title: A novel approach to investigation of the pathogenesis of pterygium based on assessment of promoter hyper-methylation and expression profile of CTLA4 gene: A credible report of CTLA4 gene expression in human eye tissue.
abstract: BACKGROUND
  Pterygium is the human eye lesion whose prevalence in the general population is estimated about 2%. The disease, in extreme phase, can lead to visual disturbance and eventually causes complete loss of vision due to the lesion growth over the papillary axis. Pterygium invasive tissue is a tumor-like tissue that is initially identified and then is attacked by cytotoxic T cells. Cytotoxic T lymphocyte associated antigen 4 (CTLA4), as a modulator molecule of the adaptive immune system, plays a critical role in maintaining peripheral T cell tolerance by diminishing its responsiveness and increasing its activation threshold. The aim of this study is to investigate the association between some epigenetic changes of the CTLA4 gene, such as promoter methylation and gene expression, and pathogenesis of pterygia.
  
  
  METHODS
  Genomic DNA was extracted from 75 formalin-fixed, paraffin-embedded tissues of pterygia and 70 specimens of normal conjunctiva from eyes without pterygium as the control group, collected from Sistan and Baluchestan population. CTLA4 gene promoter methylation was carried out by methylation-specific PCR technique. The gene expression analysis was done on extracted total RNA from 20 healthy and 23 pterygium tissue samples using Real-Time PCR technique.
  
  
  RESULTS
  Promoter methylation changes of CTLA4 gene were not statistically different in patients with pterygium in comparison with healthy controls (OR=1.614; 95% CI=0.57-4.75; P value=0.37). However, gene expression level of CTLA4 was remarkably different in patients and healthy controls (Mean±SD: 1.343±0.133 and 2.027±0.219, respectively; P value=0.009).
  
  
  CONCLUSIONS
  This is a credible evidence of CTLA4 gene expression in human eye tissue. This first hand attempt of investigating the association of epigenetic changes of the CTLA4 gene and pathogenesis of pterygia, indicated a significant intensification of the gene expression of CTLA4 in patients with pterygia. We suggest that increasing CTLA4 gene expression can be a trigger which promotes pterygium enlargement. However, further studies on more populations with larger sample sizes need to be done to verify this hypothesis in the future.
authors.affiliation: Department of Biology, University of Sistan and Baluchestan, Zahedan, Iran.
authors.forename: Mohammad Essmail
authors.initials: ME
authors.lastname: Ebrahimi
authors.affiliation: Department of Biology, University of Sistan and Baluchestan, Zahedan, Iran. Electronic address: dor_kordi@yahoo.com.
authors.forename: Dor Mohammad
authors.initials: DM
authors.lastname: Kordi-Tamandani
authors.affiliation: Department of Ophthalmology, Al-Zahra Eye Hospital, Zahedan University of Medical Sciences, Zahedan, Iran.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Arish
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D060908:CTLA-4 Antigen / Q000235:genetics* / Q000378:metabolism; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D003228:Conjunctiva / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D019175:DNA Methylation; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D011625:Pterygium / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000601:surgery
chemical_list: D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human
keywords: CTLA4 gene; Cytotoxic T cell; Epigenetic change; Gene expression; Promoter hyper-methylation; Pterygium