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 美国有超过 700 万痴呆症病例（约占人口的 3%），其中约 5% 是额颞叶痴呆症 痴呆症（FTD）。最近的研究表明，美国 5-10% 的 FTD 是由基因突变引起的 颗粒体蛋白前体 (PGRN) 基因。这伴随着患者生物体内 PGRN 水平降低 50% 液体。 PGRN 是一种由 593 个氨基酸组成的生长和存活因子。 PGRN 的克隆和测序表明 包含 17 个用于分泌的氨基酸信号肽和七个半 6 kDa grn 重复序列，具有独特的 串联重复富含半胱氨酰的颗粒蛋白 (grn) 基序。 6 Kda Grns 是通过处理生成的 PGRN 通过弹性蛋白酶刺激的蛋白水解裂解并被分泌性白细胞蛋白酶抑制剂阻断 SLPI。最近的证据表明 PGRN 和 grns 可能直接影响神经退行性疾病 过程，例如阿尔茨海默病、帕金森病和某些类型的 FTD。已建议 PGRN 和 grn 水平的平衡对于神经元保护/降解很重要，因为 炎。因此，PGRN 和 grn 水平的测量可以揭示正常的平衡 与大脑中疾病相关的过程。目前，人们对其功能和功能知之甚少。 与 PGRN 相比，大脑中特定 grns 的表达以及联系的可能机制 PGRN 和 grn 针对这些疾病。因此，开发特定的经过验证的测定系统以推进 PGRN/grn 在神经退行性疾病领域处于领先地位。 PI 是 PGRN 研究领域公认的专家 特别是在癌症方面，并开发了经过临床验证的高质量单克隆抗体和检测方法 测量患者组织和生物体液中的 PGRN。她的 PGRN 检测和试剂也用于 与两个神经退行性疾病卓越中心的合作研究。然而，没有 可靠、灵敏和特异的测定法来测量 grn 分子，从而阻碍了对其作用的理解 grns 在神经退行性疾病中的作用。 PI 请求对此 RO3 应用程序的支持，以开发 grn 特异性单克隆抗体组合。该方法将针对新切割位点 七种不同的 grns 来开发适合 7 个 grns 中每一种的夹心 ELISA 测定的 mAb。具体来说，它 建议：1) 开发针对 7 个 grn 分子中每一个的高亲和力 mAb，适用于夹心 ELISA 生物样品中的检测：2) 开发专门检测 7 种 grns 的标准化实验室检测方法 在生物体液中浓度低于 100 pg/ml，且不受 PGRN 干扰。这些研究 为 7 种 grns 中的每一种开发经过验证的夹心 ELISA 试剂盒将非常重要，这将使 研究人员调查脑脊液和其他生物中各种 grns 和 PGRN 的存在和比例 来自患有神经退行性疾病（例如 FTD）的患者的样本。提供可以检测测量的工具 并阐明这些重要生物产品的作用也可能有助于揭示 将它们用作疾病或治疗监测的治疗靶点或替代标记。