Genetics, Pathophysiology, and Treatment of Recessive Autoinflammatory Diseases

隐性自身炎症性疾病的遗传学、病理生理学和治疗

基本信息

批准号：
8565567
负责人：
Daniel Kastner
金额：
$ 112.3万
依托单位：
NATIONAL HUMAN GENOME RESEARCH INSTITUTE
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/8565567
关键词：
4 year old Admission activity Adverse event Age Anemia Animal Model Animals Apple Area Arthralgia Autoimmune Process Berlin Biochemical Biology Blood Proteins Candidate Disease Gene Cell Line Chronic Clinical Trials Coagulation Process Colchicine Collaborations Complex Cysteine Data Dermatitis Development Diagnosis Diarrhea Dideoxy Chain Termination DNA Sequencing Disease Dose Double-Blind Method Event Familial Mediterranean Fever Family Female Fever Frequencies Functional disorder Genes Genetic Goals Human Inheritance Patterns Inherited Injection Site Reaction Injection of therapeutic agent Intensive Care Units Interleukin-1 Internal Medicine KLK3 gene Laboratories Lead Life Lipodystrophy Marriage Molecular Mutation National Institute of Arthritis and Musculoskeletal and Skin Diseases Odds Ratio Panniculitis Paper Participant Patients Phenotype Placebos Play Prekallikrein Prolonged Partial Thromboplastin Time Proteins Randomized Recruitment Activity Recurrence Reporting Research Personnel Resistance Role Safety Sampling Studies Screening procedure Shock Site Son Syndrome Tail Temperature Tertiary Protein Structure Variant anakinra cohort cytokine disulfide bond early onset efficacy testing exome improved knockin animal knockout animal male man marenostrin multicatalytic endopeptidase complex mutant receptor skin lesion standard care therapy design vascular inflammation

项目摘要

During the current reporting period we have made significant progress in three areas: (1) in collaboration with Dr. Phil Hashkes, now in Jerusalem, we completed a clinical trial testing the efficacy of rilonacept, a long-acting interleukin-1 blocker, in patients with FMF who are unresponsive to or intolerant of the standard treatment, colchicine, and now have a paper in press describing these results in the Annals of Internal Medicine; (2) in collaboration with Dr. Raphaela Goldbach-Mansky of NIAMS, using a combination of candidate gene analysis and whole-exome sequencing, we have identified mutations in 4 proteasome components in patients with a new recessively inherited autoinflammatory disease characterized by atypical neutrophilic dermatitis with lipodystrophy and elevated temperature (CANDLE syndrome); and (3) we have utilized whole-exome sequencing in small families with unexplained recurrent fevers and autoinflammatory disease to identify possible recessively inherited causal variants. Rilonacept in FMF Currently there is no proven alternative for FMF patients resistant to or intolerant of colchicine. Previous data from our laboratory have implicated interleukin-1 as a key proinflammatory cytokine in FMF. We assessed the efficacy and safety of rilonacept, an interleukin-1 decoy receptor, using a double-blind, single-subject alternating treatment design. Subjects over 4 years old with at least 1 monthly attack despite adequate doses of colchicine, or who were colchicine intolerant, were recruited at 6 U.S. sites. Subjects were randomized to 1 of 4 treatment sequences that included two 3-month courses of rilonacept 2.2 mg/kg (max 160 mg) by weekly SC injection, and two 3-month courses of placebo. Eight males and six females with a mean age of 24.4 years (SD, 11.8) were randomly assigned. Among 12 participants who completed 2 or more courses, the rilonacept-placebo attack risk ratio was 0.59 (SD, 0.12) (equal-tail 95% credible interval, 0.39 to 0.85). The median number of attacks per month was 0.77 (0.18 and 1.20 attacks in the first and third quartiles, respectively) with rilonacept versus 2.00 (0.90 and 2.40, respectively) with placebo (median difference, -1.74 95% CI, -3.4 to -0.1, P = 0.027). There were more treatment courses of rilonacept without attacks (29% vs. 0%; P = 0.004), and with a decrease in attacks of greater than 50% compared with the baseline rate during screening (75% vs. 35%, P = 0.006) than with placebo. However, the duration of attacks did not differ between placebo and rilonacept (median difference, 1.2 days -0.5 and 2.4 days in the first and third quartiles, respectively; P = 0.32). Injection site reactions were more frequent with rilonacept (median difference, 0 events per patient treatment month medians of -4 and 0 in the first and third quartiles, respectively; P = 0.047), but no differences were seen in other adverse events. From this study we conclude that rilonacept reduces the frequency of FMF attacks and seems to be a treatment option for colchicine-resistant or intolerant FMF. Mutational Analysis in CANDLE During the previous reporting period our laboratory participated in a collaborative study that identified homozygous T75M PSMB8 mutations in 4 of 8 patients with CANDLE, a syndrome characterized by onset during the first year of life, recurrent fevers, purpuric skin lesions, arthralgia, progressive lipodystrophy, anemia, and delayed physical development. Nevertheless, there remained patients with the CANDLE phenotype with only a single demonstrable PSMB8 mutation, and other patients with no demonstrable PSMB8 mutations. We hypothesized that mutations in other components of the multi-molecular immunoproteasome complex or of the constitutive proteasome might also cause this phenotype. We therefore systematically performed Sanger sequencing on the genes encoding the various proteasome components, in conjunction with whole-exome sequencing to exclude other possible candidates. Preliminary data from these sequencing studies identify mutations in 3 other genes encoding various components of the proteasome in patients with the CANDLE phenotype, but not in large cohorts of appropriately matched controls. In collaboration with investigators in Berlin, we are assessing the functional consequences of these mutations. Our genetic data suggest that early-onset panniculitis is a molecular disorder of the proteasome, with possible digenic inheritance of multiple interacting subunits. Whole-exome Sequencing in Patients with Unexplained Autoinflammatory Phenotypes We have utilized whole-exome sequencing in a number of small families with unexplained, possibly recessively inherited autoinflammatory phenotypes. In one such case, we performed whole-exome analysis in a south Indian man, the son of an uncle-niece marriage, with chronic diarrhea, a prolonged partial thromboplastin time (PTT), and episodes of fever and shock requiring repeated intensive care unit admissions. This patient is homozygous for the G355C mutation in KLKB1, which encodes prekallikrein, a blood protein that plays a role in inflammation, vascular tone, and coagulation. The mutation introduces an unpaired ninth cysteine residue into the fourth apple domain of the protein, potentially causing abnormal disulfide bonding. Functional studies of the mutant protein are under way. We are also analyzing whole-exome data on several other small families with unexplained, possibly recessive disease.

在当前的报告期间，我们在三个领域取得了重大进展：（1）与现在在耶路撒冷的Phil Hashkes博士合作，我们完成了一项临床试验，对Rilonacept（长效的Interleukin-1 Blocker Rilonacept的功效）进行了临床试验，对FMF的患者对FMF的患者无反应或不适中，并批准了该标准的综合治疗，并在colchicine colchicin的范围内进行了批准，并以此为综述，这些疾病是一项针对该标准的综述。（2）与Niams的Raphaela Goldbach-Mansky博士合作，结合了候选基因分析和全外活体测序，我们在膜片遗传遗传遗传遗传自遗传性自鼻疾病的患者中已经确定了4个蛋白酶体成分的突变，该疾病由非典型嗜中性粒细胞炎与脂质症状症和高度合成体温度（Candtrorys）（Canddrorys）（Candtrorys）（Candtrorys）（Canddrorys）（Candtrorys）（Candtrorys）; （3）我们已经在具有无法解释的复发性发烧和自动炎性疾病的小家族中使用了全外活体测序，以鉴定可能遗传遗传的因果变异。 FMF中的Rilonacept 目前，对于抗秋水仙碱具有抵抗力或不耐受性的FMF患者，尚无可见的替代方法。我们实验室的先前数据已将白介素1视为FMF中的关键促炎细胞因子。我们使用双盲，单个受试者交替的处理设计评估了白介素-1诱饵受体Rilonacept的功效和安全性。尽管有足够剂量的秋水仙碱或秋水仙碱不耐受，但在4岁以上的受试者在美国6个地点招募了至少1个月攻击的受试者。受试者被随机分为4个治疗序列中的1个，其中包括两个3个月的Rilonacept 2.2 mg/kg（最大160 mg），每周注射160 mg）和安慰剂的两个3个月的课程。随机分配了八名男性和六名女性，平均年龄为24.4岁（SD，11.8）。在完成2个或更多课程的12名参与者中，Rilonacept-place攻击风险比率为0.59（SD，0.12）（相等的95％可靠间隔，0.39至0.85）。每月攻击的中位数为0.77（分别为0.18和1.20次数，分别为第一和第三四分位数），而Rilonacept分别为2.00（分别为0.90和2.40），安慰剂（中位数差异为-1.74 95％CI，-3.4至-3.4至-0.1，-0.1，p = 0.027）。没有攻击的Rilonacept的治疗课程（29％vs. 0％； P = 0.004），与筛查期间的基线率相比，攻击率降低了50％（75％vs. 35％，P = 0.006）。但是，安慰剂和rilonacept之间的攻击持续时间没有差异（中位差异为1.2天-0.5和2.4天，第一四分位数和第三四分位数； p = 0.32）。 rilonacept的注射部位反应更为频繁（中位差异，每个患者治疗月的0事件分别为-4和0，在第一和第三四分位数中为-4和0； p = 0.047），但在其他不良事件中没有差异。从这项研究中，我们得出的结论是，Rilonacept降低了FMF攻击的频率，并且似乎是耐秋水仙碱或不耐受性FMF的治疗选择。蜡烛中的突变分析在上一个报告期间，我们的实验室参加了一项合作研究，该研究确定了8例蜡烛患者中的4例纯合T75M PSMB8突变，这是一种综合征，该综合征在生命的第一年中以发作为特征，经常发烧，紫罗兰皮型皮肤病变，Arthralgia，Arthralgia，Arthralgia，渐进的脂肪流行病，身体疾病，体育疾病和延迟的身体和延迟。然而，仍有患者患有蜡烛表型，只有一个可证明的PSMB8突变，而其他没有可证明的PSMB8突变的患者。我们假设多分子免疫蛋白酶体复合物或组成型蛋白酶体的其他成分中的突变也可能导致这种表型。因此，我们系统地对编码各种蛋白酶组成分的基因进行了Sanger测序，并与全外观测序结合使用，以排除其他可能的候选者。来自这些测序研究的初步数据鉴定了在蜡烛表型患者中编码蛋白酶体各种成分的其他3个基因中的突变，但在大量适当匹配的对照组中却没有。与柏林的研究人员合作，我们正在评估这些突变的功能后果。我们的遗传数据表明，早发pansiculisis是蛋白酶体的分子障碍，可能具有多个相互作用亚基的基因遗传。无法解释的自发炎性表型的患者的全外观测序我们已经在许多无法解释的，可能遗传遗传的自身炎症表型的小型家庭中使用了全外观测序。在一个这样的案例中，我们在南印度人（南印度人的儿子）中进行了全面分析，这是一个叔叔婚姻的儿子，慢性腹泻，长时间的部分血栓形成时间（PTT），发烧和震惊的发作，需要重复护理单位。该患者是KLKB1中G355C突变的纯合子，该患者编码Prepkallikrein，这是一种在炎症，血管张力和凝结中起作用的血液蛋白。该突变将未配对的第九个半胱氨酸残基引入蛋白质的第四个苹果结构域，可能导致二硫键异常。突变蛋白的功能研究正在进行中。我们还正在分析其他几个无法解释的，可能是隐性疾病的小型家庭的全面数据。