GENE DIAGNOSIS AND GENE THERAPY OF CHOLESTEROL TRANSPORT AND CHOLESTEROL REVERSE TRANSPORT DISORDERS

胆固醇转运和胆固醇逆向转运障碍的基因诊断和基因治疗

• 批准号: 07457123
• 负责人: MABUCHI Hiroshi
• 金额: $ 3.97万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457123/
• 关键词: MOLECULAR GENETICS; LDL-RECEPTOR; FAMILIAL HYPERCHOLESTEROLEMIA; CETP DEFICIENCY; CHOLESTEROL REVERSE TRANSPORT; ATHEROSCLEROSIS; MTP DEFICIENCY; ABETALIPOPROTEINEMIA; MIP欠損症; 動脈硬化症

LDL-RECEPTOR ABNORMALITIES IN FAMILIAL HYPERCHOLESTEROLEMIA.More than 230 different mutations in the LDL receptor gene have been reported in the world. We have collected 20 homozygotes and more than 1,500 heterozygotes of FH.Nine variants of LDL receptor gene have been identified in our laboratory. Four mutants showed large deletions detected by Southern blot analysis, and 5 mutants were point mutations detected by SSCP analysis and direct sequencing of PCR products. These 9 mutants accounted for only 17.5% of FH.FH Tsuruga showed a point mutation in exon 6 (280 Asp-Tyr). FH Kanazawa-2 showed a point mutaiopn of 395 Arg-Trp. FH Morioka shwoed a point mutation from C to T (395 Arg-Trp) in exon 9.FH Nanao showed 23 stop mutant in exon 2.FH Yokote showed a mutation of 718 stop in exon 15.CHOLESTERYL-ESTER TRANSFER PROTEIN (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 PC … More R.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 68 CETP heterozygotes in 718 general subjects, and might raise the HDL-cholesterol levels and reduce coronary heart disease in the Japanese.LCATDEFICIENCY.The proband is 37 male patient, and his CHOL level was 228mg/dl, HDL-C was 34 mg/dl, his LCAT activity was 0%, and the gene analysis showed a point mutant of 30 Ser-Gly.MICROSOMAL TRANSFER PROTEIN (MTP) GENE MUTATION IN ABETALIPOPROTEINEMIA.The proband was 29 male patient, and his CHOL level was 33mg/dl, TG was 0mg/dl, and HDL-C was 28mg/dl. The gene analysis showed a point mutation in the junction of exon 10 and intron 9 (G to A), which would produce splicing abnormalities and no MTP protein. Less

家族性高脂血症的低密度脂蛋白受体异常：世界上已有230多种不同的低密度脂蛋白受体基因突变的报道。我们收集了20个FH纯合子和1500多个杂合子，在我们实验室已经鉴定出9个低密度脂蛋白受体基因的变异体。Southern杂交检测到4个突变体有较大的缺失，SSCP分析和PCR产物直接测序检测到5个突变体为点突变。这9个突变仅占FH的17.5%，FH Tsuruga在第6外显子(280 Asp-Tyr)有点突变。FH-Kanazawa-2显示395个Arg-Trp点突变。FH盛冈发现外显子9由C突变为T(395Arg-Trp)。FH Nanao发现外显子2有23个终止突变，FH Yokote发现外显子15有718个终止突变。以家族性高密度脂蛋白-胆固醇酯转移蛋白缺乏症患者基因组DNA为底物，用PC-…方法扩增在内含子14的5‘端剪接供体(+1位)上，发生了由G到A的转变，将严格保守的G-T内含子剪接供体改变为A-T。我们发现了两个新的CETP基因突变。一个剪接供体位点突变是在内含子14的+3位插入胸苷，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，导致天冬氨酸变成甘氨酸。先证者是37名男性患者，其胆固醇水平为228 mg/dl，高密度脂蛋白胆固醇水平为34 mg/dl，LCAT活性为0%，基因分析显示30个序列甘氨酸转运蛋白(MTP)基因点突变。微转移蛋白(MTP)基因突变为ABETALIPOPROTEINIA。先证者为29名男性患者，胆固醇水平为33 mg/dl，胆固醇水平为0 mg/dl，LCAT活性为0高密度脂蛋白胆固醇为28 mg/dl。基因分析显示外显子10和内含子9(G到A)交界处存在点突变，可产生剪接异常，不产生MTP蛋白。较少

项目成果

Nohara A,et al.: "Absence of familial defective apolipoprotein B-100 in Japanese patients with familiat hypercholesterolemia" Lancet. 345. 1435- (1995)

Nohara A 等人：“日本家族性高胆固醇血症患者缺乏家族性缺陷载脂蛋白 B-100”《柳叶刀》。

Kajinami K,Seki H,Takekoshi N,Mabuchi H: "Noninvasive prediction of coronary atherosclerosis by quantification of coronary artery calcification using electron beam computed tomography : comparison with electrocardiographic and thallium exercise stress tes

Kajinami K、Seki H、Takekoshi N、Mabuchi H：“使用电子束计算机断层扫描量化冠状动脉钙化来无创预测冠状动脉粥样硬化：与心电图和铊运动负荷测试的比较

Kajinami K,et al.: "Long-term probucol trcatment results in regression of xanhtoms,but in progression of coronary atherosclerosis in a heterozygous patient with familial hypercholesterolemia." Atherosclerosis. 120. 181-187 (1996)

Kajinami K 等人：“长期普罗布考治疗可导致家族性高胆固醇血症杂合子患者黄斑消退，但导致冠状动脉粥样硬化进展。”

Koizumi J,Haraki T,Yagi K,Inazu A,Kajinami K,Mabuchi H,et al.: "Clinical efficacy of fluvastatin in the long-term treatment of familial hypercholesterolemia." Am J Cardiol. 76. 47A (1995)

Koizumi J、Haraki T、Yagi K、Inazu A、Kajinami K、Mabuchi H 等：“氟伐他汀长期治疗家族性高胆固醇血症的临床疗效”。

Kajinami K,et al.: "Noninvasive prediction of coronary atherosclerosis by quantification of coronary artery calcification using electron beam conputed torrography : conparison with electrocardiographic and thallium exercise stress test results." J Am Coll

Kajinami K 等人：“通过使用电子束计算机断层扫描量化冠状动脉钙化来无创预测冠状动脉粥样硬化：与心电图和铊运动负荷测试结果进行比较。”