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（由适用提供）：阿尔茨海默氏病（AD）是21世纪的公共卫生危机。除非做过事情，否则预计到2050年，广告将花费超过1.1万亿美元，从而对社会造成了巨大的燃烧（www.alz.org）。在诊断阶段，AD病理学已经非常先进。因此，迫切需要早期（或预科前）诊断生物标志物。 AD的标志是脑组织中一系列AD特异性泛素化蛋白（例如UBI-APP）的显着积累。该项目将使用NEWL发明的方法研究早期与广告相关的UBI-蛋白血液生物标志物。通过单体分子（例如，磷酸化蛋白质）对蛋白质进行抗体的抗体的方法已经很好地确定。磷酸化位点特异性抗体已被广泛用于获取知识，诊断性疾病和在整个生命科学范围内开发治疗剂。相比之下，目前尚无方法可以使抗体以一种结合位点特异性方式识别聚合物蛋白结合物。聚合物蛋白结合定义为通过氨基酸侧链（例如蛋白质泛素化）在两个多肽之间共轭。我们致力于产生泛素与蛋白质共轭特定抗体，但没有成功的努力。通过使用最先进的定量蛋白质组学技术，我们最近发现，在早期“预科前”和“高级”阶段，大脑样品中许多UBI蛋白结合位点特异性肽片段在大脑样品中显着增加。因此，我们采用了一种新方法来制造两种泛素结合位点特异性抗体，用于在转基因AD小鼠模型和动物缺血 - 再生灌注中风模型中检测潜在的血液生物标志物。我们发现，这些转基因AD小鼠的血液样本中这些UBI-蛋白偶联点特异性表位显着增加，但是它们的水平非常低，并且在中大脑中动脉闭塞后的血液样本中基本上不变（中风模型）。该项目将采用这种新方法来生成UBI蛋白结合位点特异性抗体，以研究脑组织中潜在的AD UBI-蛋白生物标志物，并从两种不同的转基因小鼠AD模型中进行血液样本。如果实现了该提案的目标，则可以进一步应用这些创新的方法来研究脑组织，CSF和血液样本中的新人类AD生物标志物。因此，长期目标是开发对早期人类AD生物标志物进行AD研究，诊断和药物开发的临床评估。