Deconvolution of adaptive metabolic responses of the endoplasmic reticulum

描述(由申请人提供)：本提案涉及的研究领域涉及系统方法，以探索引起慢性代谢性疾病的机制，如肥胖、糖尿病和心血管疾病，并与癌症和其他退行性疾病有关。这一疾病群目前构成了最具破坏性的全球卫生问题，预计未来25年将继续以惊人的速度增长。因此，我们的建议与“关注全球健康”高度相关。此外，我们系统地解码慢性代谢性疾病中细胞器特定的蛋白质和脂类景观的方法也涉及到“应用基因组学和其他高通量技术”的研究主题，该项目的结果将成为开发细胞器疗法作为一种新的平台来筛选和开发与该倡议的翻译目标一致的疗法的强有力的指导和资源。我们的研究灵感来自于最近出现的有力支持这些慢性非传染性疾病以及许多与年龄相关的代谢和退行性疾病的证据，这些疾病都以细胞器功能障碍为特征，特别是线粒体和内质网。然而，目前对这些病理情况如何与细胞器功能障碍有关以及这些细胞器的慢性衰竭如何导致这些病理变化的了解很少或根本没有。在了解慢性细胞器适应方面的这些重大差距或缺乏，限制了对预防和治疗衰弱慢性疾病的新途径和可能性的开发。在这个项目中，我们建议将重点放在内质网上，并使用细胞器特异性蛋白质组学、脂质组学、内质网相关多聚体分析和图谱的综合平台来系统地研究这个细胞器及其功能输出，以确定受调控的多聚体的所有翻译输出，以及代谢性疾病背景下的功能扰动。我们的方法不假设任何先前关于该细胞器的动态平衡的偏见，目的是以一种全面的方式解决导致其失败的机制，而不仅仅限于该细胞器在蛋白质合成、折叠和新陈代谢中的公认功能。我们认为，这一新的新兴领域为翻译的可能性提供了巨大的机会，并需要一种系统的方法来确定该细胞器适应慢性病需求的能力中的机械瓶颈。从这个平台涌现的技术、方法和模型将产生对ER生物学的重要见解，并将应用于广泛的慢性复杂疾病。 公共卫生相关性：肥胖、胰岛素抵抗、2型糖尿病和心血管疾病等慢性代谢性疾病是对全球健康产生不利影响的最常见疾病。尽管它们给人类生活带来巨大负担，但预防和治疗机会有限，目前仍需要新的更有效的补救办法。我们的项目旨在通过关注代谢性疾病中细胞器功能障碍这一新兴而令人兴奋的概念来确定导致这些病理变化的核心机制。在这项提案中，我们将研究细胞器衰竭的原因，重点放在内质网上，并使用系统的高通量方法来鉴定来自对照组织和疾病组织的纯化细胞器中所有调节的脂类和蛋白质。通过这些实验发现的机制将有助于预防和治疗战略的设计和实施，并使一般现场试剂和技术能够探索营养暴露的生物效应和机制。拟议的研究与该倡议的几个主题非常吻合。

DESCRIPTION (provided by applicant): The research area addressed in this proposal involves systems approaches to explore the mechanisms giving rise to chronic metabolic diseases such as obesity, diabetes, and cardiovascular disease with implications to cancer and other degenerative diseases. This disease cluster currently constitutes the most devastating global health problem and projected to continue to grow at tremendous rates in the next 25 years. Hence, our proposal is highly relevant to the "focus on global health". In addition, our approach to systematically decode organelle-specific protein and lipid landscape in chronic metabolic disease also relates to research theme "applying genomics and other high throughput technologies" and the outcomes of the project will be a strong guide and resource for exploiting organelle therapy as a novel platform to screen and develop therapeutics which is consistent with the translational goals of the initiative. Our research is inspired by recently emerging evidence strongly supporting that these chronic non-communicable diseases, as well as many age-related metabolic and degenerative disorders, all feature dysfunction of cellular organelles, particularly mitochondria and endoplasmic reticulum. However, there is currently little to no understanding of how these pathological conditions relate to organelle dysfunction and how chronic failure of these organelles lead to development of these pathologies. These major gaps in understanding chronic organelle adaptation, or lack thereof, limit the exploitation of novel avenues and possibilities for prevention and treatment for debilitating chronic diseases. In this project, we propose to focus on endoplasmic reticulum and systematically study this organelle and its functional output using an integrated platform of organelle-specific proteomics, lipidomics, ER-associated polysome analysis and profiling to identify all of the translational outputs of the polysomes that are subject to regulation, as well as functional perturbations in the context of metabolic disease. Our approach does not assume any prior biases regarding the homeostasis of this organelle and aims to address the mechanisms leading to its failure in a comprehensive manner not just limited to the accepted functions of this organelle in protein synthesis, folding, and, metabolism. We believe this new emerging area offers tremendous opportunities for translational possibilities and warrants a systematic approach to define the mechanistic bottlenecks in the ability of this organelle to adapt to the demands of chronic diseases. The technologies, approaches, and the models emerging from this platform will generate important insights into the biology of ER and will have applications for a broad array of chronic complex diseases. PUBLIC HEALTH RELEVANCE: Chronic metabolic diseases, such as obesity, insulin resistance, type 2 diabetes, and cardiovascular disease are among the most common diseases with adverse effects on global health. Despite their enormous burden on human life, the preventive and therapeutic opportunities are limited and there is ongoing need for new and more effective remedies. Our project aims to identify core mechanisms that give rise to these pathologies by focusing on the emerging and exciting concept of organelle dysfunction in metabolic diseases. In this proposal we will study the causes of organelle failure focusing on endoplasmic reticulum and using systematic high-throughput approaches to identify all regulated lipids and proteins in purified organelles from control and disease-afflicted tissues. The mechanism uncovered through these experiments will be instrumental in the design and implementation of preventive and therapeutic strategies and allow the general field reagents and technologies to explore the biological effects and mechanisms of nutrient exposures. The proposed studies are in excellent match with the several themes of the initiative.