Discovering small molecules activators of stress responsive signaling

项目总监/首席调查员(Lasl，Fitst，MItldle)：Morimoto，Richard I.，PL Project 4--Kelly，Jeffery W. 项目总结(见inslrucUons)： 蛋白质平衡的维持包括平衡蛋白质的生物合成，折叠， 我们假设囊泡运输、降解等对健康衰老至关重要。既然这些要求 在亚细胞隔间上，以维持蛋白质平衡随年龄和环境压力的变化， 应激反应信号通路已经进化到快速调整亚细胞蛋白平衡网络 满足需求的能力。在这里，我们重点开发和利用基于细胞的记者 筛选可以选择性地激活压力反应信号通路的小分子。一个 建议建立双发光报告细胞系以发现手臂选择性的未折叠蛋白反应 影响内质网蛋白稳定的激活剂。先前基于细胞的热休克反应 (影响胞浆蛋白平衡)激活剂报告筛选产生大量小分子先导 ARM选择性UPR激活器屏幕也有望实现同样的效果。概述了一项计划，以辨别 应激反应信号通路激活这些导联的选择性，以建立其治疗指数， 鉴定选定激活物的转录和蛋白质组学变化，并鉴定和验证 高排名激活者的目标(S)。我们还建议利用小分子调节的不稳定 域-FOXO转录因子融合建立一种灵敏和选择性的细胞报告分析方法 FOXO信号小分子激活剂的最终筛选。FOXO影响新陈代谢和 蛋白质抑制控制。选择性的应激反应信号通路激活剂将使 该项目中的其他研究人员计划了解哪些亚细胞隔室对 保持蛋白质平衡，以利于健康衰老。细胞器选择性蛋白平衡在细胞和动物中的增强 模型，结合蛋白质平衡网络的系统生物学特征作为衰老的函数 激活剂治疗应该就细胞器特异性的影响提供一个明确的答案 蛋白质平衡对健康的影响寿命，即个体无疾病的生命周期--由于 蛋白质组的适合性。 相关性(参见inslrucUons)： 项目4中产生的分子及其在检验假设中的应用 生物体整个生命周期中的蛋白质组对健康至关重要。生命周期代表着衰老的范式转变 到目前为止，研究主要集中在基因组的不稳定性上。

Program Director/Principal Investigator (Lasl, Fitst, MItldle): Morimoto, Richard I., PL Project 4-Kelly, Jeffery W. PROJECT SUMMARY (See inslrucUons): The maintenance of protein homeostasis, or proteostasis, involves balancing protein biosynthesis, folding, vesicular trafficking, degradation, etc., which we hypothesize is critical for healthy aging. Since the demands on subcellular compartments to maintain proteostasis change with aging and due to environmental stresses, stress-responsive signaling pathways have evolved to quickly adjust subcellular proteostasis network capacity to meet demand. Herein, we focus on the development and utilization of cell-based reporter screens to discover small molecules that can activate stress-responsive signaling pathways selectively. A dual luminescence reporter cell line is proposed to discover arm-selective unfolded protein response activators affecting proteostasis in the endoplasmic reticulum. A prior cell-based heat shock response (influencing cytosolic proteostasis) activator reporter screen has generated numerous small molecule leads and the same is expected from the arm-selective UPR activator screen. A plan is outlined to discern the stress responsive signaling pathway activation selectivity of these leads, to establish their therapeutic index, to identify the transcriptional and proteomic changes of select activators, and to identify and validate the target(s) of the highly ranked activators. We also propose to utilize small molecule-regulated destabilized domain-FOXO transcription factor fusions to deveiop a sensitive and selective cell-based reporter assay for eventual screening of small molecule activators of FOXO signaling. FOXO influences metabolic and proteostatic control. Selective stress-responsive signaling pathway activators will make it possible for the other investigators in this program project to learn which subcellular compartments are most important to maintain proteostasis in for healthy aging. Organelle-selective proteostasis enhancement in cell and animal models, coupled with the systems biology characterization of the proteostasis network as a function of aging and activator treatment should provide a clear answer regarding the influence of organelle-specific proteostasis on healthspan, the period of life where individuals are disease free-owing to the enhanced fitness of the proteome. RELEVANCE (See inslrucUons): The molecules produced in Project 4 and their utilization in testing the hypothesis that maintenance of the proteome over the lifespan of an organism is critical for healthspan represents a paradigm shift in aging research, which heretofore has largely focused on genome instability.