Whole-lifespan monitoring of yeast health

酵母健康的全生命周期监测

• 批准号: 9302225
• 负责人: Kenneth Chen
• 金额: $ 4.06万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9302225
• 关键词: Acidity; Age; Aging; Aging-Related Process; American; Appearance; Cardiovascular Diseases; Cell Culture Techniques; Cell Separation; Cell division; Cells; Collection; Culture Media; Data; Daughter; Development; Devices; Diabetes Mellitus; Diffusion; Disease; Economics; Environment; Gases; Glucose; Health; Homeostasis; Immobilization; In Situ; Individual; Investigation; Iron; Kinetics; Learning; Life; Link; Liquid substance; Longevity; Malignant Neoplasms; Measurement; Measures; Mediating; Membrane; Methods; Microfluidic Microchips; Microfluidics; Microscopy; Mitochondria; Molds; Molecular; Monitor; Mothers; Nerve Degeneration; Oxidative Stress; Pathology; Periodicity; Permeability; Phenotype; Polymers; Population; Process; Regulation; Reporting; Research; Resolution; Role; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Starvation; Time; Yeasts; age related; aged; biological adaptation to stress; cell age; design; elastomeric; environmental change; experimental study; fluid flow; insight; mitochondrial dysfunction; mortality; polydimethylsiloxane; protein expression; resilience; response; senescence; social; transcription factor; transmission process; young mother

The fraction of Americans 65 and older is expected to double by 2060.13 Unless significant progress is made in the treatment of age-related diseases, this fundamental demographic shift will herald drastic economic and social challenges. The study of aging in S. cerevisiae has revealed fundamental insights into the most common causes of mortality including cancer, cardiovascular disease, neurodegeneration, and diabetes.14–16 Using a microfluidic device, we can monitor the yeast aging process at single-cell resolution via time-lapse microscopy. This device also allows us to study environmental perturbations of aged cells. Microfluidic trapping is a rare validated method for collection of longitudinal, dynamic, in situ aging data at single-cell resolution. I will exploit this device to perform whole-lifespan monitoring of yeast cell towards the following specific aims: Aim 1: Determine mechanism of iron starvation response in age-related mitochondrial dysfunction Mitochondrial dysfunction has been implicated as a driver of multiple age-related diseases,17 and changes in mitochondrial phenotype have been mechanistically tied to yeast RLS.18 Previous research has linked an early life loss of vacuolar acidity to later mitochondrial dysfunction.19 Preliminary data indicates that an iron starvation response mediates this process. I will affirm the links between vacuolar acidity, iron regulation, and mitochondrial dysfunction and determine the causal mechanism of these connections. Aim 2: Determine mechanism of age-related loss of mother-daughter lifespan asymmetry Asymmetric cell division is a hallmark of metazoan life, with important implications in both development and age-related pathology.20,21 Contrary to popular assumption in the yeast aging field, I have observed a progressive age-dependent decline in mother-daughter lifespan asymmetry that begins early in life. I hypothesize that this decline is due to a progressive decline in the septin-mediated cortical and membrane diffusion barrier. I will investigate the kinetics and the mechanism of this loss of asymmetry. Aim 3: Define role of age-related loss of fidelity in MSN2 glucose-sensing signal transduction Aging is characterized by a reduced ability to maintain homeostasis in response to environmental change.22,23 Our microfluidic device makes it possible to perturb and observe aged cells. I have found that MSN2 signaling differs between young and old cells in the same environment. Moreover, I have found that cells exposed to temporally varying environments as they age have shorter lifespans. I hypothesize that the MSN2 loses glucose-signaling transmission fidelity due to its role in reporting of increased oxidative stress with age. I will characterize this loss of information transfer and determine whether it is responsible for an age-related loss of resilience.

到 2060 年，65 岁及以上的美国人比例预计将翻一番。13 除非在 与年龄相关的疾病的治疗，这种根本性的人口变化将预示着巨大的经济和 社会挑战。对酿酒酵母衰老的研究揭示了对最常见的衰老现象的基本见解 死亡原因包括癌症、心血管疾病、神经退行性疾病和糖尿病。14–16 使用微流体装置，我们可以通过延时以单细胞分辨率监测酵母老化过程 显微镜。该设备还使我们能够研究老化细胞的环境扰动。微流体捕获 是一种罕见的经过验证的方法，用于收集单细胞纵向、动态、原位老化数据 解决。我将利用该设备对酵母细胞进行全寿命监测，以实现以下目的 具体目标： 目标 1：确定年龄相关线粒体功能障碍中铁饥饿反应的机制 线粒体功能障碍被认为是多种与年龄相关的疾病的驱动因素，17 线粒体表型与酵母 RLS 有机械联系。 18 先前的研究已将早期的 液泡酸度的丧失导致后来的线粒体功能障碍。 19 初步数据表明，铁 饥饿反应介导了这一过程。我将肯定液泡酸度、铁调节和 线粒体功能障碍并确定这些连接的因果机制。 目标 2：确定与年龄相关的母女寿命不对称性丧失的机制 不对称细胞分裂是后生动物生命的标志，对发育和发育都有重要影响。 与年龄相关的病理学。20,21 与酵母老化领域的流行假设相反，我观察到 母女寿命不对称性随着年龄的增长而逐渐下降，这种现象从生命早期就开始了。我 假设这种下降是由于脓毒症介导的皮质和膜的进行性下降所致 扩散屏障。我将研究这种不对称性丧失的动力学和机制。 目标 3：定义与年龄相关的保真度损失在 MSN2 葡萄糖传感信号转导中的作用 衰老的特点是响应环境变化而维持体内平衡的能力下降。22,23 我们的微流体装置使得扰动和观察老化细胞成为可能。我发现MSN2信令 在同一环境中，年轻细胞和老年细胞之间存在差异。此外，我发现细胞暴露于 随着年龄的增长，环境随时间变化，寿命会缩短。我假设 MSN2 输了 葡萄糖信号传输保真度，因为它在报告氧化应激随着年龄增长而增加的过程中发挥着作用。我会 描述这种信息传输损失的特征，并确定它是否导致与年龄相关的信息损失 弹力。