A role of RIP3-mediated macrophage necrosis in atherosclerosis development.

Necrotic death of macrophages has long been known to be present in atherosclerotic lesions but has not been studied. We examined the role of receptor interacting protein (RIP) 3, a mediator of necrotic cell death, in atherosclerosis and found that RIP3(-/-);Ldlr(-/-) mice were no different from RIP3(+/+);Ldlr(-/-) mice in early atherosclerosis but had significant reduction in advanced atherosclerotic lesions. Similar results were observed in Apoe(-/-) background mice. Bone marrow transplantation revealed that loss of RIP3 expression from bone-marrow-derived cells is responsible for the reduced disease progression. While no difference was found in apoptosis between RIP3(-/-);Ldlr(-/-) and RIP3(+/+);Ldlr(-/-) mice, electron microscopy revealed a significant reduction of macrophage primary necrosis in the advanced lesions of RIP3(-/-) mice. In vitro cellular studies showed that RIP3 deletion had no effect on oxidized low-density lipoprotein (LDL)-induced macrophage apoptosis, but prevented macrophage primary necrosis occurring in response to oxidized LDL under caspase inhibition or RIP3 overexpression conditions. RIP3-dependent necrosis is not postapoptotic, and the increased primary necrosis in advanced atherosclerotic lesions most likely resulted from the increase of RIP3 expression. Our data demonstrate that primary necrosis of macrophages is proatherogenic during advanced atherosclerosis development.

[1]  D. Ron,et al.  Reduced apoptosis and plaque necrosis in advanced atherosclerotic lesions of Apoe-/- and Ldlr-/- mice lacking CHOP. , 2009, Cell metabolism.

[2]  K. Moore,et al.  Macrophages in the Pathogenesis of Atherosclerosis , 2011, Cell.

[3]  R. Bogle,et al.  Regulation of chemokine expression in atherosclerosis. , 2002, Vascular pharmacology.

[4]  George Kuriakose,et al.  The endoplasmic reticulum is the site of cholesterol-induced cytotoxicity in macrophages , 2003, Nature Cell Biology.

[5]  Xiaodong Wang,et al.  Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase , 2012, Cell.

[6]  R. Evans,et al.  PPARδ regulates multiple proinflammatory pathways to suppress atherosclerosis , 2008, Proceedings of the National Academy of Sciences.

[7]  V. Dixit,et al.  Kinase RIP3 Is Dispensable for Normal NF-κBs, Signaling by the B-Cell and T-Cell Receptors, Tumor Necrosis Factor Receptor 1, and Toll-Like Receptors 2 and 4 , 2004, Molecular and Cellular Biology.

[8]  J. Thyberg,et al.  Cell death in human atherosclerotic plaques involves both oncosis and apoptosis. , 1997, Atherosclerosis.

[9]  K. Rock,et al.  How dying cells alert the immune system to danger , 2008, Nature Reviews Immunology.

[10]  M. J. Mitchinson,et al.  Toxicity of oxidised low density lipoprotein towards mouse peritoneal macrophages in vitro. , 1993, Atherosclerosis.

[11]  H. Ljunggren,et al.  CD1d-dependent Activation of NKT Cells Aggravates Atherosclerosis , 2004, The Journal of Experimental Medicine.

[12]  Na Zhang,et al.  RIP3, an Energy Metabolism Regulator That Switches TNF-Induced Cell Death from Apoptosis to Necrosis , 2009, Science.

[13]  M. Press,et al.  Quantitative ultrastructural analysis of coronary atherosclerotic involvement in two macaque species. , 1988, Experimental and molecular pathology.

[14]  William A. Boisvert,et al.  Transcriptional Repression of Atherogenic Inflammation: Modulation by PPARδ , 2003, Science.

[15]  M. Mann,et al.  Brain phosphoproteome obtained by a FASP-based method reveals plasma membrane protein topology. , 2010, Journal of proteome research.

[16]  D. Landry,et al.  Sitosterol-containing Lipoproteins Trigger Free Sterol-induced Caspase-independent Death in ACAT-competent Macrophages* , 2006, Journal of Biological Chemistry.

[17]  A. Tall,et al.  Macrophage deficiency of p38alpha MAPK promotes apoptosis and plaque necrosis in advanced atherosclerotic lesions in mice. , 2009, The Journal of clinical investigation.

[18]  D. Schrijvers,et al.  Mertk Receptor Mutation Reduces Efferocytosis Efficiency and Promotes Apoptotic Cell Accumulation and Plaque Necrosis in Atherosclerotic Lesions of Apoe−/− Mice , 2008, Arteriosclerosis, thrombosis, and vascular biology.

[19]  Joan W. Miller,et al.  Receptor interacting protein kinases mediate retinal detachment-induced photoreceptor necrosis and compensate for inhibition of apoptosis , 2010, Proceedings of the National Academy of Sciences.

[20]  A. Tall,et al.  Cholesterol-induced Apoptotic Macrophages Elicit an Inflammatory Response in Phagocytes, Which Is Partially Attenuated by the Mer Receptor* , 2006, Journal of Biological Chemistry.

[21]  F. Chan,et al.  Phosphorylation-Driven Assembly of the RIP1-RIP3 Complex Regulates Programmed Necrosis and Virus-Induced Inflammation , 2009, Cell.

[22]  Tao Wang,et al.  Receptor Interacting Protein Kinase-3 Determines Cellular Necrotic Response to TNF-α , 2009, Cell.

[23]  R. Hakem,et al.  RIP3 mediates the embryonic lethality of caspase-8-deficient mice , 2011, Nature.

[24]  I. Tabas.  Macrophage death and defective inflammation resolution in atherosclerosis , 2010, Nature Reviews Immunology.

[25]  J. Skepper,et al.  FOAM CELL APOPTOSIS AND THE DEVELOPMENT OF THE LIPID CORE OF HUMAN ATHEROSCLEROSIS , 1996, The Journal of pathology.

[26]  N. Yahagi,et al.  Targeted disruption of hormone-sensitive lipase results in male sterility and adipocyte hypertrophy, but not in obesity. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[27]  I. Tabas,et al.  Evidence That the Initial Up-regulation of Phosphatidylcholine Biosynthesis in Free Cholesterol-loaded Macrophages Is an Adaptive Response That Prevents Cholesterol-induced Cellular Necrosis , 1996, The Journal of Biological Chemistry.

[28]  Y. Furukawa,et al.  Ablation of neutral cholesterol ester hydrolase 1 accelerates atherosclerosis. , 2009, Cell metabolism.

[29]  M. Cybulsky,et al.  Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis. , 1991, Science.

[30]  J. Hogg,et al.  Ultrastructural changes in atherosclerotic plaques following the instillation of airborne particulate matter into the lungs of rabbits. , 2010, The Canadian journal of cardiology.

[31]  J. Begley,et al.  Oxidized LDL Promotes Peroxide-Mediated Mitochondrial Dysfunction and Cell Death in Human Macrophages: A Caspase-3–Independent Pathway , 2003, Circulation research.

[32]  B. Lüderitz,et al.  Apoptosis in restenosis versus stable-angina atherosclerosis: implications for the pathogenesis of restenosis. , 1998, Arteriosclerosis, thrombosis, and vascular biology.

[33]  Chuan-Qi Zhong,et al.  Programmed necrosis: backup to and competitor with apoptosis in the immune system , 2011, Nature Immunology.

[34]  Guy S. Salvesen,et al.  Catalytic activity of the caspase-8-FLIPL complex inhibits RIPK3-dependent necrosis , 2011, Nature.

[35]  Vanesa Fernández-Majada,et al.  FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation , 2011, Nature.

[36]  P. Libby.  Inflammation in atherosclerosis , 2002, Nature.