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A basic study for mitochondria-targeting liposome vector using human mitochondrial fusiogenic protein.

使用人线粒体融合蛋白进行线粒体靶向脂质体载体的基础研究。

基本信息

批准号：
14570129
负责人：
ENDO Hitoshi
金额：
$ 2.18万
依托单位：
Jichi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

The mitochondria are organelle in the cell and have the proper DNA different from the nuclear genome. The development of the new way of introducing a material into the mitochondria, which is different from the usual gene therapy to the nuclear target, is necessary for the fundamental treatment of the mitochondrial disease based on the mitochondrial DNA. In this research, it aims at the fundamental development of new liposome vector that peculiarly targets the mitochondria, using the mitochondrial fusiogenc factor as an effective component. Such a vector will make it possible that not only normal mitochondrial DNA but also the drug adjusting the mitochondrial function and/or the exogenous ribozyme regulating the mitochondrial transcripts are introduced into the mitochondria. The development of such a vector would become applied for a new drug delivery system for many diseases with dysfunction of the mitochondria in the future.We identified a mitochondrial fusiogenic protein, a human Fzo homolog, and analyzed the domain of the protein necessary for membrane fusiogenic activity. We also produced the recombinant protein of the domain, reconstituted the proteoliposome using the protein, and proved the membrane fusiogenic activity of the liposome in vitro. On the other hand, we examined the OPA1 protein which has the mitochondrial fusiogenic activity, and showed the differential sublocalization of OPA1 isoform in the human cell. This factor is one of the assistant factors of the mitochondrial fusiogenic device, which will be important for improvement for the efficiency of the liposome vector. Next step requires an examination whether the mitochondrial DNA-embedded liposome, introduced in the cell by microinjection, actually targets the mitochondria in vivo. This research is the first step to establish the fundamental technology of the gene therapy of mitochondrial diseases.

线粒体是细胞中的细胞器，具有不同于核基因组的适当DNA。开发一种不同于通常的针对核靶点的基因治疗的将材料引入线粒体的新方法对于基于线粒体DNA的线粒体疾病的根本治疗是必要的。本研究以线粒体促融合因子为有效成分，旨在初步开发特异靶向线粒体的新型脂质体载体。这样的载体将使得不仅正常的线粒体DNA而且调节线粒体功能的药物和/或调节线粒体转录物的外源核酶被引入线粒体成为可能。这种载体的开发将成为应用于许多疾病的线粒体功能障碍的新的药物输送系统在future.We确定了线粒体融合蛋白，人类FZO同源物，并分析了膜融合活性所需的蛋白质的结构域。我们还制备了该结构域的重组蛋白，利用该蛋白重构了脂质体，并在体外证实了脂质体的膜融合活性。另一方面，我们检测了具有线粒体融合活性的OPA 1蛋白，并显示了OPA 1亚型在人细胞中的差异亚定位。该因子是线粒体融合装置的辅助因子之一，对于提高脂质体载体的效率具有重要意义。下一步需要检查通过显微注射引入细胞的线粒体DNA包埋脂质体是否真的靶向体内线粒体。本研究为建立线粒体疾病基因治疗的基础技术迈出了第一步。

项目成果

期刊论文数量（71）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Establishment of Cre/LoxP recombination system in transgenic rats.

DOI：
10.1016/j.bbrc.2004.04.204
发表时间：
2004-07
期刊：
Biochemical and biophysical research communications
影响因子：
3.1
作者：
Yuki Sato;H. Endo;T. Ajiki;Y. Hakamata;T. Okada;T. Murakami;E. Kobayashi
通讯作者：
Yuki Sato;H. Endo;T. Ajiki;Y. Hakamata;T. Okada;T. Murakami;E. Kobayashi

Polymorphism, heteroplasmy, mitochondrial fusion and diabetes.

多态性、异质性、线粒体融合和糖尿病。

DOI：
发表时间：
2003
期刊：
Bioscience Rep. 23
影响因子：
0
作者：
Sato;A.;Endo;H.;Umetsu;K.;Sone;H.;Yanagisawa;Saigusa;A.;Aita;S.;Kagawa;Y.
通讯作者：
Y.

Stage-specific regulatory element of mouse Sry gene.

小鼠 Sry 基因的阶段特异性调控元件。

DOI：
发表时间：
2003
期刊：
Mol.Reprod.Dev. 64
影响因子：
0
作者：
Yokouchi;K.;Ito;M.;Nishino;K.;Yamanouchi;K.;Naito;K.;Suzawa;M.;Kato;S.;Hakamata;Y.;Endo;H.;Tojo H.
通讯作者：
Tojo H.

Hayakawa, M.: "Muscle-specific exonic splicing silencer for exon exclusion in human ATP synthase γ-subunit pre-mRNA"J.Biol.Chem.. 277. 6974-6984 (2002)

Hayakawa, M.：“用于人 ATP 合酶 γ 亚基前 mRNA 中外显子排除的肌肉特异性外显子剪接沉默子”J.Biol.Chem.. 277. 6974-6984 (2002)