April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Gene Therapy Approach Through siRNA Against p63 To Correct Ocular Surface Disorders In Ectrodactyly–Ectodermal Dysplasia–Clefting (EEC) Syndrome
Author Affiliations & Notes
  • Enzo Di Iorio
    Veneto Eye Bank Foundation, Zelarino - Venice, Italy
  • Stefano Ferrari
    Veneto Eye Bank Foundation, Zelarino - Venice, Italy
  • Diego Ponzin
    Veneto Eye Bank Foundation, Zelarino - Venice, Italy
  • Colin E. Willoughby
    Centre for Vision and Vascular Science, Queen's Univerity Belfast, Belfast, United Kingdom
  • Elisabetta Bohm
    Department of Ophthalmology, ULSS 12, Zelarino - Venice, Italy
  • Vanessa Barbaro
    Veneto Eye Bank Foundation, Zelarino - Venice, Italy
  • Footnotes
    Commercial Relationships  Enzo Di Iorio, None; Stefano Ferrari, None; Diego Ponzin, None; Colin E. Willoughby, None; Elisabetta Bohm, None; Vanessa Barbaro, None
  • Footnotes
    Support  AFM-Association Francaise contre les Myopathies-14899
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 487. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Enzo Di Iorio, Stefano Ferrari, Diego Ponzin, Colin E. Willoughby, Elisabetta Bohm, Vanessa Barbaro; Gene Therapy Approach Through siRNA Against p63 To Correct Ocular Surface Disorders In Ectrodactyly–Ectodermal Dysplasia–Clefting (EEC) Syndrome. Invest. Ophthalmol. Vis. Sci. 2011;52(14):487.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : To describe the ocular phenotype in patients with EEC syndrome, to determine the pathogenic basis of visual morbidity and finally to investigate gene silencing approaches through allele specific siRNA, in order to assess new therapeutic approaches based on transplantation of genetically corrected autologous corneal limbal stem cells.

Methods: : Genetic and ophthalmologic analyses of 19 families consisting of 25 patients with EEC syndrome were assessed. All patients with EEC syndrome resulting from p63 mutations had a complete ophthalmic examination and ocular surface assessment. Limbal stem cell deficiency was diagnosed clinically on the basis of corneal conjunctivalisation, anatomy of the limbal palisades of Vogt, and immunohistochemical analysis performed on both pannus explant and impression cytology specimens. Keratinocytes stem cells from EEC patients carrying the R279H and the R304Q mutations have been analysed to obtain informations about their proliferative potential. Gene silencing approaches have been investigated through siRNA against R279H–p63 mutant.

Results: : All EEC patients had ocular involvement and the major cause of visual morbidity was limbal stem cell deficiency (LSCD) which was seen in 13/25 (52%) of cases. Keratinocytes stem cells from EEC patients carrying the R279H and the R304Q mutations showed a rapid decrease of colony forming efficiency (CFE) and clonogenic cell number and an increased replicative senescence. In vitro organotypic cultures showed defects in EEC epithelial stem cell stratification. Allele-specific siRNA against R279H–p63 mutant provided interesting results in order to assess new therapeutic approaches based on transplantation of genetically corrected autologous corneal limbal stem cells.

Conclusions: : Our study confirms that the major cause of visual morbidity in EEC syndrome is limbal stem cell failure, resulting in a vascularised corneal pannus, leading to visual loss. Mutations in p63 gene severly compromise the ability of stem cells to give rise to a full tickness stratified and differentiated corneal tissue. The only effective management for EEC patients would therefore be to restore the limbus by grafting cultured limbal stem cells following correction of the p63 gene defect.

Keywords: gene transfer/gene therapy • cornea: epithelium • lacrimal gland 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×