Lentiviral-mediated Hematopoietic Stem Cell Gene Therapy for Canine Globoid Cell Leukodystrophy

慢病毒介导的造血干细胞基因治疗犬球状细胞脑白质营养不良

• 批准号: 9300725
• 负责人: Allison M Bradbury
• 金额: $ 5.92万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9300725
• 关键词: 2 year old; Affect; Age; Age of Onset; Allogeneic Bone Marrow Transplantation; Animal Model; Ataxia; Attenuated; Autologous; Biochemical; Biochemistry; Biodistribution; Blindness; Bone Marrow; Bone Marrow Transplantation; Brain; Breeding; Canis familiaris; Cell Transplants; Cells; Cessation of life; Child; Choline; Clinical; Clinical Trials; Collaborations; Data; Demyelinations; Deterioration; Development; Disease; Disease Progression; Disease model; Dog Diseases; Enzymes; Euthanasia; Evaluation; Faculty; Funding; Galactosylceramides; Globoid cell leukodystrophy; Green Fluorescent Proteins; Harvest; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hereditary Disease; Histologic; Human; Impaired cognition; In Vitro; Inherited; Institution; Lead; Lentivirus Vector; Life; Light; Limb structure; Lipids; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Modeling; Motor; Mus; Myelin; Nerve Degeneration; Neuraxis; Neurologic; Neurologic Dysfunctions; Neurologist; Neurotoxins; Oligodendroglia; Paralysed; Patients; Pelvis; Pennsylvania; Psychosine; Quality of life; Regimen; Research; Research Personnel; Resources; Safety; Seizures; Severities; Specialist; Stem cells; Symptoms; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Training; Translating; Transplantation; Treatment Efficacy; Tremor; Umbilical Cord Blood; United States National Institutes of Health; Universities; Vegetative States; Viral Vector; clinical application; cohort; efficacy study; enzyme activity; experience; galactosylceramidase; gene therapy; human disease; improved; infancy; innovation; lentiviral-mediated; lentivirally transduced; leukodystrophy; neuron loss; neurotoxic; novel; pre-clinical; preconditioning; premature; public health relevance; pup; safety study; skills; success; therapy development; training opportunity; vector; white matter

 DESCRIPTION (provided by applicant): Globoid cell leukodystrophy (GLD), also known as Krabbe disease, is a progressive and fatal neurodegenerative lysosomal storage disorder caused by the deficient activity of the hydrolytic enzyme β- galactocerebrosidase (GALC), which is responsible for degrading the myelin lipids galactosylceramide (GalCer) and galactosylsphingosine (psychosine). In the absence of GALC, accumulation of psychosine, a known neurotoxin, causes loss of oligodendrocytes and subsequent widespread demyelination with patient death typically occurring in the second year of life. To date the only available therapeutic intervention for children is hematopoietic stem cell transplantation (HSCT) from donor umbilical cord blood or bone marrow. When employed early, HSCT has proven capable of slowing disease progression; however, the quality of life in children is poor due to neurologic deterioration resulting in loss of cognitive and motor functions. We hypothesize that genetically enhancing HSCs prior to transplantation will increase therapeutic levels of GALC, further attenuate disease progression, and prolong survival in GLD patients. To test this hypothesis, we will take advantage of a canine model of GLD that faithfully recapitulates the clinical disease progression, neuropathological alterations, and biochemical abnormalities observed in human patients. In Aim 1 we will optimize bone marrow harvest from young pups, HSC isolation, and lentiviral transduction in normal dogs utilizing a vector encoding GFP. In Aim 2 we will assess the safety and tolerability of lentiviral HSC gene therapy, including a novel preconditioning regimen, in normal dogs using an innovative lentiviral vector encoding canine GALC. In Aim 3 we will establish the efficacy of lentiviral HSC gene therapy in GLD dogs and compare to GLD dogs treated with HSCT alone (no lentiviral transduction) and untreated GLD controls. Safety and efficacy data from current clinical trials in other infantile-onset leukodystrophies and convincing preclinical data in our canine model of GLD could lead to rapid clinical implementation of lentiviral HSC gene therapy for Krabbe disease. Training specific to this disease model will occur through regular interactions with my sponsor, a board certified veterinary neurologist and NIH-funded investigator with extensive research experience utilizing large animal models to develop therapies for numerous LSD. This training will be augmented by 1) co-sponsorship with a viral vector specialist, and 2) collaboration with an expert in canine ex vivo gene therapy. The University of Pennsylvania provides adequate resources to complete the project and ample supplemental educational and training opportunities. The expertise of my selected sponsor and co-sponsor and resources of this institution will allow development of new skill sets and prepare me to become an independent investigator capable of translating therapies from the bench to use in patients.

 描述（申请人提供）：球状细胞脑白质营养不良（GLD），也称为克拉伯病，是一种进行性、致命性的神经退行性溶酶体贮积症，由水解酶β-半乳糖脑苷酶（GALC）活性缺陷引起，该酶负责降解髓磷脂脂质半乳糖神经酰胺（GalCer）和半乳糖基鞘氨醇 （精神碱）。在缺乏 GALC 的情况下，精神嘧啶（一种已知的神经毒素）的积累会导致少突胶质细胞的损失和随后的广泛脱髓鞘，患者死亡通常发生在生命的第二年。迄今为止，儿童唯一可用的治疗干预措施是来自捐赠者脐带血或骨髓的造血干细胞移植（HSCT）。事实证明，早期使用 HSCT 能够减缓疾病进展；然而，由于神经系统恶化导致认知和运动功能丧失，儿童的生活质量很差。我们假设在移植前对 HSC 进行基因增强将提高 GALC 的治疗水平，进一步减缓疾病进展，并延长 GLD 患者的生存期。为了检验这一假设，我们将利用 GLD 的犬模型，该模型忠实地再现了在人类患者中观察到的临床疾病进展、神经病理学改变和生化异常。在目标 1 中，我们将利用编码 GFP 的载体优化幼犬的骨髓采集、HSC 分离和正常狗的慢病毒转导。在目标 2 中，我们将使用编码犬 GALC 的创新慢病毒载体，评估慢病毒 HSC 基因治疗（包括新型预处理方案）在正常犬中的安全性和耐受性。在目标 3 中，我们将确定慢病毒 HSC 基因治疗在 GLD 犬中的疗效，并与仅接受 HSCT（无慢病毒转导）治疗的 GLD 犬和未经治疗的 GLD 对照进行比较。目前其他婴儿发病的脑白质营养不良临床试验的安全性和有效性数据以及我们的犬 GLD 模型中令人信服的临床前数据可能会导致慢病毒 HSC 基因治疗克拉伯病的快速临床实施。针对这种疾病模型的培训将通过与我的资助者的定期互动进行，资助者是一位经过委员会认证的兽医神经学家和美国国立卫生研究院资助的研究人员，拥有利用大型动物模型开发多种 LSD 疗法的丰富研究经验。该培训将通过以下方式得到加强：1) 与病毒载体专家的共同赞助，以及 2) 与犬离体基因治疗专家的合作。宾夕法尼亚大学提供充足的资源来完成该项目以及充足的补充教育和培训机会。我选择的赞助商和共同赞助商的专业知识以及该机构的资源将使我能够开发新的技能，并使我成为一名能够将实验室疗法转化为患者使用的独立研究者。