Abstract Body: Peripheral arterial disease (PAD) is a major cause of limb ischemia and amputation, with limited therapeutic options that effectively restore microvascular perfusion. Although revascularization procedures improve large-artery blood flow, impaired angiogenic responses and endothelial dysfunction frequently limit tissue reperfusion, particularly in diabetes. Long non-coding RNAs (lncRNAs) have emerged as important regulators of endothelial gene expression and vascular repair, yet their therapeutic potential in PAD remains largely unexplored.
We previously identified the lncRNA that enhances endothelial nitric oxide synthase (eNOS) expression (LEENE) as a hypoxia-induced regulator of angiogenesis that promotes endothelial function through transcriptional activation of eNOS and KDR. In the current study, we investigated the therapeutic potential of LEENE in promoting ischemic limb recovery using RNA modification and monocyte-derived nanoparticles (MoNPs) that direct endothelial-specific delivery.
A chemically modified human LEENE transcript was encapsulated within MoNPs and administered in murine models of hindlimb ischemia. MoNP-mediated delivery significantly enhanced endothelial uptake of LEENE and improved post-ischemic reperfusion compared with free LEENE RNA and adenoviral LEENE (Ad-LEENE). To define cell-type–specific transcriptional responses, we performed single-cell RNA sequencing of ischemic skeletal muscle receiving LEENE through different approaches. These analyses revealed distinct endothelial transcriptional programs associated with MoNP-based delivery, including enhanced activation of angiogenic and vascular repair pathways.
Collectively, these findings demonstrate that targeted delivery of LEENE via monocyte-derived nanoparticles enhances endothelial repair and perfusion recovery in ischemic muscle. This work substantiates LEENE as a key regulator of ischemic recovery and supports MoNP as a promising platform for lncRNA-based vascular therapeutics in PAD.