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: 8729517
• 负责人: Ginette Serrero
• 金额: $ 6.9万
• 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/8729517
• 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

DESCRIPTION (provided by applicant): 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 principal investigator 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 principal investigator 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.

描述（由申请人提供）：美国有超过 700 万例痴呆症病例（约占人口的 3%），其中约 5% 为额颞叶痴呆 (FTD)。最近的研究表明，美国 5-10% 的 FTD 是由颗粒体蛋白前体 (PGRN) 基因突变引起的。这伴随着患者体液中 PGRN 水平降低 50%。 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，它包含一个用于分泌的 17 个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双半胱氨酰丰富的颗粒蛋白 (grn) 基序。 6 Kda Grns 是通过弹性蛋白酶刺激并被分泌性白细胞蛋白酶抑制剂 SLPI 阻断的蛋白水解裂解对 PGRN 进行加工而产生的。最近的证据表明，PGRN 和 grns 都可能直接影响神经退行性疾病的进程，例如阿尔茨海默病、帕金森病和某些类型的 FTD。有人提出，PGRN 和 grn 水平的平衡对于炎症引起的神经元保护/降解很重要。因此，PGRN 和 grn 水平的测量可能有助于揭示大脑中正常与疾病相关过程的平衡。目前，与 PGRN 相比，人们对大脑中特定 grns 的功能和表达以及 PGRN 和 grn 与这些疾病联系的可能机制知之甚少。因此，开发特定的经过验证的检测系统以推动 PGRN/grn 领域在神经退行性疾病领域的发展至关重要。首席研究员是 PGRN 研究（尤其是癌症领域）的公认专家，并开发了经过临床验证的高质量单克隆抗体和检测方法，用于测量患者组织和生物体液中的 PGRN。她的 PGRN 检测和试剂还用于与两个神经退行性疾病卓越中心的合作研究。然而，目前还没有可靠、灵敏和特异的方法来测量 grn 分子，从而阻碍了对 grn 在神经退行性疾病中作用的理解。首席研究员请求 RO3 申请中的支持，以开发 grn 特异性单克隆抗体组合。该方法将针对 7 个不同 grns 的新切割位点，开发适合 7 个 grns 中每一个的夹心 ELISA 测定的 mAb。具体来说，建议： 1) 开发针对每个 7 grn 分子的高亲和力 mAb，适用于生物样品中夹心 ELISA 检测： 2) 开发标准化实验室检测方法，专门检测生物液体中浓度低于 100 的 7 grn pg/ml 并且不受 PGRN 干扰。这些研究非常重要，因为为 7 种 grns 中的每一种开发了经过验证的夹心 ELISA 试剂盒，它将使研究人员能够调查各种 脑脊液和神经退行性疾病（如 FTD）患者的其他生物样本中存在 grns 和 PGRN。提供能够检测、测量和阐明这些重要生物产品的作用的工具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物的潜力。