Intra-luminal gene therapy in the porcine artery using a recombinant adeno-associated virus 9

E. S. Rezaie, N. J. Visser, P. F. Friedrich, Alexander Yong-Shik Shin, Allen Thorp Bishop

Research output: Contribution to journalArticle

Abstract

The ability to improve or restore blood flow and promote healing in ischemic tissue has many potential clinical applications. Augmentation by direct delivery of growth factors may further enhance results, but requires a method for sustained delivery. In this study, we have tested the ability of adeno-associated virus 9 (AAV9) delivered within the lumen of a porcine artery to transfect the vessel and produce a desired product. The marker chosen was green fluorescent protein (GFP) (Ke et al., 2011). In 4 farm pigs the cranial tibial artery was surgically exposed. The vessel was temporarily clamped proximally, and divided distally. A cannula was placed intraluminally, and the arterial segment was injected with 1 × 10E13 particles of AAV9.CB7.CI.GFP·WPRE.rBG. At 14 days the transfected cranial tibial artery as well as the liver, spleen and kidneys were harvested. ELISA and reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) were used to analyze the artery for GFP production. Significant GFP expression was seen in all transfected cranial tibial vessels, as determined by both GFP protein production (ELISA) and mRNA (RT-qPCR). No GFP was identified in liver, spleen or kidney, nor in the no-GFP control animal artery. Adeno-associated virus 9 is an appropriate vector for gene therapy experiments in the porcine artery model. This vector, and the intraluminal deliver method described result in robust gene expression at 2 weeks without evident systemic spill of the virus. The ability to limit delivery of the gene to an isolated segment of vessel is desirable for future research applications.

Original languageEnglish (US)
Pages (from-to)24-27
Number of pages4
JournalGene
Volume618
DOIs
StatePublished - Jun 30 2017

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Dependovirus
Green Fluorescent Proteins
Genetic Therapy
Swine
Arteries
Tibial Arteries
Reverse Transcriptase Polymerase Chain Reaction
Spleen
Enzyme-Linked Immunosorbent Assay
Kidney
Liver
Intercellular Signaling Peptides and Proteins
Viruses
Gene Expression
Messenger RNA
Genes
Proteins

Keywords

  • Adeno associated virus
  • Gene therapy
  • Porcine
  • Vascular disease

ASJC Scopus subject areas

  • Medicine(all)
  • Genetics

Cite this

Intra-luminal gene therapy in the porcine artery using a recombinant adeno-associated virus 9. / Rezaie, E. S.; Visser, N. J.; Friedrich, P. F.; Shin, Alexander Yong-Shik; Bishop, Allen Thorp.

In: Gene, Vol. 618, 30.06.2017, p. 24-27.

Research output: Contribution to journalArticle

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