The Molecular Regulation of Platelet Reprogramming by the Metabolic Milieu

项目总结(见说明)： 犹他大学分子医学血栓转化研究中心(U2M2-TRCT)的中心假设是，全身代谢环境将血小板重新编程为显示血栓前表型的细胞。作为这一方案主题的一部分，项目1将开始定义控制巨核细胞和血小板重新编程事件的分子机制。项目1得到了从肥胖和糖尿病患者和小鼠分离的血小板中的下一代RNA测序和功能数据的支持。具体地说，我们证明了代谢环境导致了血小板分子特征和功能的深刻变化。我们的论点是，这些变化不是随机的：相反，我们认为不同的分子机制重新编程了巨核细胞和血小板中的基因表达。 我们将在项目1中测试的一个控制检查点涉及到microRNAs(MiRNAs)的调节。‘ 三个特定的目标将被用来研究肥胖和糖尿病如何重新编程血小板的分子特征和功能。在第一个目标中，并与项目2协调，我们将确定饮食诱导的肥胖、肥胖之后的体重减轻以及全身代谢失衡的治疗纠正是否会改变小鼠血小板的重新编程事件。第一个目标还将纳入对患有代谢综合征的人类患者的推论研究。在目标2中，我们将确定前体miRNA的作用 在调节血小板重编程和功能的过程中，使用血小板缺乏DICER的小鼠，DICER是一种RNaseIII酶，对生产成熟的miRNAs至关重要。在最终目标中，我们将确定miRNAs如何调节从肥胖小鼠和人类分离的血小板中上调的线粒体转录/蛋白(解偶联蛋白2和丝裂原蛋白2)的表达。AIM 3生成的数据将 直接称赞项目2和项目4，项目2测试这些线粒体靶标的功能，项目3和项目4描述它们在从患有代谢综合征的人类分离的血小板中的表达模式。 因此，项目1将有助于U2M2-TRCT计划的各个方面，因为它揭示了代谢环境如何影响血小板的分子特征和功能的新见解。

PROJECT SUMMARY (See instructions): The central hypothesis of the University of Utah Molecular Medicine Translation Research Center in Thrombosis (U2M2-TRCT) is that the systemic metabolic milieu reprograms platelets into cells that display a prothrombotic phenotype. As part of this programmatic theme, project 1 will begin defining the molecular mechanisms that control reprogramming events in megakaryocytes and platelets. Project 1 is supported by next-generation RNA-sequencing and functional data in platelets isolated from obese and diabetic humans and mice. Specifically, we show that the metabolic milieu induces profound changes in the molecular signature and function of platelets. It is our contention that these changes are not random: instead, we believe that distinct molecular mechanisms reprogram gene expression in megakaryocytes and platelets. One control checkpoint, which we will test in project 1, involves regulation by microRNAs (miRNAs). ' Three specific aims will be used to examine how obesity and diabetes reprograms the molecular signature and function of platelets. In the first aim and in coordination with project 2, we will determine if diet-induced obesity, obesity followed by weight loss, and therapeutic correction of systemic metabolic imbalances alters reprogramming events in mouse platelets. The first aim will also incorporate corollary studies in human patients with metabolic syndrome. In aim 2, we will determine the role of precursor miRNA processing in regulating platelet reprogramming and function using mice whose platelets lack Dicer, an RNase III enzyme that is central to the production of mature miRNAs. In the final aim, we will determine how miRNAs regulate the expression of mitochondrial transcripts/proteins (uncoupling protein 2 and mitofusin 2) that are upregulated in platelets isolated from obese mice and humans. Data generated from Aim 3 will directly compliment project 2, which tests the functionality of these mitochondrial targets, and projects 3 and 4 that characterize their expression patterns in platelets isolated from humans with metabolic syndrome. Thus, project 1 will contribute to all facets of the U2M2-TRCT program as it reveals new insights into how the metabolic milieu influences the molecular signature and function of platelets.