Abstract Body: Diabetes is a major risk factor for peripheral artery disease (PAD). Diabetes can induce profound epigenetic changes, including those mediated by long non-coding RNAs (lncRNAs) in endothelial cells (ECs), contributing to endothelial dysfunction and PAD. Combining (single cell-)RNA-sequencing and chromatin-RNA interactome mapping assays, we have identified chromatin-associated lncRNAs that are dysregulated by diabetic conditions (e.g., high glucose and TNFa) in ECs and contribute to impaired EC function and vascular injury. One of these is lncRNA-enhancing eNOS expression (LEENE), encoded by LINC00520. LEENE is downregulated in ECs by diabetic conditions in vitro and in vivo, in human and in mouse. LEENE interacts with RNA binding proteins at pro-angiogenic gene promoters, including eNOS and KDR, and enhances their gene expression, angiogenic response, and ischemic flow recovery. To test the therapeutic potential of targeting LEENE, we have modified LEENE RNA with N1m-pseudouridylation (same as in the Moderna COVID-19 mRNA vaccine) and tested the effect of the modified LEENE RNA (mod-LEENE) in vitro and in vivo. Mod-LEENE is localized to the nucleus of human ECs and promotes the transcription of eNOS and KDR. Local intramuscular injection of mod-LEENE promotes ischemic flow recovery in a mouse model of diabetic PAD. Our findings highlight a crucial role of LEENE in EC function, tissue perfusion, and ischemic injury and suggest the therapeutic potential of lncRNA-based interventions in mitigating diabetic PAD.