Autosomal dominant OTULIN deficiency and staphylococcal disease

常染色体显性 OTULIN 缺乏症和葡萄球菌病

• 批准号: 10373489
• 负责人: Bertrand Boisson
• 金额: $ 25.43万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373489
• 关键词: Abscess; Acute Respiratory Distress Syndrome; Alleles; Antibiotics; Bacterial Infections; Bacterial Toxins; Biology; Cause of Death; Caveolins; Cell Death; Cell Survival; Cells; Cellulitis; Characteristics; Child; Chromosome 5; Chromosome Arm; Communities; Cri-du-Chat Syndrome; Defect; Dermal; Devices; Disease; Endocarditis; Epidemic; Epidemiology; Etiology; Fibroblasts; Folliculitis; Genes; Genetic; Genetic Diseases; Genetic Models; Genetic Predisposition to Disease; Genotype; Genus staphylococcus; Health; Hemolysin; Hereditary Disease; Hospitalization; Human; Immune; Immunity; Individual; Infection; Inflammation; Inherited; Innate Immune Response; Interleukin-1 Receptors; Interleukin-17; Interleukin-6; Investigation; Leukocytes; Life; Light; Link; Loss of Heterozygosity; Measures; Minority; Molecular; Mutate; Mutation; NF-kappa B; Natural Immunity; Necrotizing fasciitis; Neutropenia; Operative Surgical Procedures; Osteomyelitis; Outcome; Pathogenesis; Pathway interactions; Patients; Phagocytes; Phagocytosis; Phenotype; Pneumonia; Predisposition; Prevalence; Preventive; Prognosis; Proteins; Proteomics; Public Health; Recurrence; Reporting; Research; Resistance; Respiratory Tract Infections; Risk Factors; Role; STAT3 gene; Skin Tissue; Soft Tissue Infections; Staphylococcal Infections; Staphylococcus aureus; Staphylococcus aureus infection; Susceptibility Gene; Syndrome; TIRAP gene; TLR2 gene; Testing; Therapeutic; Ubiquitin; Ubiquitination; Vaccines; Variant; Virulence Factors; Weight; autoinflammatory; cohort; comorbidity; cytotoxicity; early onset; exome sequencing; factor A; genome-wide; induced pluripotent stem cell; inflammatory marker; innovation; insight; keratinocyte; kindred; loss of function; macrophage; methicillin resistant Staphylococcus aureus; monocyte; necrotizing pneumonia; novel; pathogen; pathogenic bacteria; prevent; receptor; recruit; transcriptomics; vaccine development; young adult; Abscess; Acute Respiratory Distress Syndrome; Alleles; Antibiotics; Bacterial Infections; Bacterial Toxins; Biology; Cause of Death; Caveolins; Cell Death; Cell Survival; Cells; Cellulitis; Characteristics; Child; Chromosome 5; Chromosome Arm; Communities; Cri-du-Chat Syndrome; Defect; Dermal; Devices; Disease; Endocarditis; Epidemic; Epidemiology; Etiology; Fibroblasts; Folliculitis; Genes; Genetic; Genetic Diseases; Genetic Models; Genetic Predisposition to Disease; Genotype; Genus staphylococcus; Health; Hemolysin; Hereditary Disease; Hospitalization; Human; Immune; Immunity; Individual; Infection; Inflammation; Inherited; Innate Immune Response; Interleukin-1 Receptors; Interleukin-17; Interleukin-6; Investigation; Leukocytes; Life; Light; Link; Loss of Heterozygosity; Measures; Minority; Molecular; Mutate; Mutation; NF-kappa B; Natural Immunity; Necrotizing fasciitis; Neutropenia; Operative Surgical Procedures; Osteomyelitis; Outcome; Pathogenesis; Pathway interactions; Patients; Phagocytes; Phagocytosis; Phenotype; Pneumonia; Predisposition; Prevalence; Preventive; Prognosis; Proteins; Proteomics; Public Health; Recurrence; Reporting; Research; Resistance; Respiratory Tract Infections; Risk Factors; Role; STAT3 gene; Skin Tissue; Soft Tissue Infections; Staphylococcal Infections; Staphylococcus aureus; Staphylococcus aureus infection; Susceptibility Gene; Syndrome; TIRAP gene; TLR2 gene; Testing; Therapeutic; Ubiquitin; Ubiquitination; Vaccines; Variant; Virulence Factors; Weight; autoinflammatory; cohort; comorbidity; cytotoxicity; early onset; exome sequencing; factor A; genome-wide; induced pluripotent stem cell; inflammatory marker; innovation; insight; keratinocyte; kindred; loss of function; macrophage; methicillin resistant Staphylococcus aureus; monocyte; necrotizing pneumonia; novel; pathogen; pathogenic bacteria; prevent; receptor; recruit; transcriptomics; vaccine development; young adult

PROJECT SUMMARY Staphylococcus aureus is a major bacterial pathogen impacting human health. Yet, while most subjects carry S. aureus, only a minority develop life-threatening staphylococcal disease. We hypothesize that severe or recurrent staphylococcal infections, at least in some patients, can result from single-gene inborn errors of immunity (IEI). We have already shown that IEIs in NF-kB- and TLR2-related genes predispose to staphylococcal infections. As a basis for this project, a cohort of 100 children and young adults with severe or recurrent S. aureus infections has been analyzed genetically using whole exome sequencing (WES), which has been pioneered in the lab for IEIs. Searching for candidate genotypes in a genome-wide manner, by testing various genetic models, we discovered heterozygous rare non-synonymous variants in OTULIN in 7 patients from 6 kindreds. OTULIN, a deubiquitinase, is known to regulate inflammation but not to govern anti-staphylococcal immunity. Bi-allelic deleterious variants in OTULIN cause a severe early-onset condition termed OTULIN-related autoinflammatory syndrome (ORAS). Interestingly, OTULIN is located on the short arm of the chromosome 5 and is often deleted in the 5p- (Cri-du-Chat) Syndrome, a genetic disease with a prevalence of 1/30,000. Patients with 5p- Syndrome are susceptible to bacterial infections. We hypothesize that autosomal dominant OTULIN deficiency operates by haploinsufficiency and underlies staphylococcal disease. Preliminary results show that the 6 patients’ alleles are loss-of-function and that expression of OTULIN in dermal fibroblasts from 4 patients carrying heterozygous OTULIN variants and from 1 patient with 5p- Syndrome (with a missing copy of OTULIN) is about half, when compared with healthy controls. Moreover, accumulation of linear ubiquitin chains was observed in dermal fibroblasts from 3 patients, including one with the 5p- Syndrome. Finally, we noticed an accumulation of Caveolin, which is known to facilitate recruitment of ADAM10, the receptor for the major staphylococcal virulence factor a- hemolysin. Consistently, we observed a high and selective susceptibility of patients’ fibroblasts to a-hemolysin cytotoxicity. The aim of the present application is therefore to explore in more depth the specific role of OTULIN in fibroblasts and monocytes from patients heterozygous for deleterious variants in OTULIN. We will investigate the impact of OTULIN on the linear ubiquitinome in dermal fibroblasts, with a special emphasis on NF-kB and markers of inflammation downstream TLR2 using transcriptomic and proteomics approaches. We will also investigate the innate immune responses in patients’ leukocyte subsets. Finally, we will investigate the intrinsic immunity of patients to bacterial toxins, including a-hemolysin, in the context of Caveolin dysregulation and ADAM10 expression. Overall, this study will give weight to the emerging notion that bacterial infection can preferentially trigger severe disease in the context of genetically deficient immunity. This project will shed light on the pathogenesis of staphylococcal disease and biology of OTULIN, while paving the way for novel preventive and therapeutic measures for OTULIN-mutated patients, including those with the 5p- Syndrome.

项目摘要 金黄色葡萄球菌是影响人类健康的主要细菌病原体。但是，尽管大多数受试者都带有S。 金黄色葡萄酒，只有少数派发展威胁生命的葡萄球菌疾病。我们假设这种严重或经常性 至少在某些患者中，葡萄球菌感染可能是由单基因天生的免疫力（IEI）引起的。 我们已经证明了NF-KB和TLR2相关基因中的IEI易感葡萄球菌感染。作为 该项目的基础，由100名患有严重或复发性金黄色葡萄球菌感染的儿童和年轻人组成 已经使用整个外显子组测序（WES）对一般分析，该测序已在实验室中率先进行 IEIS。通过测试各种遗传模型，以全基因组的方式搜索候选基因型，我们 在7种幼虫的7例患者中，在耳鼻蛋白中发现了杂合稀有的非同义变体。耳鼻蛋白，a 众所周知，去泛素酶调节炎症，但不能控制抗稳定局的免疫。 Bi-learleric 耳鼻蛋白的有害变体导致严重的早期发作状态，称为耳鼻蛋白相关自身炎症 综合征（ORAS）。有趣的是，耳鼻蛋白位于5号染色体的短臂上，经常被删除 在5p-（CRI-DU-CHAT）综合征中，一种遗传疾病的患病率为1/30,000。 5p-综合征患者 容易受到细菌感染的影响。我们假设常染色体显性耳鼻喉缺乏症的操作是 单倍症和葡萄球菌疾病的基础。初步结果表明，这6位患者的等位基因是 功能丧失和在4例携带杂合子的患者的皮肤成纤维细胞中耳鼻蛋白的表达 耳鼻喉蛋白变异和1名患有5p-综合征（含有耳纤维的副本）的患者约为一半 与健康对照相比。此外，在真皮中观察到线性泛素链的积累 来自3例患者的成纤维细胞，其中1例患有5p-综合征。最后，我们注意到可爱林的积累， 已知可以支持ADAM10的募集，ADAM10是主要葡萄球菌病毒因子A-的接收器 血素蛋白。一致地，我们观察到患者成纤维细胞对A-蛋白酶的高易感性 细胞毒性。因此，本应用的目的是更深入地探索耳鼻蛋白的特定作用 在来自杂合子的患者的成纤维细胞和单核细胞中，对于耳纤维蛋白的有害变体。我们将调查 耳鼻蛋白对真皮成纤维细胞中泛素组的影响，特别强调了NF-KB和 使用转录组和蛋白质组学方法下游TLR2的炎症标记。我们也会 研究患者白细胞子集的先天免疫反应。最后，我们将研究固有的 在可爱素失调和 ADAM10表达。总体而言，这项研究将使新兴通知细菌感染可以 在遗传缺陷的免疫力下，优先引发严重疾病。这个项目将熄灭 关于葡萄球菌疾病的发病机理和耳鼻蛋白的生物学，同时粘贴了新的预防性 以及耳鼻喉素氧化患者（包括患有5p-综合征的患者）的热度量。