Perinatal Delivery of Angiotensin Type 2 Receptor (AT2R) Antisense cDNA Increases Blood Pressure in Adult Normotensive Rats
71 Recent observations suggest that AT 2 R have a role in the counter-regulatory actions of angiotensin II (Ang II) in cardiovascular protection and angiogenesis. The objective of the current investigation was to provide evidence for this cardioprotective action of AT 2 R by utilizing antisense gene transfer technology in normotensive rats. A retroviral vector containing full length AT 2 receptor antisense cDNA (LNSV-AT 2 R-AS) or missense (LNSV-AT 2 R-MS) was constructed. The LNSV-AT 2 R-AS viral particles were highly efficient in the transduction of AT 2 R-AS in vitro . The efficacy and effectiveness of this transduction was demonstrated by the long-lasting expression of AT 2 R-AS transcript and a decrease in AT 2 R binding. In vivo administration of LNSV-AT 2 R-AS resulted in similar findings. Five-day old normotensive Sprague Dawley rats received a single intracardiac bolus (25 μl) administration of LNSV-AT 2 R-AS viral particles (1x10 9 cfu/ml), which resulted in a robust expression of AT 2 R-AS transcript in tissues such as heart, kidney, adrenals and brain as early as five days post-delivery. Control rats received either LNSV alone or LNSV-AT 2 R-MS under identical conditions. The expression of AT 2 R-AS was persistent through adulthood indicating a high degree of transgene transduction in vivo . Mean blood pressure (BP) was elevated in the adult LNSV-AT 2 R-AS-treated rats when compared to the age-matched LNSV-AT 2 R-MS or control rats (123±5 mmHg vs. 100±9 mmHg). In addition, the pressor responses produced by Ang I and Ang II were enhanced in the LNSV-AT 2 R-AS- treated rats. For example, administration of 0.1 μg/Kg Ang I elicited a maximal increase in BP of 35±6 mmHg in the LNSV-AT 2 R-AS-treated rats compared to an increase of 23±6 mmHg in the LNSV-AT 2 R-MS. These observations demonstrate for the first time, that persistant inhibition of AT 2 R in normotensive rats influence cardiovascular responsiveness. Collectively, these data suggest that use of the AT 1 receptor antagonist-based therapy, with the resultant increase in Ang II levels, might provide additional benefit to the hypertensive patient via increased AT 2 R stimulation.