Gaucher disease:Treatment of neurodegenerative disease

戈谢病：神经退行性疾病的治疗

• 批准号: 8723918
• 负责人: Dao Pan
• 金额: $ 41.03万
• 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/8723918
• 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; Cessation of life; Chimeric Proteins; Chronic; Data; Development; Disease; Enhancers; Enzymes; Erythrocytes; Fibroblasts; Gaucher Disease; Gene Transfer; Generations; Glucosylceramides; Goals; 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; Neurons; 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; 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; novel; novel strategies; novel therapeutic intervention; novel therapeutics; overexpression; preclinical evaluation; promoter; prototype; public health relevance; 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（由申请人提供）：神经病性戈谢病 (nGD) 是由 GBA1/Gba1 的有害突变以及由此产生的酸活性缺陷引起的nGD 是一种典型的溶酶体贮积病 (LSD)，由于 i) GCase 无法发挥作用，因此有效的治疗干预对 nGD 的 CNS 表现产生了重大影响。穿过血脑屏障 (BBB)，ii) GCase 在血清 pH 值下快速变性，以及 iii) 野生型表达的膜相关 GCase 的无能性我们从载脂蛋白E中鉴定出两种受体结合肽（Rp），它们可以促进蛋白质通过低密度脂蛋白受体超家族穿过血脑屏障并进入许多中枢神经系统细胞类型。 (LDLRf) 此外，我们还证明了在长期造血干细胞治疗后缺乏酶的 LSD 小鼠模型中中枢神经系统代谢校正和神经功能缺损的一致正常化的治疗潜力。使用慢病毒载体 (LV) 进行 (HSC) 介导的基因治疗 重要的是，我们证明在 GCase 的羧基末端添加 myc 标签不会改变酶的结构、活性或稳定性。巨核细胞能够过量产生溶酶体酶并将其包装到血小板中，以对酶缺陷细胞进行交叉校正。最后，我们基于这些强大的能力开发了模拟急性和慢性人类 nGD 的可行 nGD 小鼠模型。初步数据和 nGD 中 CNS 治疗的巨大未满足的医疗需求，我们将测试以下假设：Rp 与 GCase 融合将使修饰的 GCase 通过 LDLRf 转胞吞到具有广泛 CNS 细胞分布的 CNS 中，从而可以实现协同 CNS 益处通过保护性储存库（即血小板和巨噬细胞）连续生产 GCase-Rp 来实现该项目的总体目标是开发一种新的治疗方法。 LDLRf 介导的转胞吞作用，通过 LV 介导的基因转移至 HSC 中，跨 BBB 传递蛋白质，并在保护性库中进行谱系限制性表达，用于治疗 CNS 表现并基本纠正 nGD 中的内脏疾病。对于 LDLRf 介导的 CNS 递送，开发谱系限制表达系统，通过保护性仓库持续生成靶向蛋白质，并评估蛋白质生物分布和治疗益处nGD 小鼠模型。该项目旨在满足高效 BBB 转胞吞系统的主要未满足的医疗需求，该系统将治疗性大分子广泛分布到许多 CNS 细胞类型，以治疗多种 CNS 疾病。显着适用于神经退行性疾病，包括其他 LSD、帕金森病和阿尔茨海默病。