Distinct Transcriptomic Adaptations in Coronary and Cerebral Arterioles in Female Ossabaw Swine with Cardiometabolic Heart Failure Treated with a RSK3/mAKAPβ Gene Therapy

Cardiogenic dementia is associated with an increased risk of mortality in heart failure patients. Cognitive decline is more prevalent in females compared to males, of which cardiac function is the strongest predictor. We have previously demonstrated cerebrovascular insufficiency in female Ossabaw swine with cardiometabolic heart failure (CM-HF), and further shown improved cardiac function and ventricular-vascular interactions by a novel RSK3/mAKAPβ targeted gene therapy. The RSK3/mAKAPβ anchoring disruptor peptide (RBD) displaces RSK3 on mAKAPβ by adeno-associated virus (AAV)-based expression of RBD (AAV9sc.RBD). The goal of this study was to examine the effects of RBD gene therapy on transcriptomic signaling in both coronary and cerebral arterioles in an Ossabaw swine model of CM-HF. Female Ossabaw pigs were assigned to HF control (HF; n=4-5) and HF AAV9sc.RBD-treated (HF+RBD; n=4-7). Animals were fed a Western diet (3 mo. old) and aortic banded (6 mo. old) prior to terminal experiments (12 mo. old). Peripheral intravenous saline containing 3 x 10 14 viral genomes (vg) of AAV9sc.RBD biologic was infused immediately after aortic banding. Wet lung weight was decreased in HF+RBD animals compared to HF (283±19 vs. 423±64 g; unpaired t-test, p<0.05) and was associated with improved diastolic function evident by a decrease in the end diastolic pressure-volume relationship (EDPVR: 0.039±0.004 vs. 0.022±0.003 mmHg/mL; unpaired t-test, p<0.05). RBD also increased stroke volume (40±2 vs. 50±2 mL; unpaired t-test, p<0.05) and improved hemodynamic coupling between the left ventricle and peripheral vasculature. RBD increased endothelial-dependent dilation to bradykinin (BK) compared to HF (Two-way ANOVA, Group x Dose interaction; p<0.05) in isolated second order (2A) pial arterioles, which was unaltered in the presence of L-NAME (nitric oxide synthase inhibitor) and indomethacin (prostaglandin synthesis inhibitor). RNA sequencing identified 26 differentially expressed genes (DEGs) in cerebral arterioles and 11 DEGs in coronary arterioles. Ingenuity pathway analysis (IPA) on cerebral arterioles predicted inhibition of upstream regulator POR (z-score > -2), a gene associated with decreased fibrosis and organismal death, as well as increased sterol synthesis and quantity of leukocytes. Whereas gene enrichment of the coronary arterioles predicted cytokine (TNF, z-score=1.96, and IL1B, z-score=1.98) and growth factor activation (TGFB1, z-score=1.99). These findings demonstrate RBD improvement in coronary and cerebral vascular function occurs through distinct transcriptomic adaptations involving pathways associated with fibrosis, leukocyte recruitment, cytokine production, and growth factor activation. MU Tier 1 Sequencing Grant and Department of Defense (DOD) Grant W81XWH-18-1-0179 This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.