Gaucher disease:Treatment of neurodegenerative disease

戈谢病：神经退行性疾病的治疗

• 批准号: 9069618
• 负责人: Dao Pan
• 金额: $ 41.45万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9069618
• 关键词: Acids; Acute; Alzheimer' s Disease; Amino Acids; Apolipoprotein E; Area; Behavior; Blood; Blood - brain barrier anatomy; Blood Circulation; Blood Platelets; Brain; Cell Line; Cell Lineage; Cells; Chimeric Proteins; Chronic; Data; Development; Disease; Enhancers; Enzymes; Erythrocytes; Fibroblasts; Gaucher Disease; Gene Transfer; Generations; Glucosylceramides; Goals; Health; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hepatic; Histopathology; Human; Hybrids; Injection of therapeutic agent; L-Iduronidase; Lentivirus Vector; Lipoprotein Receptor; Liver; Low Density Lipoprotein Receptor; Lysosomal Storage Diseases; Mediating; Medical; Megakaryocytes; Membrane; Metabolic; Microglia; Modeling; Mus; Mutation; Nerve; Neurodegenerative Disorders; Neurologic; Neuronopathic Gaucher Disease; Parkinson Disease; Peptide Receptor; Peptides; Pharmaceutical Preparations; Platelet Activation; Production; Protein Engineering; Proteins; Rare Diseases; Resolution; Resources; Serum; Specificity; System; Testing; Therapeutic; Therapeutic Intervention; Time; Translations; Variant; Visceral; base; brain cell; brain parenchyma; cell type; cellular transduction; density; enzyme reconstitution; enzyme structure; enzyme therapy; gene therapy; glucosidase; glucosylsphingosine; improved; in vivo; macromolecule; macrophage; monocyte; mouse model; nervous system disorder; neuron loss; novel; novel strategies; novel therapeutic intervention; novel therapeutics; overexpression; preclinical evaluation; promoter; prototype; receptor binding; small molecule; success; trafficking; transcytosis; transgene expression; uptake

DESCRIPTION (provided by applicant): Neuronopathic Gaucher disease (nGD) is caused by deleterious mutations in GBA1/Gba1 and the resultant defective activity of acid �-glucosidase (GCase). nGD is a prototype lysosomal storage disease (LSD) that displays generalized neuronal death in central nerve system (CNS). Effective therapeutic interventions have had major effects on the CNS manifestations of nGD due to i) the inability of GCase to cross the blood brain barrier (BBB), ii) the rapid denaturation of GCase at serum pH, and iii) the inability f the membrane associated GCase expressed at wild-type levels in cells to be secreted in sufficient amounts for metabolic cross- correction. We identified two receptor-binding peptides (Rp) from apolipoprotein E that can facilitate protein delivery across the BBB and into many CNS cell types via the low-density lipoprotein receptor superfamily (LDLRf). Also, we demonstrated the therapeutic potential with CNS metabolic correction and concordant normalization of neurological deficits in an enzyme-deficient LSD murine model after long-term hematopoietic stem cells (HSC)-mediated gene therapy using a lentiviral vector (LV). Importantly, we show that carboxy terminal addition of the myc-tag to GCase does not alter the enzyme's structure, activity or stability. Moreover, we showed for the first time that megakaryocytes are capable of over-producing lysosomal enzymes and packaging them into platelets for cross-correction of enzyme-deficient cells. Finally, we developed viable nGD mouse models that mimic acute and chronic human nGD. Based on these strong preliminary data and the great unmet medical need for CNS therapy in nGD, we will test the hypothesis that fusion of Rp to GCase will enable the modified GCase to transcytose into the CNS with wide CNS cell distribution via the LDLRf, so that synergistic CNS benefits can be achieved from continuous production of GCase-Rp through protective depots, i.e., platelets and macrophages. The overall goal of the project is to develop a novel therapeutic approach utilizing LDLRf-mediated transcytosis for protein delivery across the BBB via LV- mediated gene transfer into HSC with lineage-restricted expression in protective depots for the treatment of the CNS manifestations and essential correction of the visceral disease in nGD. We will assess various GCase-Rp for LDLRf-mediated CNS delivery, develop lineage-restricted expression systems for sustained and targeted protein generation via protective depots, and evaluate protein bio-distribution and therapeutic benefits in nGD mouse models. This project aims at a major unmet medical need for efficient BBB transcytosis systems with broad distribution of the therapeutic macromolecules to many CNS cell types for the treatment of a wide variety of CNS diseases. The approaches developed in the studies have general and significant applicability to neurodegenerative diseases including other LSDs, and Parkinson and Alzheimer diseases.

描述（由适用提供）：神经性高刺病（NGD）是由GBA1/GBA1中有害的突变引起的，以及酸 - 葡萄糖苷酶（GCASE）的缺陷活性。 NGD是一种原型溶酶体储存疾病（LSD），在中枢神经系统（CNS）中显示全身神经元死亡。有效的治疗干预措施对NGD的CNS表现产生了重大影响。i）GCASE无法跨越血脑屏障（BBB），ii）GCASE在血清pH下的GCASE的快速变性，以及III），无力f膜相关的g case在野生型中表达的GCASE在野生型中表达的次数与次数交叉分泌，以弥补次数为中等。我们通过低密度脂蛋白受体超级家族（LDLRF）从载脂蛋白E中鉴定出了两种受体结合肽（RP），它们可以促进蛋白质递送到许多CNS细胞类型中。此外，我们证明了使用肺炎病毒载体（LV），在长期造血干细胞（HSC）介导的基因治疗后，在酶缺陷型LSD鼠模型（HSC）介导的基因治疗后，在酶缺陷型LSD鼠模型（HSC）介导的LSD鼠模型中的神经系统缺乏症具有治疗潜力。重要的是，我们表明，羧基终端将myc-tag添加到gcase上不会改变酶的结构，活性或稳定性。此外，我们首次表明，巨核细胞能够过度产生溶酶体酶并将其包装到血小板中以进行杂交缺陷酶细胞。最后，我们开发了模仿急性和慢性人类NGD的可行NGD小鼠模型。基于这些强大的初步数据和NGD中CNS治疗的巨大未满足医学需求，我们将测试以下假设：RP与GCASE融合将使经过修改的GCASE通过LDLRF通过LDLRF进行较大的CNS细胞分布到CNS中，从而可以通过CNENES forecter forder forter forter farecter forter fartecter farecter farcase protipter flate flate flatel flatel flatel flatel flatel rp rp rp rp rp rp rp。该项目的总体目的是利用LDLRF介导的转胞胞病症开发一种新型的治疗方法通过受保护的沉积物产生蛋白质，并在NGD小鼠模型中评估了蛋白质生物分布和治疗益处。该项目的目的是对有效的BBB跨经细胞增多症系统的主要未满足医疗需求，其治疗性大分子对许多CNS细胞类型进行了广泛分布，以治疗多种CNS疾病。研究中开发的方法对包括其他LSD以及帕金森氏症和阿尔茨海默氏病在内的神经退行性疾病具有一般性的适用性。