Proteostasis Regulator Pharmacology Core D

蛋白质稳态调节剂药理学核心 D

基本信息

批准号：
10432030
负责人：
JEFFERY W KELLY
金额：
$ 32.08万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10432030
关键词：
Aging Animal Model Antioxidants Autophagocytosis Biogenesis Bioinformatics Biological Caenorhabditis elegans Cell Culture Techniques Cells Chronic Client Collaborations Coupled Data Defect Degenerative Disorder Degradation Pathway Disease Docking Dose Drug Kinetics Enhancers Enzymes Genetic Genetic Transcription Goals Heat-Shock Response Human Immune system Kinetics Label Lead Link Lipids Mass Spectrum Analysis Metabolism Mitochondria Mus Neurodegenerative Disorders Oligonucleotides Oligosaccharides Optics Organelles Organism Pathology Pathway interactions Pharmaceutical Preparations Pharmacodynamics Pharmacology Phenotype Physiologic pulse Proteins Proteome Proteomics Publishing Regimen Regulation Reporter Reporting Sampling Signal Pathway Signal Transduction Stress System Technology Testing Time Ubiquitin Validation Yeasts base biological adaptation to stress disease phenotype drug candidate drug metabolism environmental stressor experimental study extracellular inhibitor interest liquid chromatography mass spectrometry multicatalytic endopeptidase complex programs protein degradation proteostasis response small molecule stoichiometry transcription factor transcriptome sequencing validation studies

项目摘要

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.

核心 D 与项目 1-4 以及核心 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）能力中与衰老相关的缺陷（包括导致病理的那些）通过使用小分子对 PN 进行药理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质 PN 能力、未折叠蛋白反应应激反应信号激活剂调节分泌途径 PN 能力，以及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活性转录因子，我们假设这种进化的解决方案将有更大的机会产生有效的生物反应来纠正蛋白质稳态缺陷，其中给定的 PN 通路中相互作用和竞争的所有成分亚细胞区室在适当的化学计量中上调。这些应激反应信号路径导致强大的新兴功能，但人们只了解了部分。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；研究药物对药物的作用）有机体）、选择性和小分子蛋白质稳态调节剂的作用机制，这些调节剂通过多种生物体中应激反应信号通路的激活。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者（使用核心 B）、靶向 RNAseq，然后是质量基于光谱分析的蛋白质组学（核心 C 活动），与生物信息学相结合以验证蛋白质稳态监管机构。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））在多种生物体中，这将有助于我们建立合理的给药方案。 PK将会使用液相色谱-质谱法进行评估。在目标 2 中，我们还寻求建立小分子蛋白质稳态调节剂参与增强蛋白质、脂质和通过激活自噬或泛素蛋白酶体系统。我们将生成PK 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）通过 m-TOR 独立机制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，人类、小鼠和酵母细胞以及线虫中的 PK 和 PD 数据很重要，因为：(1) 它使我们能够测试并因此推荐合理的蛋白质稳态调节剂给药方案，以及（2）这些数据使我们能够正确解释模式生物中的实验，尤其是负面数据。我们将提供这些为项目的 PI 以及致力于补充衰老的实验室提供经过验证的蛋白质稳态调节剂范式和衰老相关疾病来辨别哪些蛋白质稳态调节因子可以纠正与衰老相关的疾病蛋白质稳态缺陷。