ADENOVIRAL GENE REPLACEMENT OF HEPATIC LIPASE IN HL-DEFICIENT MICE

肝脂酶(HL)是一种内皮结合酶，它能介导 IDL和高密度脂蛋白中存在的甘油三酯和磷脂的水解以及 因此，它在正常的脂蛋白代谢中起着核心作用。病人 HL缺乏症可能会增加发生 早产儿心血管疾病。 为了评估更换内皮细胞结合的可行性 在HL缺乏症的动物模型中，我们产生了脂解酶 含人HL基因和巨细胞病毒载体的重组腺病毒载体 启动子(r-HL-Adv)。HL缺陷(HL-Def)小鼠静脉注射 10~9 pfu重组HL-ADV(N=6)或对照组(N=3)。 输注前血脂值(mg/dl：TC=146加/减29，PL=283加/减56， TG=70+/-34，CE=80PLUS/-41，FC=66+/-33)呈上升趋势 TC(NL=101加/减8，P小于0.001)，PL(NL=181加/减15， P&lt；0.001)和FC(NL=35加/减3，P小于0.005) 年龄/性别匹配的正常小鼠(NL，N=13)；HL-Def小鼠的FPLC图谱 显示出高密度脂蛋白和磷脂含量相对于 控制小鼠。小鼠肝素后4天血浆免疫印迹分析 注射r-HL-ADV后，显示正常大小的HL的存在。 几乎所有表达的HL都在肝素后血浆中检测到，峰值 重组HL-DEF小鼠9193nmoles/min/ml.HL-Def小鼠的活动水平 ADV组TC(-23%，P<0.001)、PL(第4-5天)下降幅度最大 (-30%，P<0.002)和FC(-35%，P<0.05) 注射空白对照的小鼠血脂无明显变化。等离子体分析技术 FPLC显示高密度脂蛋白胆固醇和磷脂水平显著下降，形成 重组人HL-ADV中低密度脂蛋白大小的胆固醇和磷脂富集物 治疗过的小鼠。因此，使用重组腺病毒进行基因转移导致了 在HL缺陷小鼠中成功地替换HL以及 HL缺陷型脂蛋白表型的正常化。

Hepatic lipase (HL) is an endothelial bound enzyme which mediates the hydrolysis of triglycerides and phospholipids present in IDL and HDL and thus, plays a central role in normal lipoprotein metabolism. Patients with HL deficiency may be at increased risk for the development of premature cardiovascular disease. In order to evaluate the feasibility of replacing an endothelial bound lipolytic enzyme in an animal model for HL deficiency, we have generated a recombinant adenovirus vector containing the human HL cDNA and the CMV- promoter (r-HL-AdV). HL deficient (HL-Def) mice were injected IV with either ten to the ninth pfu of r-HL-AdV (N=6) or control vehicle (N=3). Pre-infusion lipid values (mg/dl: TC=146plus/minus29, PL=283plus/minus56, TG=70plus/minus34, CE=80plus/minus41, FC=66plus/minus33) showed increased TC (NL=101plus/minus8, pless than 0.001), PL (NL=181plus/minus15, p<0.001), and FC (NL=35plus/minus3, pless than 0.005) when compared to age/sex matched normal mice (NL, N=13); FPLC profile of HL-Def mice demonstrated a marked increase in HDL-C and PL content relative to control mice. Immunoblot analysis of mouse post-heparin plasma 4 days post-infusion of r-HL-AdV demonstrated the presence of normal sized-HL. Virtually all expressed HL was detected in post-heparin plasma with peak activity levels of 9,193 nmoles/min/ml. HL-Def mice injected with r-HL- AdV had a peak (day 4- 5) reduction in TC (-23%, pless than 0.001), PL (-30%, pless than 0.002) and FC (-35%, pless than 0.05) whereas animals injected with control vehicle had no lipid changes. Plasma analysis on FPLC showed dramatic decreases in HDL-C and PL as well as the formation LDL-sized cholesterol and phospholipid enriched particles in r-HL-AdV treated mice. Thus, gene transfer using recombinant adenovirus resulted in successful replacement of HL in HL-deficient mice as well as normalization of the HL-deficient lipoprotein phenotype.