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

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.

项目概要 金黄色葡萄球菌是影响人类健康的主要细菌病原体，而大多数受试者都携带金黄色葡萄球菌。 金黄色葡萄球菌，只有少数会患上危及生命的葡萄球菌病，我们与严重或复发性的葡萄球菌病作斗争。 至少在某些患者中，葡萄球菌感染可能是由单基因先天性免疫错误 (IEI) 引起的。 我们已经证明 NF-kB 和 TLR2 相关基因中的 IEI 容易引起葡萄球菌感染。 该项目的基础是 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的队列 使用全外显子组测序 (WES) 进行基因分析，该技术在实验室中首创 通过测试各种遗传模型，我们以全基因组的方式搜索候选基因型。 在来自 6 个亲属的 7 名患者中发现了 OTULIN 杂合的罕见非同义变异。 已知去泛素酶可以调节炎症，但不能控制抗葡萄球菌免疫。 OTULIN 的有害变异会导致严重的早发性疾病，称为 OTULIN 相关自身炎症 综合征（ORAS）提示，OTULIN 位于 5 号染色体的短臂上，经常被删除。 5p- (Cri-du-Chat) 综合征，一种遗传性疾病，5p- 综合征患者的患病率为 1/30,000。 我们认为常染色体显性 OTULIN 缺乏症是由细菌感染引起的。 单倍体不足和葡萄球菌病的基础 初步结果表明，这 6 名患者的等位基因是。 4 例携带杂合子的患者真皮成纤维细胞功能丧失和 OTULIN 表达 OTULIN 变体和 1 名 5p-综合征患者（缺失 OTULIN 副本）的比例约为一半，当 此外，与健康对照相比，在真皮中观察到线性泛素链的积累。 来自 3 名患者（其中一名患有 5p-综合征）的成纤维细胞最后，我们注意到 Caveolin 的积累， 已知它可以促进 ADAM10 的募集，ADAM10 是主要葡萄球菌毒力因子 a 的受体 一致地，我们观察到患者的成纤维细胞对α-溶血素具有高度和选择性的敏感性。 因此，本申请的目的是更深入地探讨OTULIN的具体作用。 我们将研究 OTULIN 杂合子患者的成纤维细胞和单核细胞中的有害变异。 OTULIN 对真皮成纤维细胞线性泛素组的影响，特别强调 NF-kB 和 我们还将使用转录组学和蛋白质组学方法来确定 TLR2 下游炎症标志物。 研究患者白细胞亚群的先天免疫反应最后，我们将研究内在的免疫反应。 在 Caveolin 失调的情况下，患者对细菌毒素（包括α-溶血素）的免疫力 总体而言，这项研究将证实细菌感染可以影响这一新兴概念。 在遗传性免疫力缺陷的情况下优先引发严重疾病该项目将揭示这一点。 研究葡萄球菌疾病的发病机制和 OTULIN 的生物学，同时为新型预防措施铺平道路 OTULIN 突变患者（包括 5p-综合征患者）的治疗措施。