Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

IF 4.8 2区医学 Q1 Biochemistry, Genetics and Molecular Biology

Neoplasia Pub Date : 2025-03-14 DOI:10.1016/j.neo.2025.101153

İldeniz USLU-BIÇAK , Meliha NALÇACI , Selçuk SÖZER

{"title":"Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression","authors":"İldeniz USLU-BIÇAK , Meliha NALÇACI , Selçuk SÖZER","doi":"10.1016/j.neo.2025.101153","DOIUrl":null,"url":null,"abstract":"<div><div>Philadelphia chromosome-negative myeloproliferative neoplasms (Ph<sup>-</sup>MPNs) are clonal disorders marked by high morbidity and mortality, driven by uncontrolled myeloid proliferation from hematopoietic stem/progenitor cells (HSCs) and associated with a significant risk of thrombosis. This study explored the relationship between <em>JAK2</em>V617F and protease-activated receptor 1 (PAR1) by examining <em>PAR1</em> expression and activation across various hematopoietic stem/progenitor cell (HSPC) subgroups, assessing their contribution to the hypercoagulable state in Ph<sup>-</sup>MPNs.</div><div>We investigated the effects of thrombin, a PAR1 antagonist (vorapaxar), and a JAK2 inhibitor (ruxolitinib) on Ph<sup>-</sup>MPN cells. Mononuclear cells (MNCs) were isolated from Ph-MPN patients (<em>n</em> = 18), cord blood (CB) samples (<em>n</em> = 5) and healthy volunteers (<em>n</em> = 11). Specific subpopulations were sorted and analyzed for PAR1 expression and <em>JAK2</em>V617F status using qRT-PCR. <em>PAR1</em> expression changes, along with other PAR pathway-related genes, were assessed post-treatment.</div><div>Our results revealed that most PAR1<sup>+</sup> cells (∼95 %) co-expressed CD34<sup>+</sup>, with a smaller JAK2V617F<sup>+</sup> PAR1<sup>+</sup> population lacking CD34. PAR1 expression was significantly higher in Ph-MPN MNCs compared to CB (<em>p</em> = 0.0005), particularly in EMP, HSC/EPC, and EPC subsets. Thrombin treatment reduced surface PAR1 expression, while PAR1 antagonist treatment further decrease the expression level. Combined PAR1 antagonist and ruxolitinib treatment significantly downregulated <em>PAR1</em> expression (<em>p</em> < 0.0001), and several PAR-pathway-related genes were notably downregulated after treatment.</div><div>This study highlights that elevated PAR1 expression in primitive hematopoietic subpopulations is linked to disease progression and thrombosis in Ph<sup>-</sup>MPNs, suggesting PAR1 as a potential therapeutic target. Combining PAR1 antagonists with JAK2 inhibitors shows promise in reducing PAR1 expression and mitigating thrombotic events in Ph<sup>-</sup>MPN patients.</div></div>","PeriodicalId":18917,"journal":{"name":"Neoplasia","volume":"63 ","pages":"Article 101153"},"PeriodicalIF":4.8000,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Neoplasia","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1476558625000326","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}

引用次数: 0

Abstract

Philadelphia chromosome-negative myeloproliferative neoplasms (Ph^-MPNs) are clonal disorders marked by high morbidity and mortality, driven by uncontrolled myeloid proliferation from hematopoietic stem/progenitor cells (HSCs) and associated with a significant risk of thrombosis. This study explored the relationship between JAK2V617F and protease-activated receptor 1 (PAR1) by examining PAR1 expression and activation across various hematopoietic stem/progenitor cell (HSPC) subgroups, assessing their contribution to the hypercoagulable state in Ph^-MPNs.

We investigated the effects of thrombin, a PAR1 antagonist (vorapaxar), and a JAK2 inhibitor (ruxolitinib) on Ph^-MPN cells. Mononuclear cells (MNCs) were isolated from Ph-MPN patients (n = 18), cord blood (CB) samples (n = 5) and healthy volunteers (n = 11). Specific subpopulations were sorted and analyzed for PAR1 expression and JAK2V617F status using qRT-PCR. PAR1 expression changes, along with other PAR pathway-related genes, were assessed post-treatment.

Our results revealed that most PAR1⁺ cells (∼95 %) co-expressed CD34⁺, with a smaller JAK2V617F⁺ PAR1⁺ population lacking CD34. PAR1 expression was significantly higher in Ph-MPN MNCs compared to CB (p = 0.0005), particularly in EMP, HSC/EPC, and EPC subsets. Thrombin treatment reduced surface PAR1 expression, while PAR1 antagonist treatment further decrease the expression level. Combined PAR1 antagonist and ruxolitinib treatment significantly downregulated PAR1 expression (p < 0.0001), and several PAR-pathway-related genes were notably downregulated after treatment.

This study highlights that elevated PAR1 expression in primitive hematopoietic subpopulations is linked to disease progression and thrombosis in Ph^-MPNs, suggesting PAR1 as a potential therapeutic target. Combining PAR1 antagonists with JAK2 inhibitors shows promise in reducing PAR1 expression and mitigating thrombotic events in Ph^-MPN patients.

Abstract Image

查看原文本刊更多论文

靶向PAR1激活jak2v617f驱动的费城阴性骨髓增生性肿瘤：揭示其在血栓形成和疾病进展中的作用

费城染色体阴性骨髓增生性肿瘤（ph - mpn）是一种克隆性疾病，其特点是高发病率和死亡率，由造血干细胞/前体细胞（hsc）的髓细胞增殖失控驱动，并与血栓形成的显著风险相关。本研究通过检测PAR1在不同造血干细胞/祖细胞（HSPC）亚群中的表达和激活，探讨了JAK2V617F与蛋白酶激活受体1 （PAR1）之间的关系，评估了它们对ph - mpn高凝状态的贡献。我们研究了凝血酶、PAR1拮抗剂（vorapaxar）和JAK2抑制剂（ruxolitinib）对Ph-MPN细胞的影响。从Ph-MPN患者（n = 18）、脐带血（n = 5）和健康志愿者（n = 11）中分离单个核细胞（MNCs）。对特定亚群进行分类，并使用qRT-PCR分析PAR1表达和JAK2V617F状态。治疗后评估PAR1表达变化以及其他PAR通路相关基因。我们的研究结果显示，大多数PAR1+细胞（约95%）共表达CD34+，而较小的JAK2V617F+ PAR1+群体缺乏CD34。PAR1在Ph-MPN跨国公司中的表达明显高于CB (p = 0.0005)，特别是在EMP、HSC/EPC和EPC亚群中。凝血酶处理降低了表面PAR1的表达，而PAR1拮抗剂处理进一步降低了表达水平。PAR1拮抗剂联合鲁索利替尼治疗可显著下调PAR1的表达(p <；0.0001)，治疗后几个par通路相关基因明显下调。这项研究强调，原始造血亚群中PAR1表达升高与ph - mpn的疾病进展和血栓形成有关，表明PAR1是一个潜在的治疗靶点。PAR1拮抗剂与JAK2抑制剂联合使用有望降低Ph-MPN患者的PAR1表达并减轻血栓事件。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Neoplasia 医学-肿瘤学

CiteScore

9.20

自引率

2.10%

发文量

审稿时长

26 days

期刊介绍： Neoplasia publishes the results of novel investigations in all areas of oncology research. The title Neoplasia was chosen to convey the journal’s breadth, which encompasses the traditional disciplines of cancer research as well as emerging fields and interdisciplinary investigations. Neoplasia is interested in studies describing new molecular and genetic findings relating to the neoplastic phenotype and in laboratory and clinical studies demonstrating creative applications of advances in the basic sciences to risk assessment, prognostic indications, detection, diagnosis, and treatment. In addition to regular Research Reports, Neoplasia also publishes Reviews and Meeting Reports. Neoplasia is committed to ensuring a thorough, fair, and rapid review and publication schedule to further its mission of serving both the scientific and clinical communities by disseminating important data and ideas in cancer research.