Development of a MicroRNA Regulated Dendritic Cell Vaccine for Type 1 Diabetes

开发 MicroRNA 调控的 1 型糖尿病树突状细胞疫苗

• 批准号: 8683166
• 负责人: Robert Arthur Chong
• 金额: $ 4.77万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-19 至 2016-04-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8683166
• 关键词: 3' Untranslated Regions; Adopted; American; Antigen Presentation; Antigens; Apoptosis; Area; Autoimmune Diseases; Autoimmune Process; Beta Cell; Blood Glucose; CD8B1 gene; Cell Maturation; Cells; Chronic; Clinic; Data; Dendritic Cell Vaccine; Dendritic Cells; Development; Diabetes Mellitus; Diagnosis; Disease; Environment; Functional RNA; Future; Gene Delivery; Genetic; Genetic Translation; Goals; Hematopoietic; Histocompatibility Antigens Class I; Home environment; Human; Hyperglycemia; Hypoglycemia; IL2RA gene; Immune; Immune response; Immune system; In Vitro; Inbred NOD Mice; Incidence; Inflammatory; Insulin; Insulin-Dependent Diabetes Mellitus; Knowledge; Laboratories; Laboratory Study; Lead; Libraries; Life; MHC Class II Genes; Maintenance; Methods; MicroRNAs; Monitor; Onset of illness; Pancreas; Patients; Phenotype; Predisposition; Preparation; Process; Proinsulin; RNA Binding; Regulation; Regulatory T-Lymphocyte; Research; Rest; Risk; Site; Stimulus; Study models; Symptoms; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic Studies; Tissues; Transgenes; Transplantation; Vaccination; Vaccines; Viral Vector; Work; base; cell type; diabetic; expression vector; genome-wide; high throughput screening; immunogenic; improved; in vivo; islet; killings; mouse model; novel; novel vaccines; pathogen; prevent; public health relevance; response; sensor; transgene expression; vaccine development; vector

DESCRIPTION (provided by applicant): Type 1 Diabetes (T1D) is an autoimmune disorder whereby insulin-producing beta cells are destroyed by the immune system. Diabetogenic CD8+ T cells that recognize islet specific antigens home to the pancreas and initiate an attack on these beta cells. This leads to devastating chronic hyperglycemia and occasional, and life threatening, hypoglycemia. Genetic studies have identified several susceptibility loci that are predictive of T1D, and careful monitoring in the clinic can identify at risk patients who are in a pre-diabetic state. However, despite this knowledge, there exists no reliable method to prevent the initial autoimmune attack on the beta cells or future attacks against transplanted ones. The goal of the work proposed here is to develop a tolerogenic vaccine that is able to interfere with this process and induce regulatory T cells (Tregs) that are protective of islet specific antigens. To do so, this project is aimed at exogenously expressing these antigens in resting dendritic cells (DCs), which are capable of directing a pro-tolerogenic response in the absence of maturation or pro-inflammatory stimuli. To that end, a key challenge is to be able to restrict antigen expression from DCs when they become activated, but maintain robust expression in ones that have not. Our laboratory has recently developed a new microRNA (miRNA) based targeting strategy that is capable of doing just that. By placing target sites for miRNAs that become active upon DC maturation into a transgene, it is possible to completely eliminate expression in activated cells. Therefore, I plan to use viral vectors to deliver islet-specific antigens (Proinsulin and GAD65) to non-activated DCs, and restrict their expression in activated ones by incorporating target sites for miR-155, a miRNA that becomes highly active in mature DCs. I will also use a high-throughput method to identify new miRNAs that can achieve this same goal. These constructs will then be tested to see if they can induce tolerance to Proinsulin or GAD65 and prevent diabetes onset in a NOD mouse model of the disease.

描述（由申请人提供）：1 型糖尿病 (T1D) 是一种自身免疫性疾病，产生胰岛素的 β 细胞被免疫系统破坏。识别胰岛特异性抗原的致糖尿病 CD8+ T 细胞回到胰腺并发起对这些 β 细胞的攻击。这会导致毁灭性的慢性高血糖和偶尔危及生命的低血糖。遗传学研究已经确定了几个可预测 T1D 的易感位点，并且在临床中仔细监测可以识别处于糖尿病前期状态的高危患者。然而，尽管有这些知识，仍然没有可靠的方法来防止最初对β细胞的自身免疫攻击或未来对移植细胞的攻击。这里提出的工作目标是开发一种耐受性疫苗，能够干扰这一过程并诱导保护胰岛特异性抗原的调节性 T 细胞 (Treg)。为此，该项目旨在在静息树突细胞（DC）中外源表达这些抗原，这些抗原能够在没有成熟或促炎刺激的情况下引导促耐受反应。为此，一个关键的挑战是能够在 DC 被激活时限制其抗原表达，但在未激活的 DC 中保持强劲的表达。我们的实验室最近开发了一种新的基于 microRNA (miRNA) 的靶向策略，能够做到这一点。通过将 DC 成熟后变得活跃的 miRNA 的靶位点放入转基因中，可以完全消除活化细胞中的表达。因此，我计划使用病毒载体将胰岛特异性抗原（胰岛素原和 GAD65）递送至未激活的 DC，并通过整合 miR-155（一种在成熟 DC 中变得高度活跃的 miRNA）的靶位点来限制其在激活 DC 中的表达。 。我还将使用高通量方法来识别可以实现相同目标的新 miRNA。然后将测试这些构建体，看看它们是否可以在 NOD 小鼠模型中诱导对胰岛素原或 GAD65 的耐受性并预防糖尿病的发作。