Genetics Of The Dominantly Inherited Periodic Fever Synd

显性遗传性周期性发热综合征的遗传学

• 批准号: 7319625
• 负责人: Daniel L Kastner
• 金额: --
• 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/7319625
• 关键词: Genetics; Dominantly; Inherited; Periodic; Fever

During the last several years our laboratory has studied three different dominantly-inherited autoinflammatory disorders. The first of these illnesses is the TNF receptor-associated periodic syndrome (TRAPS), which is characterized by oftentimes prolonged attacks of fever, serositis, migratory rash and myalgia, arthritis, periorbital edema, conjunctivitis, and, in some patients, systemic amyloidosis. In 1999 our group identified the first six mutations in the gene encoding the 55 kDa tumor necrosis factor receptor (TNFRSF1A) in seven families with dominantly-inherited recurrent fevers, and therefore 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 of the immune response. In 2002 we and a French group independently discovered dominantly-inherited de novo mutations in a second gene, CIAS1 (also known as NALP3 or PYPAF1), 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 phenotypically milder conditions, familial cold autoinflammatory syndrome (FCAS) and Muckle-Wells syndrome (MWS), are caused by mutations in the same gene. CIAS1 encodes a protein, cryopyrin, that participates in a macromolecular complex called the inflammasome to regulate the activation of interleukin-1 (IL-1) beta. A third 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 mono- or pauciarticular 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. Results of the Last Year Functional studies of mutant TNFRSF1A molecules: In collaboration with the laboratory of Dr. Richard Siegel, we have continued analyses of the functional consequences of TRAPS-associated TNFRSF1A mutations. In transfection systems, mutant receptors were found to have a number of functional abnormalities, including (1) impaired activation-induced ectodomain cleavage; (2) diminished binding to TNF; (3) lack of association with wild-type receptors through the pre-ligand assembly domain (PLAD); (4) impaired ability to signal apoptosis or NF kappa B activation; and (5) abnormal intracellular trafficking, with retention in the endoplasmic reticulum (ER). Current studies focus on the response of mouse embryonic fibroblasts from TRAPS knockin mice to ER stress. Studies of cytokine production by peripheral blood mononuclear cells from TRAPS patients show heightened production in response to low doses of LPS, again suggesting exaggerated inflammatory responses to subclinical stimuli. Mutational analysis in patients with clinical FCAS, MWS, or NOMID: In collaboration with Dr. Hal Hoffman, we screened new referrals for CIAS1 mutations. Of the 22 NOMID/CINCA patients, 5 had novel CIAS1 mutations (V262A, V351L, F443L, and 2 with L264F), 7 were carriers for previously described mutations, and 10 were mutation-negative. Of the 12 MWS patients, 1 had a novel mutation (F523C), 8 had previously-described mutations, and 3 were mutation-negative. Among the 18 FCAS patients, 3 were carriers for 2 novel mutations (C259W and Y563N), 13 were carriers for previously described mutations, and 2 were mutation-negative. Several mutation-negative patients were screened for mutations in 12 candidate genes chosen for similarity to CIAS1 or involvement in the caspase-1/IL-1 beta signaling pathway; no pathogenic changes were found. We also assessed the frequency of 3 known CIAS1 variants of uncertain significance, V198M, R488K, and Q703K, in the general population. Although the allele frequencies of the first two variants were less than 1%, suggesting that they may be reduced-penetrance mutations, the allele frequency of Q703K was 5%, indicating that it is unlikely to be a pathogenic change. Computational modeling of the significant disease-causing mutations suggested a structure in which the NACHT domains of 6 cryopyrin molecules form a hexagonal assembly, with nearly all of the mutations on a common surface. Studies of IL-1 inhibition in the cryopyrinopathies: In collaboration with Dr. Raphaela Goldbach-Mansky, we have conducted therapeutic trials of an FDA-approved recombinant IL-1 receptor antagonist, anakinra, in 18 patients with NOMID. The results are more thoroughly presented under project Z01 AR041138-04 OCD, but will be briefly summarized here. In the anakinra trial, a total of 18 NOMID patients were treated with anakinra; in 11, the drug was briefly withdrawn after 3 months until a flare occurred. Clinical and laboratory parameters improved dramatically while on study drug, withdrawal resulted in relapse within days, and retreatment led to rapid improvement. There were highly significant improvements in a disease-specific diary score at 3 and 6 months of treatment, as well as in the erythrocyte sedimentation rate, C-reactive protein, and serum amyloid A. There was also evidence of reduced inflammation of the central nervous system, including decreased headache, reduced opening pressure on lumbar puncture, and decreased cochlear and leptomeningeal enhancement on magnetic resonance imaging (MRI). Before treatment peripheral blood leukocytes from patients spontaneously produced high levels of IL-1 beta, and this decreased progressively with treatment. Gene expression profiling also revealed aberrantly elevated expression of IL-1 beta and genes downstream of IL-1 beta prior to treatment, which decreased on anakinra and returned with drug withdrawal. We are continuing the anakinra trial to study long-term safety and efficacy. We are also currently studying a longer-acting investigational soluble IL-1 receptor fusion protein in patients with FCAS/MWS. Generation of new animal models: We are currently in the process of developing new animal models for the cryopyrinopathies and PAPA syndrome. In collaboration with Dr. Hal Hoffman, we are developing models for FCAS (CIAS1 L3533P), MWS (A352V), and NOMID (D303N, Y570C). We are also generating PSTPIP1 knockout mice, and the PSTPIP1 E250Q PAPA knockin mouse. Conclusions and Significance Studies on the functional consequences of TNFRSF1A mutations suggest a more complex pathophysiologic model of TRAPS than previously appreciated, in which constitutive cellular activation may play a prominent role. Our data on CIAS1 mutations expand the mutational spectrum of the cryopyrinopathies, and suggest hypotheses for the structural implications of CIAS1 mutations. Therapeutic studies of IL-1 inhibitors in NOMID patients confirm the importance of cryopyrin in IL-1 regulation, and represent a major advance in the treatment of these patients. During the next year we will continue studies of the possible role of the ER stress response in TRAPS, continue therapeutic and molecular pathophysiologic studies of patients with cryopyrinopathies, and focus on new animal models as they become available.

在过去的几年中，我们的实验室研究了三种不同的自身炎症性疾病。这些疾病中的第一种是与TNF受体相关的周期综合征（陷阱），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，以及在某些患者中，全身性干性病。在1999年，我们的小组确定了编码55 kDa肿瘤坏死因子受体（TNFRSF1A）的基因中的前六个突变，其中七个具有巨大亲发复发的家族，因此提出了名称？陷阱？对于这种临床状况。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，可能导致免疫反应的稳态受损。 在2002年，我们和一个法国群体独立发现了第二个基因CIAS1（也称为Nalp3或Pypaf1）中的主要DE突变，其中约50％的患者被称为新生儿多系统炎症性疾病（NomID）或慢性婴儿神经系统性皮肤疾病和空意性（Cincane）和空意（Cinccinal）（cincancular and cincaular and cincaular and cincanculor）。 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。两种表型温和的条件：家族性冷自身炎症综合征（FCA）和粘液 - 韦尔斯综合征（MWS）是由同一基因突变引起的。 CIAS1编码一种参与称为炎性体的大分子复合物的蛋白质，冷冻蛋白，以调节白介素-1（IL-1）β的激活。 第三种主要结构的自身炎症疾病，表示脓虫性腺坏疽和痤疮（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中的突变引起的（PSTPIP1）。爸爸的特征是单或pauciar骨无菌性化脓性关节炎发作，如果不治疗，可能具有破坏性，形成开放的，化脓性的溃疡（胸腺肿瘤）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。 去年的结果 突变TNFRSF1A分子的功能研究：与Richard Siegel博士的实验室合作，我们继续分析与陷阱相关的TNFRSF1A突变的功能后果。在转染系统中，发现突变受体具有多种功能异常，包括（1）激活诱导的外域裂解； （2）与TNF的结合减少； （3）通过配体组装域（plad）与野生型受体缺乏关联； （4）信号凋亡或NF Kappa B激活的能力受损； （5）异常的细胞内运输，并保留在内质网（ER）。当前的研究集中于小鼠胚胎成纤维细胞从陷阱敲击蛋白小鼠对ER应激的反应。周围血液单核细胞对陷阱患者的细胞因子产生的研究表明，对低剂量LP的产生增强，再次表明对亚临床刺激的炎症反应夸大。 临床FCA，MWS或NOMID患者的突变分析：与Hal Hoffman博士合作，我们筛选了CIAS1突变的新推荐。在22名Nomid/CINCA患者中，有5例具有新颖的CIAS1突变（V262A，V351L，F443L和2例L264F），有7个是用于先前描述的突变的携带者，而10个是突变阴性的。在12个MWS患者中，有1例具有新的突变（F523C），其中8例具有先前描述的突变，而3例是突变阴性。在18名FCA患者中，有3例是2个新型突变（C259W和Y563N）的载体，13例是先前描述的突变的载体，而2例是突变阴性。将几名突变阴性患者筛选为12个候选基因的突变，以与CIAS1相似或参与CASPASE-1/IL-1β信号传导途径。未发现致病性变化。我们还评估了一般人群中3个已知的CIAS1变体的频率，V198M，R488K和Q703K的频率。尽管前两个变体的等位基因频率小于1％，这表明它们可能是渗透突变，但Q703K的等位基因频率为5％，表明这不太可能是一种致病性的变化。严重引起疾病的突变的计算模型表明了一种结构，其中6个冷冻蛋白分子的NACHT结构域形成六边形组件，几乎所有突变都在公共表面上。 Cryopinopathies中IL-1抑制作用的研究：与Raphaela Goldbach-Mansky博士合作，我们对FDA批准的重组IL-1受体拮抗剂Anakinra，Anakinra进行了治疗试验。结果Z01 AR041138-04 OCD更彻底地提出了结果，但将在此处简要概述。在Anakinra试验中，总共有18名Nomid患者接受了Anakinra治疗。 3个月后，该药物在11点被短暂撤回，直到发生爆发。临床和实验室参数在学习药物的过程中显着改善，戒断导致几天内复发，而撤退导致了快速改善。在3个月和6个月的治疗中，以及红细胞沉降率，C反应蛋白和血清淀粉样蛋白A的疾病特异性日记评分方面的表现也有很大显着的改善。也有证据表明，中枢神经系统的炎症降低，包括降低的腹部凝聚力和较低的CONCTURE和cORCHINENINGENECENENENING SENCONENERING SENENENERING SERMENTING SENSENEN和降低。在治疗外周血白细胞之前，患者会自发产生高水平的IL-1β，并且随着治疗而逐渐减少。基因表达分析还显示，在治疗之前，IL-1β和IL-1β下游基因的表达异常升高，该基因在Anakinra上降低并随药物戒断而返回。我们正在继续进行Anakinra试验，以研究长期的安全性和功效。我们目前还正在研究FCAS/MWS患者的长效研究可溶性IL-1受体融合蛋白。 新动物模型的产生：我们目前正在开发用于冷冻病和爸爸综合征的新动物模型。与Hal Hoffman博士合作，我们正在开发FCAS（CIAS1 L3533P），MWS（A352V）和NOMID（D303N，Y570C）的模型。我们还生成PSTPIP1基因敲除小鼠和PSTPIP1 E250Q PAPA敲除鼠标。 结论和意义 关于TNFRSF1A突变功能后果的研究表明，陷阱的病理生理模型比以前所欣赏的更复杂的病理生理模型，其中本构细胞激活可能起重要作用。我们关于CIAS1突变的数据扩大了冷冻膜病的突变谱，并提出了CIAS1突变的结构含义的假设。 NOMID患者IL-1抑制剂的治疗研究证实了冷冻蛋白在IL-1调节中的重要性，并且代表了这些患者治疗的主要进步。在第二年，我们将继续研究核心应力反应在陷阱中的可能作用，继续对冷冻培养基病患者的患者进行治疗和分子病理生理学研究，并在新动物模型中侧重于可用。