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

• 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
• 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

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.

项目摘要 原发性免疫缺陷（PID）是由先天性误差引起的一组单基因疾病 （IEI）赋予严重感染，自身免疫性和癌症的风险。 IEI的遗传和免疫学研究 有助于了解人类免疫的机制以及管理和治疗PID患者的机制。他们的 研究对于了解与遗传变异相关的更多常见疾病也很重要 基因。但是，尽管研究基础科学和临床的PID具有相关性和重要性 观点，一半的PID患者缺乏遗传诊断。通过研究患有未知PID的患者 遗传病因赋予对严重分枝杆菌疾病的敏感性，我们发现了一个胡说八道突变 在serpinb1中。使用种群遗传学，生物化学和分子生物学的组合，我们表明 这是Serpinb1完全缺陷的第一种情况。 serpinb1是细胞内丝氨酸 蛋白酶自杀抑制剂。它在小鼠中的不存在会导致不受控制的细胞内蛋白酶活性，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，我们的患者不显示任何中性粒细胞 异常，但是我们深入的免疫特征显示出频率的降低 激活的T细胞和CD4+记忆T细胞。我们的发现表明Serpinb1先前未知 在T细胞和抗菌免疫中的功能。我们的数据还表明Serpinb1协调一个 细胞内蛋白酶介导的途径对于T细胞激活和存活必不可少。考虑到我们的发现， 应用将检验以下假设：Serpinb1对于人类T细胞激活至关重要，并且通过控制 细胞内蛋白酶活性，对人类疾病。利用该患者提供的独特机会 为了了解人serpinb1的功能，我们提出了三种互补方法。在AIM 1中，我们 将表征serpinb1缺乏对T细胞激活和CD4+ T细胞的免疫学后果 分化。我们还将研究T细胞中SERPINB1缺乏的转录后果。因此， 我们将发现SERPINB1在人T细胞中的先前未知功能。在AIM 2中，我们将确定并 表征serpinb1在T细胞中精心策划的途径。缺乏serpinb1释放 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 用于治疗目的。此外，表征SERPINB1依赖性的抗菌机制 在缺乏遗传诊断的患者中，将导致在其途径中鉴定出额外的IEI。在AIM 3中，我们 将执行全现象的关联研究（PHEWAS），以识别遗传变异之间的关联 SERPINB1途径和其他人类疾病。这种反向遗传方法将扩大我们将会 从对罕见疾病的研究中学习，使一群更广泛的患者受益。我们提出的研究众多 达到了解SERPINB1介导的免疫力，PID，抗 - 分枝杆菌免疫，以及Serpinb1途径对人类疾病的贡献。