Transthyretin's Regulatory Role in Beta-Amyloid Aggregation and Toxicity

运甲状腺素蛋白在 β-淀粉样蛋白聚集和毒性中的调节作用

• 批准号: 7614921
• 负责人: Jeffrey A Johnson
• 金额: $ 41.27万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7614921
• 关键词: Acids; Address; Adenoviruses; Affect; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Amyloid deposition; Amyloidosis; Animal Model; Asses; Astrocytes; Behavior; Benzoxazoles; Biochemical; Biological Assay; Blood; Brain; Carrier Proteins; Cell Culture Techniques; Cells; Cerebral cortex; Cerebrospinal Fluid; Cessation of life; Characteristics; Circular Dichroism; Complex; Data; Defect; Deposition; Diflunisal; Dissociation; Electron Microscopy; Engineering; Evaluation; Fluorescence; Fluorescence Resonance Energy Transfer; Goals; Heart; Hippocampus (Brain); Human; In Vitro; Infection; Inherited; Injection of therapeutic agent; Investigation; Kinetics; Ligand Binding; Ligands; Link; Maps; Mass Spectrum Analysis; Measures; Modeling; Modification; Molecular; Molecular Sieve Chromatography; Mus; Mutation; Nature; Neurofibrillary Tangles; Neurons; Pathology; Patients; Plasma; Prealbumin; Presenile Alzheimer Dementia; Principal Investigator; Production; Protein Precursors; Proteins; Recombinants; Resistance; Resveratrol; Role; Senile Plaques; Structure; Surface Plasmon Resonance; System; Testing; Tg2576; Thyroxine; Tissues; Toxic effect; Transgenic Mice; Variant; abeta accumulation; age related; amyloidogenesis; base; cell injury; crosslink; dentate gyrus; dimer; gel electrophoresis; in vivo; in vivo Model; insight; interest; lateral ventricle; light scattering; monomer; mouse model; mutant; neuron loss; neurotoxic; neurotoxicity; novel; overexpression; prevent; programs; protein folding; research study; sound; tool

Transthyretin's Regulatory Role in Beta-Amyloid Aggregation and Toxicity Alzheimer’s disease (AD) has been linked to deposition of beta-amyloid (Aβ) as amyloid plaques in the brain. Transgenic mice expressing the human Aβ precursor protein (APP) produce high levels of Aβ and develop amyloid plaques, but they do not suffer the extensive neuronal cell death that is characteristic of AD. Recent studies have uncovered a possible explanation: these transgenic mice greatly increase the synthesis of the transport protein transthyretin (TTR), and TTR appears to protect the mice from the neurotoxic effects of Aβ. The long-term goals of this project are to answer three questions that arise from these intriguing results: (1) how does TTR exert its protective activity? (2) why does this natural protective activity fail in AD? (3) can it be restored? Our hypothesis is that subtle changes in TTR tertiary and/or quaternary structure strongly modulate TTR- Aβ interactions. In specific aim 1, several TTR mutants that differ in their tertiary and/or quaternary structure and stability will be produced and characterized. Each mutant will be screened for its ability to interfere with Aβ aggregation, and to inhibit Aβ toxicity in an in vitro cell culture model. The data will be analyzed to identify correlations between TTR structure and stability with its ability to alter Aβ aggregation and toxicity. In specific aim 2, a detailed examination of the interaction between TTR (both wildtype and selected mutants) and Aβ will be undertaken. Biophysical and biochemical tools such as circular dichroism, fluorescence, static and dynamic light scattering, crosslinking, and kinetic modeling will be employed. From these data will emerge molecular-level mechanistic insights into the nature of TTR-Aβ association and the means by which TTR affects Aβ aggregation kinetics. The goal of specific aim 3 is to identify small ligands that stabilize TTR and determine their influence on TTR’s ability to modulate Aβ aggregation and toxicity. In aim 4, TTR (wildtype and mutants) along with TTR-binding ligands will be tested for protection against Aβ toxicity in ex vivo and in vivo mouse models. This will be achieved by using a newly developed assay in which stereotactic injection of Aβ into mouse brain leads to loss of CA1 and dentate gyrus neurons, and by infection of astrocytes with adenovirus-TTR constructs. The project spans from characterization of the structure and stability of TTR (and mutants), through in vitro assessment of TTR’s effect on Aβ aggregation and toxicity, to in vivo evaluation of TTR efficacy at preventing neuronal cell death. These studies will provide a sound and rational basis for developing novel pharmacological approaches to preventing AD by enhancement of the natural defenses provided by TTR.

thyretin在β-淀粉样蛋白和阿尔茨海默氏病（AD）中的作用已与TA-amyloidβ）联系在一起，因为淀粉样蛋白斑块中的淀粉样蛋白斑块中的转基因小鼠表达了人类Aβ前体蛋白（APP）。 ，但没有遭受巨大的细胞死亡tath tath tath tath tath tath tath tath tath tath tath tath tath：这些转基因小鼠大大增加了转运蛋白转卫生蛋白（TTR）的合成，而TTR似乎可以保护小鼠免受Aβ的神经毒性的影响 - 期限）TTR在AD中如何发挥保护作用？在体外培养模型中，将产生和/或稳定性，并在体外培养模型中进行表征。 ，将对TTR和Aβ之间的相互作用进行详细检查。 TTR-Aβciation和TTR影响Aβ的动力学的平均值。为了在体内和体内模型中的Aβ进行保护，这将通过新开发的测定方法来实现，在该测定中，Antero的立体定位会导致小鼠脑中的CA1和齿状回跨越TTR的结构和稳定性的跨度（和杂种），通过对TTR评估的体外评估，以防止神经元细胞死亡。 TTR提供的自然防御。