy to Increase the Endothelial Cell Density of Corneal Grafts. |
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| المؤلفون: | Kampik D; Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom.; University Hospital of Würzburg, Department of Ophthalmology, Würzburg, Germany., Basche M; Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom., Georgiadis A; Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom., Luhmann UFO; Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom., Larkin DF; Moorfields Eye Hospital, London, United Kingdom., Smith AJ; Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom., Ali RR; Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom.; NIHR Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom. |
| المصدر: | Investigative ophthalmology & visual science [Invest Ophthalmol Vis Sci] 2019 Jul 01; Vol. 60 (8), pp. 3170-3177. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Association For Research In Vision And Ophthalmology (Arvo) Country of Publication: United States NLM ID: 7703701 Publication Model: Print Cited Medium: Internet ISSN: 1552-5783 (Electronic) Linking ISSN: 01460404 NLM ISO Abbreviation: Invest Ophthalmol Vis Sci Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Brookline Ma : Association For Research In Vision And Ophthalmology (Arvo) Original Publication: St. Louis, Mosby. |
| مواضيع طبية MeSH: | Corneal Transplantation* , Gene Expression Regulation* , Gene Transfer Techniques*, Endothelium, Corneal/*metabolism , RNA, Messenger/*genetics , Zonula Occludens-1 Protein/*genetics, Cell Count ; Cells, Cultured ; Contact Inhibition ; Corneal Diseases/genetics ; Corneal Diseases/pathology ; Corneal Diseases/surgery ; Endothelium, Corneal/pathology ; Humans ; Signal Transduction ; Zonula Occludens-1 Protein/biosynthesis |
| مستخلص: | 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. |
| معلومات مُعتمدة: | United Kingdom DH_ Department of Health |
| المشرفين على المادة: | 0 (RNA, Messenger) 0 (Zonula Occludens-1 Protein) |
| تواريخ الأحداث: | Date Created: 20190724 Date Completed: 20191212 Latest Revision: 20191217 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1167/iovs.18-26260 |
| PMID: | 31335954 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1552-5783 |
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| DOI: | 10.1167/iovs.18-26260 |