AHA Scientific Sessions 2026 Abstract · Nov 2, 2026
Inhibition of G Protein-Coupled Receptor 35 Improves Ischemic Recovery in Aged Mice Through Regulation of Long Non-Coding RNAs in Endothelial Cells
By Li H, Shvartsman S, Wang JM
Introduction: Aging is a crucial factor in peripheral artery disease (PAD) and is associated with impaired endothelial function. Recent evidence indicates that a poorly characterized G protein-coupled receptor (GPR), GPR35, is elevated in endothelial cell (EC) aging and potentially contributes to EC dysfunction, but the underlying mechanisms remain largely unknown. Long non-coding RNAs (lncRNAs) play vital roles in regulating cellular function, but limited information exists on how lncRNAs are modulated in PAD-related limb ischemia in aging. Hypothesis: We hypothesized that GPR35 inhibition promotes EC angiogenesis, thereby enhancing revascularization in aged animals with hind limb ischemia through modulating lncRNA expression. Methods and Results: In a hind limb ischemia induced by unilateral femoral artery ligation, using Laser Doppler imaging, we discovered that the injection of adenovirus carrying shRNA against mouse Gpr35 (Ad-shGpr35, using Ad-Scramble as control) at the ischemic site significantly accelerated the recovery of blood perfusion in aged mice, catching up with the recovery rate in young mice (male C57BL/6, aged: 24 months; young: 3 months, n = 5-6). This was consistent with CD31 immunohistology, suggesting denser capillary formation in the recovering muscle. Mouse ECs were isolated from Gpr35 global knockout (Gpr35 KO ) and their age- and gender-matched wild-type control (Gpr35 WT ) litters. Compared to Gpr35 WT ECs, Gpr35 KO ECs exhibited much more robust proliferation, migration, and 3D tube formation (n =5). Human skeletal muscle microvascular ECs (HSMMVECs) were cultured and infected with an adenovirus carrying the human GPR35 gene or green fluorescent protein control. The Transwell assay showed that overexpression of GPR35 impaired the migration of HSMMVECs. The analyses of lncRNA and mRNA profiles in Gpr35 KO and Gpr35 WT ECs were performed using Arraystar Mouse lncRNA microarray v3.0. The results indicated 343 lncRNAs were significantly upregulated and 177 were significantly downregulated in Gpr35 KO ECs. Several endothelial-enriched lncRNAs with pro-angiogenic properties, including Neat1, Gas5, and STEEL, were upregulated in Gpr35 KO ECs as identified through microarray and confirmed by real-time PCRs. Notably, Stmn1 and its pseudogene, lncRNA Gm11223, were both among the top upregulated genes in Gpr35 KO ECs. Gene Ontology analyses revealed that GPR35 deletion upregulated genes involved in cell division, cell cycle, DNA metabolic processes, and mismatch repair. Conclusion: This study provides novel evidence that suppressing GPR35 improves EC angiogenesis and subsequently enhances ischemic recovery in aged mice, which is related to a shift in lncRNA profile, suggesting a potential role for GPR35 in modulating the lncRNA/mRNA axis in PAD in the aged population.