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: 7096113
• 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 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7096113
• 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）： 该资助提案的主要目标是使用一种具有强凋亡活性的新型 Fas 配体分子 (SA-FasL) 和一种独特的方法（称为 ProtExTM），在抗原呈递细胞上快速有效地在蛋白质水平上展示 SA-FasL (APC) 或胰岛作为免疫调节方法来预防和/或治疗胰岛素依赖性 I 型糖尿病 (T1D)。 T1D 是一种慢性自身免疫性疾病，影响着全世界很多人。全胰腺或纯化胰岛移植有望成为 T1D 患者实现血糖正常的有效方法。然而，免疫排斥限制了移植物的长期存活。胰岛细胞破坏主要由针对独特胰腺 β 细胞抗原的 T 细胞介导。在此应用中，我们建议使用 SA-FasL 诱导致病性自身反应性和同种反应性 T 细胞凋亡，以预防和治疗糖尿病。 FasL 诱导的细胞凋亡是激活诱导细胞死亡的主要机制，负责免疫稳态，对于自身抗原的耐受也很重要。文献中有证据表明，FasL不仅通过诱导抗原激活的淋巴细胞凋亡来诱导耐受，而且通过促进免疫调节淋巴细胞的生成和功能来维持耐受。因此，我们假设 SA-FasL 的免疫调节将有效消除致病性自身反应性和同种反应性淋巴细胞，并诱导维持耐受性的免疫调节机制。 我们最近生成了 FasL 分子的修饰形式 SA-FasL，具有有效的细胞凋亡活性，并开发了一种新方法，可以在短时间内 (-2小时）。在初步研究中，我们证明SA-FasL也可以有效地展示在胰岛表面，且无毒性，并且这些胰岛在移植到同系糖尿病小鼠中后恢复了正常血糖。在此应用中，我们建议使用 SA-FasL 与 ProtExTM 技术来预防糖尿病前期 NOD 中的糖尿病，并通过同种异体胰岛移植治疗糖尿病动物的糖尿病。通过用 SA-FasL 作为可溶性蛋白或展示在用胰岛抗原脉冲的 APC 上治疗糖尿病前期小鼠，可以预防 NOD 中的糖尿病。 NOD 中的糖尿病将通过移植展示 SA-FasL 的 C57BL/6 同种异体胰岛（有或没有 FasL-APC）来治疗。 SA-FasL 背景下对自身/同种异体抗原的识别预计将导致致病性自身/同种异体反应性 T 细胞的物理/功能消除，并产生维持耐受性的保护性免疫机制。诱导耐受后将进行一系列研究来描述相关机制。在细胞或组织上快速、持久地展示功能蛋白为自身免疫、移植和疫苗领域提供了一种全新的干预手段。这种方法具有简单性、安全性和有效性，使其成为治疗多种免疫疾病的基因疗法的临床相关替代方案。