Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas

S. Nair, D. T. Curiel, V. Rajaratnam, C. Thota, Ayman Al-Hendy

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

STUDY QUESTIONIs targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus -human somatostatin receptor subtype 2- arginine, glycine and aspartate-thymidine kinase), given in combination with ganciclovir (GCV) against immortalized human leiomyoma cells (HuLM) a potential therapy for uterine fibroids?SUMMARY ANSWERAd-SSTR-RGD-TK/GCV, a targeted adenovirus, effectively reduces cell growth in HuLM cells and to a significantly greater extent than in human uterine smooth muscle cells (UtSM).WHAT IS KNOWN ALREADYUterine fibroids (leiomyomas), a major cause of morbidity and the most common indication for hysterectomy in premenopausal women, are well-defined tumors, making gene therapy a suitable and potentially effective non-surgical approach for treatment. Transduction of uterine fibroid cells with adenoviral vectors such as Ad-TK/GCV (herpes simplex virus thymidine kinase gene) decreases cell proliferation.STUDY DESIGN, SIZE, DURATIONAn in vitro cell culture method was set up to compare and test the efficacy of a modified adenovirus vector with different multiplicities of infection in two human immortalized cell lines for 5 days.PARTICIPANTS/MATERIALS, SETTING, METHODSImmortalized human leiomyoma cells and human uterine smooth muscle cells were infected with different multiplicities of infection (MOI) (5-100 plaque-forming units (pfu)/cell) of a modified Ad-SSTR-RGD-TK vector and subsequently treated with GCV. For comparison, HuLM and UtSM cells were transfected with Ad-TK/GCV and Ad-LacZ/GCV. Cell proliferation was measured using the CyQuant assay in both cell types. Additionally, western blotting was used to assess the expression of proteins responsible for regulating proliferation and apoptosis in the cells.MAIN RESULTS AND THE ROLE OF CHANCETransduction of HuLM cells with Ad-SSTR-RGD-TK/GCV at 5, 10, 50 and 100 pfu/cell decreased cell proliferation by 28, 33, 45, and 84%, respectively (P < 0.05) compared with untransfected cells, whereas cell proliferation in UtSM cells transfected with the same four MOIs of Ad-SSTR-RGD-TK/GCV compared with that of untransfected cells was decreased only by 8, 23, 25, and 28%, respectively (P < 0.01). Western blot analysis showed that, in comparison with the untargeted vector Ad-TK, Ad-SSTR-RGD-TK/GCV more effectively reduced expression of proteins that regulate the cell cycle (Cyclin D1) and proliferation (PCNA, Proliferating Cell Nuclear Antigen), and it induced expression of the apoptotic protein BAX, in HuLM cells.LIMITATIONS, REASONS FOR CAUTIONResults from this study need to be replicated in an appropriate animal model before testing this adenoviral vector in a human trial.WIDER IMPLICATIONS OF THE FINDINGSEffective targeting of gene therapy to leiomyoma cells enhances its potential as a non-invasive treatment of uterine fibroids.

Original languageEnglish (US)
Pages (from-to)2398-2406
Number of pages9
JournalHuman Reproduction
Volume28
Issue number9
DOIs
StatePublished - Jan 1 2013

Fingerprint

Leiomyoma
Genetic Therapy
Ganciclovir
Myometrium
Adenoviridae
Smooth Muscle Myocytes
Cell Proliferation
Thymidine Kinase
Aspartate Kinase
Western Blotting
Cell Cycle Proteins
Cyclin D1
Proliferating Cell Nuclear Antigen

Keywords

  • Ad-SSTR-RGD-TK
  • adenovirus vectors
  • gene therapy
  • uterine leiomyomas

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology

Cite this

Nair, S., Curiel, D. T., Rajaratnam, V., Thota, C., & Al-Hendy, A. (2013). Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas. Human Reproduction, 28(9), 2398-2406. https://doi.org/10.1093/humrep/det275

Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas. / Nair, S.; Curiel, D. T.; Rajaratnam, V.; Thota, C.; Al-Hendy, Ayman.

In: Human Reproduction, Vol. 28, No. 9, 01.01.2013, p. 2398-2406.

Research output: Contribution to journalArticle

Nair, S, Curiel, DT, Rajaratnam, V, Thota, C & Al-Hendy, A 2013, 'Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas', Human Reproduction, vol. 28, no. 9, pp. 2398-2406. https://doi.org/10.1093/humrep/det275
Nair, S. ; Curiel, D. T. ; Rajaratnam, V. ; Thota, C. ; Al-Hendy, Ayman. / Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas. In: Human Reproduction. 2013 ; Vol. 28, No. 9. pp. 2398-2406.
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T1 - Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas

AU - Nair, S.

AU - Curiel, D. T.

AU - Rajaratnam, V.

AU - Thota, C.

AU - Al-Hendy, Ayman

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N2 - STUDY QUESTIONIs targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus -human somatostatin receptor subtype 2- arginine, glycine and aspartate-thymidine kinase), given in combination with ganciclovir (GCV) against immortalized human leiomyoma cells (HuLM) a potential therapy for uterine fibroids?SUMMARY ANSWERAd-SSTR-RGD-TK/GCV, a targeted adenovirus, effectively reduces cell growth in HuLM cells and to a significantly greater extent than in human uterine smooth muscle cells (UtSM).WHAT IS KNOWN ALREADYUterine fibroids (leiomyomas), a major cause of morbidity and the most common indication for hysterectomy in premenopausal women, are well-defined tumors, making gene therapy a suitable and potentially effective non-surgical approach for treatment. Transduction of uterine fibroid cells with adenoviral vectors such as Ad-TK/GCV (herpes simplex virus thymidine kinase gene) decreases cell proliferation.STUDY DESIGN, SIZE, DURATIONAn in vitro cell culture method was set up to compare and test the efficacy of a modified adenovirus vector with different multiplicities of infection in two human immortalized cell lines for 5 days.PARTICIPANTS/MATERIALS, SETTING, METHODSImmortalized human leiomyoma cells and human uterine smooth muscle cells were infected with different multiplicities of infection (MOI) (5-100 plaque-forming units (pfu)/cell) of a modified Ad-SSTR-RGD-TK vector and subsequently treated with GCV. For comparison, HuLM and UtSM cells were transfected with Ad-TK/GCV and Ad-LacZ/GCV. Cell proliferation was measured using the CyQuant assay in both cell types. Additionally, western blotting was used to assess the expression of proteins responsible for regulating proliferation and apoptosis in the cells.MAIN RESULTS AND THE ROLE OF CHANCETransduction of HuLM cells with Ad-SSTR-RGD-TK/GCV at 5, 10, 50 and 100 pfu/cell decreased cell proliferation by 28, 33, 45, and 84%, respectively (P < 0.05) compared with untransfected cells, whereas cell proliferation in UtSM cells transfected with the same four MOIs of Ad-SSTR-RGD-TK/GCV compared with that of untransfected cells was decreased only by 8, 23, 25, and 28%, respectively (P < 0.01). Western blot analysis showed that, in comparison with the untargeted vector Ad-TK, Ad-SSTR-RGD-TK/GCV more effectively reduced expression of proteins that regulate the cell cycle (Cyclin D1) and proliferation (PCNA, Proliferating Cell Nuclear Antigen), and it induced expression of the apoptotic protein BAX, in HuLM cells.LIMITATIONS, REASONS FOR CAUTIONResults from this study need to be replicated in an appropriate animal model before testing this adenoviral vector in a human trial.WIDER IMPLICATIONS OF THE FINDINGSEffective targeting of gene therapy to leiomyoma cells enhances its potential as a non-invasive treatment of uterine fibroids.

AB - STUDY QUESTIONIs targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus -human somatostatin receptor subtype 2- arginine, glycine and aspartate-thymidine kinase), given in combination with ganciclovir (GCV) against immortalized human leiomyoma cells (HuLM) a potential therapy for uterine fibroids?SUMMARY ANSWERAd-SSTR-RGD-TK/GCV, a targeted adenovirus, effectively reduces cell growth in HuLM cells and to a significantly greater extent than in human uterine smooth muscle cells (UtSM).WHAT IS KNOWN ALREADYUterine fibroids (leiomyomas), a major cause of morbidity and the most common indication for hysterectomy in premenopausal women, are well-defined tumors, making gene therapy a suitable and potentially effective non-surgical approach for treatment. Transduction of uterine fibroid cells with adenoviral vectors such as Ad-TK/GCV (herpes simplex virus thymidine kinase gene) decreases cell proliferation.STUDY DESIGN, SIZE, DURATIONAn in vitro cell culture method was set up to compare and test the efficacy of a modified adenovirus vector with different multiplicities of infection in two human immortalized cell lines for 5 days.PARTICIPANTS/MATERIALS, SETTING, METHODSImmortalized human leiomyoma cells and human uterine smooth muscle cells were infected with different multiplicities of infection (MOI) (5-100 plaque-forming units (pfu)/cell) of a modified Ad-SSTR-RGD-TK vector and subsequently treated with GCV. For comparison, HuLM and UtSM cells were transfected with Ad-TK/GCV and Ad-LacZ/GCV. Cell proliferation was measured using the CyQuant assay in both cell types. Additionally, western blotting was used to assess the expression of proteins responsible for regulating proliferation and apoptosis in the cells.MAIN RESULTS AND THE ROLE OF CHANCETransduction of HuLM cells with Ad-SSTR-RGD-TK/GCV at 5, 10, 50 and 100 pfu/cell decreased cell proliferation by 28, 33, 45, and 84%, respectively (P < 0.05) compared with untransfected cells, whereas cell proliferation in UtSM cells transfected with the same four MOIs of Ad-SSTR-RGD-TK/GCV compared with that of untransfected cells was decreased only by 8, 23, 25, and 28%, respectively (P < 0.01). Western blot analysis showed that, in comparison with the untargeted vector Ad-TK, Ad-SSTR-RGD-TK/GCV more effectively reduced expression of proteins that regulate the cell cycle (Cyclin D1) and proliferation (PCNA, Proliferating Cell Nuclear Antigen), and it induced expression of the apoptotic protein BAX, in HuLM cells.LIMITATIONS, REASONS FOR CAUTIONResults from this study need to be replicated in an appropriate animal model before testing this adenoviral vector in a human trial.WIDER IMPLICATIONS OF THE FINDINGSEffective targeting of gene therapy to leiomyoma cells enhances its potential as a non-invasive treatment of uterine fibroids.

KW - Ad-SSTR-RGD-TK

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KW - uterine leiomyomas

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