CORRECTING MITOCHONDIRAL GENE MUTATIONS IN HUMAN OOCYTES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 我们建议研究是否将来自患者卵子的细胞核DNA（在细胞质中携带线粒体（mt）DNA突变）移植到由健康供体捐赠的去核卵子中，从而用健康的mtDNA替代线粒体缺陷，但保留患者的核基因组。接下来，我们将通过评估受精，体外胚胎发育和胚胎干细胞（ES）分离来检查重建卵子的发育能力。该提案中产生的结果很有可能为线粒体疾病的遗传干预提供可靠的新方法，并提高mtDNA突变母亲生下健康无症状孩子的机会。我们最近的研究证明了这种方法在恒河猴中的可行性和有效性。我们发现，染色体转移后产生的重构卵母细胞几乎是同质的，能够支持正常受精，并有能力进行足月发育。这些研究在OHSU妇产科生殖内分泌和不育科的机构内进行。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We propose to investigate if the nuclear DNA from a patient's egg, carrying mitochondrial (mt)DNA mutations in the cytoplasm, transplanted into an enucleated egg donated by a healthy donor, thereby replacing the mitochondrial defect with healthy mtDNA but retaining the patient's nuclear genome. Next, we will examine the developmental competence of reconstructed eggs by assessing fertilization, in vitro embryo development and isolation of embryonic stem (ES) cells. It is highly likely that results generated in this proposal will provide a reliable, new approach for genetic intervention in the case of mitochondrial diseases and improve the chances of a mother with mtDNA mutation having a healthy, asymptomatic child. Our recent studies demonstrate the feasibility and efficacy of this approach in the rhesus monkey. We showed that reconstructed oocytes produced after chromosome transfer are nearly homoplasmic, capable of supporting normal fertilization and competent for full term development. These studies are performed within the facilities of the Division of Reproductive Endocrinology and Infertility, Department of Obstetrics & Gynecology, OHSU.