Genetics Of The Dominantly Inherited Periodic Fever Syndromes

显性遗传性周期性发热综合征的遗传学

• 批准号: 8175276
• 负责人: Daniel Kastner
• 金额: $ 38.8万
• 依托单位: NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8175276
• 关键词: Acne; Acute suppurative arthritis due to bacteria; Amyloidosis; Animal Model; Animals; Arthritis; Aseptic Meningitis; Binding; Biology; Cell Adhesion; Cell Death; Cell Line; Cell membrane; Cells; Child; Chimeric Proteins; Chronic; Chronic Disease; Classification; Clinical; Conjunctivitis; Cutaneous; Data Set; Defect; Development; Disease; Edema; Etanercept; Exanthema; Familial Mediterranean Fever; Familial amyloid nephropathy with urticaria and deafness; Family; Fever; Fibroblasts; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Goals; Homeostasis; Human; Immune response; Inflammation; Inflammatory; Inherited; Interleukin-1; Interleukin-1 beta; Knock-out; Lead; Location; Macromolecular Complexes; Manuscripts; Mental Retardation; Modeling; Molecular; Mononuclear; Mus; Mutant Strains Mice; Mutate; Mutation; Myalgia; Myeloid Cells; Names; Neonatal; Neurologic; Oxidative Stress; Paper; Papilledema; Pathogenesis; Pathway interactions; Patients; Pattern; Peripheral; Peripheral Blood Mononuclear Cell; Proline; Protein Serine/Threonine Phosphatase; Proteins; Publishing; Pyoderma Gangrenosum; Reactive Oxygen Species; Recombinant Interleukin-1; Recurrence; Regulation; Relative (related person); Resistance; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Sampling Studies; Sensorineural Hearing Loss; Septic Shock; Serositis; Skin Ulcer; Sterility; Surface; Syndrome; TNF gene; TNFRSF1A gene; Transcript; Treatment Failure; Tumor Necrosis Factor Receptor; Urticaria; Uveitis; anakinra; autocrine; base; cell motility; cell type; clinical remission; cytokine; disease phenotype; human MPP1 protein; infancy; inhibitor/antagonist; knockin animal; long bone; marenostrin; mutant; new technology; partial response; receptor; receptor binding; transcriptional coactivator p75

Background During the last several years we have studied five different dominantly-inherited autoinflammatory disorders. The first of these illnesses is the TNF receptor-associated periodic syndrome (TRAPS), which is characterized by prolonged attacks of fever, serositis, migratory rash and myalgia, arthritis, periorbital edema, conjunctivitis, and, in some patients, systemic amyloidosis. In 1999 we identified the first mutations in the gene (TNFRSF1A) encoding the 55 kDa tumor necrosis factor receptor (TNFR1) in families with dominantly-inherited recurrent fevers, and proposed the name TRAPS for this clinical condition. Initial mechanistic studies indicated a defect in the activation-induced shedding of the p55 (but not p75) TNF receptor, possibly leading to impaired homeostasis. Findings we published in the last year suggest an important additional mechanism of inflammation in these patients (see below). In 2002 we discovered dominantly-inherited de novo mutations in a second gene, initially named CIAS1 but now officially denoted NLRP3, in about 50% of patients with a disorder known as neonatal onset multisystem inflammatory disease (NOMID) or chronic infantile neurologic cutaneous and articular (CINCA) syndrome. Manifestations of NOMID/CINCA may include daily fevers, an urticaria-like skin rash, chronic aseptic meningitis, uveitis, papilledema, sensorineural hearing loss, mental retardation, patellar and epiphyseal long bone overgrowth, and systemic amyloidosis. Two milder conditions, familial cold autoinflammatory syndrome (FCAS) and Muckle-Wells syndrome (MWS), are caused by mutations in the same gene. NLRP3 encodes a protein, cryopyrin, that participates in a macromolecular complex called the inflammasome to regulate the activation of interleukin-1 (IL-1) beta. Collectively, all three diseases are known as the cryopyrin-associated periodic syndromes (CAPS). CAPS patients respond dramatically to inhibitors of IL-1 beta. A fifth dominantly-inherited autoinflammatory disorder, denoted the syndrome of pyogenic arthritis with pyoderma gangrenosum and acne (PAPA), is caused by mutations in a protein known as proline serine threonine phosphatase interacting protein (PSTPIP1). PAPA is characterized by episodes of sterile pyogenic arthritis, which can be destructive if not treated, formation of open, purulent ulcers of the skin (pyoderma gangrenosum), and severe cystic acne. In 2003 our group discovered that PSTPIP1 binds pyrin, the protein mutated in familial Mediterranean fever (FMF), and that disease-associated mutations in PSTPIP1 lead to more avid binding to pyrin, and increased IL-1 beta activation, relative to healthy controls. Progress During the Last Year Mechanism of inflammation in TRAPS: In May of this year we published a paper in PNAS that was the culmination of a several-year collaborative effort with Dr. Richard Siegel's group involving transfected cells, TRAPS knockin mice, and cells from TRAPS patients. Previous studies from our groups and others had shown that although there is the above-noted defect in ectodomain-cleavage of TRAPS-mutant TNF receptors, this is seen to a variable degree with different mutations and is cell-type dependent. Moreover, these mutant receptors bind TNF less well than wild-type receptors, and thus even if they do persist on the cell membrane after activation, it is not clear that retention of these receptor molecules would lead to repetitive stimulation, as we had originally hypothesized. Taken in the context of longer-term treatment failures with etanercept, the soluble p75:Fc fusion protein, in TRAPS, the pathogenesis of TRAPS has been an enigma. In the manuscript we published this year, we demonstrated that mutant TNFR1 accumulates intracellularly in peripheral blood mononuclear cells of TRAPS patients and in multiple cell types from two independent lines of knockin mice harboring TRAPS-associated TNFR1 mutations. Mutant TNFR1 did not function as a surface receptor for TNF but rather enhanced activation of MAPKs and secretion of proinflammatory cytokines upon stimulation with LPS. Enhanced inflammation depended on autocrine TNF secretion and WT TNFR1 in mouse and human myeloid cells but not in fibroblasts. Heterozygous TNFR1-mutant mice were hypersensitive to LPS-induced septic shock, whereas homozygous TNFR1-mutant mice resembled TNFR1-deficient mice and were resistant to septic shock. Thus WT and mutant TNFR1 act in concert from distinct cellular locations to potentiate inflammation in TRAPS. These findings establish a mechanism of pathogenesis in autosomal dominant diseases where full expression of the disease phenotype depends on functional cooperation between WT and mutant proteins and also may explain partial responses of TRAPS patients to TNF blockade. Gene expression profiling in CAPS: During the last year we completed a gene expression study in CAPS and have prepared a manuscript for submission. To understand the pathogenesis of CAPS we compared gene expression patterns in peripheral blool mononuclear cells (PBMCs) from 22 patients with active disease and 14 healthy children. We collected 16 paired samples from CAPS patients before and after anakinra (recombinant IL-1 receptor antagonist) treatment to identify transcripts responsive to IL-1 beta inhibition. We identified a gene expression signature that clearly distinguished CAPS patients from healthy controls. Many of the differentially expressed genes (DEGs) include transcripts related to regulation of innate and adaptive immune responses, oxidative stress, cell death, cell adhesion, and motility. Several DEGs, including transcripts related to regulation of reactive oxygen species (ROS), were validated by quantitative RT-PCR and by a functional study with primary cells from MWS patients and healthy controls. Using 17 CAPS samples and 66 non-CAPS samples, we created a set of gene expression-based models that differentiates CAPS patients from controls and from patients with other systemic inflammatory conditions. The CAPS-specific gene expression signature correctly classified all 17 samples from an independent dataset. This classifier also correctly identified 15 of 16 post-anakinra CAPS samples despite the fact that these CAPS patients were in clinical remission, thus validating the concept of gene expression profiling in the classification of inflammatory diseases and raising the possibility of IL-1 independent pathways in CAPS pathogenesis.

背景 在过去的几年中，我们研究了五种不同的自身炎症性疾病。 这些疾病中的第一个是TNF受体相关的周期综合征（诱捕症），其特征是发烧，血清炎，迁移性皮疹和肌痛，关节炎，周围性水肿，结膜炎，结膜炎，以及在某些患者中，全身性淀粉样蛋白病。 在1999年，我们确定了编码55 kDa肿瘤坏死因子受体（TNFR1）的基因（TNFRSF1A）中的第一个突变。 最初的机械研究表明，p55（但不是p75）TNF受体的激活诱导的脱落缺陷，可能导致稳态受损。 我们在去年发表的研究结果表明，这些患者的炎症有一种重要的额外机制（见下文）。 2002年，我们在第二个基因中发现了从头突变，最初命名为CIAS1，但现在正式表示NLRP3，大约有50％的患者被称为新生儿发作多系统炎症性疾病（Nomid）或慢性婴儿神经学和心肌（cincabal（Cinca）综合征。 Nomid/CINCA的表现可能包括每日发烧，类似荨麻疹的皮疹，慢性无菌脑膜炎，葡萄膜炎，乳头毛症，感觉神经性听力损失，智力低下，tell骨和epiphyseal长骨过度生长和全身性淀粉样蛋白病。 两种温和的条件：家族性冷自身炎症综合征（FCAS）和粘液 - 孔综合征（MWS）是由同一基因突变引起的。 NLRP3编码一种蛋白质冷冻蛋白，该蛋白参与称为炎性体的大分子复合物，以调节白介素-1（IL-1）β的激活。 总的来说，这三种疾病都被称为冷冻蛋白相关的周期性综合征（CAPS）。 CAPS患者对IL-1β的抑制剂做出了巨大反应。 第五个主要亲属的自发性疾病，表示脓虫性腺肿和痤疮（PAPA）的化脓性关节炎综合征是由称为脯氨酸丝氨酸丝氨酸苏氨酸磷酸酶相互作用的蛋白质相互作用的蛋白质引起的（PSTPIP1）。 爸爸的特征是无菌的化脓性关节炎发作，如果不治疗，可能具有破坏性，形成开放的皮肤化学溃疡（脓皮肾上腺坏疽）和严重的囊肿痤疮。 2003年，我们的小组发现PSTPIP1结合了pyrin，在家族性地中海发烧中突变的蛋白质（FMF），并且PSTPIP1中与疾病相关的突变导致与吡啶的结合更加狂热，而IL-1 Beta激活增加了。 去年的进步 陷阱中的炎症机制：今年5月，我们在PNAS中发表了一篇论文，这是与理查德·西格尔（Richard Siegel）博士的涉及转染细胞，陷阱敲击小鼠和陷阱患者的细胞进行了数年的合作努力的高潮。 我们小组和其他人的先前研究表明，尽管陷阱突变的TNF受体的外域裂解存在上述缺陷，但这被视为具有不同突变的可变程度，并且依赖于细胞类型。 此外，这些突变受体的结合比野生型受体的结合良好，因此，即使激活后它们确实持续在细胞膜上，也不清楚这些受体分子的保留会导致重复刺激，因为我们最初假设。 在依然切齿中长期治疗失败的背景下，可溶性p75：FC融合蛋白在陷阱中，陷阱的发病机理一直是一个谜。 在我们今年发表的手稿中，我们证明了突变体TNFR1在陷阱患者的外周血单核细胞中累积，以及来自两种具有陷阱相关TNFR1突变的敲蛋白小鼠的多种细胞类型的细胞类型。 突变体TNFR1不充当TNF的表面受体，而是在用LPS刺激后增强了MAPK的激活和促炎细胞因子的分泌。 增强的炎症取决于小鼠和人髓样细胞中的自分泌TNF分泌和WT TNFR1，但不在成纤维细胞中。 杂合子TNFR1突变小鼠对LPS引起的败血性休克过敏，而纯合TNFR1突变小鼠类似于TNFR1缺陷型小鼠，并且对败血性休克有抗性。 因此，WT和突变体TNFR1从不同的细胞位置到陷阱中的炎症进行协同作用。 这些发现在常染色体显性疾病中建立了一种发病机理的机制，在这些疾病中，疾病表型的完全表达取决于WT和突变蛋白之间的功能合作，并且也可以解释陷阱患者对TNF阻滞的部分反应。 CAPS中的基因表达分析：在去年，我们在CAPS中完成了一项基因表达研究，并准备了一个手稿以提交。 为了了解帽的发病机理，我们比较了22例活性疾病和14名健康儿童的外周白单核细胞（PBMC）中的基因表达模式。 我们在Anakinra（重组IL-1受体拮抗剂）治疗前后从CAPS患者中收集了16个配对样品，以鉴定对IL-1β抑制作用的转录本。 我们确定了一个基因表达特征，该特征将CAPS患者与健康对照区分开。 许多差异表达的基因（DEG）包括与先天和适应性免疫反应，氧化应激，细胞死亡，细胞粘附和运动性有关的转录本。 通过定量RT-PCR以及对MWS患者和健康对照组的原代细胞的功能研究来验证了几个DEG，包括与活性氧（ROS）有关的转录本。 我们使用17个盖子样品和66个非CAP样品，创建了一组基于基因表达的模型，这些模型将CAPS患者与对照组和具有其他全身性炎症状况的患者区分开来。 CAPS特异性基因表达签名正确分类了来自独立数据集的所有17个样本。 尽管这些CAPS患者处于临床缓解状态，但该分类器还正确地确定了16个Anakinra帽样品中的15个，从而验证了炎症性疾病分类中基因表达分析的概念，并提高了CAPS病原体中IL-1独立途径的可能性。