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: 10016586
• 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/10016586
• 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; 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; Neuraxis; Nimodipine; Norepinephrine; Opioid; Outcome; Pathogenesis; Pathologic; Pathway interactions; Patient Care; Patients; Pharmacology; 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; base; 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; respiratory; reuptake; transcriptome sequencing

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，但可卡因仍然是公众广泛滥用的非法药物， 退伍军人。阿片类药物过量主要导致呼吸衰竭，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种显著的CV风险。 人口和退伍军人。众所周知，可卡因刺激交感神经系统 (SNS)通过抑制神经末梢的去甲肾上腺素（NE）再摄取;然而，最近的证据表明， 提示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CV系统的作用的机制在很大程度上仍然未知。 为此，我们最近在小鼠的睾丸中进行了小RNA和RNA测序， 用可卡因、甲基碘化可卡因（CM，不进入中枢神经系统）治疗， 或生理盐水，以确定介导CV效应的潜在microRNA（miR）-mRNA途径。 可卡因涉及9条miR-mRNA通路。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和与CV生理学的相关性。他们 是：1）↑miR-30 c-↓苹果酸酶1（ME 1）-↑活性氧（ROS）活性， 在HTN和血管老化（主动脉僵硬）中至关重要; 2）↓miR-423-↑ Cacna 2d 2 （编码电压依赖性钙通道的α2δ-2亚基）-↑钙内流导致 细胞内钙离子浓度（[Ca 2 +]i）增加，这在控制血管生成中至关重要。 平滑肌细胞（SMC）收缩性和血压（BP）。我们彻底调查了 miR-30 c通路，最近在高血压杂志上发表了我们的研究结果。初步 研究中，我们发现可卡因和CM诱导的miR-423- 5 p表达沉默， 随后Cacna 2d 2的上调导致[Ca 2 +]i增加，这增强了心肌细胞的收缩力。 培养的SMC。此外，miR-423- 5 p过表达可改善可卡因诱导的血压。 体内升高。有趣的是，miR-423- 5 p与心力衰竭和冠状动脉粥样硬化相关。 动脉疾病其在HTN发病机制中的作用仍不清楚。根据我们公布的 工作和初步研究中，我们假设miR-423-Cacna 2d 2轴发挥作用， 可卡因诱导HTN中钙内流和细胞内钙的调节作用 血管细胞内钙离子浓度（[Ca 2 +]i）。此外，最近的研究支持一个强有力的 这两个生物过程-Ca 2+信号和ROS之间的串扰。我们有飞行员数据 显示两个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，这两个途径协同工作，介导 可卡因引起的CV后果。我们将彻底表征↓miR-423- 5 p- ↑ Cacna 2d 2-↑ [Ca 2 +]i轴及其与↑miR-30 c-↓ ME 1-↑ROS通路的相互作用 通过使用互补的和垂直的方式来介导可卡因对CV系统的影响， 整合的体外、离体和体内模型。此外，我们将测量NE及其代谢产物 水平，以及在体内实验中使用哌唑嗪（一种β-受体阻滞剂）来阻断NE的作用， 目的是表征miR-mRNA轴和SNS在介导 可卡因对心血管的影响