CORRECTING MITOCHONDIRAL GENE MUTATIONS IN HUMAN OOCYTES

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 我们建议调查是否将患者卵子中携带线粒体(Mt)DNA突变的核DNA移植到健康捐赠者捐赠的去核卵子中，从而用健康的mtDNA取代线粒体缺陷，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分离来检验重建卵子的发育能力。这项提议产生的结果极有可能为线粒体疾病的遗传干预提供一种可靠的新方法，并提高携带mtDNA突变的母亲生下健康的、无症状的孩子的机会。我们最近的研究证明了这种方法在恒河猴身上的可行性和有效性。我们发现，染色体移植后产生的重构卵母细胞几乎是同质的，能够支持正常受精，并能够完全发育。这些研究是在俄亥俄州立大学妇产科生殖内分泌科和不孕科的设施内进行的。

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.