Progranulin: A Novel Gene in Gaucher Diseases

颗粒体蛋白前体：戈谢病的一个新基因

• 批准号: 10011889
• 负责人: Chuanju Liu
• 金额: $ 47.91万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011889
• 关键词: Acute; Alleles; Amino Acids; Animal Model; Appearance; Binding; Biological; Blood - brain barrier anatomy; Brain; C-terminal; Cells; Characteristics; Chronic; Clinical; Complex; Crossbreeding; Cytoplasm; Data; Defect; Dependence; Development; Disease; Disease model; Event; Fibroblasts; GRN gene; Gaucher Disease; Genes; Genetic Diseases; Glucosylceramides; Heat-Shock Proteins 70; Inflammatory Arthritis; Knockout Mice; Lead; Life; Lung Inflammation; Lysosomal Storage Diseases; Lysosomes; Mediating; Medical; Modeling; Molecular; Mutation; Neuraxis; Neurologic; Neuropathy; Onset of illness; Organ; Ovalbumin; PGRN gene; Paper; Pathogenesis; Patients; Pharmaceutical Preparations; Phenotype; Play; Point Mutation; Protein Disulfide Isomerase; Proteomics; Recombinants; Reporting; Research; Role; Science; Serum; Site; Structure; Symptoms; Testing; Therapeutic; Therapeutic Effect; Tissues; Tubular formation; Variant; Visceral; asthma model; base; cell type; disease phenotype; disease-causing mutation; enzyme replacement therapy; glucosylceramidase; insight; macrophage; mouse model; new therapeutic target; novel; novel diagnostics; novel therapeutic intervention; novel therapeutics

Project Summary Gaucher disease (GD) is a genetic disease caused by mutations in the GBA1 gene which result in reduced enzymatic activity of β-glucocerebrosidase (GCase); however, patients with the same GCase mutations may have significant variably in disease presentation, ranging from a life-threatening manifestation to almost asymptomatic. It is believed that the modifier genes are responsible for the extraordinarily diverse phenotypes among patients harboring identical GBA1 mutations. Thus, identification of new modifier genes in GD will provide invaluable information in understanding the pathogenesis of GD and in searching for novel diagnostic and therapeutic targets for GD. Our previous report that progranulin (PGRN) is therapeutic against inflammatory arthritis (Tang, et al. Science, 2011) prompted us to determine whether PGRN also played a role in lung inflammation using ovalbumin (OVA)-challenged asthma model, which led to the unexpected discovery of GRN (the gene encoding PGRN) as a novel gene in GD. PGRN null mice display typical features of GD, including “wrinkled tissue paper” appearance of Gaucher cells in multiple organs, and tubular-like lysosome transformation in macrophages. In addition, serum PGRN levels of GD patients are significantly lower when compared to healthy controls, and lower PGRN level was significantly associated with the GRN variants identified in GD patients (Jian, et al, EBioMedicine, 2016a). PGRN binds directly to GCase, and its deficiency results in aggregation in the cytoplasm and defects in the lysosomal localization of GCase. Additionally, 98 C- terminal amino acids of PGRN, referred to as Pcgin, are required and sufficient for the binding to GCase. Pcgin effectively ameliorates the disease phenotype in GD patient fibroblasts and OVA-challenged GD model (Jian, et al, EBioMedicine, 2016b). There is an urgent unmet medical need for treating neuropathic GD, since current enzyme replacement therapy for GD cannot penetrate into brain. Excitingly, our preliminary data revealed that Pcgin could cross blood brain barrier and might be also therapeutic against neuropathic GD. In an effort to elucidate the molecular mechanism underlying PGRN- and Pcgin-mediated therapeutic effect against GD, we performed an unbiased proteomics screen, which led to the isolation of protein disulfide-isomerase A3 (PDIA3) as a PGRN and Pcgin co-bound molecule. In addition, loss of PDIA3 abolished Pcgin's therapeutic effects in GD patient fibroblasts. The hypothesis of this application is that PGRN is a novel modifier in GD and its derivative Pcgin is therapeutic against GD. The Specific Aims are: (1) To determine the therapeutic role of PGRN and its derivative Pcgin in GD, in particular against neuropathic GD; and (2) To elucidate the cellular and molecular mechanisms by which PGRN and Pcgin regulate GCase and GD, with special focus on their interactions with PDIA3 in GD. Completion of the proposed research will not only present PGRN as a novel modifier of GD, provide new insight into the molecular events in the pathogenesis of GD, but could also lead to the development of novel biologics for treating GD, in particular neuropathic GD.

项目摘要 Gaucher病（GD）是由GBA1基因突变引起的遗传疾病，导致降低 β-葡萄糖核苷酶（GCASE）的酶活性；但是，具有相同GCASE突变的患者可能 在疾病表现中具有显着的不同，从威胁生命的表现到几乎 无症状。据信，修饰符基因负责异常多样的表型 在具有相同GBA1突变的患者中。那就是GD中新的修饰基因基因的识别 在了解GD的发病机理和寻找新型诊断方面提供了宝贵的信息 和GD的治疗靶标。我们先前的报告，即程序Nulin（PGRN）是针对的治疗 炎症性关节炎（Tang等人，Science，2011年）促使我们确定PGRN是否也起作用 在使用椭圆蛋白（OVA）挑战哮喘模型的肺注射中，这导致了意外发现 GRN（编码PGRN的基因）作为GD中的新基因。 PGRN NULL小鼠显示GD的典型特征， 包括多个器官中的Gaucher细胞的“皱纹薄纸”外观，以及管状溶酶体 巨噬细胞的转化。此外，GD患者的血清PGRN水平显着降低 与健康对照组相比，较低的PGRN水平与GRN变体显着相关 在GD患者中鉴定（Jian等，Ebiomedicine，2016a）。 PGRN直接与GCASE结合及其缺乏症 结果导致细胞质的聚集和GCASE溶酶体定位的缺陷。另外，98 c- PGRN的末端氨基酸（称为PCGIN）是必需的，足以与GCASE结合。 PCGIN 有效地改善了GD患者成纤维细胞和OVA挑战的GD模型中的疾病表型（Jian， 等人，EbioMedicine，2016b）。由于目前 GD的酶替代疗法无法渗透到大脑中。令人兴奋的是，我们的初步数据表明 PCGIN可能会越过血脑屏障，也可能对神经性GD进行征服。努力 阐明PGRN和PCGIN介导的针对GD的分子机制，我们 进行了无偏的蛋白质组学筛选，这导致蛋白质二硫化物异构酶A3（PDIA3）的分离 作为PGRN和PCGIN共同结合的分子。此外，PDIA3的损失消除了PCGI的治疗作用 GD患者成纤维细胞。该应用的假设是PGRN是GD中的一种新颖的修饰符 衍生物Pcgin对GD具有治疗性。具体目的是：（1）确定 PGRN及其GD中的衍生物PCGIN，特别是针对神经性GD； （2）阐明细胞 PGRN和PCGIN调节GCASE和GD的分子机制，特别关注其 与GD中的PDIA3相互作用。拟议的研究的完成不仅将把PGRN作为一种新颖 GD的修饰符为GD发病机理中的分子事件提供了新的见解，但也可能导致 用于治疗GD的新生物制剂的发展，特别是神经性GD。