An Innovative Approach to Study Alzheimer Disease Blood Biomarkers

研究阿尔茨海默病血液生物标志物的创新方法

• 批准号: 9251737
• 负责人: Bingren Hu
• 金额: $ 19.3万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9251737
• 关键词: Age; Age-Months; Alzheimer' s Disease; Alzheimer' s disease model; Amino Acids; Amyloid beta-Protein; Animal Disease Models; Animals; Antibodies; Biological Assay; Biological Markers; Biological Sciences; Blood; Blood Proteins; Blood specimen; Brain; Brain Injuries; Clinical; Diagnosis; Diagnostic; Disease Progression; Disease model; Enzyme-Linked Immunosorbent Assay; Epitopes; Exhibits; Hematological Disease; Human; Immunohistochemistry; Ischemia; Knowledge; Methods; Middle Cerebral Artery Occlusion; Modeling; Modification; Mus; Neurodegenerative Disorders; Parkinson Disease; Pathologic; Pathology; Patients; Peptide Fragments; Peptides; Phosphorylation Site; Polymers; Prevention therapy; Proteins; Proteomics; Public Health; Reperfusion Therapy; Research; Sampling; Side; Site; Societies; Stroke; Symptoms; Technology; Therapeutic Agents; Tissue Sample; Tissues; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Ubiquitin; Ubiquitination; Western Blotting; age group; base; brain tissue; cost; diagnostic biomarker; disease diagnosis; disorder control; drug development; innovation; macromolecule; mind control; mouse model; neocortical; novel; novel strategies; polypeptide; protein E; protein biomarkers; public health relevance; single molecule; success; targeted biomarker; tau Proteins; therapy development

 DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is the public health crisis of the 21st century. Unless something is done, AD is projected to cost over $1.1 trillion by 2050, thus placing a huge burden on society (www.alz.org). At the diagnostic stage, AD pathologies are already quite advanced. Therefore, early (or pre-pathological) diagnostic biomarkers are desperately needed. A hallmark of AD is significant accumulation of an array of AD-specific ubiquitinated proteins (ubi- proteins, e.g., ubi-APP) in brain tissue. This project will use a newl invented method to study early AD-related ubi-protein blood biomarkers. Methods of making antibodies against proteins post-translationally modified by a monomeric molecule, e.g., phosphorylated proteins, are already very well established. Phosphorylation site- specific antibodies have been widely used in acquiring knowledge, diagnosing disease, and developing therapeutic agents throughout the whole spectrum of life sciences. In comparison, there is no method currently available for making antibodies recognizing polymeric protein conjugates in a conjugation site-specific manner. Polymeric protein conjugation is defined as covalent conjugation between two polypeptides via amino acid side chains such as protein ubiquitination. Extensive efforts have been devoted to generating ubiquitin-to-protein conjugation site-specific antibodies, but without success. By using a state-of-the-art quantitative proteomic technology, we recently found that many ubi-protein conjugation site-specific peptide fragments are dramatically increased in brain samples from a transgenic AD mouse model at both early "pre- pathological" and "advanced" stages. We therefore employed a new method to make two ubiquitin conjugation site-specific antibodies for detecting potential blood biomarkers in a transgenic AD mouse model and an animal ischemia-reperfusion stroke model. We found that these ubi-protein conjugation site-specific epitopes were significantly increased in blood samples from the transgenic AD mice, but their levels were very low and basically unchanged in the blood samples from animals after middle cerebral artery occlusion (stroke model). This project will employ this new method to generate ubi-protein conjugation site-specific antibodies to study potential AD ubi- protein biomarkers in brain tissue and blood samples from two different transgenic mouse AD models. If the objectives of this proposal are achieved, these innovative approaches can be further applied to study new human AD biomarkers in brain tissue, CSF and blood samples. Therefore, the long-term objectives are to develop clinical assays of early human AD biomarkers for AD research, diagnosis and drug development.

 描述（由申请人提供）：阿尔茨海默病 (AD) 是 21 世纪的公共卫生危机，如果不采取措施，预计到 2050 年 AD 的费用将超过 1.1 万亿美元，从而给社会带来巨大负担 (www.alz.在诊断阶段，AD 病理学已经相当先进，因此，迫切需要早期（或病理前）诊断生物标志物是一系列 AD 特异性的显着积累。脑组织中的泛素化蛋白（ubi-蛋白，例如 ubi-APP）将使用一种新发明的方法来研究早期 AD 相关的泛素蛋白血液生物标志物的制备方法，以对抗由单体翻译后修饰的蛋白质。分子，例如磷酸化蛋白质，已经非常成熟地被广泛用于获取知识、诊断疾病和开发治疗药物。相比之下，目前尚无可用的方法来制备以缀合位点特异性方式识别聚合蛋白缀合物的抗体，聚合蛋白缀合被定义为两个多肽之间通过氨基酸侧链的共价缀合，例如蛋白质泛素化。一直致力于生成泛素与蛋白质缀合位点特异性抗体，但没有成功。通过使用最先进的定量蛋白质组技术，我们最近发现了许多抗体。在转基因AD小鼠模型的早期“病理前”和“晚期”阶段，泛蛋白缀合位点特异性肽片段显着增加，因此我们采用了一种新方法来制备两种泛素缀合位点特异性抗体。用于检测转基因 AD 小鼠模型和动物缺血再灌注中风模型中潜在的血液生物标志物，我们发现来自转基因的血液样本中这些泛蛋白缀合位点特异性表位显着增加。 AD小鼠中，但在大脑中动脉闭塞（中风模型）后的动物血液样本中其水平非常低且基本没有变化，本项目将采用这种新方法生成ubi蛋白缀合位点特异性抗体来研究潜在的AD ubi。 - 来自两种不同转基因小鼠 AD 模型的脑组织和血液样本中的蛋白质生物标志物 如果该提案的目标得以实现，这些创新方法可以进一步应用于研究脑组织、脑脊液和血液样本中的新的人类 AD 生物标志物。长期目标是发展用于 AD 研究、诊断和药物开发的早期人类 AD 生物标志物的临床测定。