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: 8150939
• 负责人: Jannette M Dufour
• 金额: $ 21.81万
• 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/8150939
• 关键词: Allogenic; Allografting; Asorbicap; Beta Cell; Biological Models; Blood Glucose; Cell Survival; Cell Transplantation; Cell physiology; Cells; Cellular biology; Chronic; Data; Detection; Diabetes Mellitus; Diabetic mouse; Disease; Engineering; Factor VIII; Family suidae; Goals; Green Fluorescent Proteins; Hemophilia A; Homologous Transplantation; Human; Immune; Immune system; Immunodeficient Mouse; Immunology; Immunosuppression; Immunosuppressive Agents; In Vitro; Inbred C3H Mice; Insulin; Measures; Molecular Genetics; Monitor; Mus; Pharmaceutical Preparations; Production; Property; Proteins; Publishing; Research; Research Personnel; Rodent; SCID Mice; Single-Gene Defect; Source; System; Testing; Therapeutic; Tissue Transplantation; Tissues; Transgenic Mice; Transgenic Organisms; Transplantation; Xenograft procedure; base; cellular engineering; design; diabetic; expectation; gene therapy; in vivo; innovation; insulin secretion; novel; public health relevance; sertoli cell; therapeutic protein

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(申请人提供)：Sertoli细胞是免疫特权细胞，当移植到异体或异种屏障时，在不使用免疫抑制药物的情况下具有长期存活的能力。当大多数其他细胞被免疫排斥时，支持细胞仍能在移植中存活，这表明支持细胞可以被改造成基因治疗的载体。我这项研究的长期目标是使用经过基因工程的、免疫特权的Sertoli细胞作为无限的组织来源，可以作为一种载体来输送治疗相关蛋白质，作为治疗多种疾病的一种手段。因此，这项应用的目的是评估使用支持细胞工程产生功能水平的胰岛素作为糖尿病小鼠有效的长期治疗策略的可行性。假设免疫优势的Sertoli细胞可以通过基因工程稳定表达基础水平的胰岛素，这些细胞可以作为同种或异种移植存活在糖尿病小鼠身上，而不需要免疫抑制。这一假设是基于我们的初步数据，这些数据表明，未经修饰的Sertoli细胞在不使用免疫抑制药物的情况下作为同种或异种移植细胞长期存活，稳定表达GFP的Sertoli细胞在同种异体移植中存活，同时继续表达GFP。这些具体目标旨在检验这一假设。在目标1中，将检测表达胰岛素的支持细胞的功能。稳定表达胰岛素的小鼠和猪Sertoli细胞将在体外和体内移植到糖尿病免疫缺陷的SCID小鼠体内，以检测胰岛素的产生和分泌功能水平。在目标2中，将检查这些表达胰岛素的Sertoli细胞在跨越免疫屏障移植时存活(保持免疫特权)的能力。表达胰岛素的Sertoli细胞将作为同种移植物(小鼠Sertoli细胞)或异种移植(猪Sertoli细胞)移植到糖尿病、免疫功能良好的C3H小鼠体内。功能胰岛素的产生和支持细胞的存活将被检测。人们预计，能够稳定输送基础水平胰岛素的Sertoli细胞能够在移植到糖尿病小鼠后长期降低血糖水平。这一贡献意义重大，因为它将提供证据，证明基因工程的、免疫特权的Sertoli细胞可以用作移植的无限组织供应。 与公共卫生的相关性：有许多疾病有可能通过基因疗法来治疗；例如，为治疗血友病而输送因子VIII。然而，还没有开发出一种高效和安全的输送系统。我们将探索一种新颖而创新的概念，即免疫特权支持细胞可以被改造成输送治疗性蛋白质。从长远来看，这些细胞可以提供无限的转基因组织，可以在不使用慢性免疫抑制作为治疗多种疾病的手段的情况下进行移植。