Vector Targeting for Fracture Repair

用于骨折修复的矢量靶向

• 批准号: 6533045
• 负责人: Alan ROBERT Davis
• 金额: $ 7.53万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-28 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6533045
• 关键词: adeno associated virus group; antibody; binding proteins; biotechnology; bone fracture; bone morphogenetic proteins; bone regeneration; flow cytometry; gene delivery system; gene therapy; laboratory mouse; osteoblasts; osteogenesis; protein binding; protein engineering; protein tyrosine phosphatase; receptor binding; tissue /cell culture; transfection /expression vector; virus protein; virus receptors

DESCRIPTION (provided by applicant): Many investigators have shown that adenoviral (Ad) vectors expressing the bone morphogenetic protein BMP2 can enhance fracture repair by eliciting bone formation that closely resembles the normal process, in contrast to results obtained with direct injection of the recombinant protein. Delivery of BMP2 via an Ad vector is therefore an attractive strategy for improving bone repair, which currently is delayed or unsuccessful in 5-10% of fracture cases. A major challenge is to produce a safe osteoblast-targeted vector that does not recognize cells expressing the widespread adenovirus, CAR, the usual route by which adenoviruses infect mammalian cells. This proposal seeks to increase the specificity and safety of Ad vectors by modifying the viral fiber protein (Aim 1) so that it no longer binds to CAR, but instead recognizes the OST-PTP protein on osteoblasts (Aim 2). These objectives will be pursued by ablating critical amino acid sequences in the fiber DG loop and monitoring cells for evidence of infection through the CAR-binding pathway, and by constructing vectors whose fiber carries both the CAR-binding sequence deletion and an OST-PTP-specific antibody sequence and then assessing their ability to target osteoblasts in vitro and in vivo. Vectors emerging from these studies will be prime candidates for insertion of BMP2 or other therapeutic genes with relevance to bone disease. Given the applicant's experience in adenoviral vector design and translational research in university and pharmaceutical settings, the prospects for a successful outcome of this project appear high.

描述（由申请人提供）：许多研究人员已经表明， 表达骨形态发生蛋白BMP 2的腺病毒（Ad）载体可以 通过诱导骨形成来增强骨折修复， 正常的过程，与直接注射的结果相反， 重组蛋白因此，通过Ad载体递送BMP 2是一种有效的方法。 改善骨修复的有吸引力的策略，目前被延迟或 5-10%的骨折病例不成功。一个主要的挑战是生产一个安全的 成骨细胞靶向载体，不识别表达 广泛分布的腺病毒，CAR，腺病毒感染的通常途径 哺乳动物细胞该提案旨在增加以下方面的特异性和安全性： 通过修饰病毒纤维蛋白（Aim 1）使其不再 与CAR结合，但识别成骨细胞上的OST-PTP蛋白（Aim 2）。这些目标将通过切除关键氨基酸序列来实现 在纤维DG环和监测细胞感染的证据，通过 CAR结合途径，并通过构建其纤维携带CAR结合途径的载体， CAR结合序列缺失和OST-PTP特异性抗体序列， 然后评估它们在体外和体内靶向成骨细胞的能力。 从这些研究中出现的载体将是插入 BMP 2或其他与骨骼疾病相关的治疗基因。鉴于 申请人在腺病毒载体设计和转化研究方面的经验 在大学和制药环境中， 这个项目的成果似乎很高。