Dual Genome Omics: Adapting Genomics Techniques to Reveal Molecular Pathology in Mitochondrial Disease

项目总结 线粒体疾病是罕见的，通常是破坏性的遗传性疾病，影响大脑、肌肉和其他 器官由线粒体的遗传故障引起，线粒体是产生细胞大部分能量的细胞结构。 关键的是，没有经过验证的实验室测试来跟踪线粒体疾病的进展，也没有FDA批准的 治疗。为了改变这一点，研究线粒体紊乱存在许多障碍，其中我们 在这个项目中的地址。首先，因为它们通常是受影响最严重的，所以对线粒体的早期研究 疾病主要集中在患者的活组织检查样本上，例如需要手术才能获得的骨骼肌。基座 在临床和研究数据上，我们正在使用血液代替，使我们能够收集更多的样本，而不需要 痛苦而昂贵的手术。其次，需要两套基因--核基因和线粒体基因 制造线粒体；而大多数用于研究基因及其功能状态的方法只能用于 检查一种类型。我们正在使用改进的技术来研究是否存在损坏的 线粒体基因和核基因的功能状态同时存在。使用这两个新的 方法，我们发现了受损线粒体基因的一种新的细胞类型特异性模式。在这 我们的目标是确定受损线粒体基因的细胞类型特异性模式是否与疾病密切相关 进展，这些模式是如何产生的，以及受损的线粒体基因数量如何与 线粒体在单细胞中的功能。如果成功，这些实验将提高我们对 基因变化导致线粒体疾病的症状，并可能导致关于如何发展的理论 线粒体疾病严重程度的临床实验室测试，以及潜在的如何治疗线粒体的理论 精神错乱。

PROJECT SUMMARY Mitochondrial disorders are rare, often devastating genetic diseases affecting the brain, muscle, and other organs caused by genetic failure of mitochondria, cellular structures that produce most of the cell’s energy. Critically, there are no proven lab tests to track mitochondrial disease progression and no FDA-approved therapies. Many hurdles exist to studying mitochondrial disorders in order to change this, two of which we address in this project. First, because they are often most severely affected, earlier studies of mitochondrial disease have focused on patient biopsy samples such as skeletal muscle that require surgery to obtain. Based on clinical and research data, we are using blood instead, allowing us to collect many more samples without painful and expensive procedures. Second, two sets of genes—nuclear and mitochondrial—are needed to make mitochondria; and most methods used to study genes and their functional status can only be used to examine one type. We are using modified techniques to allow us to study the presence of damaged mitochondrial genes and functional status of nuclear genes at the same time. Using these two new approaches, we have discovered a new cell type-specific pattern of damaged mitochondrial genes. In this proposal, we aim to determine if cell type-specific patterns of damaged mitochondrial genes track with disease progression, how these patterns arise, and how the amount of damaged mitochondrial genes related to mitochondrial function in single cells. If successful, these experiments will improve our understanding of how genetic changes cause the symptoms of mitochondrial disease and may lead to theories of how to develop clinical lab tests for mitochondrial disease severity and, potentially, theories of how to treat mitochondrial disorders.