Administrative Supplement (Undergraduate Summer Research Experiences) - Mitochondrial respiratory chain disease mechanistic and therapeutic modeling

项目总结(母公司赠款)。线粒体呼吸链(RC)疾病是高度病态的 能量缺乏性疾病在所有年龄段和 系统，由两个基因组中350个不同基因的致病变异引起。没有解药，FDA- 对于RC疾病，既没有经过批准的，也没有经过临床试验验证的治疗方法。作为一刀切的疗法，单一疗法 不太可能使所有患者受益，治疗性建模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。具体地说，临床前 在人类患者细胞和简单动物模型中进行转译RC疾病研究可能是有效的 确定有效的治疗线索和特定的机械靶点，以有意义地改善整体健康。使用 我们独一无二的基于核基因的RC疾病模型集合(针对NDUFS2、NUBPL、SURF1、 FBXL4、C12ORF65、DLD)分布于线虫(线虫、无脊椎动物)、线虫(线虫、无脊椎动物)中 (斑马鱼、脊椎动物)和人类患者成纤维细胞，我们在其中验证了一套新的 在方法学方面，我们很有能力进一步利用多物种建模方法。事实上，我们 在RC疾病模型中建立了高效的研究计划，以交叉验证多个 机械洞察力，并确定有希望的新的治疗线索的原发RC疾病。NIGMS R35 Mira 支持将重点放在这个基础和翻译研究项目上，并进行示范教学 由国际公认的调查员在过去14年中建立的机会，以推进 通过确定中枢疾病机制和主导治疗候选药物的精确线粒体医学 对于不同的RC疾病亚型。具体地说，这个翻译研究计划将重点放在利用RC 疾病患者细胞和简单的动物模型，研究两个主要主题的关键问题。 主题1是病理生理学研究，涉及4个项目领域：(I)了解机制基础 不同的器官病理生理学在不同的RC疾病中起主导作用，(Ii)发展敏感 纳米传感器非侵入性地量化体内线粒体功能，(Iii)破译功能 线粒体蛋白的新的翻译后修饰，包括N-糖基化，在RC中的意义 疾病，以及(Iv)确定哪个中央营养感知信号网络节点(S)及其 调节细胞蛋白毒性应激的下游生化通路，以特定的治疗为靶点 RC疾病亚群。主题2是治疗建模，涉及3个项目领域：(5)识别最佳 营养疗法(大量营养素、维生素、辅因子)以改善RC的健康和整体功能 疾病，(Vi)利用翻译动物和细胞模型进行高通量药物筛选和铅 复合验证，以有效识别针对不同RC疾病的高效、安全和精确的治疗方法 亚组，以及(Vii)确定初级RC疾病治疗是否将改善患有以下疾病的人的健康 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.