Xiu Liu1,2, Jia-Wei Guo2,3, Xiao-Chun Lin2, Yong-Hua Tuo4, Wan-Li Peng2, Su-Yue He5, Zhao-Qiang Li6, Yan-Chen Ye7,8, Jie Yu9, Fei-Ran Zhang1,2, Ming-Ming Ma2, Jin-Yan Shang2, Xiao-Fei Lv2, An-Dong Zhou10, Ying Ouyang11, Cheng Wang1, Rui-Ping Pang5, Jian-Xin Sun12, Jing-Song Ou8,13, Jia-Guo Zhou1,2,14,15,16, Si-Jia Liang1,2. 1. Program of Kidney and Cardiovascular Diseases, the Fifth Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, 74 Zhongshan 2 Rd, Guangzhou 510080, China. 2. Department of Pharmacology, Cardiac and Cerebral Vascular Research Center, Zhongshan School of Medicine, Sun Yat-Sen University, 74 Zhongshan 2 Rd, Guangzhou 510080, China. 3. Department of Pharmacology, School of Medicine, Yangtze University, 1 Nanhuan Rd, Jingzhou 434023, China. 4. Department of Neurosurgery, the Second Affiliated Hospital of Guangzhou Medical University, 250 Changgang East Rd, Guangzhou 510260, China. 5. Department of Physiology, Pain Research Center, Zhongshan School of Medicine, Sun Yat-Sen University, 74 Zhongshan 2 Rd, Guangzhou 510080, China. 6. Guangdong Provincial Key Laboratory of Tumor Immunotherapy, Cancer Research Institute, Southern Medical University, 1023 Shatai South Rd, Guangzhou 510515, China. 7. Division of Vascular Surgery, the First Affiliated Hospital, Sun Yat-Sen University, 58 Zhongshan 2 Rd, Guangzhou 510080, China. 8. National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, the First Affiliated Hospital, Sun Yat-Sen University, 58 Zhongshan 2 Rd, Guangzhou 510080, China. 9. Department of General Surgery, Zhujiang Hospital, Southern Medical University, 253 Industrial Rd, Guangzhou 510282, China. 10. Department of Clinical Medicine, the Second Clinical Medical School, Guangdong Medical University, 1 Xincheng Rd, Dongguan 523808, China. 11. Department of Pediatrics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, 107 Yanjiang West Rd, Guangzhou 510120, China. 12. Center for Translational Medicine, Thomas Jefferson University, 1020 Locust St., Rm. 368G, Philadelphia PA 19107, USA. 13. Division of Cardiac Surgery, Heart Center, the First Affiliated Hospital, Sun Yat-Sen University, 58 ZhongShan 2 Rd, Guangzhou 510080, China. 14. Department of Cardiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, 107 Yanjiang West Rd, Guangzhou 510120, China. 15. Guangdong Province Key Laboratory of Brain Function and Disease, Zhongshan School of Medicine, Sun Yat-Sen University, 74 Zhongshan 2 Rd, Guangzhou 510080, China. 16. Key Laboratory of Cardiovascular diseases, School of Basic Medical Sciences, Guangzhou Medical University, 1 Xinzao Rd, Guangzhou 511436, China.
Abstract
AIMS: Our previous study demonstrated that Ca2+ influx through the Orai1 store-operated Ca2+ channel in macrophages contributes to foam cell formation and atherosclerosis via the calcineurin-ASK1 pathway, not the classical calcineurin-nuclear factor of activated T-cell (NFAT) pathway. Moreover, up-regulation of NFATc3 in macrophages inhibits foam cell formation, suggesting that macrophage NFATc3 is a negative regulator of atherogenesis. Hence, this study investigated the precise role of macrophage NFATc3 in atherogenesis. METHODS AND RESULTS: Macrophage-specific NFATc3 knockout mice were generated to determine the effect of NFATc3 on atherosclerosis in a mouse model of adeno-associated virus-mutant PCSK9-induced atherosclerosis. NFATc3 expression was decreased in macrophages within human and mouse atherosclerotic lesions. Moreover, NFATc3 levels in peripheral blood mononuclear cells from atherosclerotic patients were negatively associated with plaque instability. Furthermore, macrophage-specific ablation of NFATc3 in mice led to the atherosclerotic plaque formation, whereas macrophage-specific NFATc3 transgenic mice exhibited the opposite phenotype. NFATc3 deficiency in macrophages promoted foam cell formation by potentiating SR-A- and CD36-meditated lipid uptake. NFATc3 directly targeted and transcriptionally up-regulated miR-204 levels. Mature miR-204-5p suppressed SR-A expression via canonical regulation. Unexpectedly, miR-204-3p localized in the nucleus and inhibited CD36 transcription. Restoration of miR-204 abolished the proatherogenic phenotype observed in the macrophage-specific NFATc3 knockout mice, and blockade of miR-204 function reversed the beneficial effects of NFATc3 in macrophages. CONCLUSION: Macrophage NFATc3 up-regulates miR-204 to reduce SR-A and CD36 levels, thereby preventing foam cell formation and atherosclerosis, indicating that the NFATc3/miR-204 axis may be a potential therapeutic target against atherosclerosis. Published on behalf of the European Society of Cardiology. All rights reserved.
AIMS: Our previous study demonstrated that Ca2+ influx through the Orai1 store-operated Ca2+ channel in macrophages contributes to foam cell formation and atherosclerosis via the calcineurin-ASK1 pathway, not the classical calcineurin-nuclear factor of activated T-cell (NFAT) pathway. Moreover, up-regulation of NFATc3 in macrophages inhibits foam cell formation, suggesting that macrophage NFATc3 is a negative regulator of atherogenesis. Hence, this study investigated the precise role of macrophage NFATc3 in atherogenesis. METHODS AND RESULTS: Macrophage-specific NFATc3 knockout mice were generated to determine the effect of NFATc3 on atherosclerosis in a mouse model of adeno-associated virus-mutant PCSK9-induced atherosclerosis. NFATc3 expression was decreased in macrophages within human and mouse atherosclerotic lesions. Moreover, NFATc3 levels in peripheral blood mononuclear cells from atherosclerotic patients were negatively associated with plaque instability. Furthermore, macrophage-specific ablation of NFATc3 in mice led to the atherosclerotic plaque formation, whereas macrophage-specific NFATc3 transgenic mice exhibited the opposite phenotype. NFATc3 deficiency in macrophages promoted foam cell formation by potentiating SR-A- and CD36-meditated lipid uptake. NFATc3 directly targeted and transcriptionally up-regulated miR-204 levels. Mature miR-204-5p suppressed SR-A expression via canonical regulation. Unexpectedly, miR-204-3p localized in the nucleus and inhibited CD36 transcription. Restoration of miR-204 abolished the proatherogenic phenotype observed in the macrophage-specific NFATc3 knockout mice, and blockade of miR-204 function reversed the beneficial effects of NFATc3 in macrophages. CONCLUSION: Macrophage NFATc3 up-regulates miR-204 to reduce SR-A and CD36 levels, thereby preventing foam cell formation and atherosclerosis, indicating that the NFATc3/miR-204 axis may be a potential therapeutic target against atherosclerosis. Published on behalf of the European Society of Cardiology. All rights reserved.