Abstract 18998: Deletion of the Duffy Antigen Receptor for Chemokines Impairs Blood Flow Recovery in the Mouse Hindlimb Ischemia Model: Role of CCL11

Abstract

Introduction: Peripheral Artery Disease (PAD) is characterized by obstruction of the arteries in the lower extremities. Impaired angiogenesis plays a major role in the progression of PAD. Pro-angiogenic and anti-angiogenic chemokines are modulated by binding to the Duffy antigen receptor for chemokines (DARC), a non-signaling receptor expressed primarily on erythrocytes. Incidence and severity of PAD is greater in Blacks, ~70% of whom do not express DARC on their erythrocytes due to a common gene variant. Here, we tested the hypothesis that hematopoietic-specific deletion of DARC impairs blood flow recovery in mouse hindlimb ischemia model. Methods: Twelve week old female DARC Flox/Vav1-Cre+ (DARC HKO ) and littermate Vav1-Cre- (wild type, WT) mice underwent unilateral femoral artery ligation and excision. Ischemic muscle tissue and plasma were analyzed at early ischemic phase (day three post ligation) and late ischemic phase (day fourteen post ligation). Blood flow recovery was measured weekly via laser Doppler imaging. Results: DARC HKO mice had poor blood flow recovery compared to WT mice (p<0.05), along with reduced capillary formation and increased inflammation, necrosis and fibrosis in the ischemic gastrocnemius muscles. Amongst the various DARC-bound chemokines assayed, levels of CCL11 (a pro-angiogenic strong DARC-bound chemokine) were significantly reduced in the plasma of DARC HKO mice, both at early and late ischemic phases (p<0.01). CCL11 levels strongly correlated with capillary formation in ischemic hindlimb. CCL11 levels in ischemic muscle tissue were significantly reduced in the DARC HKO mice in the early, but not late, ischemic phase. Conclusions: Loss of DARC on hematopoietic cells has a negative impact on the recovery of perfusion and angiogenesis in the murine hindlimb ischemia model, possibly due to reduced levels of CCL11 consequent to DARC deletion. These findings may be relevant to ethnic differences in susceptibility to PAD and suggest that therapies directed towards increasing CCL11 may be beneficial in Blacks with PAD.