Wang Lai, Tian Fang, Arias Ana, Yang Mingjie, Sharifi Behrooz G, Shah Prediman K
Division of Cardiology, Oppenheimer Atherosclerosis Research Center, Cedars-Sinai Heart Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
Division of Cardiology, Oppenheimer Atherosclerosis Research Center, Cedars-Sinai Heart Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA
J Cardiovasc Pharmacol Ther. 2016 May;21(3):320-8. doi: 10.1177/1074248415610216. Epub 2015 Oct 22.
Apolipoprotein A-1 (Apo A-I) Milano, a naturally occurring Arg173to Cys mutant of Apo A-1, has been shown to reduce atherosclerosis in animal models and in a small phase 2 human trial. We have shown the superior atheroprotective effects of Apo A-I Milano (Apo A-IM) gene compared to wild-type Apo A-I gene using transplantation of retrovirally transduced bone marrow in Apo A-I/Apo E null mice. In this study, we compared the effect of dietary lipid lowering versus lipid lowering plus Apo A-IM gene transfer using recombinant adeno-associated virus (rAAV) 8 as vectors on atherosclerosis regression in Apo A-I/Apo E null mice. All mice were fed a high-cholesterol diet from age of 6 weeks until week 20, and at 20 weeks, 10 mice were euthanized to determine the extent of atherosclerosis. After 20 weeks, an additional 20 mice were placed on either a low-cholesterol diet plus empty rAAV (n = 10) to serve as controls or low-cholesterol diet plus 1 single intravenous injection of 1.2 × 10(12)vector genomes of adeno-associated virus (AAV) 8 vectors expressing Apo A-IM (n = 10). At the 40 week time point, intravenous AAV8 Apo A-IM recipients showed a significant regression of atherosclerosis in the whole aorta (P< .01), aortic sinuses (P< .05), and brachiocephalic arteries (P< .05) compared to 20-week-old mice, whereas low-cholesterol diet plus empty vector control group showed no significant regression in lesion size. Immunostaining showed that compared to the 20-week-old mice, there was a significantly reduced macrophage content in the brachiocephalic (P< .05) and aortic sinus plaques (P< .05) of AAV8 Apo A-IM recipients. These data show that although dietary-mediated cholesterol lowering halts progression of atherosclerosis, it does not induce regression, whereas combination of low-cholesterol diet and AAV8 mediated Apo A-I Milano gene therapy induces rapid and significant regression of atherosclerosis in mice. These data provide support for the potential feasibility of this approach for atherosclerosis regression.
载脂蛋白A-1（Apo A-I）米兰型是一种天然存在的Apo A-1的Arg173突变为Cys的突变体，已在动物模型和一项小型2期人体试验中显示出可减轻动脉粥样硬化。我们通过在Apo A-I/Apo E基因敲除小鼠中移植逆转录病毒转导的骨髓，证明了与野生型Apo A-I基因相比，Apo A-I米兰型（Apo A-IM）基因具有更强的抗动脉粥样硬化作用。在本研究中，我们比较了使用重组腺相关病毒（rAAV）8作为载体进行饮食降脂与降脂加Apo A-IM基因转移对Apo A-I/Apo E基因敲除小鼠动脉粥样硬化消退的影响。所有小鼠从6周龄开始喂食高胆固醇饮食直至20周龄，在20周时，处死10只小鼠以确定动脉粥样硬化的程度。20周后，另外20只小鼠被置于低胆固醇饮食加空rAAV（n = 10）作为对照，或低胆固醇饮食加单次静脉注射1.2×10¹²个表达Apo A-IM的腺相关病毒（AAV）8载体基因组（n = 10）。在40周时间点，与20周龄小鼠相比，静脉注射AAV8 Apo A-IM的受体小鼠在整个主动脉（P<0.01）、主动脉窦（P<0.05）和头臂动脉（P<0.05）中动脉粥样硬化有显著消退，而低胆固醇饮食加空载体对照组在病变大小上没有显著消退。免疫染色显示，与20周龄小鼠相比，AAV8 Apo A-IM受体小鼠的头臂动脉（P<0.05）和主动脉窦斑块（P<0.05）中的巨噬细胞含量显著减少。这些数据表明，虽然饮食介导的胆固醇降低可阻止动脉粥样硬化的进展，但不会诱导其消退，而低胆固醇饮食与AAV8介导的Apo A-I米兰型基因治疗相结合可诱导小鼠动脉粥样硬化快速且显著消退。这些数据为这种方法用于动脉粥样硬化消退的潜在可行性提供了支持。
