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：
10183113
负责人：
JEFFERY W KELLY
金额：
$ 32.18万
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 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10183113
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/antagonist 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的总体目的与项目1-4和核心B和C合作是测试假设可以部分逆转蛋白技术网络（PN）容量中的衰老依赖性缺陷（包括导致病理学的那些）通过使用小分子的PN进行药理调节蛋白质调节剂。我们将专注于热激响应应力响应信号传导途径激活器调节胞质PN容量，展开的蛋白质反应应力响应信号传导激活剂，调节分泌途径PN容量，并在 AIM 1。由于应力响应信号通路会产生主动转录因子，因此我们假设从这种进化的解决方案中有效的生物学反应将有更大的机会纠正蛋白质的缺陷，其中给定的所有相互作用和竞争PN途径的组成部分亚细胞室在适当的化学计量中上调。这些应力响应信号传导途径导致强大的紧急功能仅被部分理解。在AIM 1中，我们将进一步开发一个技术平台来验证药效学（PD；有机体），小分子蛋白抑制剂调节剂的选择性和作用机理多种生物中应力响应信号通路的激活。我们最初将采用基于细胞的应力响应信号通路激活的记者（用核心B），有针对性的RNASEQ，然后是质量基于光谱法的蛋白质组学（核心C活性），耦合到生物信息学以验证蛋白质抗体监管机构。我们还将评估药代动力学（PK；对有机体对药物所做的工作的研究（代谢））在多种生物中，这将有助于我们建立合理的给药方案。 PK会使用液相色谱 - 质谱法评估。在AIM 2中，我们还试图建立与蛋白质，脂质和细胞器，通过激活自噬或泛素蛋白酶体系统。我们将生成PK 和其他人报告的蛋白质调节剂的PD数据激活自噬溶酶体途径（降解蛋白质，寡糖，脂质和寡核苷酸）通过M-TOR独立机制并使蛋白质调节剂激活泛素蛋白酶体系统（降解蛋白质）。共同人，鼠和酵母细胞以及秀丽隐杆线虫中的PK和PD数据很重要，因为：（1）它使我们允许我们为了测试并建议对蛋白质调节剂进行合理的给药方案，以及（2）这些数据允许我们正确解释模型生物的实验，尤其是负数据。我们将提供这些经过验证的蛋白质调节器到项目的PI，以及从事互补衰老的实验室范式和与衰老相关的疾病，以识别哪种蛋白质调节剂正确衰老连接蛋白技术缺陷。