Activating P2Y 1 receptors improves function in arteries with repressed autophagy

• Published in: The FASEB journal Vol. 36 Suppl 1
• Main Authors: Cho, Jae Min, Park, Seul-Ki, Kwon, Oh Sung, La Salle, D T, Cerbie, James, Fermoyle, Caitlin C, Morgan, David, Nelson, Ashley, Bledsoe, Amber, Bharath, Leena P, Tandar, Megan, Kunapuli, Satya P, Richardson, Russell S, Babu, Pon Velayutham Anandh, Mookherjee, Sohom, Kishore, Bellamkonda K, Wang, Fei, Yang, Tianxin, Boudina, Sihem, Trinity, Joel D, Symons, J D
• Format: Journal Article
• Language: English
• Published: United States 01.05.2022
• ISSN: 1530-6860
• DOI: 10.1096/fasebj.2022.36.S1.R4362

The importance of endothelial cell (EC) autophagy to vascular homeostasis in the context of health and disease is evolving. Earlier we reported that intact EC autophagy is requisite to maintain shear-stress-induced nitric oxide (NO) generation via glycolysis-dependent purinergic signaling to eNOS. Here we illustrate the translational and functional significance of these findings. First, we assessed translational relevance using older male humans and mice that exhibit blunted EC autophagy and impaired arterial function vs. adult controls. Active hyperemia evoked by rhythmic handgrip exercise elevated radial artery shear rate similarly from baseline in adult (23±1 y) and older (68±2 y) subjects for 60-min. Compared to baseline, indexes of autophagy initiation, p-eNOS activation, and NO generation, occurred in radial artery ECs obtained from adult but not older volunteers. Regarding mice, indexes of autophagy and p-eNOS activation were robust in ECs from adult (7 ± 1 months) but not older (23 ± 1 months) animals that completed 60-min treadmill-running. Further, results concerning the extracellular acidification rate (ECAR; Seahorse Bioanalyzer) indicate glycolysis and glycolytic capacity were elevated in response to 20 dyn/cm laminar shear stress x 45-min in primary arterial ECs obtained from adult but not older mice. We next questioned whether the inability to initiate EC autophagy, glycolysis, and p-eNOS precipitates dysfunction in arteries from older vs. adult mice. Compromised intraluminal flow-mediated vasodilation displayed by arteries from older vs. adult mice was recapitulated in vessels from adult mice by : (i) NO synthase inhibition; (ii) acute autophagy impairment using 3-methyladenine (3-MA); (iii) EC Atg3 depletion (Atg3 mice); (iv) purinergic 2Y -receptor (P2Y -R) blockade; and (v) germline depletion of P2Y -Rs. Importantly, P2Y -R activation using 2-methylthio-ADP (2-Me-ADP) improved vasodilatory capacity in arteries from : (i) adult mice treated with 3-MA; (ii) adult Atg3 mice; and (iii) older animals with repressed EC autophagy. Arterial dysfunction concurrent with pharmacological, genetic, and age-associated EC autophagy disruption is improved by activating P2Y -Rs.