Cell and Gene Replacement Strategies for Arginase Deficiency

精氨酸酶缺乏症的细胞和基因替代策略

• 批准号: 9289701
• 负责人: Gerald S Lipshutz
• 金额: $ 34.15万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9289701
• 关键词: ARG2 gene; Address; Adolescent; Affect; Age; Ammonia; Animals; Arginine; Behavior; Behavioral; Biochemical; Birth; Cell Therapy; Cells; Clinical; Congenital neurologic anomalies; Cytokinesis; DNA Sequence Alteration; Data; Development; Differentiated Gene; Disease; Drug Metabolic Detoxication; Encapsulated; Engraftment; Enzymes; Extrahepatic; Gene Expression; Gene Transduction Agent; Genes; Genomics; Goals; Grant; Growth; Growth Factor; Hepatic; Hepatocyte; Human; Hydrogels; Hyperammonemia; Hyperargininemia; Immune response; Impairment; Infiltration; Injectable; Investigation; Kidney; Knockout Mice; Lead; Learning; Life; Liver; Longevity; Mammals; Mediating; Metabolism; Methods; Mus; Muscle; Muscle Cells; Nanotechnology; Neonatal; Nervous System Trauma; Neurons; Newborn Infant; Nitrogen; One-Step dentin bonding system; Pathway interactions; Patients; Periodicity; Peripheral; Phenotype; Plasma; Primary carcinoma of the liver cells; Production; Prostate; Protein Isoforms; Psyche structure; Recombinants; Research; Risk; Site; Spastic; Stem cell transplant; Stem cells; Technology; Testing; Time; Tissue Engineering; Tissues; Urea; Viral Vector; adeno-associated viral vector; arginase; base; behavior test; clinical application; effective therapy; enzyme deficiency; experimental study; gene correction; gene replacement; gene therapy; genome editing; improved; induced pluripotent stem cell; knockout animal; liver cell proliferation; mouse model; muscle form; nanoencapsulated; novel strategies; novel therapeutics; null mutation; promoter; scaffold; spasticity; urea cycle

Project Summary/Abstract The urea cycle is the major pathway for detoxification of ammonia in mammals. Arginase 1 deficiency is thought to be the least common of the urea cycle disorders and results in hyperargininemia. In humans, deficiency of this enzyme is characterized clinically by progressive mental impairment, spasticity, growth retardation, and periodic episodes of hyperammonemia. This proposal is two-fold: 1) to continue to advance gene-based therapies for arginase deficiency utilizing appropriate murine models; viral vectors and genomic correction technology will be applied to examine if animals can be corrected behaviorally and biochemically; and 2) to evaluate an iPSC-derived cell therapy approach with hepatocytes placed on bioactive scaffolds to supply urea cycle function. Preliminary data: Our research group has (amongst other findings): 1) constructed and characterized the arginase 1 knockout mouse; 2) demonstrated long-term survival and rescue with recombinant adeno-associated viral vectors; 3) demonstrated that only low-level ureagenesis is necessary for long-term survival; 4) shown that, using an array of behavioral tests, that treated arginase knockout animals lack nervous system abnormalities and there is no difference in learning or behavior when compared to littermates; 5) shown that peripheral metabolism can result in control of circulating plasma arginine; and 6) loss of arginase gene expression results in abnormalities of intrinsic excitability and the dendritic arbor of neurons. In Aim 1, long-term expression of arginase 2 in muscle by viral vector gene therapy will be performed to examine for biochemical and phenotypic correction in a murine model of arginase deficiency. This approach may avoid neutralizing immune responses in patients with null mutations. In Aim 2, studies will examine if an auxiliary liver grown on scaffolds can supply the minimal urea cycle function necessary to lead to phenotypic correction of hyperargininemia. This approach may be successful for other urea cycle disorders. With successful completion of the proposed studies it is expected that a new therapy with gene and cell replacement will be one step closer for patients afflicted with arginase deficiency.

项目概要/摘要 尿素循环是哺乳动物氨解毒的主要途径。精氨酸酶 1 缺乏症被认为 是最不常见的尿素循环障碍，并导致高精氨酸血症。在人类中，缺乏 这种酶的临床特征是进行性精神障碍、痉挛、生长迟缓和 周期性发作的高氨血症。该建议有两个方面：1）继续推进基于基因的研究 利用适当的小鼠模型治疗精氨酸酶缺乏症；病毒载体和基因组校正 将应用技术来检查动物是否可以在行为和生化方面得到纠正； 2) 到 评估 iPSC 衍生细胞治疗方法，将肝细胞放置在生物活性支架上以供应尿素 循环功能。初步数据：我们的研究小组（除其他发现外）：1）构建并 精氨酸酶 1 敲除小鼠的特征； 2）通过重组证明了长期存活和救援 腺相关病毒载体； 3）证明只有低水平的尿素生成对于长期而言是必要的 生存； 4）表明，通过一系列行为测试，经过治疗的精氨酸酶敲除动物缺乏紧张感 系统异常，与同窝出生的孩子相比，学习或行为没有差异； 5) 如图所示 外周代谢可导致循环血浆精氨酸的控制； 6) 精氨酸酶基因丢失 表达导致神经元内在兴奋性和树突状结构的异常。在目标 1 中，长期 将通过病毒载体基因治疗在肌肉中表达精氨酸酶 2，以检查生化和 精氨酸酶缺乏的小鼠模型的表型校正。这种方法可以避免中和免疫 无效突变患者的反应。在目标 2 中，研究将检查是否在支架上生长辅助肝脏 可以提供导致高精氨酸血症表型纠正所需的最小尿素循环功能。这 该方法可能对其他尿素循环障碍有效。随着拟议研究的成功完成 预计一种基因和细胞替代的新疗法将为患有该病的患者迈出一步 精氨酸酶缺乏症。