Generating and Investigating Antigen-Deficient Islets in Autoimmune Diabetes

自身免疫性糖尿病中抗原缺陷胰岛的生成和研究

基本信息

批准号：
10352040
负责人：
THOMAS DELONG
金额：
$ 23.33万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-22 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10352040
关键词：
Amino Acid Sequence Antigens Autoantigens Autoimmune Diabetes Beta Cell C-Peptide CD4 Positive T Lymphocytes CRISPR/Cas technology Cathepsins Cellular Assay Clone Cells Colorado Core Facility Data Development Diabetes Mellitus Disease Enzymes Epitope spreading Family Generations Genes Histology Human Hybrids Immune system Inbred NOD Mice Incidence Infiltration Insulin Insulin-Dependent Diabetes Mellitus Interferon Type II Islets of Langerhans Islets of Langerhans Transplantation Isoleucine Leucine Mass Spectrum Analysis Measures Mediating Modeling Mus Mutate Mutation Non obese Onset of illness Organ Donor Organelles Outcome Pathogenicity Patients Peptide Hydrolases Peptides Process Proinsulin Reagent Research Personnel Residual state Role T-Lymphocyte T-Lymphocyte Epitopes Transplant Recipients Tumor-infiltrating immune cells Universities autoreactive T cell autoreactivity cohort diabetic experimental study immunoregulation induced pluripotent stem cell insulin dependent diabetes mellitus onset islet mouse genetics non-diabetic non-genomic peripheral blood prevent success therapeutic target

项目摘要

PROPOSAL SUMMARY Hybrid Insulin Peptides (HIPs) are a new family of autoantigens in Type 1 Diabetes (T1D). These peptides form through covalent fusion of pro-insulin fragments to other beta-cell peptides. HIPs contain non-genomic amino acid sequences, making them plausible targets for autoreactive T cells. We demonstrated that HIP- reactive CD4 T cells trigger diabetes in non-obese diabetic (NOD) mice, are present in residual islets of organ donors with T1D, and can be detected at significantly elevated levels in the peripheral blood of recent-onset T1D patients. We also verified the presence of HIPs in human and murine islets by mass spectrometry. Our data indicate that the protease Cathepsin D is responsible for the generation of HIPs that are targeted by diabetes-triggering CD4 T cell clones. Here we will generate NOD mice carrying a mutation that prevents the generation of HIPs. We predict that HIP-deficient beta-cells of these mice cannot be recognized by pathogenic CD4 T cells and the mice will be protected from disease onset. We will analyze the islets of these mice to assess HIP-content and T cell infiltration. We will also study disease incidence in these mice and determine whether pathogenic CD4 T cell clones can transfer disease into these mice. Beta-cells, deficient of disease- critical autoantigens, could allow us to reverse autoimmune diabetes in NOD mice and humans without the need for immune-modulation or shielding of beta-cells through encapsulation.

提案摘要杂化胰岛素肽（臀部）是1型糖尿病（T1D）中的新型自身抗原家族。这些肽通过将前胰岛素片段与其他β细胞肽的共价融合形成。臀部含有非基因组氨基酸序列，使其成为自动反应性T细胞的合理靶标。我们证明了嘻哈反应性CD4 T细胞触发非肥胖糖尿病（NOD）小鼠中的糖尿病，存在于器官的残留胰岛中具有T1D的供体，可以在最近发作的外周血中以显着升高的水平检测到 T1D患者。我们还通过质谱验证了人类和鼠胰岛中臀部的存在。我们的数据表明，蛋白酶组织蛋白酶D负责产生由糖尿病触发CD4 T细胞克隆。在这里，我们将产生携带突变的点头小鼠，以防止臀部产生。我们预测这些小鼠的髋关节缺陷β细胞无法通过致病性识别 CD4 T细胞和小鼠将受到疾病发作的保护。我们将分析这些小鼠的胰岛评估髋关节和T细胞浸润。我们还将研究这些小鼠的疾病发病率，并确定致病性CD4 T细胞克隆是否可以将疾病转移到这些小鼠中。 β细胞，疾病缺陷 - 关键的自动抗原，可以使我们能够在没有的小鼠和人类中逆转自身免疫性糖尿病需要通过封装对β细胞进行免疫调节或屏蔽。