Whole-lifespan monitoring of yeast health

除非在以下方面取得重大进展，否则65岁及以上美国人的比例预计将在2060.13之前翻一番 对于与年龄相关的疾病的治疗，这一根本的人口结构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 死亡原因包括癌症、心血管疾病、神经变性和糖尿病。14-16 使用微流控装置，我们可以通过时间推移以单细胞分辨率监测酵母的老化过程 显微镜。这个装置还可以让我们研究老化细胞的环境扰动。微流控捕集 是一种在单细胞上收集纵向、动态、原位老化数据的罕见的有效方法 决议。我将利用这个设备对以下酵母细胞进行全寿命监测 具体目标： 目的1：探讨老年性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年龄相关疾病的驱动因素，17 线粒体表型与酵母RLS有机械联系。18以前的研究表明，早期的 由于后来的线粒体功能障碍而导致的空泡酸度的寿命损失。19初步数据表明，铁 饥饿反应在这一过程中起中介作用。我将肯定液泡酸度、铁调节和 线粒体功能障碍，并确定这些连接的原因机制。 目的2：确定与年龄相关的母女寿命不对称丧失的机制 不对称的细胞分裂是后生动物生命的一个标志，在发育和 与年龄相关的病理学。20，21与酵母老化领域的普遍假设相反，我观察到 从生命早期开始的母女寿命不对称的进行性年龄依赖性下降。我 假设这种下降是由于Septin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。 目的3：确定与年龄相关的保真度丧失在MSN 2葡萄糖敏感信号转导中的作用 衰老的特点是在应对环境变化时维持体内平衡的能力降低。 我们的微流控设备使干扰和观察衰老细胞成为可能。我发现MSN2信令 在相同的环境中，年轻细胞和老年细胞之间的差异。此外，我还发现细胞暴露在 随着年龄的增长，随时间变化的环境寿命更短。我假设MSN2会输掉 葡萄糖信号传递的保真度，因为它在报告随年龄增加的氧化应激中所起的作用。这就做 描述这种信息传输丢失的特征，并确定它是否对与年龄相关的 恢复力。

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.