A path to thrombosis in primary myelofibrosis

原发性骨髓纤维化的血栓形成途径

• 批准号: 10064585
• 负责人: KATYA RAVID
• 金额: $ 47.67万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-24 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064585
• 关键词: Acute; Adhesions; Affect; Aldehydes; Binding; Blood Platelets; Bone Marrow; Cardiovascular system; Carotid Arteries; Cell surface; Cells; Chronic; Chronic Kidney Failure; Cicatrix; Collaborations; Collagen; Collagen Receptors; Collagen Type I; Complement; Data; Deposition; Disease; Dyes; Elastin; Engineering; Enzymes; Event; Extracellular Matrix; Flowmeters; Future; Gene Mutation; Genes; Human; Image; Incidence; Integrin alpha2; Integrins; Investigation; Kidney; Knockout Mice; LOX gene; Laboratories; Lasers; Lead; Light; Link; Lysine; Malignant Neoplasms; Measurement; Mediating; Megakaryocytes; Membrane Proteins; Mus; Mutagenesis; Myelofibrosis; Myeloproliferative disease; Nitrogen; Oxidases; Oxides; Pathologic; Pathology; Pathway interactions; Patients; Pharmacology; Physiologic pulse; Platelet Activation; Platelet-Derived Growth Factor Receptor; Primary Myelofibrosis; Property; Protein-Lysine 6-Oxidase; Proteins; Proteomics; Publishing; Receptor Activation; Research; Risk; Role; Sampling; Serum; Shapes; Stenosis; Structure; Tensile Strength; Testing; Thrombosis; Thrombus; Tissues; Transgenic Mice; Up-Regulation; Western Blotting; Work; calreticulin; cohort; crosslink; drug development; extracellular; ferric chloride; human disease; in vivo; inhibitor/antagonist; innovation; intravital microscopy; knock-down; loss of function; mouse model; novel; overexpression; oxidation; platelet function; receptor; response; thrombotic

Abstract Controlling thrombosis is central to management of various pathologies. Studies proposed here will identify a new, unexpected path to thrombosis, involving the enzyme Lysyl Oxidase (LOX). The proposed work looks at LOX effects on platelet function and thrombosis in primary myelofibrosis (PMF), a pathology hallmarked by proliferation and clustering of megakaryocytes, and myelofibrosis in bone marrow. Data from the Swedish Cancer Register from 1980 to 2009 showed that in a cohort of 11,155 patients with myeloproliferative neoplasms and 44, 620 matched healthy controls, the risk of arterial thrombosis was significantly 4.9-fold increased (4.8-5.0 p<0.001) in the patients compared to matched controls, highlighting the importance of studying regulators implicated in controlling thrombosis associated with this pathology. LOX is known to oxidize peptidyl lysines, leading to cross-linked matrix proteins in the bone marrow niche. Our published studied identified a link between LOX upregulation in megakaryocytes and bone marrow fibrotic progression in a mouse model of myelofibrosis. Importantly, more recently we found LOX expression to be vastly upregulated in platelets of patients with PMF (of which the majority tested carry the JAK2V617F or calreticulin gene mutations), compared to matching controls, and platelets isolated from PMF patients have greater adhesion to type I collagen, compared to controls. Further, transgenic mouse platelets engineered to overexpress LOX, at a level similar to cells from a myelofibrotic mouse model, show increased adhesion to monomeric collagen, and significantly greater propensity for arterial thrombosis in vivo. Considering LOX known activity, we hypothesized that it influences platelets through lysine oxidation of at least one of the collagen receptors, further supported by preliminary studies involving Oxy-Western blot analysis. Thus, we propose two specific aims of research: Aim 1. To explore the mechanism by which LOX affects platelet response to collagen using proteomics approaches, and mouse and human MPN samples; Aim 2. To determine the relevance of LOX- regulated thrombosis in vivo in myelofibrotic mice, and the role of collagen receptors in this effect. Pursuing these aims will be facilitated by: 1) a new transgenic mouse line produced in our lab, in which platelet LOX level is upregulated under wild type background (free of MPN), and 2) the availability of a new LOX inhibitor, and Lox gene loss of function studies. This work is innovative in that we are the first to reveal a link between LOX and collagen receptors activation. Further, these receptors were not suspected before to be regulated by oxidation. Our research is significant in that it will shed new light on basic mechanisms of platelet activation, in general, and implicate LOX in thrombotic events related to PMF, thus, recognizing the LOX pathway as new potential target for future anti-thrombosis drug development.

抽象的 控制血栓形成对于各种病理的管理至关重要。这里提出的研究将确定 血栓形成的新的，出乎意料的途径，涉及酶赖氨酸氧化酶（LOX）。拟议的工作着眼于 LOX对原发性骨髓纤维化（PMF）中血小板功能和血栓形成的影响，这种病理学标志 巨核细胞的增殖和聚类和骨髓中的骨髓纤维化。来自瑞典的数据 1980年至2009年的癌症记录显示，在11,155例骨髓增生性的患者中 肿瘤和44、620匹配的健康对照，动脉血栓形成的风险显着4.9倍 与匹配的对照相比，患者的增加（4.8-5.0 p <0.001），强调了重要性 研究调节剂涉及控制与该病理相关的血栓形成。洛克斯已知可以氧化 肽基赖氨酸，导致骨髓生态位的交联基质蛋白。我们发表的研究了 确定了巨核细胞中的LOX上调与骨髓纤维化进展之间的联系 骨髓纤维化的小鼠模型。重要的是，最近我们发现Lox表达在 PMF患者的血小板（大多数测试携带JAK2V617F或钙网蛋白基因 突变），与匹配对照组相比，从PMF患者中分离出的血小板具有更大的附着力 与对照组相比，I型胶原蛋白。此外，在过表达Lox的转基因小鼠血小板，在 与来自骨髓纤维化小鼠模型的细胞相似的水平，显示对单体胶原蛋白的粘附增加， 并明显更大的体内动脉血栓形成倾向。考虑LOX已知活动，我们 假设它通过至少一种胶原蛋白受体的赖氨酸氧化来影响血小板， 涉及Oxy-Western印迹分析的初步研究进一步支持。因此，我们提出了两个特定的 研究目的：目标1。探索LOX使用使用的机制影响血小板对胶原蛋白的反应 蛋白质组学方法以及小鼠和人类MPN样品；目标2。确定Lox-的相关性 调节骨髓纤维化小鼠体内血栓形成，以及胶原受体在这种作用中的作用。追求 这些目的将通过：1）在我们的实验室中产生的新的转基因小鼠系，其中血小板Lox 水平在野生型背景下被上调（不含MPN），2）新LOX抑制剂的可用性， 和LOX基因丧失功能研究。这项工作具有创新性，因为我们是第一个揭示在 LOX和胶原受体激活。此外，在以前没有怀疑这些受体 氧化。我们的研究很重要，因为它将为血小板激活的基本机制提供新的启示 一般并暗示与PMF相关的血小板事件中的LOX，因此将LOX途径识别为新的 未来抗凝血药物开发的潜在目标。

项目成果

The role of extracellular matrix stiffness in megakaryocyte and platelet development and function.

• DOI: 10.1002/ajh.25008
• 发表时间: 2018-03
• 期刊: American journal of hematology
• 影响因子: 12.8
• 作者: Leiva O;Leon C;Kah Ng S;Mangin P;Gachet C;Ravid K
• 通讯作者: Ravid K

Lysyl oxidase inhibition in primary myelofibrosis: A renewed strategy.

• DOI: 10.46439/stemcell.1.005
• 发表时间: 2020-12
• 期刊: Archives of stem cell and therapy
• 作者: Piasecki A;Leiva O;Ravid K
• 通讯作者: Ravid K

Editorial: Novel Treatment Strategies for Myeloproliferative Neoplasms.

• DOI: 10.3389/fonc.2021.762928
• 发表时间: 2021
• 期刊: Frontiers in oncology
• 影响因子: 4.7
• 作者: Lim KH;Matsuura S;Xu B
• 通讯作者: Xu B

Novel lysyl oxidase inhibitors attenuate hallmarks of primary myelofibrosis in mice.

• DOI: 10.1007/s12185-019-02751-6
• 发表时间: 2019-12
• 期刊: International journal of hematology
• 影响因子: 2.1
• 作者: Leiva O;Ng SK;Matsuura S;Chitalia V;Lucero H;Findlay A;Turner C;Jarolimek W;Ravid K
• 通讯作者: Ravid K

Cardiovascular Disease in Myeloproliferative Neoplasms: JACC: CardioOncology State-of-the-Art Review.

• DOI: 10.1016/j.jaccao.2022.04.002
• 发表时间: 2022-06
• 期刊: JACC: CARDIOONCOLOGY
• 影响因子: 11.1
• 作者: Leiva, Orly;Hobbs, Gabriela;Ravid, Katya;Libby, Peter
• 通讯作者: Libby, Peter