Krabbe disease therapy integrating gene transfer with lectin-enhanced enzyme delivery to treat pathologies of the CNS

克拉伯病疗法将基因转移与凝集素增强酶递送相结合，以治疗中枢神经系统疾病

• 批准号: 10547167
• 负责人: Walter Acosta
• 金额: $ 39.05万
• 依托单位: BIOSTRATEGIES, LC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10547167
• 关键词: Address; Affect; Age; Age-Months; Animal Disease Models; Animals; Binding; Biodistribution; Blood - brain barrier anatomy; Body Weight; Bone Marrow Transplantation; Brain; Brain imaging; Cells; Cessation of life; Child; Chimeric Proteins; DNA cassette; Demyelinations; Dependovirus; Development; Disease; Disease Progression; Disease model; Dose; Drug Administration Routes; Drug Delivery Systems; Ensure; Enzymes; Episome; Family; Feasibility Studies; Galactosamine; Galactose; Gangliosidosis GM1; Gene Delivery; Gene Expression; Gene Transfer; Genes; Genetic Diseases; Globoid cell leukodystrophy; Glycolipids; Goals; Health system; Hereditary Disease; Human; IGF Type 2 Receptor; Immune; Impairment; Infant; Infusion procedures; L-Iduronidase; Lead; Learning; Lectin; Longevity; Lysosomal Storage Diseases; Lysosomes; Mammalian Cell; Measures; Mediating; Membrane Glycoproteins; Memory; Metabolic; Modeling; Mucopolysaccharidosis I; Mus; Myelin; Neonatal; Nervous system structure; Neurodegenerative Disorders; Neurologic; Neurologic Symptoms; Neurons; Oligodendroglia; Organ; Pathologic; Pathology; Patients; Peripheral; Peripheral Nerves; Peripheral Nervous System; Persons; Pharmaceutical Preparations; Phase; Plants; Production; Psychosine; Public Health; Rare Diseases; Recombinant adeno-associated virus (rAAV); Refractory; Route; Safety; Schwann Cells; Serotyping; Site; Skeletal system; Small Business Innovation Research Grant; Symptoms; Systems Development; Technology; Technology Transfer; Testing; Therapeutic; Tissues; Transfection; Translating; Treatment Efficacy; Treatment Protocols; Tropism; Viral; Virus; adeno-associated viral vector; base; cost; cytotoxic; dosage; effective therapy; experimental study; galactosylceramidase; gene therapy; in vivo; innovation; mouse model; nervous system disorder; neurological pathology; next generation; novel; phase 2 study; pre-clinical research; progressive neurodegeneration; public health relevance; receptor; symptom treatment; therapeutic enzyme; therapeutically effective; transplantation therapy; treatment duration; uptake

Krabbe disease (globoid cell leukodystrophy) is a rare inherited disorder caused by a deficiency in the lysosomal enzyme galactosylceramidase (GALC) leading to destruction of the protective coating (myelin) of nerve cells in the brain and throughout the nervous system. This fatal neurodegenerative disease affects about 1 in 100,000 people in the US. In most cases, Krabbe symptoms develop in infants before 6 months of age, often leading to death by age 2. Bone marrow transplantation is currently the only approved treatment option for Krabbe patients, but only slows disease progression, and only when initiated in pre-symptomatic children. More recent preclinical research has focused on gene transfer utilizing adeno-associated virus (AAV) for long term expression of the normal GALC enzyme in the “twitcher” mouse model of Krabbe disease. Although promising, this approach still shows deficiencies in distributing effective therapeutic doses to critical parts of the brain and nervous system. BioStrategies LC has developed a lectin-based therapeutic delivery module (RTB) that enables fused lysosomal enzymes to effectively target the CNS and treat neurological symptoms following weekly ERT infusions, as demonstrated in several lysosomal disease animal models. In this SBIR project, we propose to merge the benefits of AAV-mediated gene transfer technologies that provide continuous long-term in vivo enzyme production with those of the RTB delivery technology to ensure effective treatment of Krabbe’s progressive neurodegeneration. Gene expression cassettes fusing GALC with BioStrategies’ RTB lectin will be packaged into recombinant adeno-associated virus for production and subsequent administration to twitcher mice. Impacts on neurological disease progression and lifespan of mice receiving AAV-vectored GALC-RTB fusions will be compared with untreated mice and those administered AAV-vectored GALC alone. Specific aims of this Phase I feasibility study are 1) to construct GALC-RTB AAV vectors and demonstrate their ability to produce fully functional GALC-RTB fusion product in mammalian cells and mice; and 2) to compare therapeutic efficacy of AAV-mediated gene transfer of GALC-RTB versus GALC in the Krabbe mouse model as measured by lifespan and imaging of brain and peripheral nerve pathologies. Phase I milestones will show that our lectin fusion, GALC-RTB, can be expressed in the Krabbe twitcher model via AAV gene transfer and demonstrate targeting of expressed GALC-RTB to hard-to-treat cells of CNS and PNS tissue in twitcher (GALC-/-) mice, enhancing enzyme biodistribution and lifespan. Based on these proof-of concept results, Phase II studies will assess the effects of dosage levels, timing, and routes of drug administration on efficacy and safety of long-term treatment aimed at further moving this product to an IND. The long-term goal of this project is to develop and bring this novel “delivery-enhanced” AAV gene therapy to Krabbe patients. The feasibility established here will also support expanding the “RTB lectin-fused” gene therapy treatment to other rare diseases having significant CNS and PNS tissue impairment.

Krabbe病（球样细胞脑白质营养不良）是一种罕见的遗传性疾病， 溶酶体酶半乳糖神经酰胺酶（GALC）导致破坏的保护涂层（髓鞘）， 大脑和整个神经系统中的神经细胞。这种致命的神经退行性疾病影响 在美国大约是十万分之一。在大多数情况下，Krabbe症状在婴儿6个月前出现， 年龄，往往导致2岁死亡。骨髓移植是目前唯一批准的治疗方法 Krabbe患者的选择，但仅减缓疾病进展，并且仅在症状前 孩子最近的临床前研究集中在利用腺相关病毒（AAV）的基因转移上 用于在Krabbe病的"抽搐"小鼠模型中长期表达正常GALC酶。 尽管这种方法很有前途，但在向关键人群分配有效治疗剂量方面仍然存在缺陷 大脑和神经系统的一部分。BioStrategies LC开发了一种基于凝集素的治疗递送 模块（RTB），使融合的溶酶体酶能够有效靶向CNS并治疗神经系统疾病 在几种溶酶体疾病动物模型中证明，每周ERT输注后出现的症状。 在这个SBIR项目中，我们建议将AAV介导的基因转移技术的益处合并， 用RTB递送技术提供连续的长期体内酶生产，以确保 有效治疗克拉伯氏进行性神经变性融合GALC与 BioStrategies的RTB凝集素将被包装到重组腺相关病毒中用于生产， 随后给予抽搐小鼠。对小鼠神经系统疾病进展和寿命的影响 将接受AAV-载体的GALC-RTB融合体的小鼠与未处理的小鼠和施用AAV-载体的GALC-RTB融合体的小鼠进行比较。 单独的AAV载体GALC。本I期可行性研究的具体目标是：1）构建GALC-RTB AAV 载体，并证明它们在哺乳动物细胞中产生全功能GALC-RTB融合产物的能力 以及2）比较AAV介导的GALC-RTB与GALC的基因转移的治疗功效 在Krabbe小鼠模型中，如通过寿命和脑和外周神经病理学成像测量的。 第一阶段的里程碑将表明，我们的凝集素融合，GALC-RTB，可以表达在Krabbe抽搐， 通过AAV基因转移建立模型，并证明表达的GALC-RTB靶向难以治疗的CNS细胞 和PNS组织中，增强酶的生物分布和寿命。基于这些 根据概念验证结果，II期研究将评估药物剂量水平、时间和途径的影响 长期治疗的有效性和安全性，旨在进一步将本品转为IND。 该项目的长期目标是开发并将这种新型的"递送增强" AAV基因疗法应用于 克拉布病人。这里建立的可行性也将支持扩大"RTB凝集素融合"基因 治疗具有显著CNS和PNS组织损害的其他罕见疾病。