MOLECULAR GENETICS OF CHOLESTEROL TRANSPORT AND CHOLESTEROL REVERSE TRANSPORT DISORDERS

胆固醇转运和胆固醇反向转运障碍的分子遗传学

• 批准号: 04454235
• 负责人: MABUCHI Hiroshi
• 金额: $ 3.9万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454235/
• 关键词: MOLECULAR GENETICS; LDL-RECEPTOR; FAMILIAL HYPERCHOLESTEROLEMIA; CETP DEFICIENCY; CHOLESTEROL REVERSE TRANSPORT; ATHEROSCLEROSIS

LDL-RECEPTOR ABNORMALITIES IN FAMILIAL HYPERCHOLESTEROLEMIA.More than 150 different mutations in the LDL receptor gene have been reported in the world. We have collected 14 homozygotes and more than 1,300 heterozygotes of FH.Seven variants of LDL receptor gene have been identified in our laboratory. Four mutants showed large deletions detected by Southern blot analysis, and three mutants were point mutations detected by SSCP analysis and direct sequencing of PCR products. These seven mutants of 85 patients from 31 families accounted for only 15.5% of FH.FH Tonami-1 produces a mutant precursor of about 100 KDal which has no O-linked sugar domain. FH Tonami-2 has a variant of LDL receptor gene with 10Kb deletion eliminating exons 2 and 3. FH Tsuruga homozygote and FH Kanazawa-2 compound heterozygotes showed severe coronary heart disease. FH Morioka showed a point mutation from C to T in exon 9. Another compound heterozygote showed a new mutant named FH-Nanao.CHOLESTERYL-ESTER TRANSFER PR … More OTEIN (CETP) DEFICIENCY IN FAMILIAL HYPER-HDL-CHOLESTEROLEMIA.The genomic DNA of patients with CETP deficiency was used as a substrate for amplification of the CETP gene by PCR.At the 5' splice donor of intron 14 (position+1) there was a G to A change altering the strictly conserved G-T intron splice donor to A-T.We found two novel mutants of CETP gene. One splice donor site mutant is a thymidine insertion in +3 position in intron 14, which will, again, result in splicing defect. Another new mutant is a missense mutation in exon 15, producing change of aspartic acid into glycine. This mutant is also highly frequent, almost 1 in 10. Thus, these two common mutants produce at least 20 CEPT heterozygotes in 214 general subjects, and might raise the HDL-cholesterol levels and reduce coronary heart disease in the Japanese.DOUBLE HETEROZYGOTES OF FH AND CETP DEFICIENCY.We have screened CETP gene abnormalities in 348 FH patients, and found 16 double heterozygotes of LDL receptor gene and CETP gene, and 1 heterozygous FH patient combined with homozygous CETP deficiency. These double heterozygotes were often complicated by coronary heart disease, and 4 patients showed myocardial infarction and 4 showed angina pectoris. Thus, half of the 16 patients showed definite coronary heart disease. From these findings we suggest that atherogenicity of hyper-LDL-cholesterolemia in FH is more powerful than antiatherogenicity of hyper-HDL-cholesterolemia in CETP deficiency. Less

家族性高胆固醇血症中的ldl受体异常。世界上已经报道了150多种不同的LDL受体基因突变。我们收集了14个FH纯合子和1300多个FH杂合子。在我们的实验室已经鉴定出七种LDL受体基因变异。4个突变体通过Southern blot检测到大缺失，3个突变体通过SSCP分析和PCR产物直接测序检测到点突变。这7个突变体来自31个家族的85例患者，仅占FH的15.5%。FH Tonami-1产生约100 KDal的突变前体，没有o -连接糖结构域。FH Tonami-2具有低密度脂蛋白受体基因的10Kb缺失，消除了外显子2和3。FH鹤贺纯合子和FH金泽-2复合杂合子表现为严重冠心病。FH Morioka在第9外显子中出现了从C到T的点突变。另一个复合杂合子显示了一个新的突变体FH-Nanao。胆固醇-酯转移：家族性高hdl -胆固醇血症中更多的蛋白（CETP）缺乏。以CETP缺乏症患者的基因组DNA为底物，用PCR扩增CETP基因。在14号内含子的5'剪接供体（+1位置）发生了从G到a的变化，将严格保守的G- t内含子剪接供体转变为a - t。我们发现了两个新的CETP基因突变体。一个剪接供体位点突变是在14号内含子的+3位置插入胸腺嘧啶，这将再次导致剪接缺陷。另一个新的突变是外显子15的错义突变，使天冬氨酸变成甘氨酸。这种突变也非常频繁，几乎是十分之一。因此，这两种常见突变体在214名普通受试者中产生至少20个CEPT杂合子，并可能提高日本人的高密度脂蛋白胆固醇水平并减少冠心病。fh和cetp缺乏症双杂合子。我们对348例FH患者进行CETP基因异常筛查，发现LDL受体基因与CETP基因双杂合子16例，杂合子合并CETP纯合子缺失1例。这些双杂合子常并发冠心病，其中4例出现心肌梗死，4例出现心绞痛。因此，16例患者中有一半表现为明确的冠心病。根据这些发现，我们认为FH患者的高ldl -胆固醇血症的动脉粥样硬化性比CETP缺乏症患者的高hdl -胆固醇血症的抗动脉粥样硬化性更强。少

项目成果

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