The Secretory Chaperone 7B2 as an Endogenous Regulator of Amyloid Pathology

分泌伴侣 7B2 作为淀粉样蛋白病理学的内源性调节剂

基本信息

批准号：
8568474
负责人：
IRIS LINDBERG
金额：
$ 19.96万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8568474
关键词：
APP-PS1 Address Affect Age-Months Aging Alzheimer&apos s Disease Alzheimer&apos s disease model American Amyloid Amyloid beta-Protein Amyloid deposition Animals Behavioral Binding Biochemical Biochemistry Bioinformatics Biological Markers Brain California Cell Culture Techniques Cells Characteristics Chemicals Cognition Cognitive Cognitive deficits Control Groups Cytosol Data Deposition Development Disease Endocrine Enzyme-Linked Immunosorbent Assay Exhibits Family Fractionation Genotype Heat shock proteins Heat-Shock Response Hippocampus (Brain)Homeostasis Human Immune Sera Immunofluorescence Immunologic Immunohistochemistry Impaired cognition Incidence Knock-out Knockout Mice Knowledge Laboratories Learning Measures Mediating Molecular Chaperones Mouse Strains Mus Mutant Strains Mice Mutation Neurodegenerative Disorders Neurofibrillary Tangles Neurons Neurosecretory Systems Pathogenesis Pathology Pathway interactions Peptides Phenotype Pilot Projects Population Prevention Procedures Production Proteins Radioimmunoassay Recombinants Role Senile Plaques Slice Testing Therapeutic Thioflavin T Time Tissues Toxic effect Transgenic Mice Transgenic Organisms Ultracentrifugation Universities Work alpha-Crystallins amyloid pathology behavior test brain tissue cognitive function cytotoxicity density immunoreactivity in vitro testing in vivo interest member morris water maze mouse model mutant novel overexpression presenilin public health relevance research study

项目摘要

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) affects 5.2 million Americans over 65, a number expected to increase along with the general aging of the US population. While remarkable progress has been made in the last decade in defining the toxic effects of the amyloid species thought to be involved in loss of cognitive function, much remains to be learned regarding amyloid plaque pathogenesis. Recent data support the idea that chaperones, proteins which control folding homeostasis, contribute to proper neuronal function in a variety of ways. Our laboratory has recently shown that 7B2, a small secreted neuronal protein, is associated with brain amyloid plaques in tissues from both AD humans and from mouse AD models. These data are supported by five independent bioinformatics studies indicating that this protein represents a potential CSF biomarker for neurodegenerative disease. Although present in the secretory pathway and not the cytosol, 7B2 exhibits many biochemical characteristics similar to those of small heat shock protein chaperones. Like alpha crystallin, a member of the heat shock chaperone family, 7B2 can potently block the fibrillation of beta amyloid 1-42 in in vitro tests, and also blocks the cytotoxicity of beta amyloid added to Neuro 2A cell cultures. The current R21 proposal is to test the hypothesis that brain 7B2 levels can control the extent of amyloid plaque deposition and affect cognition in a known mouse model of Alzheimer's. We propose to cross existing mouse strains that either over- or underexpress 7B2 with the APP/PS1 Alzheimer's disease model mouse. Cognitive abilities will be assessed at 6 and 12 months of age in a Morris water maze. Plaque pathology will be quantitated in cortical and hippocampal slices in 6 animals of each genotype; 7B2 immunoreactivity will be measured in concert. Amyloid oligomerization state will be examined in brains of separate animals using chemical and centrifugal separation followed by ELISA and also correlated with genotype. Collectively, these experiments constitute a direct test of our hypothesis that 7B2 levels are negatively correlated with amyloid plaque and oligomer formation.

描述（由申请人提供）：阿尔茨海默病 (AD) 影响着 520 万 65 岁以上的美国人，随着美国人口的普遍老龄化，这一数字预计还会增加。尽管在过去十年中，在确定淀粉样蛋白种类的毒性作用（被认为与认知功能丧失有关）方面取得了显着进展，但关于淀粉样蛋白斑的发病机制，仍有许多问题需要了解。最近的数据支持这样的观点：伴侣蛋白（控制折叠稳态的蛋白质）以多种方式促进神经元的正常功能。我们的实验室最近表明，7B2（一种小型分泌性神经元蛋白）与 AD 人类和小鼠 AD 模型组织中的脑淀粉样斑块相关。这些数据得到了五项独立生物信息学研究的支持，表明该蛋白质代表了神经退行性疾病的潜在脑脊液生物标志物。虽然 7B2 存在于分泌途径而不是细胞质中，但它表现出许多与小热休克蛋白伴侣相似的生化特征。与热休克伴侣家族成员 α 晶状体蛋白一样，7B2 在体外测试中可以有效阻断 β 淀粉样蛋白 1-42 的纤维颤动，并且还可以阻断添加到 Neuro 2A 细胞培养物中的 β 淀粉样蛋白的细胞毒性。目前的 R21 提议是在已知的阿尔茨海默病小鼠模型中测试以下假设：大脑 7B2 水平可以控制淀粉样斑块沉积的程度并影响认知。我们建议将过度表达或表达不足的 7B2 的现有小鼠品系与 APP/PS1 阿尔茨海默病模型小鼠进行杂交。认知能力将在 6 个月和 12 个月大时在莫里斯水迷宫中进行评估。将在每种基因型的 6 只动物的皮质和海马切片中定量斑块病理学； 7B2 免疫反应性将同时测量。将使用化学和离心分离，然后进行 ELISA，在不同动物的大脑中检查淀粉样蛋白寡聚状态，并与基因型相关。总的来说，这些实验构成了对我们假设的直接检验，即 7B2 水平与淀粉样蛋白斑和寡聚体形成呈负相关。