TARGETABLE RETROVIRAL VECTORS--TISSUE ENGINEERING

靶向逆转录病毒载体--组织工程

• 批准号: 3779555
• 负责人: L BALTRUCKI
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3779555
• 关键词: Retroviridae; biotechnology; chimeric proteins; congenital blood protein disorder; disease /disorder model; gene expression; gene therapy; genetic disorder; glucuronosyltransferase; glycoproteins; hereditary hyperbilirubinemia; human tissue; liver disorder; mixed tissue /cell culture; molecular cloning; receptor binding; transfection /expression vector

This project involves the development of retroviral vectors capable of targeting the asialoglycoprotein receptor (ASG-R). It represents the "targeting" portion of a larger effort which has been one of the laboratory's major goals, ie. the development of targetable/injectable vectors, that would be both resistant to inactivation by serum complement and capable of "homing" to specific cell surface receptors, such as the ASG-R. Our strategy for targeting has involved the construction of chimeric envelope glycoproteins, in which the native receptor binding domain of gp70, has been replaced by peptide sequences derived from alpha-1-acid glycoprotein. Stable transduction of HepG2 cells mediated by the ASG-R, has been achieved with vector particles bearing these "chimeric" envelope glycoproteins. We are currently working to improve the efficiency of transduction to the levels that would permit these vectors to be used in vivo. In this regard, vectors expressing human bilirubin-UDP-glucuronyl transferase, from various mouse albumin enhancers have been constructed. The Gunn rat, our animal model for Crigler-Najjar syndrome type I, is the test system for these vectors in vivo. Tissue engineering, when combined with a gene transfer technology, represents another useful approach for delivering therapeutic proteins. Protein C deficiency is another hereditary liver disease, which is invariably fatal in the neonatal period. Vectors which express human Protein C in a variety of cell lines have been constructed. The next stage of this project involves the development of "artificial tissues", that consist of transduced cells which have been established on a three- dimensional matrix in vitro. These tissue substrates will be implanted into rats and evaluated for structure, vascularity, and their ability to secrete biologically-active protein C.

该项目涉及逆转录病毒载体的开发， 靶向去唾液酸糖蛋白受体（ASG-R）。它代表 “瞄准”是更大努力的一部分， 实验室的主要目标，即。靶向/注射剂的开发 载体，这将是抵抗灭活血清补体 并且能够“归巢”到特定的细胞表面受体，例如 助理秘书长 我们的目标定位战略包括建立 嵌合包膜糖蛋白，其中天然受体结合 gp 70的结构域，已被来自 α-1-酸性糖蛋白。 HepG 2细胞介导的稳定转导 由ASG-R，已经实现了与载体颗粒轴承这些 “嵌合”包膜糖蛋白。 我们目前正在努力改善 将转导效率提高到允许这些 在体内使用的载体。 在这方面，表达人类的载体 白蛋白-UDP-葡萄糖醛酸基转移酶，来自各种小鼠白蛋白 已经构建了增强子。 古恩大鼠，我们的动物模型， Crigler-Najjar综合征I型，是这些载体的测试系统， vivo. 当组织工程与基因转移技术相结合时， 代表了用于递送治疗性蛋白质的另一种有用的方法。 蛋白C缺乏症是另一种遗传性肝病， 在新生儿期总是致命的。 表达人类的载体 已经构建了多种细胞系中的蛋白C。 下一 该项目的第一阶段涉及“人造组织”的开发， 它由转导的细胞组成，这些细胞建立在三个- 体外三维基质。 这些组织基质将被植入 并评估其结构、血管分布及其 分泌生物活性蛋白C。