Nanoparticle-mediated delivery of a base editor for in utero treatment of Canavan Disease

纳米颗粒介导的碱基编辑器递送用于子宫内治疗卡纳万病

• 批准号: 10727872
• 负责人: David E. Pleasure
• 金额: $ 42.39万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10727872
• 关键词: Address; Affect; Age; Age Months; Anticonvulsants; Aspartoacylase; Birth; Brain; Canavan Disease; Caring; Cells; Central Nervous System Diseases; Characteristics; Child; Citrates; Clinical; Complex; DNA Sequence Alteration; Data; Development; Disease; Electroporation; Encapsulated; Enzymes; Fetus; Formulation; Genes; Genetic; Genetic Diseases; Genome; Goals; Guide RNA; Head Movements; Healthcare; Hospitals; Hour; Immune system; In Vitro; Incidence; Infant; Injections; Intraventricular; Intraventricular Injections; Life; Lipids; Lithium; Macrocephaly; Mediating; Medical; Messenger RNA; Methods; Molecular Diagnosis; Motor; Motor Skills; Mus; Muscle hypotonia; Mutation; N-acetylaspartic acid; Neonatal; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurologic Deficit; Non-Viral Vector; Oligodendroglia; Onset of illness; Other Genetics; Patients; Phenotype; Point Mutation; Population; Proliferating; Proteins; RNA; Reagent; Safety; Solid; Spinal Cord; Symptoms; System; Technology; Therapeutic; Time; Tissues; Toxic effect; Transfection; Treatment Cost; autosome; base; base editor; brain cell; carrier testing; design; dietary manipulation; enzyme activity; experimental study; fetal; follower of religion Jewish; functional outcomes; gene therapy; genome editing; in utero; infancy; innovation; leukodystrophy; lipid nanoparticle; mRNA delivery; mouse model; nanoparticle; nanoparticle delivery; nervous system disorder; obstetric care; population based; prenatal; prenatal therapy; prevent; prime editing; prime editor; progenitor; reduce symptoms; stem; stem cells; technology platform

ABSTRACT Canavan disease is a rare genetic neurological disorder characterized by a vacuolar (spongiform) leukodystrophy caused by ASPA mutations that diminish brain aspartoacylase activity. Neonatal/infantile Canavan disease is the most common and most severe form of the condition. Affected infants appear normal for the first few months of life, but problems with development become noticeable by age 3 to 5 months. These infants usually do not develop motor skills such as turning over, controlling head movement, and sitting without support. Other characteristics of this condition include hypotonia, macrocephaly, and irritability. There is no cure, nor is there a standard course of treatment. The symptomatic support for this neurodegenerative disorder includes dietary manipulations, administration of lithium citrate and anticonvulsants, and AAV-mediated brain parenchymal ASPA gene therapy, yet, these treatments do not reverse or prevent the progression of neurological deficits in affected infants and children. To address this unmet medical need, we propose to develop an innovative treatment comprised of an in utero delivery of solid lipid nanoparticle (LNP)/mRNA complexes by intraventricular injection that will target and edit oligodendrocyte before birth, correct ASAP mutations, and restore aspartoacylase expression in patients influenced by point mutations. Gene editing at the in utero stage has great potential to cure neurological disorders, including Canavan disease before symptoms are unset. Our preliminary studies demonstrated that in utero intraventricular delivery LNP mRNA complexes can efficiently deliver mRNA to the brain and spinal cord tissue. These experiments are the first demonstration of mRNA based nonviral gene editing in utero and open up numerous therapeutic applications, given the tremendous versatility of mRNA. The central hypothesis of this proposal is that: LNPs will deliver mRNA for gene editing enzymes in utero and will rescue mice from Canavan disease at and after birth. This hypothesis is based upon our preliminary data demonstrating that LNPs containing CRE can safely and efficiently transfect oligodendrocyte in the brain of Ai9 mice after in utero intraventricular delivery. The central objective of this study is to develop LNP formulations that can efficiently edit oligodendrocyte in utero and develop a strategy for treating Canavan disease.

抽象的 Canavan病是一种罕见的遗传神经系统疾病，其特征是液泡（海绵状） 白细胞营养不良是由ASPA突变引起的，这些突变会减少大脑的天冬氨酸活性。 新生儿/婴儿Canavan疾病是该疾病中最常见，最严重的形式。 在生命的头几个月中，受影响的婴儿看起来正常，但是发育问题成为 明显到3至5个月。这些婴儿通常不会发展运动技能，例如翻转， 控制头部运动，坐着没有支撑。这种情况的其他特征包括 肌畸形，脑畸形和烦躁。无法治愈，也没有标准的治疗方法。 对这种神经退行性疾病的有症状支持包括饮食操纵， 给药柠檬酸锂和抗惊厥药，以及AAV介导的脑实质ASPA基因 然而，这些治疗方法并不能逆转或阻止神经系统缺陷的进展 受影响的婴儿和儿童。为了满足这种未满足的医疗需求，我们建议开发创新 由固体脂质纳米颗粒（LNP）/mRNA复合物在子宫递送中的处理 脑室内注射将在出生前靶向和编辑少突胶质细胞，纠正ASAP突变， 并恢复受点突变影响的患者的天冬氨酸表达。基因编辑 子宫阶段具有治愈神经系统疾病的巨大潜力，包括Canavan疾病 症状是不安的。我们的初步研究表明，在子宫内室内递送LNP中 mRNA复合物可以有效地将mRNA传递到大脑和脊髓组织。这些实验 是子宫内基于mRNA的非病毒基因编辑的首次演示，并开放了许多 鉴于mRNA的多功能性，治疗应用。中心假设 建议是：LNP将在子宫内提供基因编辑酶的mRNA，并将从 出生后及之后的Canavan病。该假设基于我们的初步数据证明 含有CRE的LNP可以安全有效地将少突胶质细胞转移到AI9小鼠的大脑中 在子宫内室内递送后。这项研究的核心目的是开发LNP配方 这可以有效地编辑子宫内的少突胶质细胞，并制定治疗Canavan疾病的策略。