Establishment of genetic diagnostics, preventive and development of treatment for atherogenic hypertriglyceridemia

我们总结了从2004年到2006年的3年研究计划所获得的结果。(1)脂蛋白脂酶（LPL）基因突变的电化学阵列（ECA）芯片检测系统的改进及其在临床样品中的应用（Ikeda）：我们开发了一种新的同时多突变检测（SMMD）系统，使用ECA芯片和二茂铁萘二酰亚胺作为指示剂（参考文献# 3和4）。该系统可同时检测单核苷酸多态性、插入、缺失、易位和短串联重复序列等多种基因突变。通过使用SMMD系统，我们从高脂血症患者中新发现了两个LPL基因突变。(2)通过改善其生活方式以达到健康的生活方式来治疗杂合型LPL基因突变受试者中出现的高脂血症（Tamasawa和Ikeda）：杂合型LPL缺乏受试者（携带者）在以下情况下易于出现轻度高脂血症（IV型高脂蛋白血症）： ...更多信息 伴有环境因素，如高胰岛素血症状态和/或高酒精摄入，其刺激肝脏中的甘油三酯合成，而携带者是血脂正常的，只要他们没有环境因素（参考文献# 2）。我们的研究表明，通过让携带者改变到更健康的生活方式，携带者的高脂血症变成了正常的血脂状态。(3)基于LPL基因信息的蛋白酶抗性LPL分子的开发及其在治疗中的应用（Takagi和Ikeda）：因为天然LPL分子被血流中的蛋白酶灭活，所以通过将靶氨基酸交换为甘氨酸来构建蛋白酶抗性LPL分子。与天然LPL分子相比，蛋白酶抗性LPL分子表现出30%的甘油三酯水解功能。由于已明确蛋白酶抗性LPL分子与人血液混合后，该蛋白酶抗性LPL分子在37℃下可稳定至少6小时，因此证实了蛋白酶抗性LPL分子能够用作高血糖治疗的可能性。为了判断抗蛋白酶LPL分子对高脂血症的治疗效果，有必要检测肝甘油三酯脂酶（HTGL）的分子特征。因此，建立了用于测定HTGL质量的ELISA系统（参考# 1）。少

We summarize results obtained from the research program of 3 years through 2004 to 2006.(1) Improvement of detection system using electrochemical array (ECA) chip for lipoprotein lipase (LPL) gene mutation and its application to clinical samples (Ikeda) : We developed a new simultaneous multiple mutation detection (SMMD) system using an ECA chip and ferrocenylnapthalene diimide, as the indicator (Ref.# 3 and 4). This SMMD system can simultaneously identify several genetic mutations such as single nucleotide polymorphism, insertion, deletion, translocation and short tandem repeat. By using the SMMD system, we newly identified two LPL gene mutations from subjects with hypertriglyceridemia.(2) Treatment for hypertriglyceridemia developed in subjects with heterozygous LPL gene mutation by improving their lifestyle to a healthful one (Tamasawa and Ikeda) : Subjects (carriers) with heterozygous LPL deficiency are prone to develop mild hypertriglyceridemia (type IV hyperlipoproteinemia) when … More complicated with environmental factors such as a hyperinsulinemic state and/or a high alcohol intake, which stimulate triglyceride synthesis in the liver, while carriers are normolipidemic provided that they do not have the environmental factors (Ref.# 2). Our research revealed that hypertriglyceridemia in the carriers became normolipidemic state by getting the carriers to change to a more healthful lifestyle.(3) Development of a protease-resistant LPL molecule based on LPL gene information and its application to treatment (Takagi and Ikeda) : Because a native LPL molecule was inactivated by proteases in bloodstream, the protease-resistant LPL molecule was constructed by exchanging a target amino acid to a glycine. The protease-resistant LPL molecule exhibited 30% of the triglyceride hydrolyzing function in comparison to that of the native LPL molecule. Since it was clarified that this protease-resistant LPL molecule was steady for at least six hours at 37℃ after the protease-resistant LPL molecule was mixed with human blood, the possibility that the protease-resistant LPL molecule was able to be used as a hypertriglyceridemic treatment was confirmed. To judge the treatment effect of the protease-resistant LPL molecule to hypertriglyceridemia, it was necessary to detect a molecular characteristic of hepatic triglyceride lipase (HTGL). Therefore, ELISA system for the determination of HTGL mass was established (Ref.# 1). Less