Identification and functional characterization of arginine vasopressin receptor 1A: atypical chemokine receptor 3 heteromers in vascular smooth muscle

Recent observations suggest that atypical chemokine receptor (ACKR) 3 and chemokine (C-X-C motif) receptor (CXCR) 4 regulate human vascular smooth muscle function through hetero-oligomerization with alpha(1)-adrenoceptors. Here, we show that ACKR3 also regulates arginine vasopressin receptor (AVPR) 1A. We observed that ACKR3 agonists inhibit arginine vasopressin (aVP)-induced inositol trisphosphate (IP3) production in human vascular smooth muscle cells (hVSMCs) and antagonize aVP-mediated constriction of isolated arteries. Proximity ligation assays, co-immunoprecipitation and bioluminescence resonance energy transfer experiments suggested that recombinant and endogenous ACKR3 and AVPR1A interact on the cell surface. Interference with ACKR3 : AVPR1A heteromerization using siRNA and peptide analogues of transmembrane domains of ACKR3 abolished aVP-induced IP3 production. aVP stimulation resulted in beta-arrestin 2 recruitment to AVPR1A and ACKR3. While ACKR3 activation failed to cross-recruit beta-arrestin 2 to AVPR1A, the presence of ACKR3 reduced the efficacy of aVP-induced beta-arrestin 2 recruitment to AVPR1A. AVPR1A and ACKR3 co-internalized upon agonist stimulation in hVSMC. These data suggest that AVPR1A : ACKR3 heteromers are constitutively expressed in hVSMC, provide insights into molecular events at the heteromeric receptor complex, and offer a mechanistic basis for interactions between the innate immune and vasoactive neurohormonal systems. Our findings suggest that ACKR3 is a regulator of vascular smooth muscle function and a possible drug target in diseases associated with impaired vascular reactivity.