Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice

Xuchu Que, Ming Yow Hung, Calvin Yeang, Ayelet Gonen, Thomas A. Prohaska, Xiaoli Sun, Cody Diehl, Antti Määttä, Dalia E. Gaddis, Karen Bowden, Jennifer Pattison, Jeffrey G. MacDonald, Seppo Ylä-Herttuala, Pamela L. Mellon, Catherine C. Hedrick, Klaus Ley, Yury I. Miller, Christopher K. Glass, Kirk L. Peterson, Christoph J. BinderSotirios Tsimikas, Joseph L. Witztum

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Oxidized phospholipids (OxPL) are ubiquitous, are formed in many inflammatory tissues, including atherosclerotic lesions, and frequently mediate proinflammatory changes 1. Because OxPL are mostly the products of non-enzymatic lipid peroxidation, mechanisms to specifically neutralize them are unavailable and their roles in vivo are largely unknown. We previously cloned the IgM natural antibody E06, which binds to the phosphocholine headgroup of OxPL, and blocks the uptake of oxidized low-density lipoprotein (OxLDL) by macrophages and inhibits the proinflammatory properties of OxPL 2-4. Here, to determine the role of OxPL in vivo in the context of atherogenesis, we generated transgenic mice in the Ldlr -/- background that expressed a single-chain variable fragment of E06 (E06-scFv) using the Apoe promoter. E06-scFv was secreted into the plasma from the liver and macrophages, and achieved sufficient plasma levels to inhibit in vivo macrophage uptake of OxLDL and to prevent OxPL-induced inflammatory signalling. Compared to Ldlr -/- mice, Ldlr -/- E06-scFv mice had 57-28% less atherosclerosis after 4, 7 and even 12 months of 1% high-cholesterol diet. Echocardiographic and histologic evaluation of the aortic valves demonstrated that E06-scFv ameliorated the development of aortic valve gradients and decreased aortic valve calcification. Both cholesterol accumulation and in vivo uptake of OxLDL were decreased in peritoneal macrophages, and both peritoneal and aortic macrophages had a decreased inflammatory phenotype. Serum amyloid A was decreased by 32%, indicating decreased systemic inflammation, and hepatic steatosis and inflammation were also decreased. Finally, the E06-scFv prolonged life as measured over 15 months. Because the E06-scFv lacks the functional effects of an intact antibody other than the ability to bind OxPL and inhibit OxLDL uptake in macrophages, these data support a major proatherogenic role of OxLDL and demonstrate that OxPL are proinflammatory and proatherogenic, which E06 counteracts in vivo. These studies suggest that therapies inactivating OxPL may be beneficial for reducing generalized inflammation, including the progression of atherosclerosis, aortic stenosis and hepatic steatosis.

Original languageEnglish
Pages (from-to)301-306
Number of pages6
JournalNature
Volume558
Issue number7709
DOIs
Publication statusPublished - Jun 14 2018

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Phospholipids
Macrophages
Atherosclerosis
Peritoneal Macrophages
Inflammation
Aortic Valve
Liver
Cholesterol
Serum Amyloid A Protein
Single-Chain Antibodies
Phosphorylcholine
Antibodies
Aortic Valve Stenosis
Apolipoproteins E
Transgenic Mice
Lipid Peroxidation
Immunoglobulin M
oxidized low density lipoprotein
Diet
Phenotype

ASJC Scopus subject areas

  • General

Cite this

Que, X., Hung, M. Y., Yeang, C., Gonen, A., Prohaska, T. A., Sun, X., ... Witztum, J. L. (2018). Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice. Nature, 558(7709), 301-306. https://doi.org/10.1038/s41586-018-0198-8

Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice. / Que, Xuchu; Hung, Ming Yow; Yeang, Calvin; Gonen, Ayelet; Prohaska, Thomas A.; Sun, Xiaoli; Diehl, Cody; Määttä, Antti; Gaddis, Dalia E.; Bowden, Karen; Pattison, Jennifer; MacDonald, Jeffrey G.; Ylä-Herttuala, Seppo; Mellon, Pamela L.; Hedrick, Catherine C.; Ley, Klaus; Miller, Yury I.; Glass, Christopher K.; Peterson, Kirk L.; Binder, Christoph J.; Tsimikas, Sotirios; Witztum, Joseph L.

In: Nature, Vol. 558, No. 7709, 14.06.2018, p. 301-306.

Research output: Contribution to journalArticle

Que, X, Hung, MY, Yeang, C, Gonen, A, Prohaska, TA, Sun, X, Diehl, C, Määttä, A, Gaddis, DE, Bowden, K, Pattison, J, MacDonald, JG, Ylä-Herttuala, S, Mellon, PL, Hedrick, CC, Ley, K, Miller, YI, Glass, CK, Peterson, KL, Binder, CJ, Tsimikas, S & Witztum, JL 2018, 'Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice', Nature, vol. 558, no. 7709, pp. 301-306. https://doi.org/10.1038/s41586-018-0198-8
Que, Xuchu ; Hung, Ming Yow ; Yeang, Calvin ; Gonen, Ayelet ; Prohaska, Thomas A. ; Sun, Xiaoli ; Diehl, Cody ; Määttä, Antti ; Gaddis, Dalia E. ; Bowden, Karen ; Pattison, Jennifer ; MacDonald, Jeffrey G. ; Ylä-Herttuala, Seppo ; Mellon, Pamela L. ; Hedrick, Catherine C. ; Ley, Klaus ; Miller, Yury I. ; Glass, Christopher K. ; Peterson, Kirk L. ; Binder, Christoph J. ; Tsimikas, Sotirios ; Witztum, Joseph L. / Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice. In: Nature. 2018 ; Vol. 558, No. 7709. pp. 301-306.
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abstract = "Oxidized phospholipids (OxPL) are ubiquitous, are formed in many inflammatory tissues, including atherosclerotic lesions, and frequently mediate proinflammatory changes 1. Because OxPL are mostly the products of non-enzymatic lipid peroxidation, mechanisms to specifically neutralize them are unavailable and their roles in vivo are largely unknown. We previously cloned the IgM natural antibody E06, which binds to the phosphocholine headgroup of OxPL, and blocks the uptake of oxidized low-density lipoprotein (OxLDL) by macrophages and inhibits the proinflammatory properties of OxPL 2-4. Here, to determine the role of OxPL in vivo in the context of atherogenesis, we generated transgenic mice in the Ldlr -/- background that expressed a single-chain variable fragment of E06 (E06-scFv) using the Apoe promoter. E06-scFv was secreted into the plasma from the liver and macrophages, and achieved sufficient plasma levels to inhibit in vivo macrophage uptake of OxLDL and to prevent OxPL-induced inflammatory signalling. Compared to Ldlr -/- mice, Ldlr -/- E06-scFv mice had 57-28{\%} less atherosclerosis after 4, 7 and even 12 months of 1{\%} high-cholesterol diet. Echocardiographic and histologic evaluation of the aortic valves demonstrated that E06-scFv ameliorated the development of aortic valve gradients and decreased aortic valve calcification. Both cholesterol accumulation and in vivo uptake of OxLDL were decreased in peritoneal macrophages, and both peritoneal and aortic macrophages had a decreased inflammatory phenotype. Serum amyloid A was decreased by 32{\%}, indicating decreased systemic inflammation, and hepatic steatosis and inflammation were also decreased. Finally, the E06-scFv prolonged life as measured over 15 months. Because the E06-scFv lacks the functional effects of an intact antibody other than the ability to bind OxPL and inhibit OxLDL uptake in macrophages, these data support a major proatherogenic role of OxLDL and demonstrate that OxPL are proinflammatory and proatherogenic, which E06 counteracts in vivo. These studies suggest that therapies inactivating OxPL may be beneficial for reducing generalized inflammation, including the progression of atherosclerosis, aortic stenosis and hepatic steatosis.",
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T1 - Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice

AU - Que, Xuchu

AU - Hung, Ming Yow

AU - Yeang, Calvin

AU - Gonen, Ayelet

AU - Prohaska, Thomas A.

AU - Sun, Xiaoli

AU - Diehl, Cody

AU - Määttä, Antti

AU - Gaddis, Dalia E.

AU - Bowden, Karen

AU - Pattison, Jennifer

AU - MacDonald, Jeffrey G.

AU - Ylä-Herttuala, Seppo

AU - Mellon, Pamela L.

AU - Hedrick, Catherine C.

AU - Ley, Klaus

AU - Miller, Yury I.

AU - Glass, Christopher K.

AU - Peterson, Kirk L.

AU - Binder, Christoph J.

AU - Tsimikas, Sotirios

AU - Witztum, Joseph L.

PY - 2018/6/14

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N2 - Oxidized phospholipids (OxPL) are ubiquitous, are formed in many inflammatory tissues, including atherosclerotic lesions, and frequently mediate proinflammatory changes 1. Because OxPL are mostly the products of non-enzymatic lipid peroxidation, mechanisms to specifically neutralize them are unavailable and their roles in vivo are largely unknown. We previously cloned the IgM natural antibody E06, which binds to the phosphocholine headgroup of OxPL, and blocks the uptake of oxidized low-density lipoprotein (OxLDL) by macrophages and inhibits the proinflammatory properties of OxPL 2-4. Here, to determine the role of OxPL in vivo in the context of atherogenesis, we generated transgenic mice in the Ldlr -/- background that expressed a single-chain variable fragment of E06 (E06-scFv) using the Apoe promoter. E06-scFv was secreted into the plasma from the liver and macrophages, and achieved sufficient plasma levels to inhibit in vivo macrophage uptake of OxLDL and to prevent OxPL-induced inflammatory signalling. Compared to Ldlr -/- mice, Ldlr -/- E06-scFv mice had 57-28% less atherosclerosis after 4, 7 and even 12 months of 1% high-cholesterol diet. Echocardiographic and histologic evaluation of the aortic valves demonstrated that E06-scFv ameliorated the development of aortic valve gradients and decreased aortic valve calcification. Both cholesterol accumulation and in vivo uptake of OxLDL were decreased in peritoneal macrophages, and both peritoneal and aortic macrophages had a decreased inflammatory phenotype. Serum amyloid A was decreased by 32%, indicating decreased systemic inflammation, and hepatic steatosis and inflammation were also decreased. Finally, the E06-scFv prolonged life as measured over 15 months. Because the E06-scFv lacks the functional effects of an intact antibody other than the ability to bind OxPL and inhibit OxLDL uptake in macrophages, these data support a major proatherogenic role of OxLDL and demonstrate that OxPL are proinflammatory and proatherogenic, which E06 counteracts in vivo. These studies suggest that therapies inactivating OxPL may be beneficial for reducing generalized inflammation, including the progression of atherosclerosis, aortic stenosis and hepatic steatosis.

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