microRNA Regulation of The Cocaine Effects on the Cardiovascular System

microRNA 调节可卡因对心血管系统的影响

• 批准号: 10514596
• 负责人: CHUNMING DONG
• 金额: --
• 依托单位: MIAMI VA HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10514596
• 关键词: Accounting; Acetylcysteine; Adrenergic Agents; Affect; Aging; Anxiety; Aorta; Arrhythmia; Atherosclerosis; Biological Process; Blood Pressure; Blood Vessels; Calcium; Calcium Channel; Cardiac Catheterization Procedures; Cardiovascular Diseases; Cardiovascular Pathology; Cardiovascular Physiology; Cardiovascular system; Catecholamines; Cause of Death; Cell physiology; Cells; Central Nervous System; Cessation of life; Chronic; Cocaine; Cocaine Abuse; Complex; Coronary Arteriosclerosis; Data; Development; Disease; Emergency department visit; Ensure; Epidemiology; FDA approved; Fatigue; Florida; Funding; General Population; Genes; Genomic approach; Guidelines; Heart failure; Hypertension; Illicit Drugs; In Vitro; Journals; Link; Measures; Mediating; Mental Depression; Messenger RNA; MicroRNAs; Modification; Molecular; Mus; Muscle Contraction; Nerve; Nerve Endings; Nimodipine; Norepinephrine; Opioid; Outcome; Pathogenesis; Pathologic; Pathway interactions; Patient Care; Patients; Play; Post-Traumatic Stress Disorders; Prazosin; Predictive Factor; Prevalence; Publishing; Reactive Oxygen Species; Recovery; Regulation; Reporting; Research; Research Design; Resources; Respiratory Failure; Role; Saline; Severities; Signal Transduction; Small RNA; Smooth Muscle Myocytes; Soldier; Solid; Sympathetic Nervous System; Symptoms; Testing; Toxic effect; Treatment-related toxicity; United States National Institutes of Health; Up-Regulation; Vascular Endothelial Cell; Vascular Smooth Muscle; Veterans; Visit; War; Work; analog; blood pressure elevation; cardiovascular effects; cardiovascular risk factor; clinical practice; cocaine exposure; cocaine use; combat; experience; experimental study; genomic data; illicit drug use; in vitro Model; in vivo; in vivo Model; innovation; malic enzyme; novel therapeutics; opioid epidemic; opioid overdose; overexpression; pharmacologic; respiratory; reuptake; transcriptome sequencing; translational potential

Despite the opioid overdose crisis, cocaine remains a widely abused illicit drug by the public and by veterans. While opioid overdose primarily causes respiratory failure, cocaine abuse is mainly associated with cardiovascular (CV) toxicities, which include hypertension (HTN), aortic stiffness, and atherosclerosis. Indeed, cocaine abuse represents a significant CV risk for the general population and for veterans. It is known that cocaine stimulates the sympathetic nervous system (SNS) by inhibiting norepinephrine (NE) reuptake at nerve terminals; however, recent evidence suggests that inhibition of NE reuptake may not be the major driver of cocaine-induced HTN. As such, the mechanisms mediating the effects of cocaine on the CV system remain largely unknown. To that end, we recently performed small RNA and RNA sequencing in the aortas from mice treated with cocaine, cocaine methiodide (CM, which does not enter the central nervous system), or saline to identify potential microRNA (miR)—mRNA pathways that mediate the CV effects of cocaine. Nine miR—mRNA pathways were implicated. We prioritized and identified two miR- mRNA axes based on their levels of expression changes and relevance to CV physiology. They are: 1) the ↑miR-30c—↓Malic Enzyme 1 (ME1)—↑reactive oxygen species (ROS) activity, which is crucial in HTN and vascular aging (aortic stiffness); and 2) the ↓miR-423—↑Cacna2d2 (encoding the α2δ-2 subunit of voltage-dependent calcium channels) —↑calcium influx resulting in increased intracellular calcium concentration ([Ca2+]i) which is critical in controlling vascular smooth muscle cell (SMC) contractility and blood pressure (BP). We thoroughly investigated the miR-30c pathway and recently published our findings in the journal Hypertension. In preliminary studies, we showed that cocaine- and CM-induced silencing of miR-423-5p expression and subsequent upregulation of Cacna2d2 led to increased [Ca2+]i, which augmented contractility in cultured SMCs. Furthermore, miR-423-5p overexpression ameliorated cocaine-induced BP elevation in vivo. Interestingly, miR-423-5p has been associated with heart failure and coronary artery disease. Its role in the pathogenesis of HTN remains unknown. Based on our published work and preliminary studies, we hypothesize that the miR-423—Cacna2d2 axis plays an important role in cocaine-induced HTN by regulating calcium influx and intracellular calcium concentrations ([Ca2+]i) in vascular cells. In addition, recent studies support a strong crosstalk between these two biological processes—Ca2+ signaling and ROS. We have pilot data showing that modification of both miR axes largely abrogated cocaine-induced SMC contraction. Therefore, we further hypothesize that these two pathways work synergistically to mediate cocaine-induced CV consequences. We will thoroughly characterize the ↓miR-423-5p— ↑Cacna2d2—↑ [Ca2+]i axis and its interaction with the ↑miR-30c—↓ME1—↑ROS pathway in mediating the effects of cocaine on the CV system by using complimentary and vertically integrated in vitro, ex vivo, and in vivo models. In addition, we will measure NE and its metabolite levels, as well as use Prazosin (an -blocker) to block the effects of NE in the in vivo experiments, aiming to characterize the potential interplay between the miR-mRNA axes and SNS in mediating the CV effects of cocaine.

尽管阿片类药物过量危机，可卡因仍然是公众广泛滥用的非法药物， 由退伍军人。虽然阿片类药物过量主要导致呼吸衰竭，但可卡因滥用主要是 与心血管（CV）毒性有关，包括高血压（HTN），主动脉僵硬， 和动脉粥样硬化。实际上，可卡因滥用代表了普通的重大简历风险 人口和退伍军人。众所周知，可卡因会刺激交感神经系统 （SNS）通过抑制神经末端的去甲肾上腺素（NE）再摄取；但是，最近的证据 表明抑制NE再摄取可能不是可卡因诱导的HTN的主要驱动力。作为 这样的机制介导可卡因对CV系统的影响仍然很大未知。 为此，我们最近在小鼠的主动脉中进行了小的RNA和RNA测序 用可卡因，可卡因甲基碘化物（CM，不进入中枢神经系统）处理， 或盐水来鉴定潜在microRNA（miR） - 介导CV效应的MRNA途径 可卡因。九个miR - 暗示了mRNA途径。我们优先考虑并确定了两个mir- mRNA轴基于它们的表达变化水平以及与简历生理的相关性。他们 为：1）↑miR-30c - ↓苹果酶1（ME1） - ↑活性氧（ROS）活性，该活性 对于HTN和血管衰老至关重要（主动脉刚度）； 2）↓mir-423 - ↑cacna2d2 （编码电压依赖性钙通道的α2δ-2亚基） - ↑钙影响 在增加的细胞内钙浓度（[Ca2+] i）中，这对于控制血管至关重要 平滑肌细胞（SMC）收缩性和血压（BP）。我们彻底调查了 miR-30c途径，最近在《高血压》杂志上发表了我们的发现。在初步 研究，我们表明可卡因和CM诱导的miR-423-5p表达的沉默和 随后的cacna2d2上调导致[Ca2+] i增加，这增加了收缩性 培养的SMC。 miR-423-5p过表达可卡因诱导的BP 体内高程。有趣的是，miR-423-5p与心力衰竭和冠状动脉有关 动脉疾病。它在HTN发病机理中的作用仍然未知。根据我们出版的 工作和初步研究，我们假设miR-423-cacna2d2轴扮演 通过调节钙的影响和细胞内，在可卡因诱导的HTN中的重要作用 血管细胞中的钙浓度（[Ca2+] I）。此外，最近的研究支持了强大的 这两个生物学过程之间的串扰-CA2+信号传导和ROS。我们有飞行员数据 表明两个miR轴的修饰在很大程度上废除了可卡因诱导的SMC收缩。 因此，我们进一步假设这两种途径协同作用以进行调解 可卡因引起的简历后果。我们将彻底描述↓mir-423-5p- ↑cacna2d2-↑[Ca2+] i轴及其与↑mir-30c的相互作用 通过免费和垂直的方式介导可卡因对CV系统的影响 在体外，体内和体内模型进行整合。此外，我们将测量NE及其代谢物 水平以及使用prazosin（一个障碍物）来阻止NE在体内实验中的影响 旨在表征介导的miR-mRNA轴与SN之间的潜在相互作用 可卡因的简历作用。