Innate Immune Activation and Endothelial Cell Dysfunction in Acute Kawasaki Disease

急性川崎病的先天免疫激活和内皮细胞功能障碍

• 批准号: 10311990
• 负责人: JANE C BURNS
• 金额: $ 72.91万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10311990
• 关键词: Acute; Address; Aftercare; Agonist; Aneurysm; Attenuated; Binding; Biology; Blood Vessels; Cardiovascular system; Cell Wall; Cell model; Cell physiology; Child; Childhood; Clinical; Clinical Trials; Combined Modality Therapy; Coronary; Coronary artery; Cultured Cells; Development; Down-Regulation; Endothelial Cells; Endothelium; Enrollment; Etiology; FHA Domain; Functional disorder; GKLF protein; Heart Diseases; Homeostasis; Human; Immune response; Immunology; Impairment; In Vitro; Infant; Inflammasome; Inflammatory; Innate Immune Response; Innate Immune System; Interleukin Activation; Interleukin-1; Interleukin-1 Receptors; Interleukin-1 beta; Interleukins; Intravenous Immunoglobulins; Knock-out; Lactobacillus casei; Lead; Leucine-Rich Repeat; Life; Matrix Metalloproteinases; Measures; Mediating; Mediator of activation protein; Mesenchymal; MicroRNAs; Molecular; Monoclonal Antibodies; Mucocutaneous Lymph Node Syndrome; Mus; Myocardial Infarction; Myocardial Ischemia; Natural Immunity; Nucleotides; Outcome; Oxidative Stress; Participant; Pathogenesis; Pathway interactions; Patients; Pharmacology; Pilot Projects; Production; Proteins; RNA; Recombinant Interleukins; Recombinants; Research; Role; Signal Transduction; Small Interfering RNA; TNF Receptor-Associated Factors; TNF gene; Testing; Therapeutic; Time; Transcript; Translating; Vascular Diseases; Vascular Endothelial Cell; Vasculitis; Whole Blood; Work; anakinra; antagonist; atorvastatin; base; calcification; cell injury; connective tissue growth factor; coronary lesion; cytokine; data modeling; experimental study; immune activation; improved; in vitro Model; in vitro testing; in vivo; infliximab; inhibitor; knock-down; loss of function; marenostrin; mouse model; novel; novel therapeutic intervention; overexpression; pleiotropism; prevent; restoration; therapy design; treatment effect; vascular inflammation; vascular injury

More than 25% of children with Kawasaki disease (KD), the most common cause of pediatric acquired heart disease, develop coronary artery abnormalities (CAAs) despite standard therapy with intravenous immunoglobulin. Once aneurysms have formed, the damage to the arterial wall is irreversible, such as stenoses and calcification that may lead to ischemic complications. This collaborative team has established that activation of the IL-1 pathway and endothelial-to-mesenchymal transition (EndoMT) with downregulation of krüppel-like factor 4 (KLF4) are key to the pathogenesis of KD vasculitis. Our novel discovery of the central role of TIFA (TNF receptor-associated factor-interacting protein with a forkhead-associated domain) in NLRP3 inflammasome activation in endothelial cell (EC) injury is proposed as a further mechanism of KD vasculitis. We propose a bold plan to unite this research team with expertise in vascular biology, immunology, and clinical KD to unravel the mechanisms underlying vascular injury in KD and pilot a novel therapeutic approach. The guiding hypothesis is that acute KD not only activates the IL-1/TIFA/NLRP3 inflammasome in the endothelium, but also reduces EC homeostasis, both of which are critical in the pathogenesis of KD. The blockade of the IL-1-dependent innate immune response by anakinra (recombinant IL-1R antagonist) in combination with the restoration of EC function by atorvastatin will reduce vascular damage in acute KD. Three specific aims are proposed to test this hypothesis. Specific Aim 1 will delineate the molecular basis by which anakinra blocks activation of the innate immune response associated with KD vasculitis. In vitro, ex vivo, and in vivo experiments will investigate the role of IL-1, TIFA, and NLRP3 in KD-mediated EC dysfunction and vasculitis using pathway-specific siRNA knockdown, recombinant overexpression, pharmacologic agonists and inhibitors (including anakinra), as well as loss-of- function mice. Specific Aim 2 will elucidate the molecular basis by which atorvastatin restores EC function in acute KD. Mouse lines with gain- and loss-of-function of KLF4 as well as lineage tracing experiments will be performed to decipher the molecular mechanisms underlying EC dysfunction in KD vasculopathy. Specific Aim 3 will determine the synergistic effect of anakinra/atorvastatin combination therapy in LCWE-injected mice and in KD patients. LCWE mouse models, including Tifa-/- and Klf4-/-, will be utilized to test the effect of anakinra/atorvastatin combination therapy on vascular inflammation and EC function. Furthermore, acute KD patients with early CAAs will be enrolled in a pilot study of anakinra/atorvastatin combination therapy. Cytokine levels and MMP activity in sera and transcript abundance in whole blood RNA will be measured pre- and post- treatment. These sera will also be tested in vitro with cultured ECs with read-outs for EC innate immune responses and EC dysfunction. The synergistic expertise of the three teams in this multi-PI proposal provides a unique opportunity to understand molecular mechanisms underlying KD vasculopathy and rapidly translate the findings into a pilot study in children with in vitro and in vivo assessments of the treatment effect.

超过25%的儿童患有川崎病(KD)，这是儿童获得性心脏病的最常见原因 疾病，发展冠状动脉异常(CAA)，尽管标准的静脉治疗 免疫球蛋白。一旦形成动脉瘤，对动脉壁的损害是不可逆的，例如狭窄。 以及可能导致缺血性并发症的钙化。这个协作团队已经建立了激活 IL-1途径和内皮-间充质转化(EndoMT)与KRüppel样蛋白下调 第4因子(KLF4)在KD血管炎的发病机制中起关键作用。我们对TIFA(肿瘤坏死因子)核心作用的新发现 NLRP3炎症体中的受体相关因子相互作用蛋白 内皮细胞(EC)损伤中的激活被认为是KD血管炎的进一步机制。我们提出一个大胆的建议 计划将这个拥有血管生物学、免疫学和临床KD专业知识的研究团队联合起来，以揭示 KD患者血管损伤的潜在机制，并尝试了一种新的治疗方法。指导性假说 急性KD不仅能激活内皮细胞中的IL-1/TIFA/NLRP3炎性小体，而且还能降低EC 动态平衡，这两个因素在KD的发病机制中都是至关重要的。IL-1依赖的先天细胞的阻断 Anakinra(重组IL-1R拮抗剂)联合EC功能恢复的免疫应答 应用阿托伐他汀可减轻急性川崎病患者的血管损伤。为了检验这一假说，本文提出了三个具体目标。 具体目标1将描述阿纳金纳阻断天然免疫激活的分子基础。 与KD血管炎相关的反应。体外、体外和体内实验将研究IL-1的作用， TIFA和NLRP3在KD介导的内皮细胞功能障碍和血管炎中的作用 重组过表达，药理激动剂和抑制剂(包括阿纳金纳)，以及丢失- 功能正常的小鼠。具体目标2将阐明阿托伐他汀恢复血管内皮细胞功能的分子基础 急性KD。具有KLF4功能获得和丧失的小鼠品系以及血统追踪实验将被 旨在破译KD血管病变中EC功能障碍的分子机制。特定目标 3将确定阿那金拉/阿托伐他汀联合治疗在LC WE注射的小鼠和 在KD患者中。将利用包括TifA-/-和KLF4-/-在内的LCWE小鼠模型来测试TIFA-/-和KLF4-/-的作用 阿那金纳/阿托伐他汀联合治疗对血管炎症和内皮功能的影响。此外，急性KD 早期CAAS患者将参加Anakinra/阿托伐他汀联合治疗的先导性研究。细胞因子 检测治疗前后血清中的水平和基质金属蛋白酶的活性以及全血RNA中的转录丰度。 治疗。这些血清也将在体外与培养的EC进行测试，读出EC的天然免疫 反应和EC功能障碍。在这个多PI方案中，三个团队的协同专业知识提供了 独特的机会了解KD血管病变的分子机制并快速翻译 一项在儿童体内和体外进行治疗效果评估的先导性研究的结果。

项目成果

Characterization of SARS-CoV-2 and common cold coronavirus-specific T-cell responses in MIS-C and Kawasaki disease children.

• DOI: 10.1002/eji.202149556
• 发表时间: 2022-01
• 期刊: European journal of immunology
• 影响因子: 5.4
• 作者: Hsieh LE;Grifoni A;Sidney J;Shimizu C;Shike H;Ramchandar N;Moreno E;Tremoulet AH;Burns JC;Franco A
• 通讯作者: Franco A

Kawasaki Disease Patient Stratification and Pathway Analysis Based on Host Transcriptomic and Proteomic Profiles.

川崎疾病患者分层和途径分析基于宿主转录组和蛋白质组学特征。

• DOI: 10.3390/ijms22115655
• 发表时间: 2021-05-26
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Jackson H;Menikou S;Hamilton S;McArdle A;Shimizu C;Galassini R;Huang H;Kim J;Tremoulet A;Thorne A;Fischer R;de Jonge MI;Kuijpers T;Wright V;Burns JC;Casals-Pascual C;Herberg J;Levin M;Kaforou M;On Behalf Of The Perform Consortium
• 通讯作者: On Behalf Of The Perform Consortium

A machine-learning algorithm for diagnosis of multisystem inflammatory syndrome in children and Kawasaki disease in the USA: a retrospective model development and validation study.

• DOI: 10.1016/s2589-7500(22)00149-2
• 发表时间: 2022-10
• 期刊: LANCET DIGITAL HEALTH
• 影响因子: 30.8
• 作者: Lam, Jonathan Y.;Shimizu, Chisato;Tremoulet, Adriana H.;Bainto, Emelia;Roberts, Samantha C.;Sivilay, Nipha;Gardiner, Michael A.;Kanegaye, John T.;Hogan, Alexander H.;Salazar, Juan C.;Mohandas, Sindhu;Szmuszkovicz, Jacqueline R.;Mahanta, Simran;Dionne, Audrey;Newburger, Jane W.;Ansusinha, Emily;DeBiasi, Roberta L.;Hao, Shiying;Ling, Xuefeng B.;Cohen, Harvey J.;Nemati, Shamim;Burns, Jane C.
• 通讯作者: Burns, Jane C.

Autoantibodies Against Proteins Previously Associated With Autoimmunity in Adult and Pediatric Patients With COVID-19 and Children With MIS-C.

• DOI: 10.3389/fimmu.2022.841126
• 发表时间: 2022
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Burbelo PD;Castagnoli R;Shimizu C;Delmonte OM;Dobbs K;Discepolo V;Lo Vecchio A;Guarino A;Licciardi F;Ramenghi U;Rey-Jurado E;Vial C;Marseglia GL;Licari A;Montagna D;Rossi C;Montealegre Sanchez GA;Barron K;Warner BM;Chiorini JA;Espinosa Y;Noguera L;Dropulic L;Truong M;Gerstbacher D;Mató S;Kanegaye J;Tremoulet AH;Pediatric Emergency Medicine Kawasaki Group;Eisenstein EM;Su HC;Imberti L;Poli MC;Burns JC;Notarangelo LD;Cohen JI
• 通讯作者: Cohen JI

Bridging a diagnostic Kawasaki disease classifier from a microarray platform to a qRT-PCR assay.

• DOI: 10.1038/s41390-022-02148-y
• 发表时间: 2023-03
• 期刊: PEDIATRIC RESEARCH
• 影响因子: 3.6
• 作者: Kuiper, Rowan;Wright, Victoria J.;Habgood-Coote, Dominic;Shimizu, Chisato;Huigh, Daphne;Tremoulet, Adriana H.;van Keulen, Danielle;Hoggart, Clive J.;Rodriguez-Manzano, Jesus;Herberg, Jethro A.;Kaforou, Myrsini;Tempel, Dennie;Burns, Jane C.;Levin, Michael
• 通讯作者: Levin, Michael