Conditional and targeted overexpression of vascular chymase causes hypertension in transgenic mice.
We cloned a rat vascular chymase (RVCH) from smooth muscle cells (SMCs) that converts angiotensin I to II and is up-regulated in SMC from spontaneously hypertensive vs. normotensive rats. To determine whether increased activity of RVCH is sufficient to cause hypertension, transgenic mice were generated with targeted conditional expression of RVCH to SMC, with the use of the tetracycline-controlled transactivator (tTA). We confirmed conditional expression of RVCH by mRNA, protein, and chymase activity in the absence, but not in the presence, of dietary doxycycline. The systolic blood pressure (mmHg), measured by carotid artery cannulation at 10-12 weeks of age, was higher in tTA+/RVCH+ mice than in nonbinary transgenic littermates (136 +/- 4 vs. 109 +/- 3) (P < 0.05), as were the diastolic and mean pressures. Hypertension was completely reversed by doxycycline, suggesting a causal link with chymase expression. Medial thickening of mesenteric arteries from tTA+/RVCH+ mice vs. littermates (0.82 +/- 0.1 vs. 0.42 +/- 0.02) (P < 0.05) was associated with increased SMC proliferation, as judged by positive immunoreactivity, with the use of an antibody to the proliferating cell nuclear antigen. These structural changes were prevented by doxycycline. Perfusion myography of mesenteric arteries from tTA+/RVCH+ mice also revealed increased vasoconstriction in response to phenylephrine and impaired metacholine-induced vasodilatation when compared with littermate controls or with the doxycyline-treated group. Our studies suggest that up-regulation of this vascular chymase is sufficient to cause a hypertensive arteriopathy, and that RVCH may be a candidate gene and a therapeutic target in patients with high blood pressure.
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Division of Cardiovascular Research, Research Institute, The Hospital for Sick Children, Center for Cardiovascular Research, The Toronto Hospital, University of Toronto, Toronto, ON, Canada M5G 1X8.
Gros
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You
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Tsang
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Husain
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Rabinovitch
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D000818:Animals; D001011:Aorta / Q000201:enzymology; D001794:Blood Pressure / Q000235:genetics / Q000502:physiology*; D053818:Chymases; D003001:Cloning, Molecular; D006973:Hypertension / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D009924:Organ Culture Techniques; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
D012333:RNA, Messenger; D012697:Serine Endopeptidases; D053818:Chymases