AAV delivery of mineralocorticoid receptor shRNA prevents progression of cold-induced hypertension and attenuates renal damage.
The aim of this study was to determine the effect of RNA interference inhibition of mineralocorticoid receptor (MR) on cold-induced hypertension (CIH) and renal damage. Recombinant adeno-associated virus (AAV) carrying short hairpin small interference (si)RNA for MR (AAV.MR-shRNA) was constructed and tested for the ability to inhibit renal MR and to control CIH. Three groups of rats with CIH received AAV.MR-shRNA (1.25 x 10(9) particles/rat, intravenous), AAV carrying scrambled shRNA (AAV.Control-shRNA) (1.25 x 10(9) particles/rat, intravenous) and phosphate buffer solution (PBS), respectively. All rats were kept in a cold chamber (6.7 degrees C) throughout the experiment. Adeno-associated virus delivery of MR-shRNA prevented progression of CIH. Blood pressure (BP) of the AAV.MR-shRNA-treated group did not increase and remained at 145+/-3 mm Hg, whereas BP of the AAV.Control-shRNA-treated and PBS-treated group increased to 167+/-4 and 161+/-3 mm Hg, respectively, at 3 weeks after gene delivery. Thus, the antihypertensive effect of a single injection of AAV.MR-shRNA lasted for at least 3 weeks (length of the study). Adeno-associated virus carrying short hairpin siRNA for MR significantly increased urinary sodium excretion and decreased proteinuria. It also decreased serum creatinine and blood urea nitrogen, suggesting enhanced renal function. Both Western blot and immunohistochemical analysis showed that MR expression was decreased significantly in the kidney in the AAV.MR-shRNA-treated rats, confirming that renal MR is effectively inhibited by AAV.MR-shRNA. Adeno-associated virus carrying short hairpin siRNA for MR also significantly attenuated renal hypertrophy. In addition, AAV delivery of MR-shRNA prevented atrophy and dilation of renal tubules and abolished tubular deposition of proteinaceous material seen in CIH rats.


CONCLUSIONS
(1) AAV delivery of MR-shRNA effectively silenced MR in vivo. (2) RNA interference inhibition of MR may open a new avenue for the long-term control of hypertension and renal damage.
Department of Medicine, College of Medicine, University of Florida, Gainesville, 32610-0274, USA.
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Cade
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D000818:Animals; D001011:Aorta / Q000737:chemistry; D003080:Cold Temperature / Q000009:adverse effects*; D000229:Dependovirus / Q000235:genetics*; D018450:Disease Progression; D015316:Genetic Therapy / Q000379:methods*; D006973:Hypertension / Q000209:etiology* / Q000473:pathology / Q000628:therapy; D007150:Immunohistochemistry / Q000379:methods; D007668:Kidney / Q000737:chemistry / Q000473:pathology; D007674:Kidney Diseases / Q000473:pathology / Q000628:therapy; D009206:Myocardium / Q000737:chemistry; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011944:Receptors, Androgen / Q000032:analysis; D011965:Receptors, Glucocorticoid / Q000032:analysis; D018161:Receptors, Mineralocorticoid / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
D034741:RNA, Small Interfering; D011944:Receptors, Androgen; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid