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

基本信息

批准号：
7732808
负责人：
Daniel L Kastner
金额：
$ 97.79万
依托单位：
NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
依托单位国家：
美国
项目类别：
财政年份：
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/7732808
关键词：
Acne Acute suppurative arthritis due to bacteria Affect Aftercare Amyloidosis Animal Model Animals Arthritis Aseptic Meningitis Autoimmune Diseases Automobile Driving Binding Biology Birth Breeding Cell Line Cells Child Chronic Chronic Disease Clinical Collaborations Condition Conjunctivitis Cutaneous Daily Data Data Analyses Data Set Defect Development Disease Disease remission Edema Endoplasmic Reticulum Engineering Enzymes Exanthema Exhibits Familial Mediterranean Fever Familial amyloid nephropathy with urticaria and deafness Family Fever Gene Expression Gene Expression Profiling Generations Genes Genetic Goals Growth Hair Homeostasis Human Hypersensitivity Immune In Situ Individual Inflammation Inflammatory Inherited Interleukin-1 Interleukin-1 beta Interleukin-6 Knock-out Knockout Mice Laboratories Lead Leukocytes Life MAP Kinase Gene MAPK14 gene MAPK8 gene Macromolecular Complexes Manuscripts Mental Retardation Mitogen-Activated Protein Kinases Modeling Molecular Molecular Profiling Mouse Cell Line Mus Mutant Strains Mice Mutate Mutation Myalgia NADPH Oxidase NF-kappa B Names Neonatal Neurologic Papilledema Pathogenesis Pathway interactions Patients Pattern Peripheral Blood Mononuclear Cell Pharmacologic Substance Phenotype Process Production Proline Protein Serine/Threonine Phosphatase Proteins Pyoderma Gangrenosum Reaction Reactive Oxygen Species Recurrence Regulation Relative (related person)Reporting Reverse Transcriptase Polymerase Chain Reaction Role Sampling Sampling Studies Sensorineural Hearing Loss Serositis Signal Transduction Skin Skin Ulcer Staining method Stains Sterility Syndrome System TNF gene TNFRSF1A gene TXN gene Therapy Clinical Trials Tissues Transfection Tumor Necrosis Factor Receptor Urticaria Uveitis Wild Type Mouse anakinra base cytokine day human CYBA protein human MPP1 protein infancy inhibitor/antagonist knockin animal long bone macrophage marenostrin mutant neutrophil cytosol factor 40K new technology pup receptor response trafficking transcriptional coactivator p75

项目摘要

Background During the last several years we have studied five different dominantly-inherited autoinflammatory/ autoimmune 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 encoding the 55 kDa tumor necrosis factor receptor (TNFRSF1A) 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 immune homeostasis. In 2002 we 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 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. Collectively, all three diseases are known as the cryopyrin-associated periodic syndromes (CAPS). 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. 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. Studies performed during previous reporting periods have demonstrated that the aforementioned defect in TNF receptor ectodomain cleavage is unlikely to explain all or most of the autoinflammatory phenotype, and in transfection systems the mutant receptors have been found to exhibit diminished binding to TNF and abnormal intracellular trafficking, with retention in the endoplasmic reticulum (ER). During the present reporting period we have continued mechanistic studies in primary patient leukocytes, TRAPS knockin mice, and cell lines, and have recently submitted a manuscript summarizing our findings. In this manuscript we demonstrated that mutant TNFRSF1A protein accumulates intracellularly in knockin mice with two independent TRAPS-associated TNFRSF1A mutations and in TRAPS patient peripheral blood mononuclear cells. Cells harboring TRAPS-associated TNFRSF1A mutations exhibited spontaneous enhanced p38 and JNK MAP-kinase activation in response to LPS, while NF-kappa B activation was not affected. These signaling abnormalities led to increased production of pro-inflammatory cytokines in immune cells from knockin mice and TRAPS patients. Heterozygous TRAPS knockin mice exhibited hypersensitivity to LPS, while homozygous TNFRSF1A mutant mice had a phenotype identical to TNFRSF1A knockout mice. Hypersecretion of cytokines by heterozygous TNFRSF1A mutant cells was dependent on reactive oxygen species (ROS), most likely generated by NADPH oxidase (NOX) enzymes. These results define a pathway of MAPK and ROS-dependent inflammation triggered by intracellularly accumulated TNFRSF1A in TRAPS. Full expression of the inflammatory phenotype in TRAPS depends on the wild-type TNFRSF1A, although the wild-type and mutant receptors do not physically associate. Inhibiting ROS may synergize with TNF blockade in the treatment of this and potentially other autoinflammatory diseases. Gene expression profiling in cryopyrin-associated periodic syndromes (CAPS): We have continued studies begun in the previous reporting period concerning gene-expression profiling in CAPS patients before and after treatment with the IL-1 receptor antagonist, anakinra, and compared with healthy controls. CAPS patients respond dramatically to anakinra, consistent with a central role for IL-1 in clinical manifestations. To understand better the pathogenesis of CAPS we compared gene expression patterns in peripheral blood mononuclear cells (PBMCs) from 22 CAPS patients and 14 healthy children. We also collected 16 paired samples from CAPS patients before and after anakinra to identify genes that are highly responsive to IL-1beta inhibition. Despite stringent data analysis many highly differentially expressed genes were found in both data sets. Quantitative RT-PCR was performed on selected differentially expressed genes to confirm the microarray data. We validated several genes that are involved in regulation of reactive oxygen species (ROS) including p22phox, p40phox, TXN, and TXNIP. Treatment of PBMCs from CAPS patients and healthy controls with DPI, an inhibitor of NADPH oxidase, showed that ROS signaling was necessary for IL-1beta production in PBMC. A gene expression-based model was generated using a subset of 16 CAPS samples and 66 non-CAPS samples (including healthy controls and individuals with other autoinflammatory disorders) to differentiate CAPS patients from controls and other patients with systemic inflammation. The CAPS-specific gene expression model correctly classified 6/6 independent pre-anakinra CAPS samples and 11/11 independent non-CAPS samples. This classifier also correctly identified 15/16 post-anakinra CAPS samples despite the fact that the CAPS patients were in clinical remission. Studies of IL-1 inhibition in the cryopyrinopathies: In collaboration with Dr. Raphaela Goldbach-Mansky, we continue therapeutic trials of anakinra and a longer-acting IL-1 inhibitor manufactured by Regeneron pharmaceuticals and denoted IL-1 Trap. These studies are more thoroughly presented under project Z01 AR041138-06 OCD. Generation of new animal models: In collaboration with Dr. Hal Hoffman, we have engineered mice carrying the Muckle-Wells syndrome mutation, A352V. Pups expressing this mutation show profound growth retardation, hair loss, skin scaling, and die about ten days after birth. Massive neutrophilic influxes in multiple tissues, as well as intense in situ staining for IL-1beta and IL-6, suggest an uncontrolled inflammatory reaction. In culture, macrophages from knockin mice secrete IL-1beta in response to LPS, but do not require secondary stimulation with ATP as is the case for cells from wild-type mice. IL-1beta appears to be the main cytokine driving the disease process in these mice, in that knockins bred onto an IL-1beta receptor knockout background are disease-free. Treatment with the IL-1beta inhibitor, IL-1 Trap, extends the lives of knockin mice by about four days. We have also generated PSTPIP1 knockout mice, and the PSTPIP1 E250Q PAPA knockin mouse. Studies to characterize the phenotype of these mice are now under way.

背景在过去的几年中，我们研究了五种不同的自身炎症/自身免疫性疾病。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（TNFRSF1A）的基因中的第一个突变，该家族具有显着的复发性发烧，并提出了这种临床状况的名称陷阱。最初的机械研究表明，p55（但不是p75）TNF受体的激活引起的脱落缺陷，可能导致免疫稳态受损。在2002年，我们在第二个基因CIAS1（也称为NALP3或PYPAF1）中发现了大约50％的新生儿发作多发性多系统炎症性疾病（NOMID）或慢性婴儿神经学性皮肤和心肌（CINCA）综合症的患者中，发现了从头开始突变。 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。 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。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）。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。在先前的报道期间进行的研究表明，TNF受体外生蛋白蛋白裂解中的上述缺陷不太可能解释全部或大多数自身炎性表型，而在转染系统中，突变受体已发现突变受体可与TNF的结合表现出对TNF的结合，并依次进行了幼体的幼虫，并依次将其递减。在本报告期间，我们在原发性患者白细胞，陷阱蛋白小鼠和细胞系进行了持续的机械研究，最近提交了一份手稿，总结了我们的发现。在本手稿中，我们证明了突变体TNFRSF1A蛋白在敲击蛋白小鼠中积聚了两种独立的陷阱相关的TNFRSF1A突变和患者患者外周血单核细胞的细胞内积聚。具有陷阱相关的TNFRSF1A突变的细胞表现出自发增强的p38和JNK MAP-激酶激活，而NF-KAPPA B激活不受影响。这些信号异常导致敲击蛋白小鼠免疫细胞中促炎细胞因子的产生增加，并捕获患者。杂合诱捕蛋白小鼠对LPS表现出超敏反应，而纯合TNFRSF1A突变小鼠的表型与TNFRSF1A敲除小鼠相同。杂合TNFRSF1A突变细胞对细胞因子的过度分泌取决于活性氧（ROS），这很可能是由NADPH氧化酶（NOX）酶产生的。这些结果定义了由陷阱中细胞内积累的TNFRSF1A触发的MAPK和ROS依赖性炎症的途径。尽管野生型和突变受体没有物理关联，但陷阱中炎症表型的完全表达取决于野生型TNFRSF1A。抑制ROS可能会与TNF阻滞协同处理，以治疗这种和潜在的其他自发性疾病。冷冻蛋白相关的周期综合征（CAPS）中的基因表达分析：我们在上一个报告期开始进行研究，涉及与IL-1受体拮抗剂Anakinra治疗前和治疗后CAPS患者的基因表达分析，并且与健康对照相比。 CAPS患者对Anakinra的反应很大，与IL-1在临床表现中的核心作用一致。为了更好地了解帽的发病机理，我们比较了22名CAPS患者和14名健康儿童的外周血单核细胞（PBMC）中的基因表达模式。我们还收集了Anakinra之前和之后的16个配对样品，以鉴定对IL-1BETA抑制作用高度响应的基因。尽管存在严格的数据分析，但在两个数据集中都发现了许多高度差异表达的基因。对选定的差异表达基因进行定量RT-PCR以确认微阵列数据。我们验证了几个与活性氧（ROS）调节有关的基因，包括P22Phox，P40Phox，TXN和TXNIP。使用NADPH氧化酶的抑制剂DPI对来自CAPS患者的PBMC和健康对照组的治疗表明，ROS信号对于PBMC中的IL-1BETA产生是必需的。使用16个盖子样品和66个非CAPS样品（包括健康对照组和患有其他自身炎症性疾病的个体）的子集生成基于基因表达的模型，以将帽子患者与对照组和其他全身性炎症患者区分开。 CAPS特异性基因表达模型正确地分类了6/6个独立的前Anakinra Caps样品和11/11独立的非CAPS样品。尽管CAPS患者处于临床缓解状态，但该分类器还正确地确定了15/16后 - Anakinra Caps样品。对冷冻培养基病中IL-1抑制作用的研究：与Raphaela Goldbach-Mansky博士合作，我们继续对Anakinra进行治疗试验，以及由Regeneron Pharmaceuticals生产的长期作用IL-1抑制剂，并表示IL-1 TRAP。这些研究在Z01 AR041138-06 OCD下更彻底地介绍了。新动物模型的产生：与Hal Hoffman博士合作，我们设计了携带Muckle-Wells综合征突变的小鼠A352V。表达这种突变的幼崽显示出深刻的生长迟缓，脱发，皮肤缩放和出生后十天死亡。多种组织中的大量嗜中性粒细胞流入，以及IL-1Beta和IL-6的强烈原位染色表明炎症反应不受控制。在培养中，敲蛋白小鼠的巨噬细胞对LPS分泌IL-1Beta，但不需要对ATP进行次要刺激，就像来自野生型小鼠的细胞一样。 IL-1BETA似乎是驱动这些小鼠疾病过程的主要细胞因子，因为将敲打素繁殖到IL-1BETA受体敲除背景上的背景不含病。用IL-1BETA抑制剂IL-1 TRAP治疗将敲击蛋白小鼠的寿命延长了大约四天。我们还产生了PSTPIP1敲除小鼠，PSTPIP1 E250Q Papa敲除鼠标。现在正在进行表征这些小鼠表型的研究。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A recurrent rash with fever and arthropathy.

反复出现皮疹，伴有发烧和关节病。

DOI：
10.1016/j.jaad.2005.07.007
发表时间：
2006
期刊：
Journal of the American Academy of Dermatology
影响因子：
13.8
作者：
Jacob,SharonE;Cowen,EdwardW;Goldbach-Mansky,Raphaela;Kastner,Daniel;Turner,MariaL
通讯作者：
Turner,MariaL

A pilot study to evaluate the safety and efficacy of the long-acting interleukin-1 inhibitor rilonacept (interleukin-1 Trap) in patients with familial cold autoinflammatory syndrome.

DOI：
10.1002/art.23620
发表时间：
2008-08
期刊：
ARTHRITIS AND RHEUMATISM
影响因子：
0
作者：
Goldbach-Mansky, Raphaela;Shroff, Sharukh D.;Wilson, Mildred;Snyder, Christopher;Plehn, Sara;Barham, Beverly;Pham, Tuyet-Hang;Pucino, Frank;Wesley, Robert A.;Papadopoulos, Joanne H.;Weinstein, Steven P.;Mellis, Scott J.;Kastner, Daniel L.
通讯作者：
Kastner, Daniel L.

Treatment of patients with neonatal-onset multisystem inflammatory disease/chronic infantile neurologic, cutaneous, articular syndrome: comment on the article by Matsubara et al.

新生儿多系统炎症性疾病/慢性婴儿神经、皮肤、关节综合征患者的治疗：对 Matsubara 等人的文章的评论