Transplantation of Genetically Modified, Immune-privileged Steroli Cells

转基因、免疫豁免的甾体细胞移植

• 批准号: 8021991
• 负责人: Jannette M Dufour
• 金额: $ 22.72万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-28 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8021991
• 关键词: Transplantation; Genetically; Modified; Immune; privileged

DESCRIPTION (provided by applicant): Sertoli cells are immune-privileged cells that have the ability to survive long-term without the use of immunosuppressive drugs when transplanted across allogeneic or xenogeneic barriers. The ability of Sertoli cells to survive transplantation when most other cells are immunologically rejected suggests Sertoli cells could be engineered as a vehicle for gene therapy. The long-term goal of my research is to use genetically engineered, immune-privileged Sertoli cells as an unlimited source of tissue that can be used as a vehicle to deliver therapeutically relevant proteins as a means to treat multiple disorders. Therefore, the objective of this application is to evaluate the feasibility of using Sertoli cells engineered to produce functional levels of insulin as an effective long-term, therapeutic strategy in diabetic mice. It is hypothesized that immune- privileged Sertoli cells can be genetically engineered to stably express basal levels of insulin, and that these cells can survive transplantation as allografts or xenografts in diabetic mice without the need for immunosuppression. This hypothesis is based on our preliminary data, which demonstrated unmodified Sertoli cells survive long-term when transplanted as allografts and xenografts without the use of immunosuppressive drugs and that Sertoli cells stably expressing GFP survive allogeneic transplantation while continuing to express GFP. The specific aims are designed to test this hypothesis. In Aim 1, the function of insulin expressing Sertoli cells will be examined. Mouse and porcine Sertoli cells stably expressing insulin will be examined for production and secretion of functional levels of insulin in vitro and after transplantation into diabetic immunodeficient SCID mice in vivo. In Aim 2, the ability of these insulin-expressing Sertoli cells to survive (remain immune-privileged) when transplanted across immunological barriers will be examined. Insulin-expressing Sertoli cells will be transplanted into diabetic, immune-competent C3H mice as allografts (mouse Sertoli cells) or xenografts (porcine Sertoli cells). The production of functional insulin and Sertoli cell survival will be examined. It is expected that Sertoli cells engineered to stably deliver basal levels of insulin are capable of lowering blood glucose levels long-term after transplantation into diabetic mice. This contribution is significant because it will provide evidence that genetically engineered, immune-privileged Sertoli cells can be used as an unlimited supply of tissue for transplantation. PUBLIC HEALTH RELEVANCE: There are many disorders that have the potential to be treated by gene therapy; for example, delivery of factor VIII for the treatment of hemophilia. However, an efficient and safe delivery system has not yet been developed. We will explore the novel and innovative concept that immune-privileged Sertoli cells can be engineered to deliver therapeutic proteins. In the long-term these cells could provide an unlimited supply of genetically modified tissue that could be transplanted without the use of chronic immunosuppression as a means of treatment for multiple disorders.

描述（由申请方提供）：支持细胞是免疫豁免细胞，当穿过同种异体或异种屏障移植时，能够在不使用免疫抑制药物的情况下长期存活。当大多数其他细胞被免疫排斥时，支持细胞在移植中存活的能力表明支持细胞可以被工程化为基因治疗的载体。我的研究的长期目标是使用基因工程，免疫特权的支持细胞作为组织的无限来源，可用作载体，提供治疗相关的蛋白质作为治疗多种疾病的手段。因此，本申请的目的是评估使用经工程改造以产生功能水平的胰岛素的支持细胞作为糖尿病小鼠中的有效长期治疗策略的可行性。假设免疫豁免的支持细胞可以被遗传工程化以稳定表达基础水平的胰岛素，并且这些细胞可以在糖尿病小鼠中作为同种异体移植物或异种移植物移植后存活，而不需要免疫抑制。这一假设是基于我们的初步数据，这表明未修饰的支持细胞长期存活时，移植作为同种异体移植物和异种移植物，而不使用免疫抑制药物和支持细胞稳定表达GFP存活同种异体移植，同时继续表达GFP。具体目标是为了检验这一假设。在目的1中，将检查表达胰岛素的支持细胞的功能。将在体外和体内移植到糖尿病免疫缺陷SCID小鼠后，检查稳定表达胰岛素的小鼠和猪Sertoli细胞的胰岛素功能水平的产生和分泌。在目标2中，将检查这些表达胰岛素的支持细胞在穿过免疫屏障移植时存活（保持免疫豁免）的能力。将表达胰岛素的Sertoli细胞作为同种异体移植物（小鼠Sertoli细胞）或异种移植物（猪Sertoli细胞）移植到糖尿病免疫活性C3 H小鼠中。将检查功能性胰岛素的产生和支持细胞存活。预期经工程化以稳定递送基础水平的胰岛素的支持细胞能够在移植到糖尿病小鼠中后长期降低血糖水平。这一贡献是重要的，因为它将提供证据表明，基因工程，免疫特权支持细胞可用作无限供应的组织移植。 公共卫生关系：有许多疾病有可能通过基因治疗来治疗;例如，用于治疗血友病的因子VIII的递送。然而，尚未开发出有效和安全的输送系统。我们将探索新的和创新的概念，免疫特权支持细胞可以工程化提供治疗性蛋白质。从长远来看，这些细胞可以提供无限量的转基因组织供应，这些组织可以在不使用慢性免疫抑制的情况下移植，作为治疗多种疾病的手段。