Assessment of cytokines in human islets from patients with diabetes

描述(由申请人提供)：合理开发免疫介导性疾病的治疗方法是基于对发病机制的详细了解，特别是受影响器官或组织中的免疫事件。在1型(以及2型)糖尿病患者中，这一点到目前为止一直受到患者获取和获得胰腺的阻碍。然而，最近建立的胰腺器官捐赠者资料库(NPOD)极大地改变了这种情况，并提供了一个关键的机会来加深我们对人类状况的洞察-这不仅将缩小差距，而且将(并且已经)改变范式，使我们能够以更有针对性的方式设计基于免疫的干预措施。因此，在这里，我们希望评估哪些细胞因子在1型和2型糖尿病患者的胰腺中表达。我们将使用多种策略来确定可靠的结果，包括直接原位免疫组织学、原位杂交、定量聚合酶链式反应和激光显微解剖。这些研究应该确定在人类1型和2型糖尿病发病机制中表达的主要细胞因子，表达这些细胞因子的细胞，以及它们的表达是结构性的还是与疾病的某个阶段有关，或者更有可能是局部胰岛破坏的阶段。直接确定关键免疫病理因素作为未来治疗靶点的策略是有先例的，就像类风湿性关节炎的情况一样，根据Marc Feldman对类风湿性关节炎患者关节滑膜细胞中肿瘤坏死因子水平升高的初步观察，开发了肿瘤坏死因子阻滞剂。从翻译的角度来看，我们设想细胞因子阻断可以整合到旨在诱导长期耐受的联合疗法中，理想地补充诱导调节性T细胞的免疫方案。 公共卫生相关性：本申请中概述的研究旨在提供糖尿病患者胰岛中可溶炎性因子(细胞因子和趋化因子)混合物的详细特征。预计在一组保存最好的独特标本中更好地了解局部组织环境将使治疗阻断这些因素的方法变得更加合理。我们的方法是以确定肿瘤坏死因子是类风湿关节炎的关键细胞因子为模型的，这导致了一类高效的抗肿瘤坏死因子药物的引入。

DESCRIPTION (provided by applicant): Rational development of treatments for immune mediated diseases is optimally based on detailed understanding of the pathogenesis, especially immunological events in affected organs or tissues. In type 1 (as well as type 2) diabetes, this has been hampered so far by access to and availability of pancreata from patients. However, the recently established repository of pancreatic organ donors ('nPOD') has drastically changed this situation and offers a crucial opportunity to deepen our insight into the human condition - this will not only close gaps, but will (and already has) shift paradigms and enable us to design immune-based interventions in a more targeted fashion. Therefore here, we wish to assess, which cytokines are expressed in human pancreata of patients with type 1 and type 2 diabetes. We will use a combination of strategies to ascertain reliable results including direct in situ immunohistology, in situ hybridization, quantitative PCR as well as laser microdissection. These studies should identify the major cytokines expressed during pathogenesis of human type 1 and type 2 diabetes, the cells which express them and whether their expression is constitutive or linked to a certain stage of the disease or, more likely, stage of local islet destruction. The strategy of directly identifying pivotal immunopathological factors as future therapeutic targets i the affected organ has precedent, as this has been the case for rheumatoid arthritis, where TNF blockers were developed based on Marc Feldman's initial observation of heightened TNF levels in synovial cells from affected joints in rheumatoid arthritis. From a translational angle, we envision that cytokine blockade could be integrated in a combination therapy aimed at long term tolerance induction, ideally complementing immunization regimens that induce regulatory T cells. PUBLIC HEALTH RELEVANCE: Research outlined in this application aims to provide a detailed characterization of the mixture of soluble inflammatory factors ('cytokines' and 'chemokines') in pancreatic islets from diabetes patients. It is anticipated that a better understanding of the local tissue environment in a unique set of optimally preserved samples will enable a more rational approach toward therapeutic blockade of these factors. Our approach is modeled after the identification of TNF as a crucial cytokine in rheumatoid arthritis, which has led to the introduction of a highly effective class of anti-TNF drugs.