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Dissecting Hem-1 functions in B lymphocyte Development and Primary Immunodeficiency Disease

剖析 Hem-1 在 B 淋巴细胞发育和原发性免疫缺陷病中的功能

基本信息

批准号：
10385848
负责人：
BRIAN M IRITANI
金额：
$ 19.44万
依托单位：
UNIVERSITY OF WASHINGTON
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：
2021-04-06 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10385848
关键词：
Actins Adhesions Affect Antibody Formation Antibody Response Antigens Autoimmune Diseases Autoimmunity B-Cell Development B-Lymphocytes Bacterial Infections Biochemical Biological Models CDC42 gene CRISPR/Cas technology Cell physiology Cells Characteristics Child Communities Community-Acquired Infections Complex Cre-LoxP Cytokine Receptors Cytoskeleton Development Disease Exhibits Family Gene Targeting Genes Genetic Genetic Heterogeneity Genetic Transcription Goals Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Hematopoietic Hematopoietic stem cells Homologous Protein Human Humoral Immunities Immune Immune response Immune signaling Immunity Immunization Immunodeficient Mouse Immunologic Deficiency Syndromes Immunologic Receptors Impairment Individual Infection Infectious Skin Diseases Influenza A virus Inherited Integrins Knowledge Ligands Link Malignant Neoplasms Mediating Mission Modeling Molecular Mus Mutation Otitis Media Outcome Study Patients Peripheral Phenotype Point Mutation Polysaccharides Production Proteins Public Health Publishing Reporting Research Respiratory Tract Infections Role Signal Pathway Signal Transduction Site Streptococcus pneumoniae System T-Cell Receptor Testing Time United States National Institutes of Health Untranslated RNA Variant Vascular Endothelium Virus Diseases Wasps Wiskott-Aldrich Syndrome cell type chemokine congenital immunodeficiency cytokine depolymerization disease-causing mutation epigenomics fly genetic regulatory protein human model humanized mouse innovation loss of function loss of function mutation member migration mortality mouse model pathogen polymerization protein complex receptor rho rho GTP-Binding Proteins trafficking transcriptomics

项目摘要

PROJECT SUMMARY Primary immunodeficiency diseases (PID) are group of inherited conditions where components of immune signaling pathways are either missing or are dysfunctional, resulting in severe recurring infections that often progress to autoimmune disease or cancer. Of the greater than 300 PIDs that have been identified so far, the molecular basis of a significant number (~100) of PIDs have yet to be defined. Recently, 9 individuals (4 now deceased) from 4 independent families were identified with PIDs that were linked to mutations in NCKAP1L, which encodes for a conserved hematopoietic cell-specific actin regulatory protein called Hematopoietic protein-1 (Hem-1). Affected children presented with recurring respiratory and skin infections, otitis media, impaired antibody responses to pneumococcal immunization (characteristic of B cell immunodeficiency), dysregulated cytokine production, and autoimmunity. Although the cellular and molecular functions of Hem-1 orthologues in flies and worms are relatively well characterized, there is a critical knowledge gap regarding the cell specific functions of Hem-1 in primary mammalian immune cells. Our longterm goal is to overcome this knowledge gap by dissecting the cell-specific cellular and molecular functions of Hem-1 in the development and functions of adaptive and innate immune cells. The objective of this proposal is to target Hem-1 in primary murine and human B lymphocytes in a B cell-specific manner to define the roles of Hem-1 in B cell development, signaling, and protective immunity. Our Specific Aims are to: (1) utilize conditional Cre-LoxP mediated gene targeting in mice to define how B cell specific loss-of-function mutations in Hem-1 alter peripheral B cell development, antibody responses, and protective immunity to Pneumococcus and Influenza A virus, two important community acquired infections; (2) determine the impact of Hem-1 mutations on B cell signaling and transcription; (3) utilize CRISPR/Cas9 gene editing in “humanized mice” to assess the impact of loss-of-function mutations in Hem-1 on the dynamic development of primary human B lymphocytes. To demonstrate feasibility, we have generated mice with a non-coding point mutation in Hem1 (Hem1pt/pt), Hem1 null (Hem1-/-) mice, Hem1floxed (Hem1fl/fl) mice, as well as Hem1 deficient primary human hematopoietic stem cells. Utilizing these innovative mouse and human model systems, we will test our overall hypothesis that B cell specific expression of Hem-1 is essential for the development of marginal zone and B1 B cells, T- independent antibody responses, and protective B cell immunity. These studies are highly significant because they will utilize innovative approaches to define for the first time, the cellular and molecular mechanisms of how non-coding mutations in NCLAP1L disrupt B cell development, signaling, and protective humoral immunity, resulting in PID and potentially autoimmunity. Because of extensive genetic heterogeneity of the 4 human PID families, limited number of patients, and concurrent infections, the disruption of Hem1 in cell-type specific manner is critical for dissecting the mechanisms of how mutations in Hem-1 result in PID and autoimmunity.

项目摘要原发性免疫缺陷病（PID）是一组遗传性疾病，其中免疫缺陷的成分，信号通路缺失或功能失调，导致严重的复发性感染，自身免疫性疾病或癌症的进展。到目前为止，在已确定的300多个PID中，大量（约100种）PID的分子基础尚未被定义。最近，9名个人（目前有4名来自4个独立家庭的30名患者（已死亡）被鉴定为具有与NCKAP 1 L突变相关的PID，它编码一种保守的造血细胞特异性肌动蛋白调节蛋白，蛋白-1（Hem-1）。受影响的儿童出现反复呼吸道和皮肤感染，中耳炎，对肺炎球菌免疫的抗体应答受损（B细胞免疫缺陷的特征），失调的细胞因子产生和自身免疫。虽然Hem-1的细胞和分子功能虽然蝇和蠕虫中的直系同源物的特征相对较好，但关于它们的同源性存在关键的知识缺口。 Hem-1在原代哺乳动物免疫细胞中的细胞特异性功能。我们的长期目标是克服这一点通过剖析Hem-1在发育中的细胞特异性细胞和分子功能，以及适应性和先天免疫细胞的功能。该提案的目标是在小学中瞄准Hem-1 小鼠和人B淋巴细胞以B细胞特异性方式来确定Hem-1在B细胞中的作用发育、信号传导和保护性免疫。我们的具体目标是：（1）利用条件Cre-LoxP 介导的小鼠基因靶向，以确定Hem-1中的B细胞特异性功能丧失突变如何改变外周B细胞发育、抗体应答和对肺炎球菌和甲型流感的保护性免疫病毒，两种重要的社区获得性感染;（2）确定Hem-1突变对B细胞的影响（3）在“人源化小鼠”中利用CRISPR/Cas9基因编辑来评估 Hem-1功能缺失突变对原代人B淋巴细胞动态发育的影响到为了证明可行性，我们已经产生了在Hem 1中具有非编码点突变的小鼠（Hem 1 pt/pt），Hem 1 裸（Hem 1-/-）小鼠、Hem 1融合（Hem 1fl/fl）小鼠以及Hem 1缺陷型原代人造血干细胞细胞利用这些创新的小鼠和人类模型系统，我们将测试我们的总体假设，即B Hem-1的细胞特异性表达对于边缘区和B1 B细胞、T-淋巴细胞的发育至关重要。独立的抗体应答和保护性B细胞免疫。这些研究意义重大，因为他们将利用创新的方法来定义第一次，细胞和分子机制， NCLAP 1 L中的非编码突变破坏B细胞发育、信号传导和保护性体液免疫，导致PID和潜在的自身免疫。由于4种人类PID的广泛遗传异质性，家庭，有限的患者数量，并发感染，Hem 1在细胞类型特异性这种方式对于剖析Hem-1突变如何导致PID和自身免疫的机制至关重要。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Hem-1 regulates protective humoral immunity and limits autoantibody production in a B cell-specific manner.

DOI：
10.1172/jci.insight.153597
发表时间：
2022-05-09
期刊：
JCI INSIGHT
影响因子：
8
作者：
Avalos, Alan;Tietsort, Jacob T.;Suwankitwat, Nutthakarn;Woods, Jonathan D.;Jackson, Shaun W.;Christodoulou, Alexandra;Morrill, Christopher;Liggitt, H. Denny;Zhu, Chengsong;Li, Quan-Zhen;Bui, Kevin K.;Park, Heon;Iritani, Brian M.
通讯作者：
Iritani, Brian M.

Metabolism meets immunodeficiency disease.

新陈代谢遇到免疫缺陷疾病。