Protein Tranduction for Treatment of Krabbe Disease

治疗克拉伯病的蛋白质转导

• 批准号: 6798819
• 负责人: CHRISTOPHER B ECKMAN
• 金额: $ 18.64万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-20 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6798819
• 关键词: Krabbe' s disease; autosomal recessive trait; blood brain barrier; brain; chimeric proteins; confocal scanning microscopy; disease /disorder model; enzyme activity; enzyme therapy; fluorescence microscopy; gene expression; gene mutation; molecular cloning; myelinopathy; nonhuman therapy evaluation; polymerase chain reaction; protein transport

DESCRIPTION (provided by applicant): Krabbe disease is a degenerative neurological disorder primarily affecting infants and young children although rare cases of adult onset have been described. Affected individuals typically present with symptoms in the first few months of life. Disease progression is generally rapid, leading to death within 1-2 years. The disease is inherited as an autosomal recessive trait caused by mutations in the galactocerebrosidase (GALC) gene that severely affect the activity of the enzyme. Obvious therapeutic approaches include delivery of active GALC enzyme to the brain through either gene or protein therapy. While significant advances have been made in gene therapy over the years, stable expression of proteins in the brain has not yet been achieved. Enzyme replacement therapy offers another possible therapeutic approach and has been shown to be safe and effective for the treatment of peripheral clinical manifestations in another lysosomal storage disorder, Type I Gaucher's disease. For diseases such as Krabbe disease, however, the therapeutic enzyme must be delivered to the brain to have a significant clinical impact. Recently, Steven Dowdy and colleagues have made a significant advance in the delivery of macromolecules to the brain (Schwarze et al., 1999). They have shown that that even very large proteins can cross the blood-brain barrier to enter into the brain in biologically active form when coupled to an 11 amino acids protein transduction domain derived from the IRV TAT protein. In this application we propose experiments to generate TAT PTD/GALC fusion proteins and examine whether these will get to the brain and restore normal function and enhance survivability in an animal model of Krabbe disease. The lessons learned from these experiments may be useful for other diseases where delivery of a therapeutic protein may be desired, such as Alzheimer's disease and Parkinson's.

描述（由申请人提供）： Krabbe病是一种退化性神经系统疾病，主要影响婴儿和幼儿，尽管已经描述了罕见的成人发病病例。受影响的个体通常在生命的头几个月出现症状。疾病进展通常很快，在1 - 2年内导致死亡。该疾病被遗传为是由半乳脑脑化酶（GALC）基因突变引起的常染色体隐性性状，严重影响了酶的活性。明显的治疗方法包括通过基因或蛋白质疗法将活性的GALC酶传递到大脑。尽管多年来在基因治疗方面取得了重大进展，但尚未实现蛋白质在大脑中的稳定表达。酶替代疗法提供了另一种可能的治疗方法，并且已被证明可以安全有效地治疗另一种溶酶体储存障碍（I型Gaucher疾病）的外围临床表现。但是，对于Krabbe疾病等疾病，必须将治疗酶递送到大脑中，以产生重大的临床影响。最近，史蒂文·道迪（Steven Dowdy）及其同事在向大脑的大分子传递方面取得了重大进步（Schwarze等，1999）。他们已经表明，当与11个氨基酸蛋白转导域耦合时，即使是非常大的蛋白质也可以穿越血脑屏障以生物学活性形式进入大脑。在此应用中，我们建议实验生成TAT PTD/GALC融合蛋白，并检查这些实验是否会进入大脑并恢复正常功能并增强Krabbe疾病动物模型中的生存​​能力。从这些实验中学到的经验教训对于可能需要提供治疗蛋白的其他疾病，例如阿尔茨海默氏病和帕金森氏病。

项目成果

Molecular characterization of mutations that cause globoid cell leukodystrophy and pharmacological rescue using small molecule chemical chaperones.

• DOI: 10.1523/jneurosci.6383-09.2010
• 发表时间: 2010-04-21
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Lee WC;Kang D;Causevic E;Herdt AR;Eckman EA;Eckman CB
• 通讯作者: Eckman CB