IL-4 Overexpressing Dendritic Cells and Exosomes for Treatment of Type 1 Diabetes

IL-4 过表达树突状细胞和外泌体用于治疗 1 型糖尿病

• 批准号: 7676783
• 负责人: Melanie A Ruffner
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676783
• 关键词: Adenovirus Vector; Adoptive Transfer; Antigen-Presenting Cells; Autoimmune Diseases; Autoimmune Process; Binding; Cell physiology; Cells; Child; Chronic Inflammatory Infiltrate; Data; Delayed Hypersensitivity; Dendritic Cells; Diabetes Mellitus; Diabetes prevention; Diagnosis; Dioxygenases; Disease; Dose; Engineering; Equilibrium; Female; Frequencies; Gene Transduction Agent; Generations; Goals; Human; Hyperglycemia; Immune; Immune Tolerance; Inbred NOD Mice; Inflammation; Injection of therapeutic agent; Insulin; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Interleukin-4; Intravenous; Islets of Langerhans; Laboratories; Mediating; Membrane; Methods; Modeling; Modification; Mus; Natural regeneration; Onset of illness; Pancreas; Pathogenesis; Patients; Peripheral; Phagocytosis; Phenotype; Play; Population; Prevention; Process; Production; Research; Rheumatoid Arthritis; Role; Schedule; Signal Transduction; Spleen; Splenocyte; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Therapeutic Effect; Time; Transgenes; Treatment Protocols; United States; Vesicle; base; blood glucose regulation; comparative efficacy; diabetic; effective therapy; enzyme linked immunospot assay; improved; in vivo; indoleamine; islet; lymph nodes; overexpression; prevent; research study; response; standard of care; vector

DESCRIPTION (provided by applicant): Each year, more than 13,000 children in the United Stated are diagnosed with type I diabetes (T1D), in which autoimmune destruction of the pancreatic islets results in loss of endogenous insulin production. The current standard of care for these patients is insulin therapy, which restores glucose homeostasis and aims to prevent complications associated with hyperglycemia. However, new therapies to halt the autoimmune destruction of the islets could be useful because it has been demonstrated that the damaged islets can regenerate if immune tolerance is reestablished at the time of diagnosis of hyperglycemia. I propose to study methods using gene therapy vectors to make therapeutic cells in order to treat T1 D. I have tested DC infected with adenoviral vectors expressing several different transgenes, including FasL, IDO, IL-10, as well as a soluble and membrane-bound form of IL-4 (slL-4 and mblL-4), for their ability to delay onset of diabetes in 10-12 week old female NOD mice after intravenous adoptive transfer of a million cells. I have similarly screened 1 ug of exosomes, which are small membrane-bound vesicles, produced by these modified DC. My preliminary results indicate that administration of DC expressing slL-4 are capable of completely preventing disease onset, whereas a single treatment of exosomes expressing mbll_-4 delays diabetes onset by approximately 5 weeks. The goal of this research is to optimize the therapeutic potential of DC and DC-derived exosomes which have been engineered to be tolerogenic using adenoviral vectors expressing either slL-4 or mblL-4 for the treatment of T1 D, and to understand the mechanisms by which these DC and DC-derived exosomes exert their therapeutic effects. To this end, I propose to optimize these preliminary results, treating 12-week-old NOD/ShiLTJ mice with adoptive transfer of either DC-derived exosomes or parental DC that have been modified to express either slL-4 or mbll_-4. I will first determine the most effective treatment schedule, and then I will determine the most efficacious dose. After optimizing the dosing of DC and DC-derived exosomes for the prevention of diabetes onset, I will determine if this treatment schedule is effective at reversing established T1 D. I hypothesize that adoptive transfer of IL-4 transduced DC or their exosomes protects against T1D by modulating the function of endogenous T cell subsets. Since IL-4 seems to be effective I will first examine the effect of our DC and exosome therapies of Th1/Th2 balance in the NOD mouse, and then will conduct adoptive transfer studies into NOD/Scid mice as well as FACS and ELISPOT to see if the treatments are capable of generating a regulatory T cell population.

描述（由申请人提供）：每年，联合有13,000多名儿童被诊断出患有I型糖尿病（T1D），其中胰岛的自身免疫性破坏会导致内源性胰岛素产生的损失。这些患者的当前护理标准是胰岛素疗法，它恢复了葡萄糖稳态，旨在防止与高血糖相关的并发症。但是，停止胰岛自身免疫性破坏的新疗法可能是有用的，因为已经证明，如果在高血糖诊断时重新建立免疫耐受性，受损的胰岛可以再生。我建议研究使用基因疗法载体制作治疗细胞以治疗T1D的方法。我已经测试了DC感染了表达几种不同转基因的腺病毒载体，包括FASL，IDO，IL-10，IL-10，以及可溶性和膜结合的IL-4（SLL-4和MBL-4和MBL-4）的能力，他们的能力延迟了，他们的能力是他们的能力。一百万个细胞的静脉内产物转移后点头小鼠。我类似地筛选了1 ug的外泌体，这些外泌体是由膜结合的小囊泡，由这些修饰的DC产生。我的初步结果表明，表达SLL-4的DC的给药能够完全预防疾病发作，而表达MBLL_-4的外泌体进行了单一治疗，将糖尿病延迟大约5周。这项研究的目的是优化DC和DC衍生的外泌体的治疗潜力，这些外泌体使用表达SLL-4或MBLL-4的腺病毒矢量进行了耐受性，用于治疗T1 D，并了解这些DC和DC衍生的外观源代理的机制。为此，我提议优化这些初步结果，以通过DC衍生的外泌体或已修改以表达SLL-4或MBLL_-4的DC衍生外泌体或父母DC的生育转移来处理12周龄的点头/shiltj小鼠。我将首先确定最有效的治疗时间表，然后我将确定最有效的剂量。在优化了用于预防糖尿病发作的DC和DC衍生的外部剂量的剂量之后，我将确定该治疗时间表是否有效地逆转了既定的T1D。由于IL-4似乎是有效的，因此我将首先检查NOD小鼠Th1/Th2平衡的DC和外泌体疗法的影响，然后将对NOD/SCID小鼠以及FACS和ELISPOT进行收养转移研究，以查看治疗方法是否能够产生调节性T细胞群体。