Adenoviral gene transfer of eNOS

High-level expression in ex vivo expanded marrow stromal cells

Weiwen Deng, Trinity J. Bivalacqua, Natasha Chattergoon, Albert L. Hyman, James R. Jeter, Philip J. Kadowitz

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Endothelial nitric oxide synthase (eNOS) is an attractive target for cardiovascular gene therapy. Marrow stromal cells (MSCs), also known as mesenchymal stem cells, hold great promise for use in adult stem cell-based cell and gene therapy. To determine the feasibility of adenoviral-mediated eNOS gene transfer into ex vivo expanded MSCs, rat MSCs (rMSCs) were isolated, expanded ex vivo, and transduced with Ad5RSVeNOS, an adenoviral vector containing the eNOS gene under the control of the Rous sarcoma virus promoter. The presence of eNOS protein in Ad5RSVeNOS-transduced rMSCs was confirmed by immunohistochemical and Western blot analysis. Transduction efficiency was dose dependent, and eNOS transgene expression in rMSCs persisted for ≥21 days in culture. The rMSCs retained multipotential differentiation capability after adenoviral-mediated eNOS gene transfer. Furthermore, intracavernosal injection of Ad5RSVeNOS-transduced rMSCs increased the expression of eNOS in the corpus cavernosum, and stem cells were identified within corporal sinusoids. These findings demonstrate that replication-deficient recombinant adenovirus can be used to engineer ex vivo expanded rMSCs and that high-level eNOS transgene expression can be achieved, pointing out the clinical potential of using this novel adult stem cell-based gene therapy method for the treatment of cardiovascular diseases.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume285
Issue number5 54-5
StatePublished - Nov 2003
Externally publishedYes

Fingerprint

Gene transfer
Nitric Oxide Synthase Type III
Stromal Cells
Bone Marrow
Rats
Stem cells
Gene therapy
Genes
Genetic Therapy
Adult Stem Cells
Transgenes
Rous sarcoma virus
Cell- and Tissue-Based Therapy
Mesenchymal Stromal Cells
Viruses
Adenoviridae
Cardiovascular Diseases
Stem Cells
Western Blotting
Engineers

Keywords

  • Adenoviral vector
  • Differentiation
  • Gene expression
  • Gene therapy
  • Nitric oxide

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Adenoviral gene transfer of eNOS : High-level expression in ex vivo expanded marrow stromal cells. / Deng, Weiwen; Bivalacqua, Trinity J.; Chattergoon, Natasha; Hyman, Albert L.; Jeter, James R.; Kadowitz, Philip J.

In: American Journal of Physiology - Cell Physiology, Vol. 285, No. 5 54-5, 11.2003.

Research output: Contribution to journalArticle

Deng, Weiwen ; Bivalacqua, Trinity J. ; Chattergoon, Natasha ; Hyman, Albert L. ; Jeter, James R. ; Kadowitz, Philip J. / Adenoviral gene transfer of eNOS : High-level expression in ex vivo expanded marrow stromal cells. In: American Journal of Physiology - Cell Physiology. 2003 ; Vol. 285, No. 5 54-5.
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