Enhanced APOE2 Expression into Brain for Therapeutic Strategy for Alzheimer's Disease

增强 APOE2 在大脑中的表达，用于阿尔茨海默病的治疗策略

基本信息

批准号：
10514954
负责人：
Takahisa Kanekiyo
金额：
$ 19.1万
依托单位：
NORTH DAKOTA STATE UNIVERSITY
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：
2021-05-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10514954
关键词：
Aging Alzheimer&apos s Disease Anti-Inflammatory Agents Award Biodistribution Biological Biological Markers Birth Brain Brain-Derived Neurotrophic Factor Cannabidiol Cell Proliferation Cognitive Complementary DNA Curcumin Detection Drug Delivery Systems Drug Kinetics Enzyme Kinetics Enzyme-Linked Immunosorbent Assay Equipment Funding Genes Goals High Pressure Liquid Chromatography Hippocampus (Brain)Inflammatory Learning Life Liquid substance Memory Messenger RNA Metabolism Neuraxis Neurodegenerative Disorders Neurons Nucleic Acids Organ Pharmaceutical Preparations Pharmacology Polymerase Chain Reaction Prevention Proteins Quercetin RNA Reader Research Research Personnel Resveratrol Risk Factors Science Synapses Synaptic plasticity System Therapeutic Time Universities absorption base cytokine cytotoxicity design drug discovery genetic analysis in vivo instrument instrumentation mRNA Expression nanoparticle neurogenesis neuroinflammation non-viral gene delivery synaptic function synergism

项目摘要

SUMMARY: Neuroinflammation is one of the major risk factor which leads to the progression of neurodegenerative disorders. The birth of new neurons within the hippocampal region of the central nervous system continues throughout life, and the amount of neurogenesis correlates closely with the hippocampal functions of learning and memory. Neuroinflammation alone inhibits neurogenesis and that inflammatory blockade with an anti-inflammatory agent restores neurogenesis. Anti-inflammatory (AI) drug reduces the expression of various inflammatory cytokines and increased synapse number. Drugs such as curcumin, resveratrol, quercetin and cannabidiol from natural origin have demonstrated both anti-inflammatory activity and neurogenesis. ApoE2, VGF and BDNF have been found useful in maintaining cell proliferation, neuronal function and synaptic plasticity throughout life. Moreover, it has been shown that an increased level of ApoE2, VGF, and BDNF in the brain is associated with neurogenesis and cell proliferation. Therefore combination of AI and ApoE2 or BDNF would synergistically enhance neurogenesis and synaptic functions. The long-term goal of the requested supplement award is to design a non-viral gene delivery carrier for efficient delivery of drugs, shAPOE4, mRNA encoding APOE2, pAPOE2, BDNF and other genes to brain for prevention and treatment of aging-related cognitive decile including AD. The requested supplement equipment award would strengthen the science of drug discovery in the following ways: (1).Determination of AI drugs in delivery matrix and biological fluids as well as in vivo pharmacokinetics and biodistributions of drugs by HPLC: The HPLC system would help us to quantify the amount of AI drugs entrapped in nanoparticles, and study in vivo the pharmacokinetics and biodistribution of drugs in various organs. (2). Quantification of target genes in different matrices by Quantitative Real Time Polymerase chain reaction (qRT- PCR), thermocycler, and multi-mode microplate reader: The qRT-PCR provides fast and high-throughput reliable detection and quantification of target genes in different matrices. The qRT-PCR instrument will enable us to quantify the changes in mRNA expression as a function of time. The thermocycler will allow the conversion of RNA to cDNA which will further be quantified in the qRT-PCR. BioTek™ Synergy™ HTX Multi-Mode Microplate Reader will be used for various applications in our lab ranging from nucleic acid quantification, protein quantification, enzyme kinetics, biomarker quantification, ELISAs, genetic analysis, cell proliferation, cytotoxicity, and drug absorption and metabolism. The requested fund ($190,981) to procure the above equipment items would help investigate delivery of drugs, mRNA, pAPOE2, shAPOE4, BDNF and other nucleic acids through nanoparticle-based delivery systems. We anticipate that the requested equipment will significantly contribute to the field of drug discovery for AD pharmacological therapy. Thus, these proposed research equipment would have high impact in developing drug and nucleic acid based therapy for AD. In addition, the requested equipment would also be available to conduct quality research to other investigators in the department and university.

概括：神经炎症是导致神经退行性疾病进展的主要危险因素之一。中枢神经系统海马区新神经元的诞生贯穿一生，神经发生的数量与海马的学习记忆功能密切相关。单独的神经炎症会抑制神经发生，而抗炎剂则可以阻断炎症恢复神经发生。抗炎 (AI) 药物可减少各种炎症细胞因子的表达。以及增加突触数量，例如姜黄素、白藜芦醇、槲皮素和大麻二酚。已证明 ApoE2、VGF 和 BDNF 具有抗炎活性和神经发生作用。发现有助于维持整个生命周期的细胞增殖、神经功能和突触可塑性。研究表明，大脑中 ApoE2、VGF 和 BDNF 水平的增加与神经发生有关因此，AI与ApoE2或BDNF的组合将协同增强细胞增殖。所要求的补充奖的长期目标是设计一个神经发生和突触功能。用于有效递送药物的非病毒基因递送载体，shAPOE4，编码APOE2的mRNA，pAPOE2， BDNF 和其他基因对大脑可预防和治疗与衰老相关的认知十分位，包括 AD。所请求的补充设备奖将通过以下方式加强药物发现科学： (1).AI药物在输送基质和生物体液中的测定以及体内药动学和通过 HPLC 进行药物的生物分布：HPLC 系统将帮助我们量化捕获的 AI 药物的量纳米粒子，并研究药物在不同器官中的体内药代动力学和生物分布 (2)。通过定量实时聚合酶链式反应 (qRT- PCR）、热循环仪和多模式酶标仪：qRT-PCR 提供快速、高通量、可靠的 qRT-PCR 仪器使我们能够对不同基质中的目标基因进行检测和定量。量化 mRNA 表达随时间的变化，热循环仪将允许转换。 RNA 转为 cDNA，将在 qRT-PCR 中进一步定量。阅读器将用于我们实验室的各种应用，包括核酸定量、蛋白质定量定量、酶动力学、生物标志物定量、ELISA、遗传分析、细胞增殖、细胞毒性、所需资金（190,981 美元）用于采购上述设备。将有助于研究药物、mRNA、pAPOE2、shAPOE4、BDNF 和其他核酸的递送我们预计所需的设备将大大有助于因此，这些拟议的研究设备将应用于AD药物治疗的药物发现领域。对开发基于 AD 的药物和核酸疗法具有重大影响此外，所需的设备。还将可以为该系和大学的其他研究人员进行高质量的研究。