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

点击翻译按钮获取中文摘要

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.