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.

尽管存在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。阿片类药物过量主要导致呼吸衰竭，而可卡因滥用主要是 与心血管（CV）毒性相关，包括高血压（HTN）、主动脉僵硬、 和动脉粥样硬化。事实上，可卡因滥用对一般人来说是一个重大的心血管风险。 人口和退伍军人。众所周知，可卡因会刺激交感神经系统 (SNS) 通过抑制神经末梢去甲肾上腺素 (NE) 的再摄取；然而，最近的证据 表明 NE 再摄取的抑制可能不是可卡因诱导的 HTN 的主要驱动因素。作为 因此，介导可卡因对心血管系统影响的机制仍然很大程度上未知。 为此，我们最近对小鼠主动脉进行了小 RNA 和 RNA 测序 用可卡因、甲碘化可卡因（CM，不进入中枢神经系统）治疗， 或盐水来识别潜在的 microRNA (miR)——介导 CV 效应的 mRNA 通路 可卡因。涉及九个 miR-mRNA 通路。我们优先考虑并确定了两个 miR- mRNA 轴基于其表达变化水平以及与 CV 生理学的相关性。他们 是：1) ↑miR-30c—↓苹果酸酶 1 (ME1)—↑活性氧 (ROS) 活性，其中 在 HTN 和血管老化（主动脉僵化）中至关重要； 2) ↓miR-423—↑Cacna2d2 （编码电压依赖性钙通道的α2δ-2亚基）—↑导致钙内流 细胞内钙浓度 ([Ca2+]i) 增加，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 通路，并最近在《高血压》杂志上发表了我们的研究结果。在初步 研究表明，可卡因和 CM 诱导的 miR-423-5p 表达沉默 随后 Cacna2d2 的上调导致 [Ca2+]i 增加，从而增强了收缩力 培养的 SMC。此外，miR-423-5p 过表达可改善可卡因诱导的血压 体内升高。有趣的是，miR-423-5p 与心力衰竭和冠状动脉疾病相关 动脉疾病。其在 HTN 发病机制中的作用仍不清楚。根据我们发布的 通过工作和初步研究，我们假设 miR-423-Cacna2d2 轴起着 通过调节钙内流和细胞内在可卡因诱导的高血压中发挥重要作用 血管细胞中的钙浓度 ([Ca2+]i)。此外，最近的研究有力地支持了 这两个生物过程（Ca2+ 信号传导和 ROS）之间的串扰。我们有试点数据 结果表明，两个 miR 轴的修饰在很大程度上消除了可卡因诱导的 SMC 收缩。 因此，我们进一步假设这两种途径协同作用以介导 可卡因引起的心血管后果。我们将彻底表征 ↓miR-423-5p— ↑Cacna2d2—↑ [Ca2+]i 轴及其与 ↑miR-30c—↓ME1—↑ROS 通路的相互作用 通过使用互补和垂直调节可卡因对 CV 系统的影响 集成的体外、离体和体内模型。此外，我们还将测量NE及其代谢物 水平，以及在体内实验中使用哌唑嗪（α-阻滞剂）来阻断 NE 的作用， 旨在表征 miR-mRNA 轴和 SNS 在介导中的潜在相互作用 可卡因的心血管效应。