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Screening for enhancers of secreted clusterin (sCLU) and evaluation in AD models

分泌型凝聚素 (sCLU) 增强子的筛选和 AD 模型的评估

基本信息

批准号：
10195566
负责人：
Varghese John
金额：
$ 42.34万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10195566
关键词：
APP-PS1 Address Adverse effects Affect Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease patient Alzheimer’s disease biomarker Amyloid beta-Protein Apoptotic Binding Proteins Biological Assay Biological Availability Biological Markers Biological Process Brain Cardiovascular Diseases Cell Culture Techniques Cell Death Cell Line Cell Survival Cells Cellular Stress Response Central Nervous System Diseases Consensus Cytoprotection Development Devices Disease Disease Progression Drug Kinetics Effectiveness Enhancers Evaluation Failure Functional disorder Future Glioblastoma Goals Half-Life Hippocampus (Brain)Homeostasis Human Immunoassay Impaired cognition In Vitro Induced pluripotent stem cell derived neurons Inflammation Late Onset Alzheimer Disease Lead Libraries Liquid substance Literature Malignant Neoplasms Mass Spectrum Analysis Measures Metabolic Mitochondria Molecular Molecular Chaperones Molecular Target Nature Nerve Degeneration Neurons Oral Oxidative Regulation Oxidative Stress Pathology Permeability Pharmaceutical Preparations Pharmacology Phosphotransferases Photoaffinity Labels Preclinical Testing Predisposition Property Protein Isoforms Proteins Public Health Research Role Senile Plaques Signal Pathway Signal Transduction Slice Solubility Stress Testing Testis Therapeutic Toxic effect Treatment Efficacy abeta deposition base drug candidate effectiveness evaluation efficacy testing experimental study extracellular genetic risk factor genetic variant glycogen synthase kinase 3 beta high throughput screening hyperphosphorylated tau immunoregulation in vivo induced pluripotent stem cell innovation instrument lipid transport mouse model nerve stem cell neuroblastoma cell novel novel therapeutic intervention programs proteotoxicity screening small molecule sulfated glycoprotein 2 tau aggregation tau phosphorylation tau-1 treatment strategy

项目摘要

PROJECT SUMMARY/ABSTRACT This proposal provides a unique opportunity to identify isoform-specific modulators of clusterin (CLU) and evaluate its role in Alzheimer's disease (AD). Clusterin, also known as apolipoprotein J (Apo J) is a protein originally identified in 1979. Several CLU gene variants are associated with AD and the SNP rs11136000C is the third strongest genetic risk factor for Late Onset Alzheimer's disease (LOAD). There is evidence that enhancing brain levels of sCLU will slow or reverse various molecular mechanisms underlying onset of the AD pathophysiology. This is based on studies that show sCLU promoting the clearance of disease-causing amyloid beta (Aβ) plaques, while reducing cellular stress responses such as oxidative stress and inflammation known to lead to increased kinase activity such as GSK-3β and increased hyperphosphorylated tau involved in progression of AD. Interestingly, the CLU gene variants that confer increased susceptibility to LOAD also reduce sCLU levels and result in increased Aβ deposition and tau neurofibrillary tangles and faster cognitive decline relative to non-carriers. In this proposal a comprehensive research program will be undertaken to identify potent, brain permeable, small molecules that increase levels of sCLU and modulate biomarkers in AD models. This will be accomplished using high throughput screening (HTS) to identify `hits' with potent sCLU enhancing ability, drug-like properties, brain permeability in pharmacokinetics (PK) analyses to guide compound selection for further testing and mechanism-of-action (MOA) studies in iPSC derived neurons and ex-vivo models of AD. In Aim1 we will screen for compounds that increase extracellular concentrations of sCLU. UCLA's large compound library will be screened by using fully automated liquid handling devices and analytical instruments to identify molecules that increase sCLU levels. For these experiments, sensitive and readily formattable AlphaLISA based immunoassays will be optimized to detect different CLU isoforms. In Aim2, we would conduct in-vitro ADME/T testing and in-vivo pharmacokinetics (PK) analyses to guide hit selection including determination of sCLU enhancing potency, characterizing the physiochemical properties including solubility, brain permeability and metabolic stability of candidate sCLU enhancing compounds. In Aim3, we would conduct testing in induced pluripotent stem cells (iPSCs) derived neurons and ex-vivo brain slices. The goal is to evaluate the effectiveness of sCLU enhancers with sufficient oral brain bioavailability and half-life in AD patient-derived neurons as well as in ex-vivo organotypic brain slice cultures from the hippocampi of an AD mouse model. In Aim4, we would identify molecular targets and cell signaling pathways affected by prioritized hits. MOA studies involving target identification by photoaffinity-labeling/purification as well as global and phosphoproteome analyses will be accomplished using state-of-the-art mass spectrometry. Using these approaches, we will identify molecular targets of our sCLU enhancing compounds to identify novel mechanisms involved in regulating brain sCLU levels in AD.

项目总结/文摘