Proteostasis Regulator Pharmacology Core D

核心项目D与项目1-4以及核心项目B和C合作的总体目标是检验假设 有可能部分逆转蛋白质稳态网络（PN）能力的衰老依赖性缺陷， （包括导致病理学的那些）通过使用小分子药物对PN进行药理学调节 蛋白质稳态调节剂。我们将重点关注热休克反应应激反应信号通路激活剂 调节细胞溶质PN能力的未折叠蛋白应答应激应答信号传导激活剂， 调节分泌途径PN的能力，和抗氧化应激反应信号通路激活剂， 目标1.由于应激反应信号通路产生一个活性转录因子，我们假设， 这将是一个更大的机会，有效的生物反应，从这个进化的解决方案，以纠正 蛋白质稳态缺陷，其中在给定的蛋白质稳态中相互作用和竞争PN途径的所有组分都是蛋白质稳态缺陷。 亚细胞区室以适当的化学计量上调。这些应激反应信号 途径导致强大的新兴功能，只有部分理解。在目标1中，我们将进一步 开发一个技术平台来验证药效学（PD;研究药物对人体的影响）。 生物体），选择性和小分子蛋白质稳态调节剂的作用机制，通过 在多种生物体中激活应激反应信号通路。我们将首先采用基于细胞的 应激反应信号通路激活的报告基因（具有核心B），靶向RNAseq，随后是质量 基于光谱的蛋白质组学（核心C活动），结合生物信息学来验证蛋白质稳态 监管部门我们还将评估药代动力学（PK;研究生物体对药物的作用 （代谢）），这将有助于我们建立合理的给药方案。PK将是 使用液相色谱-质谱法评估。在目标2中，我们还寻求建立 涉及增强蛋白质、脂质和蛋白质降解的小分子蛋白质稳态调节剂的用途 细胞器，无论是通过激活自噬或泛素蛋白酶体系统。我们将生成PK 和其他人报告的激活自噬溶酶体途径的蛋白质稳态调节剂的PD数据 （降解蛋白质、寡糖、脂质和寡核苷酸） 以及蛋白质稳态调节剂激活泛素蛋白酶体系统（降解蛋白质）。总的来说， 人、鼠和酵母细胞以及C. elegans很重要，因为：（1）它允许我们 测试并因此推荐蛋白质抑制调节剂的合理给药方案，以及（2）这些数据 使我们能够正确地解释模型生物的实验，特别是负面数据。我们将提供这些 经验证的蛋白质稳态调节剂的项目的PI，以及实验室工作的补充老化 范例和衰老相关疾病，以辨别哪些蛋白质稳态调节剂纠正衰老相关疾病， 蛋白质稳态缺陷。

The overarching aim of Core D, in collaboration with Projects 1-4 and cores B and C, is to test the hypothesis that it is possible to partially reverse aging-dependent deficiencies in proteostasis network (PN) capacity (including those leading to pathology) by pharmacologic regulation of the PN employing small molecule proteostasis regulators. We will focus on heat shock response stress-responsive signaling pathway activators that regulate cytosolic PN capacity, unfolded protein response stress-responsive signaling activators that regulate secretory pathway PN capacity, and the antioxidant stress-responsive signaling pathway activators in Aim 1. Since stress-responsive signaling pathways generate an active transcription factor, we hypothesize that there will be a greater chance for an effective biological response from this evolved solution to correct proteostasis deficiencies, wherein all the components of interacting and competing PN pathways in a given subcellular compartment are up-regulated in the appropriate stoichiometry. These stress-responsive signaling pathways lead to powerful emergent functions that are only partially understood. In Aim 1, we will further develop a technology platform to validate the pharmacodynamics (PD; the study of what a drug does to the organism), selectivity, and mechanism of action of small molecule proteostasis regulators that function through activation of stress-responsive signaling pathways in multiple organisms. We will initially employ cell-based reporters of stress-responsive signaling pathway activation (with Core B), targeted RNAseq, followed by mass spectrometry-based proteomics (Core C activities), coupled to bioinformatics to validate the proteostasis regulators. We will also assess the pharmacokinetics (PK; the study of what the organism does to a drug (metabolism)) in multiple organisms, which will help us establish reasonable dosing regimens. PK will be assessed using liquid chromatography-mass spectrometry approaches. In Aim 2, we also seek to establish the utility of small molecule proteostasis regulators involved in enhancing the degradation of proteins, lipids and organelles, either through activation of autophagy or the ubiquitin proteasome system. We will generate PK and PD data for proteostasis regulators reported by others that activate the autophagy lysosomal pathway (degrades proteins, oligosaccharides, lipids and oligonucleotides) through an m-TOR independent mechanism and for proteostasis regulators to activate the ubiquitin proteasome system (degrades proteins). Collectively, the PK and PD data in human, murine and yeast cells and in C. elegans is important because: (1) it allows us to test and, therefore recommend, reasonable dosing regimens for proteostasis regulators, and (2) these data allow us to properly interpret experiments in model organisms, especially negative data. We will provide these validated proteostasis regulators to PIs of the projects, as well to labs working on complementary aging paradigms and aging-associated diseases to discern which proteostasis regulators correct aging-linked proteostasis deficiencies.