Establishment of personal protection method for life-style diseases : type IV hyperlipoproteinemia as a model case

生活方式病个体防护方法的建立——以IV型高脂蛋白血症为典型病例

• 批准号: 11670402
• 负责人: TAKAGI Atsuko
• 金额: $ 1.86万
• 依托单位: National Cardiovascular Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670402/
• 关键词: Lipoprotein lipase; Hypertriglyceridemia; Type IV hyperlipoproteinemia; Heterozygote; Tailor-made medicine; Life-style disease; Mutation; Single nucleotide polymorphism; へテロ接合体

The aim of this study is to establish individual methods of prevention for life-style-related diseases, especially hypertriglyceridemia as a model case, because we think it is practical to focus on an individual genetical weakness for prevention against life-style-related diseases. We have elucidated etiology of primary type IV hyperlipidemia is a genetic background of heterozygous lipoprotein lipase (LPL) deficiency and superimposing triglyceride synthesis-stimulating factor on its genetic background. It means a LPL heterozygous deficient person without triglyceride synthesis-stimulating factor doesn't manifest hypertriglyceridemia. Individuals with heterozygous LPL deficiency have to be more carefully of superimposition of triglyceride synthesis-stimulating factor, alcohol drinking for example, than persons with two alleles of a normal LPL gene. The reliable and accurate genetic diagnosis of heterozygous LPL aberration is needed for development of individual methods of prevention for … More hypertiglyceridemia. We developed and improved LPL and hepatic triglyceride lipase mass measurement methods, direct sequencing of LPL gene, PCR method which didn't overlook mutations, and hypertriglyceride-induced atherogenic small dense LDL detection method. On the basis of these developments, we accumulated LPL gene mutations in Japanese. From the subjects with low LPL mass values, Y61X, G188E, D204E, Int3-3' c(-6)t, G154V, G105R, Int8-5' t(2)c mutations were detected. These missense mutations were confirmed to lead non-functional LPL production with COS-1 in vitro-expression system. As the Int3-3' c(-6)t mutation didn't have an aberrant splicing product in vivo and in vitro, this mutation seemed to link to another mutation which led to LPL deficiency. The Int8-5' t(2)c mutation led to the utilization of a cryptic 5' donor splice site in exon8 as an alternative splice site, skipping of a 134-bp fragment of exon8. These technical developments and improvements, and an accumulation of LPL mutations would contribute to LPL gene diagnosis that makes individual methods of prevention for hypertriglyceridemia possible. Less

本研究的目的是建立预防生活方式相关疾病的个体方法，特别是高甘油三酯血症作为模型案例，因为我们认为关注个体遗传弱点来预防生活方式相关疾病是可行的。我们已经阐明了原发性IV型高脂血症的病因是杂合脂蛋白脂肪酶（LPL）缺乏和甘油三酯合成刺激因子叠加的遗传背景。这意味着没有甘油三酯合成刺激因子的LPL杂合缺陷者不会表现出高甘油三酯血症。杂合子LPL缺乏症患者必须比具有正常LPL基因的两个等位基因的人更小心甘油三酯合成刺激因子的叠加，例如饮酒。需要对杂合LPL畸变进行可靠、准确的遗传诊断，以开发预防高甘油三酯血症的个性化方法。我们开发并改进了LPL和肝脏甘油三酯脂肪酶质量测定方法、LPL基因直接测序法、不忽略突变的PCR法、高甘油三酯致动脉粥样硬化小密度LDL检测方法。在这些进展的基础上，我们积累了日本人的LPL基因突变。在低LPL质量值的人群中检测到Y61X、G188E、D204E、Int3-3′c(-6)t、G154V、G105R、Int8-5′t(2)c突变。在体外表达系统中，这些错义突变被证实导致COS-1产生非功能性LPL。由于Int3-3' c(-6)t突变在体内和体外都没有异常的剪接产物，因此该突变似乎与另一个导致LPL缺陷的突变有关。Int8-5‘ t(2)c突变导致外显子8上一个隐性的5’供体剪接位点被用作备选剪接位点，从而跳过了外显子8的134 bp片段。这些技术的发展和改进，以及LPL突变的积累将有助于LPL基因的诊断，使预防高甘油三酯血症的个体方法成为可能。少

项目成果

Tamazawa, N.: "Identification of homozygous lipoprotein lipase gene mutation in a woman with recurrent aggravation of hypertriglyceridemia induced by pregnancy (in Japanese)"The Lipid. 11. 79-84 (2000)

Tamazawa, N.：“妊娠引起的高甘油三酯血症反复加重的女性中纯合脂蛋白脂肪酶基因突变的鉴定（日语）”The Lipid。

Kimura, H.: "Development and evaluation of a direct sandwich enzyme-linked immunosorbent assay for the quantification of lipoprotein lipase mass in human plasma."Clinical Biochemistry. 32. 15-23 (1999)

Kimura, H.：“用于定量人血浆中脂蛋白脂肪酶质量的直接夹心酶联免疫吸附测定的开发和评估。”临床生物化学。

Nishimura, M.: "Development and evaluation of a direct sandwich-enzyme-linked immunosorbent assay for the quantification of human hepatic triglyceride lipase mass in human plasma."J Immunol Methods. 235. 41-51 (2000)

Nishimura, M.：“用于定量人血浆中人肝甘油三酯脂肪酶质量的直接夹心酶联免疫吸附测定的开发和评估。”J 免疫方法。

Kimura H: "Development and evaluation of a direct sandwich enzyme-linked immunosorbent assay for the quantification of lipoprotein lipase mass in human plasma"Clinn Biochem. 32・1. 15-23 (1999)

Kimura H：“用于定量人血浆中脂蛋白脂肪酶质量的直接夹心酶联免疫吸附测定的开发和评估”Clinn Biochem 32·1（1999）。

Mori A: "Improved method for direct DNA sequencing of the lipoprotein lipase gene using a DNA autosequencer"Clin Biochem. 33. 323-327 (2000)

Mori A：“使用 DNA 自动测序仪对脂蛋白脂肪酶基因进行直接 DNA 测序的改进方法”Clin Biochem。