title: Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells.
abstract: Our aim was to screen a panel of modified adenoviral gene transfer vectors to identify those which can sustain high gene expression in human endometrial cells.
  
  
  
  Normal endometrial stromal cell cultures were established from endometrial lining of hysterectomy specimens performed for benign gynecologic indications. Human endometrial stromal cells were transfected by modified adenoviruses expressing luciferase reporter gene. Luciferase activity mediated by each virus was expressed as a percentage of adenovirus serotype 5 (Ad5-CMV-luc) activity. The 2-tailed Student t test was used to compare data.
  
  
  
  At a multiplicity of infection (MOI) of 10 pfu/cell, of the transductionally modified adenoviruses, adenovirus-RGD (Ad-RGD-luc) mediated highest level of endometrial cell transduction with transgene expression around 4 times higher when compared to Ad5 (P < .001). Of the transcriptionally targeted adenoviruses, adenovirus under secretory leukocyte protease inhibitor promoter (Ad-SLPI-luc) and adenovirus under heparanase promoter (Ad-heparanase-luc)-mediated luciferase activation were 5.8- and 4.3-folds higher than Ad5-CMV-luc, respectively (P = .02 and .03, respectively). At MOI of 50 pfu/cell, Ad-RGD-luc and AD-SLPI-luc mediated significantly higher gene transfer efficiency compared to Ad5-CMV-luc (P values < .001, for each virus). Ad-heparanase-luc achieved higher gene activity, but difference was not significant (P = .1). Ad-SLPI-luc, at low viral dose (10 pfu/ cell), mediated gene expression effect comparable to Ad5-CMV-luc at a high dose (50 pfu/cell), with no significant difference.
  
  
  
  We conclude that when compared to the wild-type adenovirus, Ad-RGD-luc, Ad-SLPI-luc, and Ad-heparanase-luc mediate higher reporter gene activity in endometrial cells and can work as effective gene transfer vectors in gene therapy applications to the endometrium.
authors: OB-GYN Department, Assiut University, Assiut, Egypt Center of Excellence of Stem Cells and Regenerative Medicine CESCRM, Assiut University, Assiut, Egypt essamrash@yahoo.com.
authors: Essam R
authors: ER
authors: Othman
authors: Division of Cancer Biology, Department of Radiation Oncology, Washington University Medical School, Washington, DC, USA.
authors: David T
authors: DT
authors: Curiel
authors: OB-GYN Department, Assiut University, Assiut, Egypt.
authors: Mostafa
authors: M
authors: Hussein
authors: OB-GYN Department, Assiut University, Assiut, Egypt.
authors: Ibraheem I
authors: II
authors: Abdelaal
authors: OB-GYN Department, Assiut University, Assiut, Egypt.
authors: Ahmed N
authors: AN
authors: Fetih
authors: OB-GYN Department, Georgia Regents University, Medical College of Georgia, Augusta, GA, USA.
authors: Ayman
authors: A
authors: Al-Hendy
mesh_terms: D000256:Adenoviridae / Q000502:physiology*; D002478:Cells, Cultured; D004717:Endometrium / Q000166:cytology / Q000378:metabolism*; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics; D017154:Stromal Cells / Q000378:metabolism
chemical_list: D008156:Luciferases
keywords: adenovirus; endometrium; gene transfer