A direct mechanical method for accurate and efficient adenoviral vector delivery to tissues

Khurana, Vini G.; Weiler, Deborah A.; Witt, Tyra A.; Smith, Leslie; Kleppe, Laurel S.; Parisi, Joseph E.; Simari, Robert D.; O’Brien, Timothy J.; Russell, Stephen J.; Katušić, Zvonimir S.

doi:10.1038/sj.gt.3301907

Cited by 14 publications

(14 citation statements)

References 32 publications

(132 reference statements)

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“…25,26 Our data suggest that local adenovirus-mediated delivery of DDAH genes would be sufficient to reduce endothelial ADMA concentrations and promote NO production in these situations. Several studies have reported that vascular delivery of NOS genes improves endothelial function in models of cardiovascular disease [27][28][29] and our current findings indicate that manipulation of DDAH levels could achieve similar results.…”

Section: Discussionsupporting

confidence: 81%

Adenoviral-mediated overexpression of DDAH improves vascular tone regulation

et al. 2010

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Nitric oxide (NO) is a key physiological mediator in the cardiovascular, neuronal and immune systems. Numerous reports have detailed the effects of reduced NO production on signaling in these systems and reduced NO generation has been associated with many pathological conditions. 1 In the cardiovascular system, genetic or pharmacological inhibition of NO production leads to attenuation of vascular relaxation, increased vascular resistance and increased blood pressure. Therefore, endogenous mechanisms that regulate NO production are of considerable interest. 2In the cardiovascular system, NO is synthesized by the endothelial isoform of nitric oxide synthase (eNOS) and the activity of this enzyme has been reported to be regulated at many levels including transcription, translation, post-translational modification and co-factor availability.2 In addition to these mechanisms, eNOS activity can be regulated by the level of endogenously produced asymmetrically methylated arginines which are competitive enzyme inhibitors.3 In humans, two endogenously produced asymmetric methylarginines are present; asymmetric dimethylarginine (ADMA) and N-monomethylarginine (l-NMMA). However, ADMA is found at approximately 10-fold the concentration of l-NMMA and is therefore thought to be the most functionally significant asymmetric methylarginine.3 Methylarginines are generated by the post-translational methylation of certain arginine residues in proteins catalyzed by a family of protein arginine methyltransferases (PRMTs). Adenoviral-mediated overexpression of DDAH improves vascular tone regulationBelen Torondel, Manasi Nandi*, Peter Kelly*, Beata Wojciak-Stothard, Ingrid Fleming and James Leiper Abstract Dimethylarginine dimethylaminohydrolase (DDAH) degrades asymmetric dimethylarginine (ADMA), an endogenously produced nitric oxide (NO) synthase inhibitor. In mammals, two isoforms of DDAH, DDAH1 and DDAH2, are expressed in the cardiovascular system, suggesting that ADMA concentrations are actively regulated in blood vessels, raising the possibility that cardiovascular metabolism of ADMA constitutes a novel mechanism for the regulation of NO production. The purpose of this study was to determine the role of DDAH-catalyzed asymmetric methylarginine metabolism in the regulation of vascular function. We developed adenoviral vectors for the expression of human DDAH1 and 2. Overexpression of DDAH1 or 2 in human umbilical vein endothelial cells (HUVEC) increases DDAH activity, reduces ADMA concentrations and increases NO production. Similarly, overexpression of DDAH1 or 2 in DDAH1 +/-mice carotid vessels increases NO production and attenuates the response to phenylephrine (PE), enhances acetylcholine (ACh) relaxation and attenuates the effect of exogenously applied ADMA. Finally, overexpression of either DDAH1 or 2 completely reversed the vascular dysfunction seen in DDAH1 +/-mice. These data indicate that basal concentrations of ADMA in blood vessels are sufficient to regulate NO production, that increases in the level of either DDAH1...

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Section: Discussionsupporting

confidence: 81%

Adenoviral-mediated overexpression of DDAH improves vascular tone regulation

et al. 2010

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“…In addition, a recent article described a similar process for gene transfer ex vivo to canine carotid artery rings and in vivo to rabbit carotid arteries. 19 Those authors found effective gene transfer to the superficial layers without penetration into the muscular layer with the use of trypsin-free solutions and that poloxamer-free solutions required 10 minutes of painting for effective gene transfer.…”

Section: Discussionmentioning

confidence: 99%

Targeted Modification of Atrial Electrophysiology by Homogeneous Transmural Atrial Gene Transfer

et al. 2005

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Background-Safe and effective myocardial gene transfer remains elusive. Heterogeneous ventricular gene delivery has been achieved in small mammals but generally with methods not readily transferable to the clinic. Atrium-specific gene transfer has not yet been reported. We hypothesized that homogeneous atrial gene transfer could be achieved by direct application of adenoviral vectors to the epicardial surface, use of poloxamer gel to increase virus contact time, and mild trypsinization to increase virus penetration. Methods and Results-We "painted" recombinant adenovirus encoding the reporter gene Escherichia coli ␤-galactosidase directly onto porcine atria. Investigational variables included poloxamer use, trypsin concentration, and safety. Using the painting method, we modified the atrial phenotype with an adenovirus expressing HERG-G628S, a long-QT-syndrome mutant. Our results showed that application of virus with poloxamer alone resulted in diffuse epicardial gene transfer with negligible penetration into the myocardium. Dilute trypsin concentrations allowed complete transmural gene transfer. After trypsin exposure, echocardiographic left atrial diameter did not change. Left atrial function decreased on postoperative day 3 but returned to baseline by day 7. Tissue tensile strength was affected only in the 1% trypsin group. HERG-G628S gene transfer prolonged atrial action potential duration and refractory period without affecting ventricular electrophysiology. Conclusions-We

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“…Each rabbit received the a1-PDX construct at one side and the control construct at the contralateral artery. For applying the virus peri-adventitially, the arteries were lifted and the viral constructs were spread by slight pressing with a brush as described before [10]. After a minute the arteries were put back in position and tissues were closed.…”

Section: In Vivo Adventitial Gene Deliverymentioning

confidence: 99%

“…Normal rabbit aortas were harvested and transfected periadventitially, with the a1-PDX construct or the empty virus (50 Al of 2.10 10 pfu/ml), by slight pressing with a brush [10]. The segments were cut into aortic rings for culture (3 + 7 days) as described before [8] and total protein was isolated.…”

Section: Ex Vivo Adventitial Gene Delivery and Protein Expressionmentioning

confidence: 99%