Slc7a11-Mediated Cystine/Glutamate Antiport Reprograms Macrophage Polarization and Ameliorates Atherosclerosis

Abstract

Atherosclerotic cardiovascular diseases (ASCVDs) remain the primary cause of morbidity and mortality. Macrophages are involved in the progression and regression of atherosclerosis, and macrophage amino acid metabolism is important during this process. Here, we identified that the expression of cystine/glutamate antiporter Slc7a11 was upregulated by oxidized low-density lipoprotein, and specifically enhanced in the macrophages of atherosclerotic plaques. Macrophage-specific Slc7a11 overexpression in ApoE null mice (ApoE^–/–Slc7a11^MOE) attenuated atherosclerotic lesions and increased the plaque stability under a 16-week western diet. ApoE^–/–Slc7a11^MOE displayed unchanged blood lipids, decreased inflammatory cytokines, and increased antioxidant capacity. Mechanistically, Slc7a11-mediated cystine uptake and glutathione synthesis inhibited the classically activated macrophage (M1) polarization via reducing Stat1 phosphorylation, and promoted alternatively activated macrophage (M2) polarization via enhancing Stat6 phosphorylation. Macrophage-targeting lipid nanoparticles loading with ferrostatin-1, an antioxidant reagent, promotes Slc7a11-mediated glutathione synthesis, also enhanced plaque stability and ameliorated the progression of atherosclerosis. These findings reveal a critical role of Slc7a11 in the phenotypic switch of macrophage and indicate that Slc7a11-mediated amino acid metabolism could be utilized as a novel therapeutic strategy in the prevention of ASCVDs.

Keywords

atherosclerosis / glutathione synthesis / lipid nanoparticles / macrophage polarization / Slc7a11

References

[1] P. Roy, M. Orecchioni, and K. Ley, “How the Immune System Shapes Atherosclerosis: Roles of Innate and Adaptive Immunity,” Nature Reviews Immunology 22, no. 4 (2022): 251–265.

[2] C. W. Tsao, A. W. Aday, Z. I. Almarzooq, et al., “Heart Disease and Stroke Statistics-2023 Update: A Report from the American Heart Association,”. Circulation 2023; 147(8): e93–e621.

[3] K. J. Moore and I. Tabas, “Macrophages in the Pathogenesis of Atherosclerosis,” Cell 145, no. 3 (2011): 341–355.

[4] G. R. Y. De Meyer, M. Zurek, P. Puylaert, and W. Martinet, “Programmed Death of Macrophages in Atherosclerosis: Mechanisms and Therapeutic Targets,” Nature Reviews Cardiology 21, no. 5 (2024): 312–325.

[5] K. J. Moore, S. Koplev, E. A. Fisher, et al., “Macrophage Trafficking, Inflammatory Resolution, and Genomics in Atherosclerosis: JACC Macrophage in CVD Series (Part 2),” Journal of the American College of Cardiology 72, no. 18 (2018): 2181–2197.

[6] G. Chinetti-Gbaguidi, S. Colin, and B. Staels, “Macrophage Subsets in Atherosclerosis,” Nature Reviews Cardiology 12, no. 1 (2015): 10–17.

[7] G. Chinetti-Gbaguidi, M. Baron, M. A. Bouhlel, et al., “Human Atherosclerotic Plaque Alternative Macrophages Display Low Cholesterol Handling but High Phagocytosis Because of Distinct Activities of the PPARγ and LXRα Pathways,” Circulation Research 108, no. 8 (2011): 985–995.

[8] P. Hou, J. Fang, Z. Liu, et al., “Macrophage Polarization and Metabolism in Atherosclerosis,” Cell Death & Disease 14, no. 10 (2023): 691.

[9] K. Nitz, M. Lacy, and D. Atzler, “Amino Acids and Their Metabolism in Atherosclerosis,” Arteriosclerosis, Thrombosis, and Vascular Biology 39, no. 3 (2019): 319–330.

[10] O. Rom, Y. Liu, A. C. Finney, et al., “Induction of Glutathione Biosynthesis by Glycine-based Treatment Mitigates Atherosclerosis,” Redox Biology 52 (2022): 102313.

[11] B. Bednarz, T. Jaxa-Chamiec, P. Maciejewski, et al., “Efficacy and Safety of Oral L-arginine in Acute Myocardial Infarction. Results of the Multicenter, Randomized, Double-blind, Placebo-controlled ARAMI Pilot Trial,” Kardiologia Polska 62, no. 5 (2005): 421–427.

[12] Y. Zhao, X. Y. Dai, Z. Zhou, G. X. Zhao, X. Wang, and M. J. Xu, “Leucine Supplementation via Drinking Water Reduces Atherosclerotic Lesions in apoE Null Mice,” Acta Pharmacologica Sinica 37, no. 2 (2016): 196–203.

[13] X. Zhang, D. Kapoor, S. J. Jeong, et al., “Identification of a Leucine-mediated Threshold Effect Governing Macrophage mTOR Signalling and Cardiovascular Risk,” Nature metabolism 6, no. 2 (2024): 359–377.

[14] P. Koppula, Y. Zhang, L. Zhuang, and B. Gan, “Amino Acid Transporter SLC7A11/xCT at the Crossroads of Regulating Redox Homeostasis and Nutrient Dependency of Cancer,” Cancer Commun (Lond) 38, no. 1 (2018): 12.

[15] X. Fang, Z. Cai, H. Wang, et al., “Loss of Cardiac Ferritin H Facilitates Cardiomyopathy via Slc7a11-Mediated Ferroptosis,” Circulation Research 127, no. 4 (2020): 486–501.

[16] Y. Ye, A. Chen, L. Li, et al., “Repression of the Antiporter SLC7A11/Glutathione/Glutathione Peroxidase 4 Axis Drives Ferroptosis of Vascular Smooth Muscle Cells to Facilitate Vascular Calcification,” Kidney International 102, no. 6 (2022): 1259–1275.

[17] Y. Zheng, F. B. Hu, M. Ruiz-Canela, et al., “Metabolites of Glutamate Metabolism Are Associated with Incident Cardiovascular Events in the PREDIMED PREvención con DIeta MEDiterránea (PREDIMED) Trial,” Journal of the American Heart Association 5, no. 9 (2016): e003755.

[18] A. Lehn-Stefan, A. Peter, J. Machann, et al., “Elevated Circulating Glutamate Is Associated with Subclinical Atherosclerosis Independently of Established Risk Markers: A Cross-Sectional Study,” Journal of Clinical Endocrinology and Metabolism 106, no. 2 (2021): e982–e989.

[19] Y. Wei, B. Lan, T. Zheng, et al., “GSDME-mediated Pyroptosis Promotes the Progression and Associated Inflammation of Atherosclerosis,” Nature Communications 14, no. 1 (2023): 929.

[20] C. Ma, Y. Li, M. Tian, et al., “Gsα Regulates Macrophage Foam Cell Formation during Atherosclerosis,” Circulation Research 134, no. 7 (2024): e34–e51.

[21] K. Kim, D. Shim, J. S. Lee, et al., “Transcriptome Analysis Reveals Nonfoamy Rather than Foamy Plaque Macrophages Are Proinflammatory in Atherosclerotic Murine Models,” Circulation Research 123, no. 10 (2018): 1127–1142.

[22] M. Rojo de la Vega, E. Chapman, and D. D. Zhang, “NRF2 and the Hallmarks of Cancer,” Cancer Cell 34, no. 1 (2018): 21–43.

[23] X. Liu, J. W. Guo, X. C. Lin, et al., “Macrophage NFATc3 Prevents Foam Cell Formation and Atherosclerosis: Evidence and Mechanisms,” European Heart Journal 42, no. 47 (2021): 4847–4861.

[24] A. Canfrán-Duque, N. Rotllan, X. Zhang, et al., “Macrophage-Derived 25-Hydroxycholesterol Promotes Vascular Inflammation, Atherogenesis, and Lesion Remodeling,” Circulation 147, no. 5 (2023): 388–408.

[25] L. B. Ivashkiv, “IFNγ: Signalling, Epigenetics and Roles in Immunity, Metabolism, Disease and Cancer Immunotherapy,” Nature Reviews Immunology 18, no. 9 (2018): 545–558.

[26] L. Varinou, K. Ramsauer, M. Karaghiosoff, et al., “Phosphorylation of the Stat1 Transactivation Domain Is Required for Full-fledged IFN-gamma-dependent Innate Immunity,” Immunity 19, no. 6 (2003): 793–802.

[27] P. Koppula, L. Zhuang, and B. Gan, “Cystine Transporter SLC7A11/xCT in Cancer: Ferroptosis, Nutrient Dependency, and Cancer Therapy,” Protein Cell 12, no. 8 (2021): 599–620.

[28] S. J. Dixon, K. M. Lemberg, M. R. Lamprecht, et al., “Ferroptosis: An Iron-dependent Form of Nonapoptotic Cell Death,” Cell 149, no. 5 (2012): 1060–1072.

[29] W. S. Yang, R. SriRamaratnam, M. E. Welsch, et al., “Regulation of Ferroptotic Cancer Cell Death by GPX4,” Cell 156, no. 1-2 (2014): 317–331.

[30] Q. Chen, M. Gao, Z. Li, et al., “Biodegradable Nanoparticles Decorated With Different Carbohydrates for Efficient Macrophage-targeted Gene Therapy,” Journal of Controlled Release: Official Journal of the Controlled Release Society 323 (2020): 179–190.

[31] A. A. Alfaifi, R. S. Heyder, E. R. Bielski, et al., “Megalin-targeting Liposomes for Placental Drug Delivery,” Journal of Controlled Release 324 (2020): 366–378.

[32] S. H. Shah, J. R. Bain, M. J. Muehlbauer, et al., “Association of a Peripheral Blood Metabolic Profile with Coronary Artery Disease and Risk of Subsequent Cardiovascular Events,” Circulation: Cardiovascular Genetics 3, no. 2 (2010): 207–214.

[33] X. Peng, B. Sun, C. Tang, et al., “HMOX1-LDHB Interaction Promotes Ferroptosis by Inducing Mitochondrial Dysfunction in Foamy Macrophages During Advanced Atherosclerosis,” Developmental Cell 60, no. 7 (2025): 1070–1086.

[34] S. Benhmammouch, C. Borowczyk, C. Pierrot-Blanchet, et al., “Slc7a7 licenses Macrophage Glutaminolysis for Restorative Functions in Atherosclerosis,” Nature Metabolism 7, no. 9 (2025): 1924–1938.

[35] K. Nitz, M. Lacy, M. Bianchini, et al., “The Amino Acid Homoarginine Inhibits Atherogenesis by Modulating T-Cell Function,” Circulation Research 131, no. 8 (2022): 701–712.

[36] I. Tabas and K. E. Bornfeldt, “Macrophage Phenotype and Function in Different Stages of Atherosclerosis,” Circulation Research 118, no. 4 (2016): 653–667.

[37] G. J. Koelwyn, E. M. Corr, E. Erbay, and K. J. Moore, “Regulation of Macrophage Immunometabolism in Atherosclerosis,” Nature Immunology 19, no. 6 (2018): 526–537.

[38] J. Zhao, L. Ling, W. Zhu, et al., “M1/M2 re-polarization of Kaempferol Biomimetic NPs in Anti-inflammatory Therapy of Atherosclerosis,” Journal of Controlled Release: Official Journal of the Controlled Release Society 353 (2023): 1068–1083.

[39] H. Su, J. Huang, S. Weng, B. Zhang, T. Zhang, and Y. Xu, “Glutathione Synthesis Primes Monocytes Metabolic and Epigenetic Pathway for β-glucan-trained Immunity,” Redox Biology 48 (2021): 102206.

[40] Y. Zhang, S. Choksi, K. Chen, Y. Pobezinskaya, I. Linnoila, and Z. G. Liu, “ROS Play a Critical Role in the Differentiation of Alternatively Activated Macrophages and the Occurrence of Tumor-associated Macrophages,” Cell Research 23, no. 7 (2013): 898–914.

[41] L. Formentini, F. Santacatterina, C. Núñez de Arenas, et al., “Mitochondrial ROS Production Protects the Intestine From Inflammation Through Functional M2 Macrophage Polarization,” Cell Reports 19, no. 6 (2017): 1202–1213.

[42] B. Tang, J. Zhu, Y. Wang, et al., “Targeted xCT-mediated Ferroptosis and Protumoral Polarization of Macrophages Is Effective Against HCC and Enhances the Efficacy of the Anti-PD-1/L1 Response,” Advanced Science (Weinheim, Baden-Wurttemberg, Germany) 10, no. 2 (2023): e2203973.

[43] B. Tang, Y. Wang, W. Xu, et al., “Macrophage xCT Deficiency Drives Immune Activation and Boosts Responses to Immune Checkpoint Blockade in Lung Cancer,” Cancer Letters 554 (2023): 216021.

[44] R. Xu, Y. Huang, D. Zhu, and J. Guo, “Iron Promotes Slc7a11-deficient Valvular Interstitial Cell Osteogenic Differentiation: A Possible Mechanism by Which Ferroptosis Participates in Intraleaflet Hemorrhage-induced Calcification,” Free Radical Biology & Medicine 184 (2022): 158–169.

[45] M. Munder, K. Eichmann, and M. Modolell, “Alternative Metabolic States in Murine Macrophages Reflected by the Nitric Oxide Synthase/Arginase Balance: Competitive Regulation by CD4+ T Cells Correlates With Th1/Th2 Phenotype,” Journal of Immunology 160, no. 11 (1998): 5347–5354.

[46] P. K. Langston, M. Shibata, and T. Horng, “Metabolism Supports Macrophage Activation,” Frontiers in Immunology 8 (2017): 61.

[47] Y. Zhu, Z. Zhou, X. Du, et al., “Cancer Cell-derived Arginine Fuels Polyamine Biosynthesis in Tumor-associated Macrophages to Promote Immune Evasion,” Cancer Cell 43, no. 6 (2025): 1045–1060. e7.

[48] Y. Liu, M. Ge, X. Xiao, et al., “Sarcosine Decreases in Sarcopenia and Enhances Muscle Regeneration and Adipose Thermogenesis by Activating Anti-inflammatory Macrophages,” Nature Aging 5, no. 9 (2025): 1810–1827.

[49] A. E. Rodriguez, G. S. Ducker, L. K. Billingham, et al., “Serine Metabolism Supports Macrophage IL-1β Production,” Cell Metabolism 29, no. 4 (2019): 1003–1011. e4.

[50] J. Van den Bossche, J. Baardman, N. A. Otto, et al., “Mitochondrial Dysfunction Prevents Repolarization of Inflammatory Macrophages,” Cell Reports 17, no. 3 (2016): 684–696.

[51] C. Tang, H. Wang, L. Guo, et al., “Multifunctional Nanomedicine for Targeted Atherosclerosis Therapy: Activating Plaque Clearance Cascade and Suppressing Inflammation,” ACS Nano 19, no. 3 (2025): 3339–3361.

[52] C. Tang, H. Wang, L. Guo, et al., “CpG-Conjugated Silver Nanoparticles as a Multifunctional Nanomedicine to Promote Macrophage Efferocytosis and Repolarization for Atherosclerosis Therapy,” ACS Applied Materials & Interfaces 15, no. 45 (2023): 52162–52179.

[53] B. Bay, R. Tanner, M. Gao, et al., “Residual Cholesterol and Inflammatory Risk in Statin-treated Patients Undergoing Percutaneous Coronary Intervention†,” European Heart Journal 46, no. 32 (2025): 3167–3177.

[54] P. M. Ridker, D. L. Bhatt, A. D. Pradhan, et al., “Inflammation and Cholesterol as Predictors of Cardiovascular Events Among Patients Receiving Statin Therapy: A Collaborative Analysis of Three Randomised Trials,” Lancet 401, no. 10384 (2023): 1293–1301.

[55] S. M. Nidorf, A. T. L. Fiolet, A. Mosterd, et al., “Colchicine in Patients With Chronic Coronary Disease,” New England Journal of Medicine 383, no. 19 (2020): 1838–1847.

[56] A. T. L. Fiolet, T. S. J. Opstal, A. Mosterd, et al., “Efficacy and Safety of Low-dose Colchicine in Patients With Coronary Disease: A Systematic Review and Meta-analysis of Randomized Trials,” European Heart Journal 42, no. 28 (2021): 2765–2775.

[57] J. Shen, E. Xie, S. Shen, et al., “Essentiality of SLC7A11-mediated Nonessential Amino Acids in MASLD,” Sci Bull (Beijing) 69, no. 23 (2024): 3700–3716.

[58] H. Xu, J. Zhu, S. Smith, et al., “Notch-RBP-J Signaling Regulates the Transcription Factor IRF8 to Promote Inflammatory Macrophage Polarization,” Nature Immunology 13, no. 7 (2012): 642–650.

[59] B. J. van Vlijmen, A. M. van den Maagdenberg, M. J. Gijbels, et al., “Diet-induced Hyperlipoproteinemia and Atherosclerosis in Apolipoprotein E3-Leiden Transgenic Mice,” Journal of Clinical Investigation 93, no. 4 (1994): 1403–1410.

[60] X. Chen, F. Jia, Y. Li, et al., “Nitric Oxide-induced Stromal Depletion for Improved Nanoparticle Penetration in Pancreatic Cancer Treatment,” Biomaterials 246 (2020): 119999.

RIGHTS & PERMISSIONS

2026 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.