Administrative Supplement - Mitochondrial respiratory chain disease mechanistic and therapeutic modeling

项目概要（赠款）。线粒体呼吸链（RC）疾病是高度病态的 在所有年龄段具有显著异质性病因和表型的能量缺乏症， 系统，由两个基因组中超过350个不同基因的致病性变异引起。无药可救，药监局- 对于RC疾病，不存在批准的，也不存在临床试验验证的疗法。由于一刀切，单一疗法是 治疗模型不太可能使所有患者受益，但对于开发有意义的精确药物至关重要。 改善不同分子、生化或临床RC疾病亚型的健康状况。具体而言，临床前 在人类患者细胞和简单动物模型中的转译RC疾病研究可以有效地 确定有效的治疗线索和具体的机制目标，以有意义地改善整体健康。与 我们独特的“匹配的”基于核基因的RC疾病模型集（NDUFS 2，NUBPL，SURF 1， FBXL 4、C12 ORF 65、DLD）在3个进化上不同的种间的同源性。elegans（worm，invertebrate），D. rerio （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，我们处于有利地位，以进一步利用多物种建模方法。我确 在RC疾病模型中建立了一个高效的研究计划，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 支持将使这个基础和转化研究计划的重点与示范教学 机会，由国际公认的研究人员在过去14年中建立，以促进 通过识别中心疾病机制和领先的治疗候选药物， 不同的RC疾病亚型。具体来说，这个转化研究计划将集中在利用RC 疾病患者细胞和简单的动物模型，以研究两个总体主题的关键问题。 主题1是病理生理学研究，涉及4个项目领域：（i）了解机制基础 不同的器官病理生理学在不同的RC疾病中占主导地位，（ii）发展敏感的 纳米传感器，以非侵入性地量化体内线粒体功能，（iii）解密功能性线粒体功能， RC中线粒体蛋白新的翻译后修饰（包括N-糖基化）的意义 （iv）确定哪个或哪些中枢营养传感信号网络（NSSN）节点，以及它们的 调节细胞蛋白毒性应激的下游生物化学途径，以治疗性靶向特定的 RC疾病亚群。主题二是治疗模式，涉及三个项目领域：（v）认识最佳治疗模式 营养疗法（宏量营养素、维生素、辅助因子），以改善RC的健康和整体功能 （vi）利用转化动物和细胞模型进行高通量药物筛选， 化合物验证，以有效地识别针对不同RC疾病的高效、安全和精确的疗法 亚组，和（vii）确定主要RC疾病治疗是否会改善具有以下特征的疾病的健康状况： 继发性（如21三体）或急性RC功能障碍。

PROJECT SUMMARY (PARENT GRANT). 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.