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 尽管采用静脉注射标准治疗，但仍会出现冠状动脉异常（CAA） 免疫球蛋白。一旦动脉瘤形成，动脉壁的损伤是不可逆转的，例如狭窄 钙化可能导致缺血性并发症。该协作团队已确定激活 IL-1 通路和内皮间质转化 (EndoMT) 的影响与 krüppel 样下调 因子 4 (KLF4) 是 KD 血管炎发病机制的关键。我们对 TIFA (TNF) 核心作用的新发现 NLRP3 炎性体中的受体相关因子相互作用蛋白（具有叉头相关结构域） 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。我们提出了一个大胆的 计划联合这个具有血管生物学、免疫学和临床 KD 专业知识的研究团队来解开这个问题 川崎病中血管损伤的机制并试点一种新的治疗方法。指导性假设 急性KD不仅激活内皮细胞中的IL-1/TIFA/NLRP3炎症小体，而且降低EC 体内平衡，两者在 KD 的发病机制中都至关重要。阻断 IL-1 依赖性先天性 阿那白滞素（重组 IL-1R 拮抗剂）的免疫反应与 EC 功能的恢复相结合 阿托伐他汀可减少急性川崎病的血管损伤。提出了三个具体目标来检验这一假设。 具体目标 1 将描述阿那白滞素阻断先天免疫激活的分子基础 与 KD 血管炎相关的反应。体外、离体和体内实验将研究 IL-1、 使用途径特异性 siRNA 敲低，TIFA 和 NLRP3 治疗 KD 介导的 EC 功能障碍和血管炎， 重组过度表达、药理学激动剂和抑制剂（包括阿那白滞素）以及丢失- 功能小鼠。具体目标 2 将阐明阿托伐他汀恢复 EC 功能的分子基础 急性KD。 KLF4 功能获得和丧失的小鼠品系以及谱系追踪实验将 旨在破译 KD 血管病变中 EC 功能障碍的分子机制。具体目标 图3将确定阿那白滞素/阿托伐他汀联合疗法在LCWE注射小鼠中的协同效应 在 KD 患者中。 LCWE小鼠模型，包括Tifa-/-和Klf4-/-，将用于测试效果 阿那白滞素/阿托伐他汀联合治疗对血管炎症和 EC 功能的影响。此外，急性 KD 患有早期 CAA 的患者将参加阿那白滞素/阿托伐他汀联合治疗的试点研究。细胞因子 将在治疗前和治疗后测量血清中的 MMP 水平和活性以及全血 RNA 中的转录本丰度。 治疗。这些血清还将使用培养的 EC 进行体外测试，并读出 EC 先天免疫 反应和 EC 功能障碍。这个多 PI 提案中三个团队的协同专业知识提供了 了解川崎病血管病变分子机制并快速转化该疾病的独特机会 对儿童进行的一项试点研究的结果对治疗效果进行了体外和体内评估。

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