Protein Tranduction for Treatment of Krabbe Disease

治疗克拉伯病的蛋白质转导

• 批准号: 6662500
• 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 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6662500
• 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病是一种退行性神经系统疾病，主要影响婴儿和幼儿，尽管已描述了罕见的成人发病病例。受影响的个体通常在生命的最初几个月出现症状。疾病进展通常很快，导致1-2年内死亡。这种疾病是一种常染色体隐性遗传性状，由半乳糖苷酶（GALC）基因突变引起，严重影响酶的活性。明显的治疗方法包括通过基因或蛋白质疗法将活性GALC酶递送至大脑。虽然多年来基因治疗取得了重大进展，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，并已被证明是安全和有效的治疗外周临床表现的另一种溶酶体贮积症，I型戈谢病。然而，对于诸如克拉布病之类的疾病，治疗酶必须被递送到大脑才能产生显著的临床影响。最近，Steven Dowdy及其同事在向大脑递送大分子方面取得了重大进展（Schwarze et al.，1999年）。他们已经表明，当与源自IRV达特蛋白的11个氨基酸的蛋白转导结构域偶联时，即使非常大的蛋白也可以穿过血脑屏障以生物活性形式进入脑。在本申请中，我们提出了达特PTD/GALC融合蛋白的实验，并检查这些融合蛋白是否会到达大脑并恢复正常功能，并提高克拉伯病动物模型的存活率。从这些实验中吸取的经验教训可能对其他疾病有用，其中可能需要递送治疗性蛋白质，例如阿尔茨海默病和帕金森病。