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Transplantation of Genetically Modified, Immune-privileged Steroli Cells

转基因、免疫豁免的甾体细胞移植

基本信息

批准号：
8291112
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
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-28 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8291112
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

项目摘要

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（由申请人提供）：支持细胞是免疫豁免细胞，当跨越同种异体或异种屏障移植时，无需使用免疫抑制药物即可长期存活。当大多数其他细胞受到免疫排斥时，支持细胞能够在移植后存活下来，这表明支持细胞可以被设计为基因治疗的载体。我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无限的组织来源，可以用作传递治疗相关蛋白质的载体，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过改造的支持细胞产生功能水平的胰岛素作为糖尿病小鼠的有效长期治疗策略的可行性。据推测，免疫特权支持细胞可以通过基因工程改造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。这一假设基于我们的初步数据，该数据证明未经修饰的支持细胞在不使用免疫抑制药物的情况下作为同种异体移植物和异种移植物移植时可以长期存活，并且稳定表达 GFP 的支持细胞在同种异体移植中存活，同时继续表达 GFP。具体目标旨在检验这一假设。在目标 1 中，将检查表达胰岛素的支持细胞的功能。将检查稳定表达胰岛素的小鼠和猪支持细胞在体外以及在体内移植到糖尿病免疫缺陷SCID小鼠后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。表达胰岛素的支持细胞将作为同种异体移植物（小鼠支持细胞）或异种移植物（猪支持细胞）移植到患有糖尿病、具有免疫能力的 C3H 小鼠中。将检查功能性胰岛素的产生和支持细胞的存活。预计，经过改造可稳定输送基础水平胰岛素的支持细胞能够在移植到糖尿病小鼠体内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、免疫特权的支持细胞可以用作移植组织的无限供应。公共卫生相关性：许多疾病有可能通过基因疗法得到治疗；例如，输送因子 VIII 用于治疗血友病。然而，高效、安全的输送系统尚未开发出来。我们将探索新颖和创新的概念，即可以对免疫特权支持细胞进行改造以提供治疗性蛋白质。从长远来看，这些细胞可以提供无限量的转基因组织，这些组织可以在不使用慢性免疫抑制的情况下进行移植，作为治疗多种疾病的手段。