Granulin specific monoclonal antibodies to investigate their expression and role

颗粒蛋白特异性单克隆抗体研究其表达和作用

• 批准号: 8624365
• 负责人: Ginette Serrero
• 金额: $ 6.97万
• 依托单位: A AND G PHARMACEUTICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8624365
• 关键词: Accounting; Acute; Affect; Affinity; Alzheimer' s Disease; Amino Acids; Anti-Inflammatory Agents; Anti-inflammatory; Behavior; Biological; Biological Assay; Biological Process; Biological Products; Biopsy; Brain; Carbohydrates; Cerebrospinal Fluid; Cloning; Collaborations; Core Protein; Dementia; Detection; Development; Diagnostic Reagent; Disease; Elastases; Enzyme-Linked Immunosorbent Assay; Equilibrium; Frontotemporal Dementia; Generations; Glycoproteins; Growth; Healthcare; Inflammation; Inflammatory; Judgment; Laboratories; Language; Leukocytes; Light; Link; Liquid substance; Malignant Neoplasms; Measurement; Measures; Memory; Methods; Monitor; Monoclonal Antibodies; Mutation; Neurodegenerative Disorders; Neurons; PGRN gene; Parkinson Disease; Patients; Peptide Hydrolases; Peptide Signal Sequences; Plasma; Population; Presenile Dementia; Principal Investigator; Process; Production; Progranulin; Protease Inhibitor; Proteins; Reagent; Research; Research Personnel; Role; Sampling; Site; Surrogate Markers; System; Tandem Repeat Sequences; Testing; Thinking; Tissues; Urine; Work; Wound Healing; advanced dementia; antileukoprotease; cross reactivity; extracellular; granulin; human SLPI protein; programs; public health relevance; therapeutic target; tool

Abstract Over 7 million cases of dementia exist in the US (~3% of the population) and of these ~5% are frontotemporal dementia (FTD). It has been demonstrated recently that 5-10% of all FTD in the US are caused by mutations in the Progranulin (PGRN) gene. This is accompanied with a 50% reduction of PGRN level in patients' biological fluids. PGRN is a 593 amino-acid growth and survival factor. Cloning and sequencing of PGRN showed that it contains a 17 amino-acid signal peptide for secretion and seven and a half 6 kDa grn repeats with a unique tandem repeat double cysteinyl rich granulin (grn) motif. The 6 Kda Grns are generated by processing of PGRN by a proteolytic cleavage stimulated by elastase and blocked by secretory leucocyte protease inhibitor SLPI. Recent evidence suggests that both PGRN and grns may directly influence neurodegenerative diseases process, such as Alzheimer's disease, Parkinson's disease and certain types of FTD. It has been suggested that the balance of PGRN and grn levels is important for neuron protection/degradation as a result of inflammation. Accordingly, the measurement of PGRN and grn levels may shed light on the balance of normal versus disease related processes in the brain. Currently, relatively little is known about the function and expression of specific grns in the brain when compared to PGRN and about the possible mechanisms linking PGRN and grn to these diseases. Thus, it is critical to develop specific validated assay systems to advance the PGRN/grn field forward in the neurodegenerative disease. The PI is a recognized expert in PGRN research particularly in cancer and has developed clinically validated high quality monoclonal antibodies and assays to measure PGRN in tissue and biological fluids in patients. Her PGRN assays and reagents are also used in collaborative studies with two centers of excellence in neurodegenerative disease. However, there are no reliable, sensitive and specific assays to measure grn molecules, thereby hampering understanding of the role of grns in neurodegenerative diseases. The PI is requesting support in this RO3 application to develop a portfolio of monoclonal antibodies specific to grn. The approach will be to target the neo-cleavage sites of the seven different grns to develop mAbs suitable for sandwich ELISA assay for each of the 7 grns. Specifically, it is proposed to: 1) Develop high affinity mAbs specific to each of 7 grn molecule suitable for sandwich ELISA detection in biological samples: 2) Develop standardized laboratory assays for specifically detecting the 7 grns in biological fluids at concentrations of less than 100 pg/ml and without interference by PGRN. These studies will be important as the development of validated sandwich ELISA kits for each of the 7 grns, it will enable researchers to investigate the presence and the ratio of various grns and PGRN in CSF and other biological samples from patients with neurodegenerative diseases such as FTD. Providing tools that can detect measure and elucidate the role of these important biological products may also be useful in revealing the potential to use them as therapeutic targets or surrogate markers for disease or therapy monitoring.

抽象的 在美国存在超过700万例痴呆症（人口约3％），其中约5％是额颞 痴呆（FTD）。最近已经证明，美国所有FTD的5-10％是由突变引起的 前植物（PGRN）基因。伴随于患者生物学的PGRN水平降低50％ 流体。 PGRN是593个氨基酸的生长和生存因子。 PGRN的克隆和测序表明它 包含一个17个氨基酸信号肽，用于分泌和7个半6 kDa grn重复，独特 串联重复双半胱氨酸富集颗粒（GRN）基序。 6 kDa GRN是通过处理生成的 由弹性蛋白酶刺激的蛋白水解裂解PGRN，并被分泌白细胞蛋白酶抑制剂阻塞 Slpi。最近的证据表明，PGRN和GRN都可能直接影响神经退行性疾病 过程，例如阿尔茨海默氏病，帕金森氏病和某些类型的FTD。有人建议 由于PGRN和GRN水平的平衡对于神经元保护/降解很重要 炎。因此，PGRN和GRN水平的测量可能会揭示正常的平衡 大脑中与疾病相关的过程。目前，关于该功能和 与PGRN相比，大脑中特定GRN的表达以及有关可能连接的机制 这些疾病的PGRN和GRN。因此，建立特定验证的测定系统以推动 神经退行性疾病中的PGRN/GRN场。 PI是PGRN研究的公认专家 特别是在癌症中，并且已经开发了临床验证的高质量单克隆抗体和测定 测量患者组织和生物液中的PGRN。她的PGRN分析和试剂也用于 与神经退行性疾病卓越的两个卓越中心的合作研究。但是，没有 可靠，敏感和特定的测定法以测量GRN分子，从而阻碍了对角色的理解 神经退行性疾病中的GRN。 PI在此RO3应用程序中请求支持以开发 特定于GRN的单克隆抗体的组合。该方法将是针对的 七个不同的GRN可为7个GRN中的每个GRN开发适合三明治ELISA测定法的mAb。具体来说，它 提出：1）开发针对适合三明治ELISA的7个GRN分子中的每个特有的高亲和力mAbs 在生物样品中检测：2）开发标准化实验室测定法以专门检测7个GRN 在浓度小于100 pg/mL的生物流体中，无PGRN干扰。这些研究 对于为7个GRN中的每个GRN中的每一个开发了经过验证的三明治ELISA套件的开发将很重要，它将启用 研究人员研究CSF和其他生物学中各种GRN和PGRN的存在和比率 来自神经退行性疾病的患者（例如FTD）的样本。提供可以检测度量的工具 阐明这些重要生物产品的作用也可能有助于揭示潜力 将它们用作治疗靶标或疾病或治疗监测的替代标记。