Delivery Of Human Tissue Kallikrein Gene Research Articles

Recombinant adeno-associated virus-mediated human kallikrein gene therapy prevents high-salt diet-induced hypertension without effect on basal blood pressure1

To investigate the effects of the expression of human kallikrein (HK) on basal level blood pressure and high-salt diet-induced hypertension. We delivered the recombinant adeno-associated viral (rAAV)-mediated HK (rAAV- HK) gene and rAAV-LacZ (as the control) to normal, adult Sprague-Dawley rats. The animals were administered a normal diet in the first 4 weeks, followed by a high-salt diet. The expression of HK in the rats was assessed by ELISA and RT- PCR. Blood pressure and Na+ and K+ urinary excretion were monitored. Under the normal diet, no obvious changes in blood pressure and Na+ and K+ urinary excretion were observed. When the high-salt diet was administered, systolic blood pressure in the control animals receiving rAAV-LacZ increased from 122.3+/-1.13 mmHg to a stable 142.4+/-1.77 mmHg 8 weeks after the high-salt diet. In contrast, there was no significant increase in the blood pressure in the rAAV-HKtreated group, in which the blood pressure remained at 121.9+/-1.73 mmHg. In the rAAV-HK-treated group, Na+ and K+ urinary excretion were higher compared to those of the control group. The morphological analysis showed that HK delivery remarkably protected against renal damage induced by a high-salt intake. Our study indicates that rAAV-mediated human tissue kallikrein gene delivery is a potentially safe method for the long-term treatment of hypertension. More importantly, it could be applied in the salt-sensitive population to prevent the occurrence of hypertension.

Prolonged hypotensive effect of human tissue kallikrein gene delivery and recombinant enzyme administration in spontaneous hypertension rats.

To evaluate the feasibility of treating hypertension by human tissue kallikrein gene (KLK1) delivery and by enzyme (rK1) administration, two recombinant vectors expressing KLK1 cDNA were constructed for gene delivery (pcDNA-KLK1) and recombinant enzyme preparation (pOV-KLK1). Expression of the pcDNA-KLK1 vector in COS-1 cells was confirmed by immunofluorescence and in spontaneous hypertension rats (SHR) by enzymatic detection. Following intramuscular or intravenous injection with the pcDNA-KLK1 vector, systolic pressure of SHR was significantly decreased, which lasted for 20 d to two months depending on dose, route and/or time of injection. Egg white containing recombinant hK1 was prepared by injection of egg-laying hens with the oviduct-specific expression vector pOV-KLK1 and administered into SHR via oral gavage. Following administration, systolic pressure of the SHR was decreased to that of normal rats, which lasted for 3-5 d depending on the dosage used. These data suggest that both hKLK1 gene delivery and recombinant enzyme administration can be used as alternative strategies for treating human hypertension.

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HomeArteriosclerosis, Thrombosis, and Vascular BiologyVol. 22, No. 10Correction Free AccessCorrectionPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessCorrectionPDF/EPUBCorrection Originally published15 Mar 2018https://doi.org/10.1161/atvb.22.10.1742Arteriosclerosis, Thrombosis, and Vascular Biology. 2002;22:1742This article corrects the followingAdenovirus-Mediated Human Tissue Kallikrein Gene Delivery Induces Angiogenesis in Normoperfused Skeletal MuscleIn “Adenovirus-Mediated Human Tissue Kallikrein Gene Delivery Induces Angiogenesis in Normoperfused Skeletal Muscle” by Emanueli et al (Arterioscler Thromb Vasc Biol. 2000;20;2379–2385), Figure 3B was provided in error by the authors.The correct figure is now provided. Download figureDownload PowerPointRepresentative picture (hematoxylin and eosin staining) showing the capillary density of adductor muscle injected with Ad.CMV-chKall. Magnification ×100. Previous Back to top Next FiguresReferencesRelatedDetailsRelated articlesAdenovirus-Mediated Human Tissue Kallikrein Gene Delivery Induces Angiogenesis in Normoperfused Skeletal MuscleCostanza Emanueli, et al. Arteriosclerosis, Thrombosis, and Vascular Biology. 2000;20:2379-2385 October 2002Vol 22, Issue 10 Advertisement Article InformationMetrics https://doi.org/10.1161/atvb.22.10.1742 Originally publishedMarch 15, 2018 PDF download Advertisement

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HomeArteriosclerosis, Thrombosis, and Vascular BiologyVol. 20, No. 11Correction Free AccessCorrectionPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessCorrectionPDF/EPUBCorrection Originally published1 Nov 2000https://doi.org/10.1161/01.ATV.20.11.2500Arteriosclerosis, Thrombosis, and Vascular Biology. 2000;20:2500This article corrects the followingAdenovirus-Mediated Human Tissue Kallikrein Gene Delivery Inhibits Neointima Formation Induced by Interruption of Blood Flow in MiceIn the June 2000 issue of Arteriosclerosis, Thrombosis, and Vascular Biology in the article by Emanueli et al (Adenovirus-mediated human tissue kallikrein gene delivery inhibits neointima formation induced by interruption of blood flow in mice; pp 1459–1466), an error appeared in the byline and affiliation notice. The correct authors and their affiliations for the aforementioned article appear below.Costanza Emanueli,*† Maria Bonaria Salis,*† Julie Chao,§ Lee Chao,§ Jun Agata,§ Kuei-Fu Lin,§ Antonella Munaò,*† Stefania Straino,* Alessandra Minasi,* Maurizio C. Capogrossi,* Paolo Madeddu*†‡*Laboratorio di Patologia Vascolare, Istituto Dermopatico dell’Immacolata, Istituto di ricovero e Cura a Carattere Scientifico, Rome, Italy; †Gene Therapy Section of the National Laboratory of the National Institute of Biostructures and Biosystems, Osilo, Italy; ‡Institute of Internal Medicine, School of Medicine, University of Sassari, Sassari, Italy; and §Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston. Previous Back to top Next FiguresReferencesRelatedDetailsCited By Kurotaki T, Sumi T, Atobe T and Hiyama J (2008) Numerical Simulation Around NACA0015 with Tonal Noise Generation 46th AIAA Aerospace Sciences Meeting and Exhibit, 10.2514/6.2008-672, 978-1-62410-128-1, Online publication date: 7-Jan-2008. Alred J, Visentine J and Albyn K (2001) Summary of Shuttle-MIR risk mitigation experiments contamination/optical observations 39th Aerospace Sciences Meeting and Exhibit, 10.2514/6.2001-97, , Online publication date: 8-Jan-2001. Okabe M and Sakakibara M (2009) Practical Measurement and Evaluation of Thermal Behavior of NC Lathe and its Application to Turning Process Modification, Key Engineering Materials, 10.4028/www.scientific.net/KEM.407-408.59, 407-408, (59-62) Related articlesAdenovirus-Mediated Human Tissue Kallikrein Gene Delivery Inhibits Neointima Formation Induced by Interruption of Blood Flow in MiceCostanza Emanueli, et al. Arteriosclerosis, Thrombosis, and Vascular Biology. 2000;20:1459-1466 November 2000Vol 20, Issue 11 Advertisement Article InformationMetrics https://doi.org/10.1161/01.ATV.20.11.2500 Originally publishedNovember 1, 2000 PDF download Advertisement

Adenovirus-mediated human tissue kallikrein gene delivery induces angiogenesis in normoperfused skeletal muscle.

We investigated whether local delivery of the tissue kallikrein gene induces angiogenesis in normoperfused mouse hindlimb muscles. Intramuscular injection of adenovirus containing the human tissue kallikrein gene under the control of a cytomegalovirus enhancer/promoter sequence resulted in local production and release of recombinant human tissue kallikrein, whereas transgene expression was absent in muscles of the contralateral hindlimb. Angiogenesis in infected muscles was documented by histological evidence of increased capillary density. In contrast, no angiogenic effect was seen either in the ipsilateral gastrocnemius or contralateral hindlimb muscles. Neovascularization was associated with a transient increase in muscular blood flow as determined by laser Doppler flowmetry. We also investigated the mechanisms of kallikrein-induced angiogenesis. We found that the angiogenic response to kallikrein was abolished by chronic blockade of the kinin B(1) or B(2) receptor or by inhibition of nitric oxide synthase. In addition, inhibition of cyclooxygenase-2 by nimesulide significantly reduced kallikrein-induced effects. These results indicate that (1) human tissue kallikrein acts as an angiogenic factor in normoperfused skeletal muscle and (2) nitric oxide and prostacyclin are essential mediators of kallikrein-induced angiogenesis. Our findings provide new insights into the role of the tissue kallikrein-kinin system in vascular biology.

Adenovirus-Mediated Human Kallikrein Gene Delivery Increases Endogenous Rat Tissue Kallikrein in Hypertensive Rat Models

P201 Our recent studies showed that adenovirus-mediated human tissue kallikrein gene delivery attenuated hypertension, cardiac hypertrophy, and renal injury in various hypertensive rat models. However, mechanisms underlying the beneficial effects of human kallikrein gene delivery are not clear. In the present study, we investigated the potential of human kallikrein gene delivery on synthesis and excretion of endogenous rat tissue kallikrein in normotensive and hypertensive rat models. Adenovirus carrying the human kallikrein cDNA under the control of cytomegalovirus (CMV) promoter/enhancer was administered via tail vein injection. Control rats received adenovirus harboring the b-galactosidase or luciferase gene under control of the CMV promoter/enhancer. A single injection of the human tissue kallikrein gene caused a prolonged blood pressure-lowering effect in spontaneously hypertensive rats, two kidney-one clip, Dahl salt-sensitive, deoxycorticosterone acetate-salt and monocrotaline-induced pulmonary hypertensive rats, while systolic blood pressure of normotensive Sprague-Dawley rats remained unaltered. Human kallikrein gene delivery significantly increased endogenous rat tissue kallikrein levels in the kidney and urine of different models of hypertensive rats as compared with their respective controls. However, urinary excretion of rat tissue kallikrein was not affected by human kallikrein gene delivery in normotensive rats. The endogenous rat tissue kallikrein mRNA expression in the kidney remained unaltered after human kallikrein gene delivery, which indicated that expression of human kallikrein in hypertensive rats increased endogenous rat kallikrein synthesis and excretion. These findings indicate that beneficial effects of human kallikrein gene delivery in hypertensive rat models are mediated, at least in part, by enhanced endogenous tissue kallikrein-kinin system.

Adenovirus-Mediated Human Kallikrein Gene Delivery Increases Endogenous Rat Tissue Kallikrein in Hypertensive Rat Models

P201 Our recent studies showed that adenovirus-mediated human tissue kallikrein gene delivery attenuated hypertension, cardiac hypertrophy, and renal injury in various hypertensive rat models. However, mechanisms underlying the beneficial effects of human kallikrein gene delivery are not clear. In the present study, we investigated the potential of human kallikrein gene delivery on synthesis and excretion of endogenous rat tissue kallikrein in normotensive and hypertensive rat models. Adenovirus carrying the human kallikrein cDNA under the control of cytomegalovirus (CMV) promoter/enhancer was administered via tail vein injection. Control rats received adenovirus harboring the b-galactosidase or luciferase gene under control of the CMV promoter/enhancer. A single injection of the human tissue kallikrein gene caused a prolonged blood pressure-lowering effect in spontaneously hypertensive rats, two kidney-one clip, Dahl salt-sensitive, deoxycorticosterone acetate-salt and monocrotaline-induced pulmonary hypertensive rats, while systolic blood pressure of normotensive Sprague-Dawley rats remained unaltered. Human kallikrein gene delivery significantly increased endogenous rat tissue kallikrein levels in the kidney and urine of different models of hypertensive rats as compared with their respective controls. However, urinary excretion of rat tissue kallikrein was not affected by human kallikrein gene delivery in normotensive rats. The endogenous rat tissue kallikrein mRNA expression in the kidney remained unaltered after human kallikrein gene delivery, which indicated that expression of human kallikrein in hypertensive rats increased endogenous rat kallikrein synthesis and excretion. These findings indicate that beneficial effects of human kallikrein gene delivery in hypertensive rat models are mediated, at least in part, by enhanced endogenous tissue kallikrein-kinin system.

Human tissue kallikrein gene delivery attenuates hypertension, renal injury, and cardiac remodeling in chronic renal failure

Tissue kallikrein cleaves kininogen substrate to produce the potent vasodilating peptide kinin, which plays important roles in cardiovascular and renal function. To explore cardiac and renal potential protective effects of kallikrein gene delivery in chronic renal failure, we delivered adenovirus carrying the human tissue kallikrein cDNA (cHK) into rats with 5/6 reduction of renal mass. Expression of human tissue kallikrein in rats was assessed by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR)/Southern blotting. Physiological parameters monitored in rats included systolic blood pressure, heart rate, and urinary excretion of protein, albumin, kinin, cGMP, cAMP, and nitrate/nitrites. Systemic and regional hemodynamics were measured by fluorescent-labeled microspheres. Heart weight and myocyte diameter were used to assess left ventricular hypertrophy. Quantitative and qualitative morphological analyses were used to evaluate histologic changes in kidney and heart sections. Active tissue kallikrein reached a peak serum level of 463 +/- 76 ng/mL following gene delivery and returned to control levels within 21 days. A maximal blood pressure reduction of 37 mm Hg was observed within one week in rats receiving kallikrein gene delivery as compared with control rats receiving adenovirus containing the luciferase gene (159 +/- 5 vs. 196 +/- 6 mm Hg, N = 15, P < 0.001), and a significant blood pressure difference continued for five weeks postgene delivery. Kallikrein gene delivery significantly decreased total urinary protein and albumin excretion and increased levels of urinary kinin, nitrite/nitrate, and cGMP levels. Cardiac output and regional blood flow were also increased, while peripheral vascular resistance decreased. Kallikrein gene transfer reduced glomerular sclerotic lesions, tubular damage, lumenal protein cast accumulation, and interstitial inflammation in the kidney. Myocardial hypertrophy and fibrosis were also attenuated in rats receiving kallikrein gene delivery. These findings indicated that kallikrein gene delivery attenuates hypertension and protects against renal injury and cardiac remodeling in the rat remnant kidney model of chronic renal failure.

Adenovirus-mediated human tissue kallikrein gene delivery inhibits neointima formation induced by interruption of blood flow in mice.

Tissue kallikrein cleaves kininogen to produce vasoactive kinin peptides. Binding of kinins to bradykinin B(2) receptors on vascular endothelial cells stimulates the release of nitric oxide and prostacyclin, thus activating the cGMP and cAMP pathways. In this study, we evaluated the effects of adenovirus-mediated human tissue kallikrein gene (Ad.CMV-cHK) delivery in a mouse model of arterial remodeling induced by permanent alteration in shear stress conditions. Mice underwent ligature of the left common carotid artery and were injected intravenously with saline or 1.8 x 10(9) plaque-forming units of Ad.CMV-cHK or control virus (Ad.CMV-LacZ). Fourteen days after surgery, morphometric analysis revealed that Ad. CMV-cHK reduced neointima formation by 52% (P<0.05) compared with Ad. CMV-LacZ. Expression of human tissue kallikrein (HK) mRNA was detected in mouse carotid artery, aorta, kidney, heart, and liver, and recombinant HK was present in the urine and plasma of mice receiving HK gene. Kallikrein gene transfer resulted in increases in urinary kinin, cGMP, and cAMP levels. The protective action of Ad. CMV-cHK on neointima formation was significantly reduced (P<0.05) in mice with knockout of the kinin B(2) receptor gene compared with wild-type control mice (J129Sv mice). In contrast, the effect of Ad. CMV-cHK was amplified (P<0.05) in transgenic mice overexpressing human B(2) receptor compared with wild-type control mice (c57/Bl6 mice). Thus, the inhibitory effect of recombinant kallikrein on structural alterations caused by the interruption of blood flow appears to be mediated by the B(2) receptor. These results provide new insight into the role of the tissue kallikrein-kinin system in vascular remodeling and suggest the application of HK gene therapy to treat restenosis and atherosclerosis.

Adenovirus-mediated kallikrein gene transfer inhibits neointima formation via increased production of nitric oxide in rat artery

Tissue kallikrein cleaves kininogen substrate to produce vasoactive kinin peptides that have been implicated to play a role in the proliferation of vascular smooth muscle cells (VSMC). In order to explore potential roles of the kallikrein–kinin system in vascular biology, we evaluated the effects of adenovirus-mediated kallikrein gene delivery on neointima formation in balloon-injured rat artery. Infection of isolated rat aortic segments with adenovirus containing the human tissue kallikrein gene resulted in a time-dependent secretion of recombinant human tissue kallikrein, and significant increases in nitric oxide (NOx) and guanosine 3′,5′-cyclic monophosphate (cGMP) levels post gene transfer. Human tissue kallikrein gene was delivered locally via adenoviral vectors into left common carotid artery after balloon angioplasty. Two weeks following gene transfer, we observed a 39% reduction in intima/media ratio at the injured vessel as compared to that of rats receiving control virus (n=8, P<.01). Delivery of Nω-nitro-l-arginine methyl ester (l-NAME), a NOx synthase inhibitor via minipump for 2 weeks, blocked the protective effect and reversed the intima/media ratio to that of control rats (n=5, P<.01). These results indicated that human tissue kallikrein gene delivery inhibits neointima formation via NO-cGMP signaling pathway. This study provides new insights into the role of the vascular kallikrein–kinin system and may have significant implications for gene therapy in treating occlusive vascular diseases.