Lentiviral-human heme oxygenase targeting endothelium improved vascular function in angiotensin II animal model of hypertension
Authors: David E Stec
Luca Vanella
Nader G Abraham
John Quilley
Komal Sodhi
Jian Cao
Rita Rezzani
Attallah Kappas
Kazuyoshi Inoue
Luigi Rodella
Lucrezia Germinario
Journal: Human Gene Therapy (2011)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: vascular dysfunction; renal dysfunction; angiotensin II; lentiviral-mediated delivery; endothelial heme oxygenase (HO)-1
Affiliations: Department of Physiology and Pharmacology, The University of Toledo, Toledo, OH 43614., Department of Pharmacology, New York Medical College, Valhalla, NY 10595., Department of Biomedical Science, University of Brescia, Brescia, 25123, Italy., Department of Physiology and Biophysics, University of Mississippi Medical Center, Jackson, MS 39215., The Rockefeller University, New York, NY 10065.

Abstract:  We examined the hypothesis that vascular and renal dysfunction caused by angiotensin II (Ang II) through increased levels of blood pressure, inflammatory cytokines, and oxidative stress in Sprague-Dawley rats can be prevented by lentiviral-mediated delivery of endothelial heme oxygenase (HO)-1. We targeted the vascular endothelium using a lentiviral construct expressing human HO-1 under the control of the endothelium-specific promoter VE-cadherin (VECAD-HO-1) and examined the effect of long-term human HO-1 expression on blood pressure in Ang II-mediated increases in blood pressure and oxidant stress. A bolus injection of VECAD-HO-1 into the renal artery resulted in expression of human HO-1 for up to 6-9 weeks. Sprague-Dawley rats were implanted with Ang II minipumps and treated with lentivirus carrying either the HO-1 or green fluorescent protein. Renal tissue from VECAD-HO-1-transduced rats expresses human HO-1 mRNA and proteins without an effect on endogenous HO-1. Infusion of Ang II increased blood pressure (p &lt; 0.001) but decreased vascular relaxation in response to acetylcholine, endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (peNOS) levels, and renal and plasma levels of adiponectin (p &lt; 0.05); in contrast, plasma tumor necrosis factor-Œ± and monocyte chemoattractant protein-1 levels increased. Ang II-treated animals had higher levels of superoxide anion and inducible nitric oxide synthase and increased urinary protein and plasma creatinine levels. Lentiviral transduction with the VECAD-HO-1 construct attenuated the increase in blood pressure (p &lt; 0.05), improved vascular relaxation, increased plasma adiponectin, and prevented the elevation in urinary protein and plasma creatinine in Ang II-treated rats. Endothelial-specific expression of HO-1 also reduced oxidative stress and levels of inflammatory cytokines resulting in increased expression of the anti-apoptotic proteins phosphorylated AKT, phosphorylated AMP-activated protein kinase, peNOS, and eNOS. Collectively, these findings demonstrate that endothelial-specific increases in HO-1 expression attenuate Ang II hypertension and the associated vascular dysfunction that is associated with increases in adiponectin and peNOS and reductions in oxidative stress and levels of inflammatory cytokines. 

Retracted: No

PubPeer comments:
 The six actin lanes in figure 1A are remarkably similar to six of the eight lanes in western blots in four other publications from this group. In four instances it is identified as actin, and in one instance it is AKT. The experiments in papers where these lanes appear involve cultured cells, rats (heart and adipose tissue) and mice. Various experimental protocols are reportedly used. Different animals, different cells and tissues, different experiments, sort of your all-purpose Western blot. Previously commented on in: https: pubpeer.com publications B3ABFE1EBACF517FD703E0BED6354F https: pubpeer.com publications BEEA22046712F49D470C06C83D6AC1 https: pubpeer.com publications 6BF6BCD78CF68B8B1DC710A72C8656  , 
 In addition to the data integrity issues raised above, could the authors please review the blot in Figure 6 s left panel. It seems as if p-eNOS band may be flipped horizontally based on the shape of the adjacent bands, and or perhaps is a duplication of the eNOS band. The penultimate author may have changed institutions since this paper was published. I will attempt to notify him when I notify the journal via email. ***  *** Acknowledgments. This work was supported by grants DK068134, HL55601, and HL34300 (to N.G.A.) and HL088421 and HL088421-S1 (to D.E.S.) from the National Institutes of Health. ,