Mitochondrial Genomes in Aging & Neurodegeneration

描述（由申请人提供）：存在大量证据支持氧化损伤线粒体基因组有助于衰老和神经退行性变病理生理的假设。线粒体基因受损和线粒体氧化应激是组织生物能量衰竭、非有丝分裂肌肉细胞和神经元过早死亡以及分裂组织中端粒功能障碍导致的细胞过早衰老的主要原因。该STTR项目进一步开发线粒体基因组转染（“mitofection”）和线粒体基因组去除（“mitoclean”）的新技术，以实现商业化。将实现三个目标。在Aim 1中，有丝分裂转染技术将根据转染参数进行优化。在Aim 2中，将证明基于rnai的基因沉默和有丝分裂联合用野生型mtDNA取代致病性mtDNA并纠正异常代谢表型的可行性。在Aim 3中，基于rnai的基因沉默将用于产生无mtdna （rho0）的人类原代神经元和成纤维细胞系，这将作为治疗开发的独特平台。这个小组已经证明了这两项主要技术的可行性。这一提议的结果将是一种独特的能力来操纵整个线粒体基因组的线粒体细胞。由此将发展线粒体基因替代疗法的方法，以逆转衰老的有害影响并防止神经退行性变。第一阶段STTR的结果将产生可以很容易地扩展到第二阶段建议的结果。“Mitofection”和“Mitoclean”都是科学上引人注目的技术，适用于多种组织，包括神经和非神经组织。Gencia和弗吉尼亚大学CSND之间的互动已经很好地建立和富有成效。

DESCRIPTION (provided by applicant): Substantial evidence exists to support the hypothesis that oxidatively-damaged mitochondrial genomes contribute to the pathophysiology of aging and neurodegeneration. Damaged mitochondrial genes and mitochondrial oxidative stress contribute to or are primarily causal in bioenergetic failure of tissues, premature cell death of non-mitotic muscle cells and neurons and premature cell senescence through telomere dysfunction in dividing tissues. This STTR project further develops novel technologies of mitochondrial genome transfection ("mitofection") and mitochondrial genome removal ("mitoclean") towards commercialization. Three Aims will be addressed. In Aim 1 mitofection technology will be optimized with respect to transfection parameters. In Aim 2, the feasibility of combined RNAi-based gene silencing and mitofection to replace pathogenic mtDNA with wild-type mtDNA and correct abnormal metabolic phenotype will be demonstrated. In Aim 3, RNAi-based gene silencing will be used to produce mtDNA-free (rho0) lines of human primary neurons and fibroblasts that will serve as unique platforms for therapeutic development. This group has already demonstrated the feasibility of both major technologies. The outcome of this proposal will be a unique ability to manipulate the entire mitochondrial genome in mitochondria of cells. From this will develop methodologies for mitochondrial gene replacement therapies to reverse deleterious effects of aging and to prevent neurodegeneration. The results of this Phase I STTR will yield results that can readily be expanded into a Phase II proposal. Both "Mitofection" and "Mitoclean" are technologies that are scientifically compelling and applicable to multiple tissues, neural and non-neural. The interaction between Gencia and University of Virginia CSND is already well established and productive.