First-in-class peptide therapeutics for mitochondrial disorders: molecular mechanism of action and optimization of design

线粒体疾病的一流肽疗法：分子作用机制和设计优化

• 批准号: 10624434
• 负责人: NATHAN N ALDER
• 金额: $ 49.48万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10624434
• 关键词: Affect; Aging; Alzheimer' s Disease; Animal Diseases; Behavior; Binding; Bioenergetics; Biological Assay; Biological Availability; Biomimetics; Biophysics; Calcium; Calorimetry; Cardiolipins; Cell Culture Techniques; Cell Line; Cell Survival; Cell model; Cell physiology; Chronic; Clinical Trials; Complex; Confocal Microscopy; Coupled; Crista ampullaris; Diabetes Mellitus; Disease; Electrostatics; Energy Metabolism; Equilibrium; Evaluation; FDA approved; Fluorescence; Foundations; Free Radical Scavenging; Gene Expression; Goals; Health; Heart Diseases; Heritability; Inner mitochondrial membrane; Interdisciplinary Study; Ions; Kidney Diseases; Kinetics; Label; Lead; Libraries; Lipid Bilayers; Lipids; Lipoproteins; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Medicine; Membrane; Membrane Structure and Function; Metabolic Diseases; Mitochondria; Mitochondrial Diseases; Modeling; Molecular; Molecular Analysis; Molecular Conformation; Molecular Mechanisms of Action; Morphology; Nerve Degeneration; Non-Insulin-Dependent Diabetes Mellitus; Organelles; Outcome; Oxidative Phosphorylation; Oxidative Stress; Pathologic; Pathology; Pattern; Peptide Conformation; Peptides; Permeability; Phase; Physiologic pulse; Play; Polymers; Process; Production; Property; Proteins; Reactive Oxygen Species; Research; Research Support; Resolution; Role; Safety; Sampling; Series; Shotguns; Side; Site; Solid; Stress; Structure; Surface; System; Techniques; Testing; Therapeutic; Thermodynamics; Treatment Efficacy; Variant; Work; Yeasts; biophysical analysis; biophysical techniques; crosslink; design; functional restoration; improved; insight; interdisciplinary approach; interfacial; islet amyloid polypeptide; lipidomics; membrane assembly; membrane model; mitochondrial dysfunction; mitochondrial membrane; molecular dynamics; nanodisk; next generation; peptide analog; peptide drug; physical property; preclinical trial; preservation; protein complex; protein expression; protein transport; rational design; stressor; virtual

PROJECT SUMMARY Mitochondria are organelles that play a dominant role in energy metabolism and many other cellular processes. Mitochondrial dysfunction is associated with primary heritable diseases and aging-related declines in health, including chronic pathologies like cancer, diabetes and neurodegeneration. There are currently no cures for mitochondrial diseases. However, Szeto-Schiller (SS) peptides have emerged among the most promising therapeutics for promoting mitochondrial health. As shown by preclinical and clinical trials, and as exemplified by the lead compound SS-31 (Elamipretide), SS peptides show exceptionally broad therapeutic efficacy in treating mitochondrial dysfunction. Using a multidisciplinary approach, our research team has conducted the first in-depth analysis of the molecular mechanism of action (MoA) of SS peptides. Our work supports a unique mechanism in which SS peptides interact with cardiolipin-rich mitochondrial membranes and modulate general physical membrane properties, thereby underpinning their broad therapeutic potential. The objective of the proposed project is twofold. The first goal is to thoroughly understand the MoA of SS peptides. To this end, we will leverage our solid foundation of mechanistic insights to test, refine, and expand our working models using computational, reductionist, mitochondrial, and cellular systems. The second goal is to identify the physicochemical properties of SS peptides that are most critical to their mechanism. To this end, we will evaluate a series of rationally designed SS peptide constructs with variations in the tetrapeptide cationic/aromatic motif, using our established functional assays. With our highly interdisciplinary research team, we will approach these goals as three separate aims. First, we will address how SS peptides interact with lipid bilayers and modulate their physical properties using a combination of computational and biophysical approaches with biomimetic model membrane systems. This will render critical information on equilibrium peptide binding models, high resolution structural information on peptide conformational dynamics and interaction with lipid groups, and how peptides modulate membrane electrostatics, lipid structural dynamics, and bilayer polymorphic changes. Second, we will evaluate the effects of SS peptides on the structure and function of membranes from yeast and mammalian models. This will establish the sites of peptide interaction in the morphologically complex mitochondrion, how peptides affect the stability and assembly of membrane complexes, the distribution of lipids within mitochondria, and lipid turnover kinetics. Finally, using mitochondrial and cellular models, we will analyze the mechanisms by which SS peptides restore function under pathological conditions including oxidative stress, high calcium load, and amyloidogenic proteins involved in type II diabetes and Alzheimer’s disease. By this multi-tiered approach, our results will yield unprecedented insights into the mechanism of this class of therapeutics with particular relevance to aging-related diseases. Further, our peptide screen will inform the design of SS peptide variants with greater efficacy and/or bioavailability.

项目摘要 线粒体是在能量代谢和许多其他细胞中起主要作用的细胞器 过程。线粒体功能障碍与主要可遗传疾病和与衰老有关的下降有关 在健康方面，包括癌症，糖尿病和神经变性等慢性病理。目前没有 线粒体疾病的治疗方法。但是，Szeto-Shiller（SS）Pepperides已经出现了最多的 有希望促进线粒体健康的有前途的疗法。如临床前和临床试验所示，以及 SS Petides以铅化合物SS-31（Elamipretide）为例 治疗线粒体功能障碍的功效。使用多学科的方法，我们的研究团队拥有 对SS肽的分子作用机理（MOA）进行了深入分析。我们的工作 支持一种独特的机制，其中SS Petides与富含Cardiodipin的线粒体膜和 调节一般的物理膜特性，从而支持其广泛的治疗潜力。这 拟议项目的目标是双重的。第一个目标是彻底了解SS肽的MOA。 为此，我们将利用我们的机械见解的坚实基础来测试，完善和扩展我们的 使用计算，还原学家，线粒体和细胞系统的工作模型。第二个目标是 确定最重要的SS Petides的物理特性。为此， 我们将评估一系列合理设计的SS胡椒构建体，静脉肽有变化 使用我们既定的功能测定法，阳离子/芳族基序。随着我们高度跨学科的研究 团队，我们将以三个单独的目标来实现这些目标。首先，我们将解决SS Petides如何互动 使用脂质双层，并使用计算和生物物理的组合来调节其物理特性 仿生模型膜系统的方法。这将提供有关平衡的关键信息 肽结合模型，有关胡椒构象动力学的高分辨率结构信息和 与脂质组的相互作用，以及辣椒如何调节膜静电，脂质结构动力学， 和双层多态性变化。其次，我们将评估SS Petides对结构的影响 酵母和哺乳动物模型的膜功能。这将在 形态上复杂的线粒体，辣椒如何影响膜的稳定性和组装 复合物，线粒体内脂质的分布以及脂质周转动力学。最后，使用线粒体 和细胞模型，我们将分析SS在病理下恢复功能的机制 包括II型糖尿病的氧化应激，高钙负荷和淀粉样蛋白蛋白在内的疾病 和阿尔茨海默氏病。通过这种多层方法，我们的结果将产生前所未有的见解 这类治疗的机制与与衰老有关的疾病特别相关。此外，我们的 肽屏幕将为SS肽变体的设计提供更高的效率和/或生物利用度的设计。