TRANSPLANT OF TRANSDUCED ENDOTHELIUM INTO CAPILLARIES

将转导内皮移植到毛细血管中

• 批准号: 2227766
• 负责人: Louis Michael Messina
• 金额: $ 14万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-09 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2227766
• 关键词: angiogenesis; blood vessel transplantation; calcium; capillary bed; cardiovascular pharmacology; cell adhesion; coagulation factor IX; cytokine; fibroblast growth factor; gene expression; gene therapy; hemophilia B; human genetic material tag; human tissue; integrins; laboratory rat; magnesium; nonhuman therapy evaluation; plasminogen activator; recombinant proteins; striated muscles; thrombosis; tissue /cell culture; vascular endothelium; xenotransplantation

The long-term objective of this research project is to develop a technique of somatic gene therapy in which endothelial cells that have undergone gene transfer ex vivo are transplanted into the capillary bed of skeletal muscle. This technique of somatic gene therapy could be used to achieve a regional or systemic effect by co-or repopulating a vascular bed with endothelial cells which constitutively or inducibly express a protective recombinant protein such as tissue plasminogen activator or Factor IX. Thus, the development of such a technique of somatic gene therapy could have a wide application to both surgical and genetic human deceases. The Specific Aims of the project are: Aim I: To identify and characterize the molecular mechanism mediating the adhesion of endothelial cells seeded onto quiescent, confluent endothelial cell monolayers. Aim II; To optimize the adhesion and incorporation of genetically transduced endothelial cells transplanted into the capillary bed of skeletal muscle. Aim III; To document long-term recombinant gene expression by endothelial cells transplanted into the capillary bed of skeletal muscle. The investigators have considerable experience with an in vitro model to study endothelial cell-endothelial cell adhesion as well as with an in vivo rt model to study transplantation of genetically modified endothelial cells into skeletal muscle capillary beds. The proposed experiments to achieve th above Aims include identification and characterization of specific adhesion molecules mediating endothelial cell-endothelial cell adhesion. These experiments would reveal fundamental new knowledge concerning endothelial cell-endothelial cell interactions. This knowledge could also be used to enhance adhesion and incorporation of transduced endothelium into the capillary wall. In a rat model the adhesion and incorporation of genetically transduced endothelium transplanted into the capillary bed will be optimized by angiogenesis induced with injection of basic fibroblast growth factor or stimulation f skeletal muscle regeneration. Finally, the optimal techniques of transplantation of transduced endothelium into host capillary beds, determined in the aforenoted studies, will be used to document the efficacy of this technique to deliver recombinant proteins into the circulation. At the completion of these studies, this technique of somatic gene therapy will be ready for evaluation in a primate model in preparations for its use in humans.

该研究项目的长期目标是开发一种技术 体细胞基因疗法，其中经过基因的内皮细胞 转移离体被移植到骨骼的毛细管床中 肌肉。 这种体细胞基因疗法的技术可用于实现 区域或全身效应，通过共同供应血管床 内皮细胞组成或诱导地表达保护性 重组蛋白，例如组织纤溶酶原激活剂或因子IX。 因此，这种躯体基因治疗技术的发展可能 在外科和遗传人类的衰落中都有广泛的应用。 这 该项目的具体目标是： 目的I：识别和表征介导的分子机制 种子上的内皮细胞的粘附到静止的，汇合的内皮 细胞单层。 目标II； 优化基因的粘附和掺入 转导的内皮细胞移植到 骨骼肌。 目标三； 记录内皮的长期重组基因表达 细胞移植到骨骼肌的毛细血管床中。 调查人员在体外模型中具有丰富的经验 研究内皮细胞 - 内皮细胞粘附以及体内 研究转基因内皮细胞移植的RT模型 进入骨骼肌毛细管床。 提出的实验以实现 以上目的包括特定的识别和表征 粘附分子介导内皮细胞 - 内皮细胞粘附。 这些实验将揭示有关有关的基本新知识 内皮细胞 - 内皮细胞相互作用。 这些知识也可以 用于增强转带内皮的粘附和掺入 进入毛细管墙。 在大鼠模型中 移植到毛细管床的遗传转导的内皮将 通过注射碱性成纤维细胞诱导的血管生成优化 生长因子或刺激F骨骼肌再生。 最后， 转导的内皮移植到宿主的最佳技术 在上述研究中确定的毛细管床将用于 记录该技术提供重组蛋白的功效 进入循环。 这些研究完成时，这项技术 体细胞基因疗法将准备在灵长类动物模型中评估 为其在人类中使用的准备。