Engineered Mitochondria for Therapeutic Donation and Mitochondrial Genome Editing

用于治疗性捐赠和线粒体基因组编辑的工程线粒体

基本信息

批准号：
10001252
负责人：
Seth Lawler Shipman
金额：
$ 283.5万
依托单位：
J. DAVID GLADSTONE INSTITUTES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-30 至 2025-05-31
项目状态：
未结题

项目摘要

Project Summary/Abstract The ~sixteen-thousand nucleotides of the mitochondrial genome play an outsized role in human health. This gene-dense, circular genome is contained, replicated, transcribed, and regulated independently from our nuclear genome. Its contents are critical to the function of nearly every cell in our body, so when a nucleotide is mutated or lost, physiological processes in cells break down. Mutations to the mitochondrial genome cause diseases often associated with degeneration of central nervous system, heart, and muscle. One in ~five-thousand people inherits a disease-causing mutation to the mitochondrial genome (e.g. Leber’s hereditary optic neuropathy), but that likely underestimates the total burden of disease, as one in ~two-hundred people carries a suspected pathogenic mutation. We do not understand the ramifications of all mitochondrial mutations, which are often tissue-specific and complex to diagnose. Moreover, the mitochondrial genome accumulates mutations throughout the life of an individual, which are thought to play a role in degenerative diseases like Parkinson’s and even normal cellular aging. Unlike the nuclear genome, where emerging technologies to re-write DNA in living cells have enabled breathtaking experimental insights and will soon enable precision genetic medicine, the mitochondrial genome has remained nearly untouched by scientists. The inner membrane of the mitochondria is impermeable to polynucleotides, so template-based repair or augmentation with new DNA is out of reach. This means we can only manage, and never truly cure an individual with a disease that stems from a mutation to their mitochondrial DNA. It also means that we cannot experimentally introduce precise mutations to the mitochondrial DNA to test the effect of particular changes under controlled conditions. The research contained in this proposal is aimed at overcoming these limitations. We will test means of engineering mitochondria to enable active transplantation of mitochondrial populations into living cells as a therapeutic in diseases of the mitochondrial genome. We will assess therapeutic efficacy of mitochondrial transplantation in models of degenerative diseases such as Leber’s hereditary optic neuropathy. We will also modify the genome of the transplanted mitochondria to introduce or fix mutations, allowing for phenotypic analysis in a controlled experimental framework. Completion of this work will yield new opportunities to treat intractable disease and a new molecular paradigm to investigate a biological system that is critical to all cells.

项目概要/摘要线粒体基因组的约一万六千个核苷酸在人类健康中发挥着巨大的作用。基因密集的环状基因组的包含、复制、转录和调节均独立于我们的细胞核基因组的内容对于我们体内几乎每个细胞的功能都至关重要，因此当核苷酸发生突变时。线粒体基因组的突变或丢失会导致疾病。通常与中枢神经系统、心脏和肌肉的退化有关，约五千人中就有一人。遗传了线粒体基因组的致病突变（例如莱伯遗传性视神经病），但是这可能低估了疾病的总负担，因为约两百人中就有一人携带疑似病毒我们不了解所有线粒体突变的后果，这些突变通常是组织特异性且诊断复杂。此外，线粒体基因组会积累突变。在个体的一生中，这被认为在帕金森氏症等退行性疾病中发挥着作用甚至正常的细胞老化。与核基因组不同的是，在活细胞中重写 DNA 的新兴技术已经使得令人惊叹的实验见解，很快将实现精准遗传医学——线粒体基因组科学家们几乎没有触及过线粒体的内膜。多核苷酸，因此基于模板的修复或使用新 DNA 的增强是不可能的，这意味着我们可以做到。只能控制，而不能真正治愈患有线粒体突变疾病的个体这也意味着我们无法通过实验向线粒体 DNA 引入精确的突变来进行测试。在受控条件下特定变化的影响。本提案中包含的研究旨在克服这些限制。改造线粒体，使线粒体群体能够主动移植到活细胞中我们将评估线粒体基因组疾病的治疗效果。我们还将对莱伯遗传性视神经病等退行性疾病模型进行移植。修改移植线粒体的基因组以引入或修复突变，从而实现表型在受控实验框架中进行分析将产生新的治疗机会。难治性疾病和新的分子范式来研究对所有细胞都至关重要的生物系统。