Adenovirus-mediated gene transfer of rat apolipoprotein B mRNA-editing protein in mice virtually eliminates apolipoprotein B-100 and normal low density lipoprotein production.
Adenovirus-mediated gene transfer of rat apolipoprotein B mRNA-editing protein in mice virtually eliminates apolipoprotein B-100 and normal low density lipoprotein production.
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DOI：
10.1016/s0021-9258(18)43892-6
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发表时间：
1994-11
期刊：
影响因子：
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通讯作者：
B. Teng;S. Blumenthal;T. Forte;N. Navaratnam;J. Scott;A. Gotto;L. Chan
中科院分区：
文献类型：
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作者：
B. Teng;S. Blumenthal;T. Forte;N. Navaratnam;J. Scott;A. Gotto;L. Chan
载脂蛋白（apo）B-100是低密度脂蛋白（LDL）中的主要蛋白质成分;它含有LDL受体的结合结构域和脂蛋白（a）中载脂蛋白（a）的附着位点。ApoB-48与apoB-100的氨基末端的一半共线，并且错过了LDL受体相互作用和脂蛋白（a）形成所需的分子部分。ApoB-48 mRNA是通过编辑apoB-100 mRNA产生的，这是一个将Gln-2153的密码子CAA变为UAA（框内终止密码子）的过程。我们使用大鼠apoB mRNA编辑酶（REPR）的克隆催化组分构建了含有REPR cDNA的复制缺陷型重组腺病毒载体（AvREPR）和含有β-半乳糖苷酶cDNA的对照载体（Av 1 LacZ 4），以研究REPR基因递送在C57 BL/6小鼠中的效果。在小鼠中静脉注射AvREPR导致肝细胞的有效转导，其中REPR mRNA和蛋白质过表达，在7天和12天达到峰值，在AvREPR给药后39天恢复到对照水平。肝脏提取物中的apoB mRNA编辑活性显示出与REPR mRNA表达平行的变化;在REPR表达峰值时，编辑的apoB mRNA在总肝脏apoB mRNA中的比例从约60%增加到90%以上。AvREPR转导动物的血浆apoB-100比例从血浆apoB总浓度的约50%降至< 10%。对照动物的血浆极低密度脂蛋白是多分散的，平均直径分别为54.9 +/- 20.6 nm（未注射对照）和54.7 +/- 16.8 nm（Av 1 LacZ 4处理）。在AvREPR给药后第12天，它们变得更小（平均直径39.3 +/- 12.7 nm），尺寸更均匀。同一天，给药动物的正常血浆LDL（26.2-25.5 nm）几乎完全消除。腺病毒介导的REPR cDNA转移是降低血浆apoB-100和正常LDL产生的有效方法。
Apolipoprotein (apo) B-100 is the major protein component in low density lipoprotein (LDL); it contains the binding domain for the LDL receptor and the attachment site for apolipoprotein(a) in lipoprotein(a). ApoB-48 is colinear with the amino-terminal half of apoB-100 and misses the part of the molecule required for LDL receptor interaction and lipoprotein(a) formation. ApoB-48 mRNA is produced by the editing of apoB-100 mRNA, a process by which the codon CAA for Gln-2153 is changed to UAA, an in-frame stop codon. We used the cloned catalytic component of the rat apoB mRNA-editing enzyme (REPR) to construct a replication-defective recombinant adenoviral vector containing REPR cDNA (AvREPR) and a control vector (Av1LacZ4) containing a beta-galactosidase cDNA to investigate the effect of REPR gene delivery in C57BL/6 mice. Intravenous injection of AvREPR in mice resulted in efficient transduction of liver cells, where REPR mRNA and protein were overexpressed, reaching a peak at 7 and 12 days, returning toward control levels at 39 days after AvREPR administration. ApoB mRNA editing activity in liver extracts showed changes parallel to those of REPR mRNA expression; the proportion of edited apoB mRNA in the total hepatic apoB mRNA increased from approximately 60% to more than 90% at the peak of REPR expression. The proportion of plasma apoB-100 in AvREPR-transduced animals decreased from approximately 50% to < 10% of total plasma apoB concentration. Plasma very low density lipoproteins were polydisperse in control animals with an average diameter of 54.9 +/- 20.6 nm (uninjected control) and 54.7 +/- 16.8 nm (Av1LacZ4-treated), respectively. They became much smaller (average diameter 39.3 +/- 12.7 nm) and more uniform in size at day 12 following AvREPR administration. On the same day, the normal plasma LDL (26.2-25.5 nm) was almost completely eliminated in treated animals. Adenovirus-mediated transfer of the REPR cDNA is an efficient method to reduce plasma apoB-100 and normal LDL production.