Regulation of Cellular Behavior in Response to Extracellular Cues

项目总结 COBRE-1P20GM144265-01 家长奖励目标： 父级奖项的目标是科布雷细胞行为调控中心 对细胞外暗示的反应。这笔科布雷奖助金支持来自大学的5名初级项目负责人 缅因州(UMaine)和芒特荒岛生物实验室(MDIBL)。父级奖励建立 运行基本研究项目并帮助发展研究社区的组织结构 围绕有凝聚力的共同研究主题。由科布雷中心资助的研究项目 对细胞外信号反应的细胞行为的调节将显著发展我们的 对外周血细胞的生理和病理生理学作用的理解和认识 发育过程中的刺激(如细胞间交流、信号通路或病毒感染)， 再生、健康或疾病。 补充奖励目标： 标题：肌肉老化背景下的蛋白质平衡、萎缩和退化 骨质疏松症是一种多因素疾病，其特征是肌肉质量、力量和 功能。与老龄化和与年龄相关的疾病有关，它是一个对质量产生负面影响的主要问题 生活成本和不断上涨的医疗保健费用。骨质疏松症的问题有两个： 蛋白质抑制剂的维持导致肌肉纤维的退化以及缺乏 再生健康的组织。在衰老过程中，肌肉萎缩与血管内皮生长因子的表达增加有关 横纹肌特异性蛋白周转因子阿特罗金-1。Madelaine实验室发现过度表达 斑马鱼中这一因子的缺失会导致肌肉纤维迅速退化，肌肉组织萎缩，并 运动功能障碍。令人惊讶的是，在饮食中，该因子的表达也增加了 限制(DR)，包括禁食。DR是延长健康寿命的最有力的衰老干预措施 并保持肌肉的长期保养。当代谢参数被重置以匹配饮食时 局限性，适应DR暂时诱导肌肉分解代谢，与增加萎缩相关- 1.有趣的是，Atrogin-1蛋白似乎在蛋白平衡和肌肉中起着核心作用 在健康和疾病的背景下的稳态。了解Atrogin-1及其相关基因 调节肌肉蛋白平衡的因素在不同的背景下可能有助于治疗的发展 限制肌肉萎缩和改善肌肉萎缩的方法。然而，脊椎动物系统的使用 像小鼠一样，研究自然衰老背景下的骨骼肌萎缩需要几年的时间 而且要付出巨大的代价。使用另一种脊椎动物模式生物将允许遗传和 在相对较短的时间内以较低的成本进行环境干预。 斑马鱼是一种成熟的强大的生物，可以进行遗传操作和建模 人类疾病，而弗氏诺氏杆菌(N.Furzeri)最近成为 研究衰老和再生。N.Furzeri是圈养繁殖的最短寿命的脊椎动物， 只活了几个月。我们建议使用这些鱼类模型来研究肌肉维护 与蛋白质平衡和再生能力在衰老和/或DR期间的变化有关 Madelaine实验室已经建立了斑马鱼肌肉加速衰老的新遗传模型，我们将 用于利用与增加肌肉退化和萎缩相关的修改。罗杰斯夫妇 实验室已经开发出一种饮食限制(DR)模型，包括在N.Furzeri间歇性禁食，该模型有效 延长雄性和雌性动物的健康寿命。这提供了一种干预策略来缓解 随着年龄增长而出现的蛋白质平衡和再生能力的丧失。这些不同的实验 条件也允许比较Atrogin-1在适应DR中的积极作用与其 在石棺减少症中的病理作用。为了生成更好地了解细胞变化所需的数据 与肌肉老化相关并支持未来的拨款申请，我们希望建立一个实质性的 我们实验室的合作努力。我们提出了以下研究目标：1)测试 鱼加速模型运动和游泳能力测定肌肉功能的研究 肌肉老化和禁食后。2)使用组织清除法和光片法评估肌肉结构的完整性 使用单细胞多组学对衰老期间和医生在位期间的骨质疏松症的遗传模型进行成像 一种表征与蛋白表达相关的肌肉蛋白平衡维持变化的方法 Atrogin-1，老化和在Dr. 本补充奖励项目符合家长奖励的范围，目的是确定 与肌肉衰老和肌肉组织内细胞变化相关的分子效应因子 基因和饮食干预。两个实验室的协作成果具有互补的技术 技能和专业知识(加速肌肉老化和寿命/健康寿命干预)将加强 本研究项目的成果。

PROJECT SUMMARY COBRE - 1P20GM144265-01 Parent award goal: The parent award goal is the COBRE Center for the Regulation of Cellular Behavior in Response to Extracellular Cues. This COBRE grant supports 5 junior project leaders from University of Maine (UMaine) and Mount Desert Island Biological Laboratory (MDIBL). The parent award builds organizational structure to run essential research programs and help grow the research community around cohesive common research themes. Research projects funded through the COBRE center for the Regulation of Cellular Behavior in Response to Extracellular Cues will significantly develop our understanding and knowledge regarding the physiological and pathophysiological roles of external stimuli (such as cell-cell communication, signaling pathway, or viral infection) during development, regeneration, health or diseases. Supplement award goal: Title: Proteostasis, atrophy and degeneration in the context of muscle aging Sarcopenia is a multifactorial disease characterized by the loss of muscle mass, strength and function. Associated with aging and age-related diseases, it is a major issue negatively affecting quality of life and the rising cost of health care. The problem with sarcopenia is two-fold: there is a loss of proteostatic maintenance leading to degeneration of muscle fibers as well as a lack of ability to regenerate healthy tissue. During aging, muscle wasting is associated with increased expression of the striated muscle-specific protein turnover factor Atrogin-1. The Madelaine lab found that overexpression of this factor in zebrafish leads to rapid degeneration of muscle fibers, atrophic muscle tissue and locomotor dysfunctions. Surprinsigly, the expression of this factor is also increased under dietary restriction (DR) involving fasting. DR is the most robust aging intervention to increase healthy lifespan and preserve long-term muscle maintenance. As metabolic parameters are reset to match dietary limitations, adaptation to DR temporarily induces muscle catabolism associated with increased atrogin- 1. Interestingly, the Atrogin-1 protein appears to have a central role in proteostasis and muscle homeostatis in the context of both health and disease. Understanding how Atrogin-1 and associated factors regulate muscle proteostasis in different contexts may help in the development of therapeutic approaches to limit muscle atrophy and ameliorate muscle wasting. However, use of vertebrate systems like mice for investigating skeletal muscle wasting in the context of natural aging require several years and comes at great cost. Use of a alternative vertebrate model organisms would allow genetic and environmental interventions at a reduced cost and within a relatively short period of time. The zebrafish is a well established powerful organism for genetic manipulations and modeling of human diseases, while Nothobranchius furzeri (N. furzeri) has recently emerged as a model for investigating both aging and regeneration. N. furzeri is the shortest-lived vertebrate bred in captivity, living only a few months. We propose to use these fish models to investigate muscle maintenance associated with changes in proteostasis and regenerative capacity during aging and/or DR. The Madelaine lab has established a new genetic model of accelerated muscle aging in zebrafish that we will use to leverage modifications associated with increase muscle degeneration and atrophy. The Rogers lab has developed a model of dietary restriction (DR) involving intermittent fasting in N. furzeri that works to increase healthy lifespan in male and female animals. This provides an intervention strategy to mitigate loss of proteostasis and regenerative capacity that occur with age. These different experimental conditions also allows for comparison of the positive role of Atrogin-1 in adaptation to DR with its pathological role in sarcopenia. In order to generate data required to better understand cellular changes associated with muscle aging and to support future grant applications, we want to establish a substantive collaborative effort on the part of our labs. We propose to carry out the following research goals: 1) Test locomotion and swimming performance to determine muscle function in fish models of accelerated muscle aging and after fasting. 2) Assess muscle structural integrity using tissue-clearing and light-sheet imaging in a genetic model of sarcopenia, during aging and under DR. 3) Use a single cell multiomics approach to characterize changes in muscle proteostatic maintenance associated with expression of Atrogin-1, aging and under DR. This supplement award project fits in the scope of the parent award with the objective to identify molecular effectors of muscle aging and cellular changes within the muscle tissue associated with genetic and dietary interventions. The collaborative effort from two labs with complementary technical skills and expertise (accelerated muscle aging and lifespan/healthspan intervention) will strengthen the outcome of this research project.