A Novel Approach to Prevent and Treat Type 1 Diabetes

预防和治疗 1 型糖尿病的新方法

• 批准号: 6821845
• 负责人: Haval Shirwan
• 金额: $ 29.4万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6821845
• 关键词: CD95 molecule; NOD mouse; T lymphocyte; apoptosis; binding proteins; biotherapeutic agent; biotin; cell proliferation; chimeric proteins; dendritic cells; diabetes mellitus therapy; disease /disorder prevention /control; enzyme linked immunosorbent assay; gene expression; genetically modified animals; homologous transplantation; immune tolerance /unresponsiveness; immunomodulators; insulin dependent diabetes mellitus; ligands; metabolism disorder chemotherapy; nonhuman therapy evaluation; pancreatic islet transplantation; pancreatic islets; therapy design /development; tissue /cell culture

DESCRIPTION (provided by applicant): The primary objective of this grant proposal is to use a novel Fas Ligand molecule (SA-FasL) with potent apoptotic activity and a unique method, designated as ProtExTM, to rapidly and effectively display SA-FasL at the protein level on antigen-presenting cells (APCs) or pancreatic islets as an immunomodulatory approach to prevent and/or treat insulin-dependent Type I diabetes mellitus (T1D). T1D is a chronic autoimmune disorder that affects a large number of people worldwide. Transplantation of whole pancreas or purified islets promises an efficient approach to achieving euglycemia in T1D patients. However, immune rejection limits long-term graft survival. Islet cell destruction is primarily mediated by T cells directed at unique pancreatic beta cell antigens. In this application, we propose to use SA-FasL to induce apoptosis in pathogenic autoreactive and alloreactive T cells for the prevention and treatment of diabetes. FasL-induced apoptosis is the main mechanism of activation-induced cell death responsible for immune homeostasis and is also important for tolerance to self-antigens. There is evidence in the literature suggesting that FasL is not only important for the induction of tolerance by inducing apoptosis in antigen-activated lymphocytes, but also for the maintenance of tolerance by facilitating the generation and function of immunoregulatory lymphocytes. Therefore, we hypothesize that immunomodulation with SA-FasL will effectively eliminate pathogenic autoreactive and alloreactive lvmphocytes and induce immunoregulatory mechanisms that will maintain tolerance. We have recently generated a modified form of FasL molecule, SA-FasL, with potent apoptotic activity and developed a novel approach to display SA-FasL at the protein level on the surface of any cell of interest within a short period of time (-2 hrs). In preliminary studies, we demonstrated that SA-FasL can also be effectively displayed on the surface of pancreatic islets without toxicity and these islets restored euglycemia upon transplantation into syngeneic diabetic mice. In this application, we propose to use SA-FasL with the ProtExTM technology to prevent diabetes in prediabetic NOD and treat diabetes in diabetic animals using allogeneic islet transplantation. Diabetes in NOD will be prevented by the treatment of prediabetic mice with SA-FasL as soluble protein or displayed on APCs pulsed with islet antigens. Diabetes in NOD will be treated by the transplantation ofC57BL/6 allogeneic islets displaying SA-FasL with and without FasL-APCs. The recognition of auto/alloantigens in the context of SA-FasL is expected to result in physical/functional elimination of pathogenic auto/alloreactive T cells and generation of protective immune mechanisms that maintain tolerance. Induction of tolerance will be followed by a series of studies to delineate the implicated mechanisms. Rapid and durable display of functional proteins on cells or tissues offers a whole new means of intervention in the areas of autoimmunity, transplantation, and vaccines. This approach possesses the simplicity, safety, and efficacy required to make it a clinically relevant alternative to gene therapy in the treatment of a broad spectrum of immune-based diseases.

描述（由申请人提供）： The primary objective of this grant proposal is to use a novel Fas Ligand molecule (SA-FasL) with potent apoptotic activity and a unique method, designated as ProtExTM, to rapidly and effectively display SA-FasL at the protein level on antigen-presenting cells (APCs) or pancreatic islets as an immunomodulatory approach to prevent and/or treat insulin-dependent Type I diabetes mellitus （T1D）。 T1D是一种慢性自身免疫性疾病，影响了全世界的许多人。对整个胰腺或纯化胰岛的移植有望在T1D患者中采用尤利克血症的有效方法。但是，免疫排斥限制了长期移植物的生存。胰岛细胞破坏主要是由针对独特胰腺β细胞抗原的T细胞介导的。在此应用中，我们建议使用SA-FASL诱导致病性自动反应性和同种异体T细胞的凋亡，以预防和治疗糖尿病。 FASL诱导的细胞凋亡是激活诱导的细胞死亡的主要机制，负责免疫稳态，对于耐受性抗原也很重要。文献中有证据表明，FASL不仅对于通过诱导抗原激活的淋巴细胞凋亡来诱导耐受性很重要，而且对于通过促进免疫调节性淋巴细胞的产生和功能来维持耐受性。因此，我们假设用SA-FASL的免疫调节将有效消除致病性自动反应性和同种异体反应性LVMPHOPYTES，并诱导可维持耐受性的免疫调节机制。 我们最近生成了一种具有有效凋亡活性的FASL分子SA-FASL的改良形式，并开发了一种新颖的方法，可以在短时间内（-2小时）内在任何感兴趣的细胞表面显示在蛋白质水平上显示SA-FASL。在初步研究中，我们证明了SA-FASL也可以有效地显示在没有毒性的胰岛表面上，这些小岛在移植到合成型糖尿病小鼠中后恢复了尤利西亚。在此应用中，我们建议将SA-FASL与Protextm技术一起使用，以防止使用同种异体胰岛移植的糖尿病动物中糖尿病前点头糖尿病并治疗糖尿病。通过用SA-FASL作为可溶性蛋白或在用胰岛抗原脉冲的APC上，将糖尿病中的糖尿病预防。 NOD中的糖尿病将通过C57BL/6的同种异体胰岛的移植来治疗，这些胰岛显示带有和没有FASL-APC的SA-FASL。在SA-FASL的背景下，对自动/同种抗原的识别有望导致致病性自动/同种异体反应性T细胞的物理/功能消除，并产生保持耐受性的保护性免疫机制。诱导耐受性将进行一系列研究，以描述所涉及的机制。在细胞或组织上快速耐用的功能蛋白显示，可以在自身免疫，移植和疫苗的区域进行全新的干预手段。这种方法具有使其成为基因治疗的临床相关替代方案所需的简单，安全性和功效，以治疗多种免疫疾病。