Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells

Essam R. Othman, David T. Curiel, Mostafa Hussein, Ibraheem I. Abdelaal, Ahmed N. Fetih, Ayman Al-Hendy

Research output: Contribution to journalArticle

Abstract

Objective: 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. Methods: 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. Results: 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. Conclusions: 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.

Original languageEnglish (US)
Pages (from-to)1109-1115
Number of pages7
JournalReproductive Sciences
Volume23
Issue number8
DOIs
StatePublished - Aug 1 2016

Fingerprint

Adenoviridae
Gene Expression
Luciferases
Stromal Cells
Reporter Genes
Genes
Secretory Leukocyte Peptidase Inhibitor
Viruses
Endometrium
Infection
Hysterectomy
Transgenes
Genetic Therapy
Cell Culture Techniques
Students
heparanase

Keywords

  • adenovirus
  • endometrium
  • gene transfer

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Othman, E. R., Curiel, D. T., Hussein, M., Abdelaal, I. I., Fetih, A. N., & Al-Hendy, A. (2016). Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells. Reproductive Sciences, 23(8), 1109-1115. https://doi.org/10.1177/1933719116630420

Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells. / Othman, Essam R.; Curiel, David T.; Hussein, Mostafa; Abdelaal, Ibraheem I.; Fetih, Ahmed N.; Al-Hendy, Ayman.

In: Reproductive Sciences, Vol. 23, No. 8, 01.08.2016, p. 1109-1115.

Research output: Contribution to journalArticle

Othman, ER, Curiel, DT, Hussein, M, Abdelaal, II, Fetih, AN & Al-Hendy, A 2016, 'Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells', Reproductive Sciences, vol. 23, no. 8, pp. 1109-1115. https://doi.org/10.1177/1933719116630420
Othman ER, Curiel DT, Hussein M, Abdelaal II, Fetih AN, Al-Hendy A. Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells. Reproductive Sciences. 2016 Aug 1;23(8):1109-1115. https://doi.org/10.1177/1933719116630420
Othman, Essam R. ; Curiel, David T. ; Hussein, Mostafa ; Abdelaal, Ibraheem I. ; Fetih, Ahmed N. ; Al-Hendy, Ayman. / Enhancing Adenoviral-Mediated Gene Transfer and Expression to Endometrial Cells. In: Reproductive Sciences. 2016 ; Vol. 23, No. 8. pp. 1109-1115.
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AB - Objective: 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. Methods: 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. Results: 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. Conclusions: 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.

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