Determining the rap1b signaling pathway to integrin activation in platelets

确定血小板中整合素激活的 rap1b 信号通路

• 批准号: 7800285
• 负责人: Luis Alberto Paniagua
• 金额: $ 3.25万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-15 至 2013-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7800285
• 关键词: Adoptive Transfer; Age; Animal Model; Animals; Autoimmune Diabetes; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Bioinformatics; Blood Platelets; CD4 Positive T Lymphocytes; CD8B1 gene; Cell physiology; Cells; Child; Congenic Strain; Cytotoxic T-Lymphocytes; Data; Dependence; Dependency; Development; Diabetes Mellitus; Disease; Disease Progression; Eotaxin; Epidemiology; Event; Exhibits; Fellowship; Functional disorder; Gene Expression; Genes; Genetic; Genomics; Health; Human; Immune; Immune System Diseases; Immune system; In Vitro; Inbred WF Rats; Incidence; Individual; Infiltration; Injection of therapeutic agent; Insulin; Insulin-Dependent Diabetes Mellitus; Integrins; Interleukin-1; Interleukins; Islets of Langerhans; Life; Major Histocompatibility Complex; Maps; Mediating; Mediator of activation protein; Mutation; Onset of illness; Pancreas; Pathogenesis; Pathway Analysis; Peripheral; Pharmaceutical Preparations; Population; Predisposition; Rat Strains; Rattus; Recruitment Activity; Regulation; Regulatory T-Lymphocyte; Relative (related person); Research; Risk; Role; Sampling; Signal Pathway; Signal Transduction; Staining method; Stains; Stress; T cell response; T-Lymphocyte; Testing; Time; Tissues; cell type; chemokine; congenic; diabetic; eosinophil; functional genomics; high risk; human subject; in vivo; inhibitor/antagonist; insight; islet; lymph nodes; mast cell; non-diabetic; prevent

DESCRIPTION (provided by applicant): Type 1 diabetes mellitus (T1DM) arises through autoimmune destruction of pancreatic ¿ cells and results in life-long dependency on daily insulin injection. Epidemiological data show that T1DM incidence is rising by 2-3% per year while onset is occurring at an earlier age. Since acquisition of relevant immune tissues from prediabetic children is nearly impossible, we have used congenic derivatives of the BioBreeding (BB) rat. The lymphopenic DRIyp/lyp animal develops spontaneous T1DM, whereas the DR+/+ does not. Evidence points to disease progression in BB rats being dependent on the absence of natural regulatory T (nTreg) cells. T1DM in BB rats also involves cytotoxic T cells, since their depletion is protective. Our preliminary studies show that BB rats are predisposed to T1 DM in that pancreatic ¿ cells of both DRIyp/lyp and DR+/+ strains recruit immune cells through expression of eotaxin, an eosinophil and mast cell recruiting chemokine, as well as interleukin 1¿ prior to insulitis in the DRIyp/lyp rat. Gene expression studies of prediabetic pancreatic lymph nodes (PLN) have shown that mast cells are terminally activated in DRIyp/lyp animals versus.negatively regulated in DR+/+ animals. We have successfully delayed T1DM in DRIyp/lyp rats with two different mast cell inhibiting drugs, confirming a pathogenic role for mast cells. Thus, we propose that nTreg cells in the DR+/+ rat can maintain the islet health by a peripheral regulatory mechanism, whereas in the absence of nTreg cells, DRIyp/lyp animals spontaneously progress to T1DM. We hypothesize that in the BB rat, innate immune cells mediate islet destruction unless nTreg cells prevent their mobilization. We will test our hypotheses through the following aims: 1) Temporally and spatially dissect the activities of nTreg cells during the prediabetic period at day 30 (prior to islet eotaxin signaling), day 40 (prior to insulitis), day 50 (early insulitis) and day 60 (late insulitis, pre-onset) using a combination of functional genomics and histological approaches. 2) Validate Aim 1 observations through in vitro and in vivo studies that characterize the action of nTreg on CD8 and CD4 T cells as well as innate cells. A role for mast cells in initiating and/or amplifying autoimmunity in T1DM has only recently been defined. These studies will utilize genomic, bioinformatic, histological, and immunological approaches to elucidate the relationship between innate cells and nTreg cells. This integrated approach will help us better understand how mast cells and nTreg cells interact and has the potential to establish a new paradigm for immune regulation, thus opening new insights into immune disease.

描述(申请人提供)：1型糖尿病(T1 DM)由胰腺细胞的自身免疫破坏而产生，导致终生依赖每日注射胰岛素。流行病学数据显示，T1 DM的发病率以每年2-3%的速度上升，而且发病年龄较早。由于从糖尿病前期儿童身上几乎不可能获得相关的免疫组织，我们使用了生物育种(BB)大鼠的同源基因衍生品。淋巴细胞减少的DRIyp/LYP动物发生自发性T1 DM，而DR+/+动物不发生。有证据表明，BB大鼠的疾病进展依赖于缺乏自然调节T(NTreg)细胞。BB大鼠的T1 DM也涉及细胞毒性T细胞，因为它们的耗尽是保护性的。我们的初步研究表明，BB大鼠易患T1 DM，DRIyp/LYP和DR+/+株的胰腺细胞在DRIyp/LYP大鼠的胰岛炎症之前通过嗜酸性粒细胞和肥大细胞募集趋化因子嗜酸性粒细胞趋化因子和白细胞介素1的表达来招募免疫细胞。对糖尿病前期胰腺淋巴结(PLN)的基因表达研究表明，肥大细胞在DRIyp/LYP动物中处于末端激活状态，而在DR+/+动物中处于负调控状态。我们用两种不同的肥大细胞抑制药物成功地延迟了DRIyp/LYP大鼠的T1 DM，证实了肥大细胞的致病作用。因此，我们认为DR+/+大鼠中的nTreg细胞可以通过外周调节机制维持胰岛的健康，而在没有nTreg细胞的情况下，DRIyp/LYP动物自发地进展为T1 DM。我们推测，在BB大鼠中，除非nTreg细胞阻止其动员，否则先天免疫细胞介导胰岛破坏。我们将通过以下目标验证我们的假设：1)结合功能基因组学和组织学方法，从时间和空间上剖析糖尿病前期30天(胰岛嗜酸粒细胞信号转导之前)、40天(胰岛素炎前)、50天(早期胰岛素炎)和60天(晚期胰岛素炎，发病前)nTreg细胞的活动。2)通过体外和体内研究证实Aim 1观察到的nTreg对CD8和CD4T细胞以及天然细胞的作用。肥大细胞在启动和/或放大T1 DM自身免疫中的作用直到最近才被确定。这些研究将利用基因组学、生物信息学、组织学和免疫学的方法来阐明天然细胞和nTreg细胞之间的关系。这种综合的方法将帮助我们更好地了解肥大细胞和nTreg细胞如何相互作用，并有可能建立一种新的免疫调节范式，从而为免疫疾病打开新的洞察力。