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

• 批准号: 10706156
• 负责人: Daniel Kastner
• 金额: $ 234.17万
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 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/10706156
• 关键词: Allogenic; Animal Model; Animals; Anti-Tumor Necrosis Factor Therapy; Antibodies; Autoimmune; Biochemical; Biology; Blood Vessels; COVID-19; Cell Differentiation process; Cell Line; Cells; Clinical; Complex; Cutaneous; Dermal; Development; Diagnosis; Disease; Endothelial Cells; Fever; Fibroblasts; Flow Cytometry; Functional disorder; Genes; Genetic; Genus staphylococcus; Goals; Graft Rejection; Hematology; Human; Immune; Immunity; Immunoglobulin Class Switching; Immunohistochemistry; Inflammation; Inflammatory; Infusion procedures; Inherited; Interferons; Journals; Lead; Leukocytes; Life; Longterm Follow-up; Manuscripts; Mediating; Medicine; Memory B-Lymphocyte; Mutation; Myeloid Cells; NF-kappa B; Names; Natural History; Necrosis; Neutropenia; New England; Pancytopenia; Paper; Pathway interactions; Patients; Penetrance; Phenotype; Polyarteritis Nodosa; Procedures; Proteins; Publishing; Pure Red-Cell Aplasia; Recurrence; Reporting; Sampling; Science; Signal Transduction; Stains; Staphylococcus aureus infection; Stroke; Stroke prevention; TNF gene; Transplant Recipients; Treatment Factor; Tumor Necrosis Factor Receptor; Ubiquitin; United States National Academy of Sciences; Universities; Vaccination; Variant; Vasculitis; Virulence Factors; adenosine deaminase deficiency; alpha Toxin; autoinflammatory; caveolin 1; cell injury; clinical heterogeneity; cohort; conditioning; cytokine; cytotoxic; exome; experience; experimental study; genome-wide analysis; graft failure; hematopoietic cell transplantation; hypogammaglobulinemia; improved; inhibitor; knockin animal; knockout animal; loss of function mutation; macrophage; medical specialties; novel; response; transcriptome

During the current reporting period we have focused on the following projects: 1) Natural history of the deficiency of adenosine deaminase 2 (DADA2) In previous reporting periods we published manuscripts in the New England Journal of Medicine (2014, 2019) first describing a new disorder we named the deficiency of adenosine deaminase 2 (DADA2), which is a recessively inherited illness caused by biallelic loss-of-function mutations in ADA2 that presents with fevers, recurrent strokes, livedo racemosa, and polyarteritis nodosa, and then showing that inhibitors of tumor necrosis factor (TNF) are very effective in preventing strokes in DADA2. We currently follow what is probably the largest cohort of DADA2 patients in the world. In early 2022 we published an observational analysis of what was then a 60-patient single-center DADA2 cohort, describing the multisystem disease that may span multiple medical specialties. In this report we summarized the broad phenotypic presentation, as well as our experience with hematopoietic cell transplantation and COVID-19. Disease manifestations could be separated into three major phenotypes: inflammatory/vascular, immune dysregulatory, and hematologic. However, most patients presented with significant overlap among these three phenotype groups. The cardinal features of the inflammatory/vascular group included cutaneous manifestations and stroke. Evidence of immune dysregulation was commonly observed, including hypogammaglobulinemia, absent to low class-switched memory B cells, and inadequate response to vaccination. Despite these findings, infectious complications were exceedingly rare in this cohort. Hematologic findings including pure red cell aplasia (PRCA), immune-mediated neutropenia, and pancytopenia were observed in half of patients. We significantly extended our experience using anti-TNF agents, with no strokes observed in 2026 patient months on TNF inhibitors. Meanwhile, hematologic and immune features had a more varied response to anti-TNF therapy. Six patients received a total of 10 allogeneic hematopoietic cell transplant (HCT) procedures, with secondary graft failure necessitating repeat HCTs in three patients, as well as unplanned donor cell infusions to avoid graft rejection. All transplanted patients had been on anti-TNF agents prior to HCT and received varying degrees of reduced-intensity or non-myeloablative conditioning. All transplanted patients are still alive and have discontinued anti-TNF therapy. The long-term follow up afforded by this large single-center study underscores the clinical heterogeneity of DADA2 and the potential for phenotypes to evolve in any given patient. 2) Treatment of the deficiency of adenosine deaminase 2 (DADA2) In early 2022 we also published a manuscript examining the pathophysiology and the underlying mechanisms of TNF-inhibitor response in DADA2 patients. In this study we used flow cytometry, intracellular cytokine staining, transcriptome analysis, immunohistochemistry, and cell differentiation experiments to define an inflammatory signature in patients with DADA2 and studied their response to TNF-inhibitor treatment. We demonstrated increased inflammatory signals and overproduction of cytokines mediated by interferon and nuclear factor kappa B pathways in patients primary cells. Treatment with TNF inhibitors led to reduction in inflammation, rescued the skewed differentiation toward the proinflammatory M1 macrophage subset, and restored integrity of endothelial cells in blood vessels. We also reported 8 novel disease-associated variants in 7 patients with DADA2. We concluded that DADA2 vasculitis is strongly related to the presence of activated myeloid cells, and the endothelial cell damage is rescued with anti-TNF treatment. 3) OTULIN haploinsufficiency In a previous reporting period we published a paper in the Proceedings of the National Academy of Sciences USA, describing what was then a new autoinflammatory disease caused by biallelic loss-of-function mutations in OTULIN, which encodes a deubiquitylase that removes linear ubiquitin chains that would ordinarily stabilize proinflammatory signaling complexes. The deficiency of OTULIN causes increased NF-kappa B signaling. During the current reporting period collaborators at the Rockefeller University performed a genome-wide analysis of the exomes of 105 patients with life-threatening staphylococcal disease and 1274 controls. They found enrichment for heterozygous OTULIN variants in patients with severe staphylococcal disease. Patients who are haploinsufficient for OTULIN suffered from episodes of life-threatening necrosis, typically triggered by S. aureus infection. OTULIN haploinsufficiency caused an accumulation of linear ubiquitin in dermal fibroblasts, but tumor necrosis factor receptor-mediated nuclear factor kappa B signaling remained intact. Blood leukocytes were unaffected. The OTULIN-dependent accumulation of caveolin-1 in dermal fibroblasts, but not leukocytes, facilitated the cytotoxic damage inflicted by staphylococcal virulence factor alpha-toxin. Naturally elicited antibodies against alpha-toxin contributed to incomplete clinical penetrance. We concluded that human OTULIN haploinsufficiency underlies life-threatening staphylococcal disease by disrupting cell-intrinsic immunity to alpha-toxin in nonleukocytic cells. A manuscript reporting these findings was published in Science during the current reporting period.

报告期内，我们重点开展了以下项目： 1) 腺苷脱氨酶 2 (DADA2) 缺乏的自然史 在之前的报告周期中，我们在《新英格兰医学杂志》（2014 年、2019 年）上发表了手稿，首先描述了一种新的疾病，我们将其命名为腺苷脱氨酶 2 缺乏症 (DADA2)，这是一种由双等位基因功能丧失引起的隐性遗传性疾病ADA2 突变导致发烧、复发性中风、总状青斑和结节性多动脉炎，然后显示肿瘤坏死因子 (TNF) 抑制剂DADA2 中对于预防中风非常有效。我们目前正在追踪可能是世界上最大的 DADA2 患者队列。 2022 年初，我们发表了对当时 60 名患者的单中心 DADA2 队列的观察分析，描述了可能跨越多个医学专业的多系统疾病。在本报告中，我们总结了广泛的表型表现，以及我们在造血细胞移植和 COVID-19 方面的经验。疾病表现可分为三种主要表型：炎症/血管、免疫失调和血液。然而，大多数患者在这三个表型组之间表现出显着的重叠。炎症/血管组的主要特征包括皮肤表现和中风。免疫失调的证据很常见，包括低丙种球蛋白血症、低类别转换记忆 B 细胞缺失以及对疫苗接种反应不足。尽管有这些发现，该队列中感染性并发症极为罕见。一半患者观察到血液学检查结果，包括纯红细胞再生障碍性贫血 (PRCA)、免疫介导的中性粒细胞减少症和全血细胞减少症。我们显着扩展了使用抗 TNF 药物的经验，在使用 TNF 抑制剂的 2026 个患者月中没有观察到中风。与此同时，血液学和免疫特征对抗 TNF 治疗的反应更加多样化。 6 名患者总共接受了 10 次同种异体造血细胞移植 (HCT) 手术，其中 3 名患者因继发性移植失败而需要重复 HCT，并进行了计划外的供体细胞输注以避免移植排斥。所有移植患者在 HCT 前均已服用抗 TNF 药物，并接受不同程度的降低强度或非清髓性预处理。所有移植患者仍然活着，并且已停止抗 TNF 治疗。这项大型单中心研究提供的长期随访强调了 DADA2 的临床异质性以及表型在任何特定患者中进化的潜力。 2) 腺苷脱氨酶2（DADA2）缺乏症的治疗 2022 年初，我们还发表了一篇手稿，研究 DADA2 患者中 TNF 抑制剂反应的病理生理学和潜在机制。在这项研究中，我们使用流式细胞术、细胞内细胞因子染色、转录组分析、免疫组织化学和细胞分化实验来定义 DADA2 患者的炎症特征，并研究他们对 TNF 抑制剂治疗的反应。我们证明了患者原代细胞中由干扰素和核因子 kappa B 通路介导的炎症信号增加和细胞因子过量产生。 TNF 抑制剂治疗可减少炎症，挽救向促炎性 M1 巨噬细胞亚群的偏向分化，并恢复血管内皮细胞的完整性。我们还报告了 7 名 DADA2 患者的 8 种新的疾病相关变异。我们得出的结论是，DADA2 血管炎与活化的骨髓细胞的存在密切相关，并且内皮细胞损伤可以通过抗 TNF 治疗来挽救。 3) OTULIN单倍体不足 在上一个报告期，我们在《美国国家科学院院刊》上发表了一篇论文，描述了一种由 OTULIN 中双等位基因功能丧失突变引起的新的自身炎症性疾病，该突变编码一种去泛素化酶，可去除线性泛素链，通常会稳定促炎信号复合物。 OTULIN 缺乏会导致 NF-kappa B 信号传导增加。 在当前报告期内，洛克菲勒大学的合作者对 105 名患有危及生命的葡萄球菌疾病的患者和 1274 名对照者的外显子组进行了全基因组分析。他们发现严重葡萄球菌疾病患者的杂合 OTULIN 变异体富集。 OTULIN 单倍剂量不足的患者会发生危及生命的坏死，通常由金黄色葡萄球菌感染引发。 OTULIN 单倍体不足导致真皮成纤维细胞中线性泛素积累，但肿瘤坏死因子受体介导的核因子 kappa B 信号传导保持完整。血液白细胞不受影响。 Caveolin-1 在真皮成纤维细胞（而非白细胞）中的 OTULIN 依赖性积累促进了葡萄球菌毒力因子 α 毒素造成的细胞毒性损伤。自然引发的抗α毒素抗体导致临床外显率不完全。我们得出的结论是，人 OTULIN 单倍体不足是通过破坏非白细胞中对 α 毒素的细胞内在免疫力而导致危及生命的葡萄球菌疾病的原因。 报告这些发现的手稿在本报告期内发表在《科学》杂志上。