Regression of Atherosclerosis in ApoE-/- Mice Via Modulation of Monocyte Recruitment and Phenotype, Induced by Weekly Dosing of a Novel "Cytotopic" Anti-Thrombin Without Prolonged Anticoagulation.
Regression of Atherosclerosis in ApoE-/- Mice Via Modulation of Monocyte Recruitment and Phenotype, Induced by Weekly Dosing of a Novel "Cytotopic" Anti-Thrombin Without Prolonged Anticoagulation.
复制标题
通过调节单核细胞募集和表型诱导载脂蛋白E-/-小鼠动脉粥样硬化的消退，每周服用新型 "细胞异位 "抗凝血酶而无需长期抗凝。
DOI：
10.1161/jaha.119.014811
复制
发表时间：
2020-07-07
影响因子：
5.4
通讯作者：
Dorling A
中科院分区：
文献类型：
--
作者：
Chen D;Li K;Festenstein S;Karegli J;Wilkinson H;Leonard H;Wei LL;Ma N;Xia M;Tam H;Wang JA;Xu Q;McVey JH;Smith RAG;Dorling A
抗凝剂在动物模型中可诱导动脉粥样硬化消退，但在临床上应用却受到出血事件的限制。在此我们测试了一种新型凝血酶抑制剂PTL060，它由共价连接到合成肉豆蔻酰静电开关的水蛭素类似物组成，可与细胞膜结合。
载脂蛋白E - / - 小鼠在移植同基因主动脉段或注射PTL060、母体水蛭素、对照生理盐水或标记的CD11b阳性细胞之前，分别喂食普通饲料或高脂饮食。来自在内皮上表达抗凝剂的转基因小鼠的主动脉移植物未发生动脉粥样硬化。在4周后评估时，单次静脉注射PTL060（而非水蛭素）与对照组相比，抑制动脉粥样硬化斑块形成超过50%。小鼠出血时间延长的时长仅为PTL060具有生物活性时长的七分之一。每周重复注射PTL060（而非水蛭素）可导致动脉粥样硬化斑块消退。我们剖析了两个相关机制。首先，在PTL060处理的小鼠中，招募到斑块中的大多数CCR2 +（C - C趋化因子受体2型 +）单核细胞表达CCR7（C - C趋化因子受体7型）、ABCA1（ATP结合盒转运体 - 1）和白细胞介素 - 10，而在对照组中，只有不到20%的CCR2 +招募细胞呈现这种表型。其次，在多次给药后，单核细胞招募数量显著减少，其中大多数为CCR2 - 阴性，且具有类似的与消退相关的表型。通过过继转移预先用PTL060包被的CD11b +细胞可诱导出与静脉注射PTL060相当的消退效果。
PTL060中肉豆蔻酰静电开关与水蛭素的共价连接使对止血和动脉粥样硬化的药效学作用解偶联，从而使得主要通过对单核细胞的作用介导的斑块消退仅伴有短暂的抗凝作用。
Anticoagulants induce atherosclerosis regression in animal models but exploiting this clinically is limited by bleeding events. Here we test a novel thrombin inhibitor, PTL060, comprising hirulog covalently linked to a synthetic myristoyl electrostatic switch to tether to cell membranes. ApoE−/− mice were fed chow or high‐fat diets, before transplantation of congenic aortic segments or injection of PTL060, parental hirulog, control saline, or labeled CD11b positive cells. Aortic transplants from transgenic mice expressing anticoagulants on endothelium did not develop atherosclerosis. A single intravenous injection of PTL060, but not hirulog inhibited atheroma development by >50% compared with controls when assessed 4 weeks later. Mice had prolonged bleeding times for only one seventh of the time that PTL060 was biologically active. Repeated weekly injections of PTL060 but not hirulog caused regression of atheroma. We dissected 2 contributory mechanisms. First, the majority of CCR2+ (C‐C chemokine receptor type 2+) monocytes recruited into plaques expressed CCR7 (C‐C chemokine receptor type 7), ABCA1 (ATP‐binding cassette transporter – 1), and interleukin‐10 in PTL060 mice, a phenotype seen in <20% of CCR2+ recruits in controls. Second, after several doses, there was a significant reduction in monocyte recruits, the majority of which were CCR2‐negative with a similar regression‐associated phenotype. Regression equivalent to that induced by intravenous PTL060 was induced by adoptive transfer of CD11b+ cells pre‐coated with PTL060. Covalent linkage of a myristoyl electrostatic switch onto hirulog in PTL060 uncouples the pharmacodynamic effects on hemostasis and atherosclerosis, such that plaque regression, mediated predominantly via effects on monocytes, is accompanied by only transient anticoagulation.