PMID- 38077163 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20240216 IS - 2666-3503 (Electronic) IS - 2666-3503 (Linking) VI - 4 DP - 2023 TI - Ceramides in peripheral arterial plaque lead to endothelial cell dysfunction. PG - 100181 LID - 10.1016/j.jvssci.2023.100181 [doi] LID - 100181 AB - BACKGROUND: Peripheral arterial atheroprogression is increasingly prevalent, and is a risk factor for major limb amputations in individuals with risk factors such as diabetes. We previously demonstrated that bioactive lipids are significantly altered in arterial tissue of individuals with diabetes and advanced peripheral arterial disease. METHODS: Here we evaluated whether sphingolipid ceramide 18:1/16:0 (C16) is a cellular regulator in endothelial cells and peripheral tibial arterial tissue in individuals with diabetes. RESULTS: We observed that C16 is the single most elevated ceramide in peripheral arterial tissue from below the knee in individuals with diabetes (11% increase, P < .05). C16 content in tibial arterial tissue positively correlates with sphingomyelin (SPM) content in patients with and without diabetes (r(2) = 0.5, P < .005; r(2) = 0.17, P < .05; respectively). Tibial arteries of individuals with diabetes demonstrated no difference in CERS6 expression (encoding ceramide synthase 6; the predominate ceramide synthesis enzyme), but higher SMPD expression (encoding sphingomyelin phosphodiesterase that catalyzes ceramide synthesis from sphingomyelins; P < .05). SMPD4, but not SMPD2, was particularly elevated in maximally diseased (Max) tibial arterial segments (P < .05). In vitro, exogenous C16 caused endothelial cells (HUVECs) to have decreased proliferation (P < .03), increased apoptosis (P < .003), and decreased autophagy (P < .008). Selective knockdown of SMPD2 and SMPD4 decreased native production of C16 (P < .01 and P < .001, respectively), but only knockdown of SMPD4 rescued cellular proliferation (P < .005) following exogenous supplementation with C16. CONCLUSIONS: Our findings suggest that C16 is a tissue biomarker for peripheral arterial disease severity in the setting of diabetes, and can impact endothelial cell viability and function. CLINICAL RELEVANCE: Peripheral arterial disease and its end-stage manifestation known as chronic limb-threatening ischemia (CLTI) represent ongoing prevalent and intricate medical challenges. Individuals with diabetes have a heightened risk of developing CLTI and experiencing its complications, including wounds, ulcers, and major amputations. In the present study, we conducted a comprehensive examination of the molecular lipid composition within arterial segments from individuals with CLTI, and with and without diabetes. Our investigations unveiled a striking revelation: the sphingolipid ceramide 18:1/16:0 emerged as the predominant ceramide species that was significantly elevated in the peripheral arterial intima below the knee in patients with diabetes. Moreover, this heightened ceramide presence is associated with a marked impairment of endothelial cell function and viability. Additionally, our study revealed a concurrent elevation in the expression of sphingomyelin phosphodiesterases, enzymes responsible for catalyzing ceramide synthesis from sphingomyelins, within maximally diseased arterial segments. These findings underscore the pivotal role of ceramides and their biosynthesis enzymes in the context of CLTI, offering new insights into potential therapeutic avenues for managing this challenging disease process. FAU - Meade, Rodrigo AU - Meade R AD - Section of Vascular Surgery, Department of Surgery, Washington University School of Medicine, St. Louis, MO. FAU - Chao, Yang AU - Chao Y AD - Section of Vascular Surgery, Department of Surgery, Washington University School of Medicine, St. Louis, MO. FAU - Harroun, Nikolai AU - Harroun N AD - Section of Vascular Surgery, Department of Surgery, Washington University School of Medicine, St. Louis, MO. FAU - Li, Chenglong AU - Li C AD - Section of Vascular Surgery, Department of Surgery, Washington University School of Medicine, St. Louis, MO. FAU - Hafezi, Shahab AU - Hafezi S AD - Section of Vascular Surgery, Department of Surgery, Washington University School of Medicine, St. Louis, MO. FAU - Hsu, Fong-Fu AU - Hsu FF AD - Division of Endocrinology, Lipid, and Metabolism, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO. FAU - Semenkovich, Clay F AU - Semenkovich CF AD - Division of Endocrinology, Lipid, and Metabolism, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO. FAU - Zayed, Mohamed A AU - Zayed MA AD - Section of Vascular Surgery, Department of Surgery, Washington University School of Medicine, St. Louis, MO. AD - Department of Surgery, Veterans Affairs St. Louis Health Care System, St. Louis, MO. AD - Department of Radiology, Washington University School of Medicine, St. Louis, MO. AD - Division of Molecular Cell Biology, Washington University School of Medicine, St. Louis, MO. AD - Department of Biomedical Engineering, Washington University, McKelvey School of Engineering, St. Louis, MO. LA - eng GR - R01 HL150891/HL/NHLBI NIH HHS/United States GR - P30 DK056341/DK/NIDDK NIH HHS/United States GR - P41 GM103422/GM/NIGMS NIH HHS/United States GR - R01 DK101392/DK/NIDDK NIH HHS/United States GR - P60 DK020579/DK/NIDDK NIH HHS/United States GR - P30 DK020579/DK/NIDDK NIH HHS/United States GR - R01 HL157154/HL/NHLBI NIH HHS/United States GR - R01 HL153262/HL/NHLBI NIH HHS/United States GR - K08 HL132060/HL/NHLBI NIH HHS/United States PT - Journal Article DEP - 20231111 PL - United States TA - JVS Vasc Sci JT - JVS-vascular science JID - 101767073 PMC - PMC10704331 OTO - NOTNLM OT - Ceramide OT - Diabetes OT - Endothelial dysfunction OT - Peripheral arterial disease OT - Sphingolipid OT - Sphingomyelinase COIS- None. EDAT- 2023/12/11 12:41 MHDA- 2023/12/11 12:42 PMCR- 2023/11/11 CRDT- 2023/12/11 06:25 PHST- 2023/07/06 00:00 [received] PHST- 2023/10/22 00:00 [accepted] PHST- 2023/12/11 12:42 [medline] PHST- 2023/12/11 12:41 [pubmed] PHST- 2023/12/11 06:25 [entrez] PHST- 2023/11/11 00:00 [pmc-release] AID - S2666-3503(23)00085-8 [pii] AID - 100181 [pii] AID - 10.1016/j.jvssci.2023.100181 [doi] PST - epublish SO - JVS Vasc Sci. 2023 Nov 11;4:100181. doi: 10.1016/j.jvssci.2023.100181. eCollection 2023.