腺相关病毒LPL(S447X)基因疗法在低密度脂蛋白受体敲除小鼠中的应用

Adeno-associated virus LPL(S447X) gene therapy in LDL receptor knockout mice.

作者信息

Rip Jaap, Sierts Jeroen A, Vaessen Stefan F C, Kastelein John J P, Twisk Jaap, Kuivenhoven Jan Albert

机构信息

Department of Vascular Medicine, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.

出版信息

Atherosclerosis. 2007 Sep;194(1):55-61. doi: 10.1016/j.atherosclerosis.2006.09.023. Epub 2006 Nov 7.

DOI:10.1016/j.atherosclerosis.2006.09.023

PMID:17087965

Abstract

BACKGROUND

Overexpression of lipoprotein lipase (LPL) protects against atherosclerosis in genetically engineered mice. We tested whether a gene therapy vector that delivers human (h) LPL(S447X) cDNA to skeletal muscle could induce similar effects.

METHODS

LDL receptor knockout (LDLr-/-) mice were injected intramuscular (i.m.) with adeno-associated virus serotype 1 (AAV1) LPL(S447X) or PBS. Four weeks later they were started on an atherogenic diet for 12 weeks. After termination, atherosclerosis was assessed and homogenates of muscle and liver tissue were analyzed.

RESULTS

AAV1-treated mice showed hLPL concentrations of 768+/-293 ng/mL in post-heparin plasma associated with 48% reductions of fasting triglycerides (TG) levels (p<0.0001). In the absence of an effect on total cholesterol (TC) levels, no effects on atherosclerosis were found. An increase in lipid content of injected muscles was accompanied by a significant decrease of TG (-20%, p<0.0001) and free cholesterol (FC) content (-24%, p<0.0001) in liver homogenates.

CONCLUSIONS

The data show that transgenic hLPL(S447X) on top of endogenous murine LPL reduces fasting TG levels in plasma but has no effect on atherosclerosis in LDLr-/- mice. While lipid accumulation in the injected muscle was anticipated, this coincided with an interesting decrease of both TG and FC in liver homogenates.

摘要

背景

脂蛋白脂肪酶（LPL）的过表达可保护基因工程小鼠免受动脉粥样硬化的影响。我们测试了一种将人（h）LPL（S447X）cDNA传递至骨骼肌的基因治疗载体是否能诱导类似的效果。

方法

给低密度脂蛋白受体敲除（LDLr-/-）小鼠肌肉内（i.m.）注射1型腺相关病毒（AAV1）LPL（S447X）或磷酸盐缓冲液（PBS）。四周后，让它们开始接受致动脉粥样化饮食12周。处死小鼠后，评估动脉粥样硬化情况，并分析肌肉和肝脏组织的匀浆。

结果

接受AAV1治疗的小鼠肝素后血浆中hLPL浓度为768±293 ng/mL，同时空腹甘油三酯（TG）水平降低了48%（p<0.0001）。在对总胆固醇（TC）水平无影响的情况下，未发现对动脉粥样硬化有影响。注射肌肉的脂质含量增加，同时肝脏匀浆中的TG（-20%，p<0.0001）和游离胆固醇（FC）含量（-24%，p<0.0001）显著降低。