The Role of Layilin as a Novel Regulator of Platelet Activation and Thromboinflammation

Layilin 作为血小板活化和血栓炎症的新型调节剂的作用

• 批准号: 10638243
• 负责人: Aaron Christopher Petrey
• 金额: $ 66.51万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10638243
• 关键词: Abnormal Platelet; Address; Adhesions; Adhesives; Affect; Agonist; Alpha Granule; American; Atherosclerosis; Autopsy; Biological Assay; Biological Response Modifiers; Biology; Blood; Blood Platelets; Blood Vessels; Blood coagulation; C Type Lectin Receptors; C-Type Lectins; CD44 Antigens; CRISPR/Cas technology; Cardiovascular Diseases; Cause of Death; Cell Adhesion; Cell surface; Cells; Clinical; Colitis; Complex; Crohn' s disease; Data; Development; Disease; Disease Outcome; Disease model; Effector Cell; Event; Fatal Outcome; Functional disorder; Gastrointestinal tract structure; Gene Expression; Gene Expression Profiling; General Population; Genetic; Genetic Transcription; Glycosaminoglycans; Guanosine Triphosphate; Hemostatic function; Homeostasis; Human; Hyaluronan; Immune; Immune response; Immunology; In Vitro; Incidence; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Integrins; Intestinal Diseases; Leukocytes; Life; Ligands; Link; Mediating; Metabolism; Molecular; Monomeric GTP-Binding Proteins; Mucous Membrane; Mus; Outcome; Pathogenesis; Pathway interactions; Patients; Platelet Activation; Play; Predisposition; Public Health; Reporting; Research; Resources; Risk; Role; Sepsis; Severities; Severity of illness; Signal Transduction; Steroids; Stream; Structure; Surface; Techniques; Testing; Thromboembolism; Thrombophilia; Thrombosis; Tissues; Travel; Treatment Protocols; Ulcerative Colitis; Vascular Endothelial Cell; Venous Thrombosis; Work; burden of illness; gut inflammation; human disease; improved; in vivo; inflammatory marker; inhibitor; innovation; intravital microscopy; mortality; mouse model; murine colitis; novel; novel strategies; novel therapeutics; pharmacologic; preservation; receptor; response; scaffold; therapeutic target; thromboinflammation; thrombotic; transcriptome sequencing; venous thromboembolism

A condition with no cure, inflammatory Bowel Disease (IBD) affects ~3 million patients in the US each year with the number ever increasing. IBD consists of two closely related disorders, Crohn’s disease (CD) and ulcerative colitis (UC), and usually manifest in the first three decades of life leading to relentless inflammatory destruction of the gastrointestinal tract in susceptible individuals. The blood of IBD patients exists in a hypercoagulable state and thromboembolism (TE), in which life-threatening blood clots break off and travel through the blood stream to block other vessels, is the most significant cause of mortality in IBD. This increased risk is between 3- and 15- fold during inactive and active disease respectively. Necropsy studies report thrombosis affects up to 40% of IBD patients, and 25% of patients suffering a thrombotic event have a fatal outcome. Current treatment regimens do not address thrombosis risk, and widespread steroid use increases risk. In addition to their classical role in hemostasis, platelets have emerged as novel regulators of the immune response that act as significant drivers of inflammation and tissue damage. Clinical reports suggest that platelet abnormalities associate with IBD activity, severity, and thrombosis risk, however molecular pathways responsible for increased platelet reactivity and thrombosis in IBD are not well studied. This proposal tests the significant and innovative hypothesis that platelets become transcriptionally altered toward hyperactivation in IBD and identified the C-type lectin layilin as a novel regulator of platelet activation and thrombosis. We will employ complementary clinical, in vitro, and in vivo approaches, a unique resource of patient tissues and expertise with human platelets and microvascular endothelial cells, along with state-of-the-art sequencing techniques, CRISPR/Cas9 technology, and intravital microscopy to rigorously test this hypothesis. Specific Aim 1 will determine how regulators of platelet activation, including layilin, are altered during active and inactive disease in IBD and contribute to disease outcomes. Specific Aim 2 will determine how layilin regulates platelet activation, define downstream activation pathways, and identify receptors and ligands mediating platelet adhesion to novel, inflammatory hyaluronan-cable matrices. Specific Aim 3 will establish how targeting pathways downstream of layilin in platelets improves inflammation and thrombosis during colitis. Successful completion of these aims will 1) determine how transcriptional changes in platelets contribute to inflammation and thrombosis in IBD, 2) determine downstream regulators of platelet activation mediated by layilin, 3) establish whether targeting dysregulated platelet activation pathways in improves inflammation and thrombosis in murine colitis. Data generated in this proposal will significantly increase our understanding of how the hyperreactive platelets contribute to the pathophysiology of IBD.

炎症性肠病（IBD）是一种无法治愈的疾病，每年在美国影响约300万患者， 数量不断增加。IBD由两种密切相关的病症组成，克罗恩病（CD）和溃疡性结肠炎（UC）。 结肠炎（UC），通常表现在生命的前三十年，导致无情的炎症破坏 对胃肠道的影响。IBD患者的血液处于高凝状态 血栓栓塞（TE），其中危及生命的血凝块破裂并通过血流 阻塞其他血管是IBD死亡的最重要原因。这种增加的风险在3- 15之间- 在非活动期和活动期分别折叠。尸检研究报告血栓形成影响高达40%的 IBD患者和25%发生血栓形成事件的患者具有致死性结局。目前的治疗方案 没有解决血栓形成的风险，广泛使用类固醇会增加风险。除了它们的经典作用之外， 止血，血小板已成为免疫反应的新型调节剂， 炎症和组织损伤临床报告提示血小板异常与IBD有关 活动性、严重程度和血栓形成风险，但导致血小板增加的分子途径 IBD中的反应性和血栓形成没有得到很好的研究。这一提议考验着重大而创新的 假设血小板在IBD中转录改变为过度活化，并鉴定了 C型凝集素layilin作为血小板活化和血栓形成的新调节剂。我们会委聘 互补的临床，体外和体内方法，独特的患者组织资源和专业知识， 人血小板和微血管内皮细胞，沿着最先进的测序技术， CRISPR/Cas9技术和活体显微镜来严格验证这一假设。具体目标1将 确定血小板活化的调节剂，包括layilin，在活动性和非活动性疾病中是如何改变的， IBD和有助于疾病的结果。具体目标2将确定layilin如何调节血小板活化， 定义下游活化途径，并鉴定介导血小板粘附的受体和配体， 炎症性透明质酸电缆基质。具体目标3将确定如何靶向下游途径 血小板中的layilin改善结肠炎期间的炎症和血栓形成。成功实现这些目标将 1)确定血小板中的转录变化如何促成IBD中的炎症和血栓形成，2） 确定layilin介导的血小板活化的下游调节因子，3）确定是否靶向 血小板活化途径失调可改善小鼠结肠炎的炎症和血栓形成。数据 这一提议产生的结果将大大增加我们对高反应性血小板如何 有助于IBD的病理生理学。