The role of SERPINB1 in T cell function and its contribution to human diseases

SERPINB1在T细胞功能中的作用及其对人类疾病的贡献

基本信息

批准号：
10659419
负责人：
Ruben Martinez Barricarte
金额：
$ 66.5万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-05 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659419
关键词：
Abnormal Neutrophil Affect Alleles Antimycobacterial Agents Apoptosis Autoimmunity Basic Science Biochemistry Biological CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CRISPR/Cas technology Cell Line Cell physiology Cells Cellular biology Cessation of life Chemicals Clinical Critical Pathways Cytometry Data Disease Electronic Health Record Frequencies Future Genes Genetic Genetic Databases Genetic Polymorphism Genetic Predisposition to Disease Genetic Transcription Hereditary Disease Human Immunity Immunologic Deficiency Syndromes Immunologics Immunophenotyping Impairment Infection Knowledge Learning Left Malignant Neoplasms Mediating Mendelian disorder Methods Molecular Biology Mus Mutation Mycobacterium Infections Nature Neutropenia Nonsense Mutation Pathway interactions Patients Peptide Hydrolases Persons Population Genetics Predisposition Proteins Rare Diseases Research Risk Role STAT3 gene Sampling Serine Protease T cell differentiation T memory cell T-Cell Activation T-Lymphocyte Testing Therapeutic Variant Work biobank cell injury congenital immunodeficiency exome sequencing experimental study follow-up genetic approach genetic disorder diagnosis genetic variant human disease immunoregulation inhibitor interleukin-23 memory CD4 T lymphocyte mycobacterial neutrophil phenome prevent response reverse genetics suicide inhibitor

项目摘要

PROJECT SUMMARY Primary immunodeficiencies (PIDs) are a group of monogenic diseases caused by inborn errors of immunity (IEI) that confer risk of severe infection, autoimmunity, and cancer. The genetic and immunological study of IEI is instrumental to understand mechanisms of human immunity and to manage and treat patients with PIDs. Their study is also important to understand more common diseases associated with genetic variants in the same genes. However, despite the relevance and importance of studying PIDs from the basic science and clinical points of view, half of the PID patients lack a genetic diagnosis. By studying a patient with a PID of unknown genetic etiology that confers susceptibility to severe mycobacterial disease, we identified a nonsense mutation in SERPINB1. Using a combination of population genetics, biochemistry, and molecular biology, we showed that this is the first case of SERPINB1 complete deficiency ever described. SERPINB1 is an intracellular serine protease suicide inhibitor. Its absence in mice causes uncontrolled intracellular protease activity, leading to neutrophil death and subsequent neutropenia. Interestingly, our patient does not display any neutrophil abnormality, but instead, our in-depth immunological characterization showed a reduction in the frequency of activated T cells and CD4+ memory T cells. Our findings suggest that SERPINB1 has a previously unknown function in T cells and antimycobacterial immunity. Our data also indicates that SERPINB1 orchestrates an intracellular protease-mediated pathway essential for T cell activation and survival. Given our findings, this application will test the hypothesis that SERPINB1 is critical for human T cell activation and, by controlling intracellular protease activity, to human diseases. Capitalizing on the unique opportunity that this patient offers to understand the function of human SERPINB1, we propose three complementary approaches. In Aim 1, we will characterize the immunological consequences of SERPINB1 deficiency on T cell activation and CD4+ T cell differentiation. We will also study the transcriptional consequences of SERPINB1 deficiency in T cells. With this, we will uncover a previously unknown function of SERPINB1 in human T cells. In Aim 2, we will identify and characterize the pathways orchestrated by SERPINB1 in T cells. The absence of SERPINB1 unleashes the action of intracellular proteases. This rare mechanism of PID will allow for chemical inhibition of these proteases for therapeutic purposes. Furthermore, characterizing the SERPINB1-dependent antimycobacterial mechanisms will lead to the identification of additional IEI in its pathway in patients lacking a genetic diagnosis. In Aim 3, we will perform phenome-wide association studies (pheWAS) to identify associations between genetic variants in the SERPINB1 pathway and additional human diseases. This reverse genetic approach will expand what we will learn from the study of a rare disease to benefit a broader group of patients. Our proposed research has far- reaching clinical and biological implications for understanding SERPINB1 mediated immunity, PIDs, anti- mycobacterial immunity, and the contribution of the SERPINB1 pathway to human disease.

项目概要原发性免疫缺陷（PID）是一组由先天性免疫缺陷引起的单基因疾病 (IEI) 会带来严重感染、自身免疫和癌症的风险。 IEI 的遗传和免疫学研究有助于了解人体免疫机制以及管理和治疗 PID 患者。他们的研究对于了解与相同基因变异相关的更常见疾病也很重要基因。然而，尽管从基础科学和临床角度研究 PID 具有相关性和重要性。观点认为，一半的PID患者缺乏基因诊断。通过研究 PID 未知的患者遗传病因导致对严重分枝杆菌疾病的易感性，我们发现了无义突变在 SERPINB1 中。结合群体遗传学、生物化学和分子生物学，我们发现这是有史以来第一例 SERPINB1 完全缺陷的病例。 SERPINB1 是一种细胞内丝氨酸蛋白酶自杀抑制剂。它在小鼠体内的缺失会导致细胞内蛋白酶活性不受控制，从而导致中性粒细胞死亡和随后的中性粒细胞减少症。有趣的是，我们的患者没有显示任何中性粒细胞异常，但相反，我们深入的免疫学特征显示，激活的 T 细胞和 CD4+ 记忆 T 细胞。我们的研究结果表明 SERPINB1 具有以前未知的功能 T 细胞和抗分枝杆菌免疫中的功能。我们的数据还表明 SERPINB1 精心策划了细胞内蛋白酶介导的途径对于 T 细胞激活和存活至关重要。鉴于我们的发现，这该应用程序将测试 SERPINB1 对于人类 T 细胞激活至关重要的假设，并通过控制细胞内蛋白酶的活性，对人类疾病的影响。利用该患者提供的独特机会为了了解人类 SERPINB1 的功能，我们提出了三种互补的方法。在目标 1 中，我们将描述 SERPINB1 缺陷对 T 细胞激活和 CD4+ T 细胞的免疫学后果差异化。我们还将研究 T 细胞中 SERPINB1 缺陷的转录后果。有了这个，我们将揭示人类 T 细胞中 SERPINB1 的一个先前未知的功能。在目标 2 中，我们将确定并表征 T 细胞中 SERPINB1 协调的通路。 SERPINB1 的缺失释放了细胞内蛋白酶的作用。这种罕见的 PID 机制将允许对这些蛋白酶进行化学抑制出于治疗目的。此外，表征 SERPINB1 依赖性抗分枝杆菌机制将导致在缺乏基因诊断的患者的途径中识别出额外的 IEI。在目标 3 中，我们将进行全表型关联研究（pheWAS），以确定基因变异之间的关联 SERPINB1 通路和其他人类疾病。这种反向遗传方法将扩展我们的研究范围从罕见疾病的研究中学习，使更广泛的患者群体受益。我们提出的研究已经远达到理解 SERPINB1 介导的免疫、PID、抗分枝杆菌免疫以及 SERPINB1 途径对人类疾病的贡献。