Purinergic modulation of the autoimmune vascular phenotype

自身免疫血管表型的嘌呤能调节

• 批准号: 10581346
• 负责人: Jason Knight
• 金额: $ 36.12万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 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 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581346
• 关键词: ADORA2A gene; Adenosine; Adjuvant; Agonist; Animal Model; Anti-Inflammatory Agents; Antibodies; Anticoagulants; Anticoagulation; Antiphospholipid Antibodies; Antiphospholipid Syndrome; Area; Autoimmune; Bioenergetics; Blood Platelets; Blood Vessels; Blood coagulation; COVID-19; Cell Respiration; Cell Surface Receptors; Cells; Chromatin; Chronic; Clinic; Clinical; Coagulation Process; Communication; Cyclic AMP; Cyclic AMP Receptors; Cyclic AMP-Dependent Protein Kinases; Data; Deterioration; Disease; Enzymes; Event; Functional disorder; Glucose; Glycogen; Goals; Grant; Homeostasis; Hyperactivity; Immune; In Vitro; Inflammation; Intervention; Knockout Mice; Lupus; Mass Spectrum Analysis; Measurement; Mediating; Metabolic; Metabolism; Modeling; Mus; Myelogenous; Neurofibrillary Tangles; Neutrophil Activation; Organ; Patients; Pentosephosphate Pathway; Phenotype; Phosphorylation; Play; Pregnancy loss; Proteins; Purine Nucleotides; Purinergic P1 Receptors; Research Personnel; Resistance; Role; Signal Transduction; Surface; System; Therapeutic; Thrombosis; Time; Work; blood damage; cohort; combat; coronavirus disease; dietary; dietary manipulation; disease phenotype; drug candidate; ecto-nucleotidase; experience; experimental study; extracellular; glucose metabolism; immunothrombosis; microbicide; neutrophil; platelet function; protective pathway; receptor; restraint; thrombotic

PROJECT SUMMARY/ABSTRACT This proposed renewal of “Purinergic modulation of the autoimmune vascular phenotype” builds upon 4 years of discovery by an early stage investigator in the areas of lupus, COVID-19, and antiphospholipid syndrome (APS). Despite the routine use of traditional anticoagulants in the latter disease, 1 in 5 APS patients is still expected to experience a breakthrough thrombotic event. Furthermore, anticoagulants do little to mitigate the chronic occlusive microangiopathy that damages APS organs over time. How to combat anticoagulant- resistant manifestations of APS is unknown. This project now endeavors to use a deeply-phenotyped patient cohort, selected in vitro systems, and the most relevant animal models to identify adjuvant therapeutic approaches for the APS clinic. Its successful completion will shed additional light on neutrophil phenotypes in APS and will provide a new understanding of the role that purinergic signaling, neutrophil-platelet interactions, and neutrophil metabolism play in thrombotic events. The goal is that by the next cycle of this grant, we will have identified the 1-2 most promising drug candidates for repurposing in our APS clinic. APS is a leading acquired cause of both thrombosis and late-term pregnancy loss. In pursuit of a mechanistic understanding of immunothrombosis in APS, our group was the first to show that neutrophil extracellular traps (NETs, tangles of chromatin and microbicidal proteins expelled from activated neutrophils via “NETosis”) are required for APS-associated thrombosis. Since our last competitive submission, we have found that extracellular adenosine generated by the ectonucleotidase CD73 restrains NET release by activating surface adenosine A2A receptors (A2AR) and thereby boosting intracellular cAMP. Key preliminary data that inform this renewal demonstrate (i) restraint of platelet-mediated neutrophil activation by the CD73-A2AR axis; (ii) exaggerated thrombosis in myeloid lineage-specific A2AR knockout mice; (iii) hyperactive glucose metabolism in APS neutrophils that normalizes with A2AR agonists; and (iv) mitigation of thrombosis in APS mice by metabolism-focused interventions. The hypothesis is that manipulation of the CD73-A2AR-cAMP axis will restore neutrophil homeostasis in APS. Specific Aim 1 will define mechanisms by which purinergic signaling influences neutrophil and platelet function in APS. This Aim will define for the first time the purinergic landscape of a thrombophilic disorder, elucidate mechanisms by which purinergic signaling regulates neutrophil-platelet communication, and potentially identify the subset of APS patients most likely to benefit from antiplatelet and/or adenosine receptor-modulating therapies. Specific Aim 2 will determine the extent to which purinergic signaling can be leveraged to normalize neutrophil metabolism in APS. This Aim is expected to provide a new understanding of the metabolic requirements of NETosis, elucidate strategies for manipulating neutrophil metabolism, and identify the subset of APS patients most likely to benefit from those strategies.

项目总结/摘要 这项关于“自身免疫性血管表型的嘌呤能调节”的更新建议建立在4年前， 早期研究人员在狼疮、COVID-19和抗磷脂综合征领域的发现 （APS）。尽管在后一种疾病中常规使用传统抗凝剂，但仍有1/5的APS患者 预期发生突破性血栓形成事件。此外，抗凝剂几乎不能减轻 慢性闭塞性微血管病，随着时间的推移损害APS器官。如何对抗抗凝剂- APS的耐药表现尚不清楚。这个项目现在努力使用一个deeep-phenotyped病人 队列、选定的体外系统和最相关的动物模型，以确定辅助治疗 APS诊所的方法。它的成功完成将进一步阐明中性粒细胞表型， APS，并将提供一个新的理解的作用，嘌呤能信号，血小板相互作用， 和中性粒细胞代谢在血栓形成事件中起作用。我们的目标是，到下一个赠款周期，我们将 已经确定了1-2个最有前途的候选药物在我们的APS诊所重新利用。 APS是血栓形成和晚期妊娠丢失的主要后天原因。为了追求一种机械的 为了了解APS中的免疫血栓形成，我们的研究小组首次表明中性粒细胞胞外陷阱 (NETs通过“NETosis”从活化的嗜中性粒细胞排出的染色质和杀微生物蛋白质缠结） APS相关血栓形成所必需的。自上次提交竞争性申请以来，我们发现， 外核苷酸酶CD 73产生的细胞外腺苷通过激活表面活性剂来抑制NET的释放 腺苷A2 A受体（A2 AR），从而增加细胞内cAMP。关键的初步数据表明， 更新证明（i）CD 73-A2 AR轴抑制血小板介导的中性粒细胞活化;（ii） 骨髓谱系特异性A2 AR敲除小鼠中的过度血栓形成;（iii）葡萄糖代谢过度活跃 在APS中性粒细胞中，A2 AR激动剂使其正常化;和（iv）通过 以代谢为重点的干预措施。假设操纵CD 73-A2 AR-cAMP轴将导致细胞凋亡。 恢复APS中的中性粒细胞稳态。具体目标1将定义嘌呤能信号传导的机制， 影响APS中中性粒细胞和血小板功能。这一目标将首次定义嘌呤能 一个血栓性疾病的景观，阐明嘌呤能信号调节的机制， 血小板-血小板通信，并可能识别最有可能从 抗血小板和/或腺苷受体调节疗法。具体目标2将决定 嘌呤能信号传导可用于使APS中的中性粒细胞代谢正常化。预计这一目标将 提供对NETosis代谢要求的新认识，阐明操纵策略 中性粒细胞代谢，并确定APS患者的子集最有可能受益于这些策略。