Characterize the Landscape and Origin of Hybrid Peptides in Beta Cells

描述 Beta 细胞中混合肽的景观和起源

基本信息

批准号：
10660635
负责人：
THOMAS DELONG
金额：
$ 65.97万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-25 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660635
关键词：
Algorithms Amino Acid Sequence Animal Model Antigens Artificial Intelligence Autoantigens Autoimmune Automobile Driving Autophagocytosis Bayesian Analysis Benchmarking Beta Cell Biochemical Bioinformatics Biological Markers Blinded CD4 Positive T Lymphocytes Cathepsins Cells Cellular Structures Clone Cells Communities Computational algorithm Computers Data Data Set Databases Defect Degradation Pathway Detection Diabetes Mellitus Disease Epitopes Expert Systems Family Fever Funding Generations Grant High temperature of physical object Homeostasis Human Hybrids Inbred NOD Mice Individual Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Lead Ligation Lysosomal Function Inhibition Marketing Mass Spectrum Analysis Mediating Metabolic stress Methodology Mus Non obese Organ Donor Pathogenesis Pathogenicity Patients Peptide Hydrolases Peptides Peripheral Blood Mononuclear Cell Play Post-Translational Protein Processing Prevention Proinsulin Proteomics Reagent Research Research Design Residual state Role Sampling Specificity Stress Subgroup T-Cell Receptor T-Lymphocyte T-Lymphocyte Epitopes Techniques Technology Testing United States National Institutes of Health aged autoreactive T cell autoreactivity cytokine diabetes pathogenesis diabetic enzyme linked immunospot assay exposed human population granule cell improved induced pluripotent stem cell innovation instrument insulin dependent diabetes mellitus onset insulin granule interest islet mass spectrometer neoantigens new technology new therapeutic target non-diabetic non-genomic novel peripheral blood predictive modeling success therapeutic target tool

项目摘要

Project Summary/Abstract In Type 1 Diabetes (T1D) autoreactive CD4 T cells mediate the destruction of insulin producing beta-cells. The reasons for this misguided attack are poorly understood. Post-translational protein modifications could provide plausible explanations for the existence of autoreactive T cells in T1D. Hybrid insulin peptides (HIPs) are a form of post translationally modified antigens that form in beta-cells through the covalent ligation between proinsulin fragments and other beta-cell peptides. The resulting HIPs contain non-genomic amino acid sequences making them plausible targets for pathogenic T cells in T1D. Various HIP-reactive CD4 T cell clones were shown to trigger diabetes in non-obese diabetic (NOD) mice, a major animal model for the study of T1D. In addition, HIP-reactive CD4 T cells were identified in residual pancreatic islets of T1D organ donors. Significantly elevated levels of HIP-reactive T cells were also be detected in the peripheral blood of recent-onset T1D patients, but not in non-diabetic control subjects. Importantly, we applied mass spectrometric analyses on islets and confidently validated the presence of several HIPs that participate in T1D pathogenies. Autophagy is a cellular mechanism that removes unnecessary or dysfunctional components of the cell through lysosomal degradation. Our data indicate that autophagy plays a critical role on HIP-content in islets. Here we propose to modulate autophagy in human islets through various mechanisms, including cytokine treatment, with the objective to increase cellular HIP content. This will facilitate our identification efforts of novel HIP through mass spectrometric analyses on islets. We will also study the role of metabolic stress that may lead to the accumulation of a new sub-group of HIPs in beta-cells that could provide a trigger for T1D. Furthermore, we will study the mechanism of HIP-formation in human islets. Understanding this mechanism may provide us with new therapeutic targets that could be used to block HIP-formation in beta-cells and remove epitopes for disease- driving T cells in T1D. Additionally, we will use ELISPOT analyses to establish the role of identified HIPs as T cell epitopes in T1D pathogenesis. Such HIP-reactive T cells could serve as T1D biomarkers and allow us to improve current disease-prediction models. Lastly, we will advance and benchmark an innovative computer algorithm for the mass spectrometric identification of hybrid and native peptides. This algorithm may allow us to identify other types of hybrid peptides that cannot be discovered through conventional methodologies. In summary, success in this project will deliver new technologies, biomarkers, and therapeutic targets for the study and prevention of T1D. Identification of novel HIPs will also deliver targets for the induction of antigen specific tolerance induction in T1D, which may be required for the reversal of T1D.

项目摘要/摘要在1型糖尿病（T1D）自动反应性CD4 T细胞中介导了产生β细胞的破坏。这种误导攻击的原因对此很众所周知。翻译后蛋白质修饰可能为T1D中的自动反应性T细胞存在提供了合理的解释。杂化胰岛素肽（臀部）是通过在β细胞中形成的翻译后修饰抗原的一种形式。促硫素片段和其他β细胞肽。所得的臀部含有非基因组氨基酸序列使其成为T1D病原T细胞的合理靶标。各种嘻哈CD4 T细胞克隆被证明会引发非肥胖糖尿病（NOD）小鼠的糖尿病，这是研究T1D的主要动物模型。此外，在T1D器官供体的残留胰岛中鉴定出髋透反应的CD4 T细胞。在最近发作的外周血中也检测到髋部反应性T细胞水平的显着升高 T1D患者，但不在非糖尿病对照受试者中。重要的是，我们对质谱分析应用胰岛和自信地证实了参与T1D病原体的几个臀部的存在。自噬是通过溶酶体去除细胞不必要或功能失调成分的细胞机制降解。我们的数据表明，自噬在胰岛中的髋关节中起着至关重要的作用。在这里我们建议通过包括细胞因子处理在内的各种机制调节人类胰岛的自噬，目标增加细胞髋关节含量。这将促进我们通过质量进行新型臀部的识别工作胰岛的光谱分析。我们还将研究代谢应力的作用，这可能导致积累在β细胞中的新臀部子组的子组，可以为T1D提供触发。此外，我们将研究人类胰岛中髋关节形成的机制。了解这种机制可能会为我们提供新的可用于阻止β细胞中髋关节形成的治疗靶标，并去除疾病的表位 - 在T1D中驱动T细胞。此外，我们将使用ELISPOT分析来确定臀部的作用 T1D发病机理中的细胞表位。这种髋关节反应性T细胞可以用作T1D生物标志物，并让我们能够改善当前的疾病预测模型。最后，我们将推进并基准一台创新的计算机杂种和天然肽的质谱鉴定算法。该算法可能使我们能够识别其他类型的混合肽，这些混合肽无法通过常规方法发现。在总结，该项目的成功将为研究提供新的技术，生物标志物和治疗目标和预防T1D。新型臀部的识别还将提供诱导抗原特异性的靶标 T1D的耐受性诱导，这可能是T1D逆转所必需的。