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。除非采取措施，否则到2050年，AD预计将花费超过1.1万亿美元，从而给社会带来巨大负担（www.alz.org）。在诊断阶段，AD病理已经相当先进。因此，迫切需要早期（或病理前）诊断生物标志物。AD的标志是大量AD特异性泛素化蛋白（ubi蛋白，例如，ubi-APP）。本项目将采用新发明的方法研究AD早期相关的泛蛋白血液生物标志物。 制备针对被单体分子（例如，磷酸化的蛋白质已经非常成熟。磷酸化位点特异性抗体已广泛用于整个生命科学领域的知识获取、疾病诊断和治疗剂开发。相比之下，目前没有方法可用于制备以缀合位点特异性方式识别聚合蛋白缀合物的抗体。聚合蛋白质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。已经进行了大量的努力来产生泛素-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学技术，我们最近发现，许多泛蛋白缀合位点特异性肽片段在早期“病理前”和“晚期”阶段的转基因AD小鼠模型的脑样品中显著增加。因此，我们采用了一种新的方法，使两个泛素结合位点特异性抗体检测潜在的血液生物标志物在转基因AD小鼠模型和动物缺血再灌注中风模型。我们发现这些泛蛋白结合位点特异性表位在转基因AD小鼠的血液样品中显著增加，但在大脑中动脉闭塞（中风模型）动物的血液样品中它们的水平很低且基本不变。该项目将采用这种新方法产生泛蛋白缀合位点特异性抗体，以研究来自两种不同转基因AD小鼠模型的脑组织和血液样品中潜在的AD泛蛋白生物标志物。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、CSF和血液样品中的新的人类AD生物标志物。因此，长期目标是开发用于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。