CD39-carrying extracellular vesicles regulate pulmonary thrombosis in Sickle Cell Disease

携带CD39的细胞外囊泡调节镰状细胞病中的肺血栓形成

基本信息

批准号：
10736531
负责人：
Prithu Sundd
金额：
$ 78.95万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-07-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10736531
关键词：
Acute Address Adenosine Diphosphate Adenosine Monophosphate Affect African American African American population Attenuated Autopsy Biochemical Blood Blood Platelets Blood flow Blood specimen Clinical Trials Development Diagnostic Disease Endothelium Erythrocytes Event Generations Genes Genetic Hemolysis Hospitalization Human Image Impairment In Situ In Vitro Intravenous Introns Lung Microfluidics Microscopy Molecular Mus Pathway interactions Patients Plasma Platelet Activation Platelet Aggregation Induction Platelet aggregation Preventive therapy Recording of previous events Respiratory Failure Risk Sickle Cell Anemia Signal Transduction Single Nucleotide Polymorphism Solid Testing Therapeutic Thrombocytopenia Thrombosis Thrombus Time Wild Type Mouse X-Ray Computed Tomography antagonist arteriole ecto-nucleotidase extracellular vesicles genetic analysis genome analysis high risk in vivo in vivo imaging intravenous administration lung imaging mouse model novel precision medicine prevent receptor whole genome

Acute Address Adenosine Diphosphate Adenosine Monophosphate Affect African American African American population Attenuated Autopsy Biochemical Blood Blood Platelets Blood flow Blood specimen Clinical Trials Development Diagnostic Disease Endothelium Erythrocytes Event Generations Genes Genetic Hemolysis Hospitalization Human Image Impairment In Situ In Vitro Intravenous Introns Lung Microfluidics Microscopy Molecular Mus Pathway interactions Patients Plasma Platelet Activation Platelet Aggregation Induction Platelet aggregation Preventive therapy Recording of previous events Respiratory Failure Risk Sickle Cell Anemia Signal Transduction Single Nucleotide Polymorphism Solid Testing Therapeutic Thrombocytopenia Thrombosis Thrombus Time Wild Type Mouse X-Ray Computed Tomography antagonist arteriole ecto-nucleotidase extracellular vesicles genetic analysis genome analysis high risk in vivo in vivo imaging intravenous administration lung imaging mouse model novel precision medicine prevent receptor whole genome

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项目摘要

PROJECT SUMMARY Sickle cell disease (SCD) is the most common hemolytic disorder affecting African Americans. In situ (de novo) acute pulmonary thrombosis is the underlying cause in ~20% of SCD patients hospitalized with respiratory failure. The current therapy for pulmonary thrombosis in SCD is primarily supportive and a preventive therapy does not exist. Autopsy and computed tomography studies have identified that occlusion of pulmonary arterioles by platelet-rich thrombi contributes to the development of pulmonary thrombosis in SCD patients. Adenosine diphosphate (ADP) released from lysed erythrocytes activates platelets by stimulating purinergic P2Y1 and P2Y12 receptors. We recently discovered that this pathway promotes pulmonary arteriole thrombosis following acute-hemolysis in wild-type mice, suggesting that ADP-induced purinergic signaling may also promote pulmonary thrombosis in a hemolytic disorder such as SCD. However, P2Y12 receptor antagonists have shown no benefit to SCD patients in recent clinical trials, and it remains to be identified why pulmonary thrombosis develops only in a sub-set but not all SCD patients. Identifying molecular and genetic mechanisms that trigger pulmonary thrombosis in SCD, would enable the development of Precision Medicine diagnostic and therapeutic approaches for these at-risk SCD patients. Based on our new preliminary findings, we hypothesize that CD39 (ecto-nucleotidase) present in circulating extracellular vesicles (CD39+-EVs) degrades excess ADP to prevent pulmonary thrombosis in SCD, however, single-nucleotide-polymorphism (SNP) rs3176891G in the CD39- encoding gene (ENTPD1) attenuates this protection and identifies SCD patients who can benefit from anti- purinergic therapy. We will test this hypothesis using our newly developed mouse model of pulmonary thrombosis triggered by intravenous (IV) administration of ADP, in vivo imaging of lung in live mice, in vitro microfluidic studies with SCD patient blood, isolation and characterization of EVs, SCD mice genetically deficient in CD39, and genetic analyses in SCD patients with vs without SNP rs3176891G. In Aim 1, we will determine whether ADP-induced platelet aggregation and pulmonary thrombosis is impaired in SCD. In Aim 2, we will determine whether circulating CD39+-EVs degrade excess ADP to prevent pulmonary thrombosis in SCD. In Aim 3, we will determine whether SNP rs3176891G promotes pulmonary thrombosis event in SCD patients by attenuating CD39+-EVs, leading to increased ADP-induced platelet aggregation. These studies will introduce a novel paradigm that CD39+-EVs prevent pulmonary thrombosis in SCD, and establish the premise for first-ever precision medicine in SCD by identifying rs3176891G as a risk for pulmonary thrombosis.

项目摘要镰状细胞疾病（SCD）是影响非洲裔美国人的最常见的溶血疾病。原位（从头开始）急性肺血栓形成是约20％的SCD患者的根本原因失败。 SCD中肺血栓形成的当前疗法主要是支持性的，是一种预防疗法不存在。尸检和计算机断层扫描研究已经确定肺动脉的阻塞富含血小板的血栓有助于SCD患者的肺血栓形成。腺苷从裂解的红细胞释放的双磷酸（ADP）通过刺激嘌呤能P2Y1和 P2Y12受体。我们最近发现，此途径促进了肺动脉血栓形成野生型小鼠的急性溶血，表明ADP诱导的嘌呤能信号传导也可能促进溶血障碍（例如SCD）中的肺血栓形成。但是，P2Y12受体拮抗剂已显示在最近的临床试验中，SCD患者没有任何好处，尚待确定为什么肺血栓形成仅在子设置中开发，但不是所有SCD患者。识别触发的分子和遗传机制 SCD中的肺血栓形成将使精确医学的诊断和治疗能力发展这些高危SCD患者的方法。根据我们的新初步发现，我们假设CD39 （Ecto-核苷酸酶）存在于循环外囊泡（CD39+-EVS）中，使多余的ADP降解以防止然而，SCD中的肺血栓形成，单核苷酸 - 多态性（SNP）rs3176891g在CD39-中编码基因（ENTPD1）削弱了这种保护，并确定可以从抗抗事中受益的SCD患者嘌呤能疗法。我们将使用我们新开发的肺动物模型检验该假设由静脉内（IV）ADP施用的血栓形成，活小鼠的肺部体内成像，体外 SCD患者血液，EV的隔离和表征，SCD小鼠的微流体研究在遗传上缺乏在CD39中，在没有SNP RS3176891G的SCD患者中进行了遗传分析。在AIM 1中，我们将确定 SCD中是否会损害ADP诱导的血小板聚集和肺血栓形成。在AIM 2中，我们将确定循环CD39+-EV是否会降低过量的ADP，以防止SCD中的肺血栓形成。目标 3，我们将确定SNP rs3176891g是否通过减弱CD39+-EV，导致ADP诱导的血小板聚集增加。这些研究将引入 CD39+-EVS预防SCD中的肺血栓形成的新型范式，并为有史以来的前提建立前提 SCD中的精密药物通过识别Rs3176891g是肺血栓形成的风险。