Mitochondrial respiratory chain disease mechanistic and therapeutic modeling

项目摘要。线粒体呼吸链（RC）疾病是高度病态的能量缺乏 在所有年龄和系统中具有显著异质性病因和表型的疾病，由以下原因引起： 两个基因组中> 350个不同基因的致病性变体。没有治愈方法，FDA批准，也没有临床试验- 对于RC疾病存在有效的疗法。由于一刀切，单一疗法不太可能使所有患者受益， 治疗建模对于开发精确的药物至关重要，这些药物可以显著改善健康状况， 分子、生物化学或临床RC疾病亚型。具体而言，临床前转化性RC疾病 在人类患者细胞和简单的动物模型中的研究可以有效地鉴定有效的治疗方法， 领导和具体的机制目标，以有意义地改善整体健康。凭借我们独特的 “匹配的”基于核基因的RC疾病模型组（对于NDUFS 2，NUBPL，SURF 1，FBXL 4，C12 ORF 65， DLD）在3个进化上不同的种间的差异。elegans（worm，invertebrate），D.鱼（斑马鱼，脊椎动物）， 和人类患者成纤维细胞，我们已经验证了一套新的方法，我们强烈 以进一步利用多物种建模方法。我确已建立了一个高度的 RC疾病模型的生产性研究计划，以交叉验证多种机制的见解，并确定 有前途的新的治疗线索，为原发性RC疾病。NIGMS R35 MIRA支持将使重点放在这一点上 基础和转化研究计划与证明教学机会，如建立在过去14 由国际公认的研究人员，以推进精密线粒体医学，通过识别 中心疾病机制和不同RC疾病亚型的主要治疗候选者。具体地说， 这项转化研究计划将侧重于利用RC疾病患者细胞和简单的动物模型 调查两个重要主题的关键问题。主题1是病理生理学研究， 涉及4个项目领域：（i）了解不同器官病理生理学 在不同的RC疾病中占主导地位，（ii）开发敏感的纳米传感器，以非侵入性地在体内定量 线粒体功能，（iii）破译新的翻译后修饰的功能意义， 包括N-糖基化，以及（iv）确定哪种中心营养素- 传感信号网络（NSSN）节点，以及它们调节细胞内信号传导的下游生物化学途径。 蛋白毒性应激，以治疗特定RC疾病亚群。主题2是治疗建模， 涉及3个项目领域：㈤确认最佳营养疗法（常量营养素、维生素、辅因子） 改善RC疾病的健康和整体功能，（vi）利用转化动物和细胞模型 用于高通量药物筛选和先导化合物验证，以有效地鉴定高效、安全、 精确治疗不同的RC疾病亚组，和（vii）确定原发性RC疾病是否 治疗将改善具有继发性（如21三体）或急性RC功能障碍的疾病的健康。

PROJECT SUMMARY. Mitochondrial respiratory chain (RC) diseases are highly morbid energy deficiency disorders with remarkably heterogeneous etiologies and phenotypes across all ages and systems, caused by pathogenic variants in > 350 different genes across both genomes. No cure, FDA-approved, nor clinical trial- validated therapies exists for RC diseases. As one-size-fits-all, single therapy is unlikely to benefit all patients, therapeutic modeling is essential to develop precision medicines that meaningful improve health in distinct molecular, biochemical, or clinical RC disease subtypes. Specifically, pre-clinical translational RC disease investigations in human patient cells and simple animal models may efficiently identify potent therapeutic leads, and specific mechanistic targets, to meaningfully improve overall health. With our unique collection of ‘matched’ nuclear gene-based RC disease model sets (for NDUFS2, NUBPL, SURF1, FBXL4, C12ORF65, DLD) across 3 evolutionarily distinct species in C. elegans (worm, invertebrate), D. rerio (zebrafish, vertebrate), and human patient fibroblasts in which we have validated a suite of novel methodologies, we are strongly situated to further harness a multi-species modeling approach. Indeed, we have established a highly productive research program in RC disease models to cross-validate multiple mechanistic insights and identify promising new therapeutic leads for primary RC diseases. NIGMS R35 MIRA support will enable focus of this basic and translational research program with demonstrated teaching opportunities, as built over the past 14 years by an internationally-recognized investigator, to advance precision Mitochondrial Medicine by identifying central disease mechanisms and lead therapeutic candidates for diverse RC disease subtypes. Specifically, this translational research program will focus on harnessing RC disease patient cell and simple animal models to investigate key questions across 2 overarching themes. Theme 1 is Pathophysiology Investigations, involving 4 project areas: (i) Understanding the mechanistic basis by which different organ pathophysiology predominates in distinct RC diseases, (ii) Developing sensitive nanosensors to non-invasively quantify in vivo mitochondrial functions, (iii) Deciphering the functional significance of novel post-translational modifications, including N-glycosylation, of mitochondrial proteins in RC disease, and (iv) Determining which central nutrient- sensing signaling network (NSSN) node(s), and their downstream biochemical pathways that regulate cellular proteotoxic stress, to therapeutically target in specific RC disease subsets. Theme 2 is Therapeutic Modeling, involving 3 project areas: (v) Recognizing the optimal nutritional therapies (macronutrients, vitamins, cofactors) to improve health and overall function in RC disease, (vi) Harnessing translational animal and cellular models for high-throughput drug screening and lead compound validation, to efficiently identify highly potent, safe, and precision therapies for distinct RC disease subgroups, and (vii) Identifying whether primary RC disease treatments will improve health in disorders with secondary (such as Trisomy 21) or acute RC dysfunction.