Defining the antigenic determinants of the adaptive immune response in IgG4-related disease

定义 IgG4 相关疾病中适应性免疫反应的抗原决定因素

基本信息

批准号：
10284753
负责人：
Cory Perugino
金额：
$ 17.5万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
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：
2021-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10284753
关键词：
Affect Alleles Antigen Receptors Antigen Targeting Antigens Apoptosis Arthritis Autoantibodies Autoantigens Autoimmune Automobile Driving B cell therapy B-Cell Activation B-Lymphocytes Bioinformatics Biological Assay Biometry Blood CD4 Positive T Lymphocytes Cell Communication Cells Center for Translational Science Activities Clinical Clinical Immunology Clinical Sciences Clonal Expansion Clone Cells Collaborations Deposition Development Disease Disseminated Malignant Neoplasm Environment Epitopes Extracellular Matrix Proteins Faculty Fibrosis Fostering Foundations Funding Future Galectin 3 General Hospitals Genes Goals HLA-DR Antigens Human Hypersensitivity IgG4 Immune Immune response Immunologic Receptors Immunologics Individual Infiltration Institutes Investigation Journals Knowledge Lead Learning Lesion Libraries Light Link Mass Spectrum Analysis Massachusetts Mediating Mentors Mentorship Mesenchymal Molecular Molecular Biology Monoclonal Antibodies Organ Paper Parents Pathogenesis Pathogenicity Patients Peptide/MHC Complex Peptides Phenotype Positioning Attribute Proteins Publishing Recombinants Reporting Research Research Personnel Rheumatology Sampling Scientist Series Severity of illness Specificity Statistical Data Interpretation Systemic Scleroderma T cell receptor repertoire sequencing T cell response T-Lymphocyte T-cell receptor repertoire Tissues Training Training Activity Transforming Growth Factor beta Translational Research Tumor-infiltrating immune cells United States United States National Institutes of Health Universities Validation Work Yeasts adaptive immune response base biobank career clinical investigation clinical phenotype clinical remission cohort cytotoxic individual patient member novel patient subsets programs repository response screening single cell sequencing single-cell RNA sequencing skills tumor

项目摘要

This mentored research program aims to develop Dr. Perugino into an independent investigator studying the immunologic mechanisms of fibrosis. The research goals entail the identification of specific self-proteins driving adaptive immune responses in the context of IgG4-related disease (IgG4-RD), a recently described immune- mediated fibrotic disease. Candidate: Dr. Perugino is a junior faculty member in the Rheumatology Unit at Massachusetts General Hospital and has been conducting bench-based research since 2015. His short-term career goals are to build a stronger foundation in molecular biology, advance his skills in statistical analysis, develop proficiency in single-cell sequencing, gain expertise in creating and screening peptide-MHC yeast display libraries, and build his professional skills towards independent investigation. These goals will be supported by a series of coursework through Harvard University and the Harvard Clinical and Translational Science Center. He has recently published four first-author papers, the first identifying galectin-3 as a novel B cell self-antigen in IgG4-RD (Journal of Allergy and Clinical Immunology, 2019), the 2nd linking the diversity of auto-antibody responses with disease severity in IgG4-RD (Arthritis & Rheumatology, 2019), a 3rd establishing CD4+ T cells with cytotoxic features as likely disease drivers in systemic sclerosis (Journal of Clinical Investigation, 2020) and a 4th establishing the effector phenotype of cytotoxic CD4+ T cells in IgG4-RD (Journal of Allergy and Clinical Immunology, 2020). This work forms the foundation for this K08 proposal. Mentorship, Training Activities, and Environment: Dr. Perugino has been under the direct mentorship of Dr. Shiv Pillai since 2015 and Dr. John Stone since 2014 acting as mentors in investigation and translational research, respectively. The training plan builds upon the skills learned under this mentorship team. Guided by his advisors and collaborators, Dr. Perugino will gain proficiency in (1) single cell RNA sequencing (Dr. Alex Shalek), (2) Ig/TCR repertoire analyses (Dr. Mark Davis), (3) peptide-MHC yeast display library development (Dr. Michael Birnbaum), immune cell interactions (Dr. Michael Brenner) and biostatistical analysis (Dr. Musie Ghebremichael). Research Program: The overarching hypothesis of this proposal is that the immune response in IgG4-RD is driven by the crosstalk between B and T cells directed at specific epitopes derived from the same protein antigen. Leveraging the identification of clonal expansions of B and T cells in IgG4-RD, the proposal entails the determination of dominant B and T cell clones, which will subsequently be used to single cell clone soluble antigen receptors and use those as probes to pull down their cognate antigens. These complementary approaches, one focused on B cells (Aim 1) and the other on T cells (Aim 2), entail the validation of B and T cell responses among the largest single-center cohort of IgG4-RD bio-samples in the United States. These studies will serve as a foundation for Dr. Perugino's future goals of identifying the HLA and antigenic determinants that distinguish the clinical phenotypes of different immune-mediated fibrotic diseases, such as IgG4-RD, as an independent investigator.

这项指导性研究计划旨在将佩鲁吉诺博士培养成为一名独立调查员，研究纤维化的免疫学机制。研究目标需要识别特定的自身蛋白驱动 IgG4 相关疾病（IgG4-RD）中的适应性免疫反应，这是最近描述的一种免疫- 介导的纤维化疾病。候选人：Perugino 博士是风湿病科的初级教员马萨诸塞州总医院自 2015 年以来一直在进行基于实验室的研究。他的短期研究职业目标是在分子生物学方面打下更坚实的基础，提高统计分析技能，提高单细胞测序的熟练程度，获得创建和筛选肽-MHC 酵母的专业知识展示图书馆，并培养他独立调查的专业技能。这些目标将是得到哈佛大学和哈佛临床与转化学院一系列课程的支持科学中心。他最近发表了四篇第一作者论文，其中第一篇将 galectin-3 鉴定为新型 B IgG4-RD 中的细胞自身抗原（过敏与临床免疫学杂志，2019），第二个链接多样性 IgG4-RD 中自身抗体反应与疾病严重程度的关系（关节炎与风湿病学，2019），第三个建立具有细胞毒性特征的 CD4+ T 细胞可能是系统性硬化症的疾病驱动因素（临床杂志调查，2020）和第四次建立 IgG4-RD 中细胞毒性 CD4+ T 细胞的效应表型（杂志过敏和临床免疫学，2020）。这项工作构成了 K08 提案的基础。指导，培训活动和环境：Perugino 博士自那时以来一直受到 Shiv Pillai 博士的直接指导 John Stone 博士自 2015 年起担任调查和转化研究导师，自 2014 年起分别担任导师。培训计划建立在该指导团队所学到的技能的基础上。在他的顾问和指导下在合作者中，Perugino 博士将熟练掌握 (1) 单细胞 RNA 测序（Alex Shalek 博士）、(2) Ig/TCR 谱分析（Mark Davis 博士），（3）肽-MHC 酵母展示库开发（Michael Birnbaum 博士），免疫细胞相互作用（Michael Brenner 博士）和生物统计分析（Musie Ghebrmichael 博士）。研究项目：该提案的总体假设是 IgG4-RD 中的免疫反应是由 B 细胞和 T 细胞之间针对源自相同蛋白质抗原的特定表位的串扰。杠杆作用 IgG4-RD 中 B 细胞和 T 细胞克隆扩增的鉴定，该提案需要确定显性 B 和 T 细胞克隆，随后将用于单细胞克隆可溶性抗原受体和使用它们作为探针来拉下它们的同源抗原。这些互补的方法，其中一种侧重于 B 细胞（目标 1）和 T 细胞（目标 2）的另一个，需要验证最大的 B 细胞和 T 细胞反应美国 IgG4-RD 生物样本单中心队列。这些研究将作为基础 Perugino 博士未来的目标是鉴定区分临床的 HLA 和抗原决定簇作为独立研究者，研究不同免疫介导的纤维化疾病的表型，例如 IgG4-RD。