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

基本信息

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
项目类别：
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

项目摘要

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.

克拉伯病（球状细胞脑白质营养不良）是一种罕见的遗传性疾病，由脑白质缺乏症引起溶酶体酶半乳糖神经酰胺酶 (GALC) 会破坏神经元的保护层（髓磷脂）这种致命的神经退行性疾病会影响大脑和整个神经系统的神经细胞。在美国，大约有十分之一的人在出生后 6 个月内出现克拉伯症状。年龄，通常导致 2 岁时死亡。骨髓移植是目前唯一批准的治疗方法 Krabbe 患者的选择，但只能减缓疾病进展，并且仅在出现症状前开始最近的临床前研究主要集中在利用腺相关病毒（AAV）进行基因转移。用于在克拉伯病“抽搐”小鼠模型中长期表达正常 GALC 酶。尽管很有希望，但这种方法在向关键患者分配有效治疗剂量方面仍然存在缺陷。 BioStrategies LC 开发了一种基于凝集素的治疗递送方法。模块（RTB），使融合溶酶体酶能够有效地靶向中枢神经系统并治疗神经系统疾病每周 ERT 输注后出现的症状，如几种溶酶体疾病动物模型所示。在这个 SBIR 项目中，我们建议融合 AAV 介导的基因转移技术的优点，通过 RTB 递送技术提供连续长期的体内酶生产，以确保将 GALC 与融合的基因表达盒有效治疗 Krabbe 进行性神经变性。 BioStrategies 的 RTB 凝集素将被包装到重组腺相关病毒中进行生产和随后对抽搐小鼠的给药对神经系统疾病进展和小鼠寿命的影响。接受 AAV 载体的 GALC-RTB 融合的小鼠将与未治疗的小鼠和接受治疗的小鼠进行比较单独使用 AAV 载体的 GALC。这一阶段可行性研究的具体目标是 1) 构建 GALC-RTB AAV。载体并证明其在哺乳动物细胞中产生全功能 GALC-RTB 融合产物的能力和小鼠；2) 比较 AAV 介导的 GALC-RTB 基因转移与 GALC 的治疗效果在 Krabbe 小鼠模型中，通过寿命以及大脑和周围神经病理学成像来测量。第一阶段的里程碑将表明我们的凝集素融合物 GALC-RTB 可以在 Krabbe twitcher 中表达通过 AAV 基因转移建立模型，并证明表达的 GALC-RTB 靶向难以治疗的中枢神经系统细胞和 Twitcher (GALC-/-) 小鼠中的 PNS 组织，增强酶的生物分布和寿命。概念验证结果，第二阶段研究将评估剂量水平、时间和药物途径的影响对长期治疗的有效性和安全性的管理旨在进一步将该产品纳入 IND。该项目的长期目标是开发这种新型“增强递送”AAV 基因疗法并将其推广到全球各地。 Krabbe 患者的可行性也将支持扩展“RTB 凝集素融合”基因。治疗具有显着中枢神经系统和三七总皂甙组织损伤的其他罕见疾病。