BREACHING THE ENDOTHELIAL BARRIER FOR GENE THERAPY

突破基因治疗的内皮屏障

基本信息

批准号：
2770667
负责人：
Jon A Wolff
金额：
$ 14.3万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-09-30 至 2000-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2770667
关键词：
animal genetic material tag bacteriophage M13 bacteriophage T7 extracellular matrix gene therapy genetic library intramuscular injections intravenous administration laboratory mouse microorganism culture muscle metabolism nonhuman therapy evaluation striated muscles transcytosis transfection /expression vector vascular endothelium virus infection mechanism virus protein

项目摘要

DESCRIPTION (Abstract of the application) Muscle has emerged as one of the most important target tissues for gene therapy. Human clinical trial are in progress to determine the ability of intramuscularly injected plasmid DNA to induce a more effective immune response against viral infections and cancer. Muscle can also be used to manufacture and secrete proteins, such as hormones and growth factors. Genetic muscle diseases, such as Duchennes muscular dystrophy, could be treated by the expression of the respective normal gene in muscle. Finally, muscle could be used as a vehicle to clear circulating toxic metabolites that accumulate in inborn errors of metabolism. Despite this promise, a clinically-viable gene transfer method has yet to be developed. The bottleneck for realizing this treatment is the efficiency of gene transfer and stability of expression from the gene vectors developed to date. The proposed studies will address the efficiency of expression by developing new methods to deliver genes via blood vessels. Several advantages flow from the intravascular delivery of genes instead of direct intramuscular injections. Since the blood vessels are in direct and close contact with every muscle cell, delivery to the cellular surface of muscle cells can be a much more efficient process. More muscle groups could be easier reached through this method than using direct muscle injections. Breaching the endothelial barrier in muscles without toxicity is a challenging endeavor. However, many natural macromolecules and cells use different "keys" (ligands) to pass from the lumen of blood vessels into the interstitium of muscle. The development of phage display libraries provides an exciting new approach for finding the "key for unlocking this door." Phage-display libraries will be used in both in vitro and in vivo endothelial cell selection systems to identify peptides that enable transendothelial movement (TEM) of phages. These ligands will then be attached to vectors in order to enable them to transfer genes into muscle cells following their intravascular injection. Our proposed studies are likely to lead to a greater understanding of TEM in muscle and the generation of critical reagents for muscle gene therapy. These ligands will be of great use for both non-viral and viral gene transfer methods to muscle and other tissues as well.

描述（应用程序的摘要）肌肉已成为基因最重要的目标组织之一治疗。正在进行人类临床试验以确定肌肉内注射的质粒DNA，诱导更有效的免疫对病毒感染和癌症的反应。肌肉也可以用来生产和分泌蛋白质，例如激素和生长因子。遗传肌肉疾病，例如肌营养不良症，可能是通过在肌肉中各自的正常基因的表达治疗。最后，肌肉可以用作清除循环有毒代谢物的车辆这积累在天生的新陈代谢错误中。尽管有这样的承诺，但临床可行的基因转移方法尚未发达。实现这种治疗的瓶颈是从基因向量的基因转移和表达的稳定性发展为日期。拟议的研究将解决表达的效率开发新方法通过血管传递基因。一些从基因的血管内输送而不是直接的优势流动肌内注射。由于血管处于直接和关闭状态与每个肌肉细胞接触，传递到肌肉的细胞表面细胞可能是一个更有效的过程。更多的肌肉群可能是与使用直接肌肉注射相比，通过这种方法更容易到达。破坏没有毒性的肌肉中的内皮屏障是具有挑战性的努力。但是，许多天然大分子和细胞使用不同的“键”（配体）从血管的腔传递到肌肉间质。噬菌体展示库的开发提供一种令人兴奋的新方法，以找到“解锁这扇门的钥匙”。噬菌体播放库将在体外和体内使用内皮细胞选择系统以识别启用的肽噬菌体的跨内皮运动（TEM）。这些配体将是连接到向量以使其能够将基因转移到肌肉中血管内注射后细胞。我们提出的研究可能会导致对TEM的更深入肌肉和肌肉基因治疗的关键试剂的产生。这些配体对非病毒和病毒基因都非常有用将方法也转移到肌肉和其他组织。