MicroRNA to decrease vascular CaV1.2 in hypertension

MicroRNA 可降低高血压患者的血管 CaV1.2

基本信息

批准号：
7785275
负责人：
PHILIP T. PALADE
金额：
$ 36.25万
依托单位：
UNIV OF ARKANSAS FOR MED SCIS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-22 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7785275
关键词：
Adverse effects American Angiotensin II Antihypertensive Agents Blood Pressure Blood Vessels Calcium Channel Calcium Channel Blockers Cardiovascular Diseases Cells Chronic Clinical Coupled DNA Sequence Dependovirus Dose Drug usage Enhancers Epidemic Event Exhibits Experimental Animal Model Foundations Genes Hypertension Hypotension In Vitro Infusion procedures Lead Mediating Membrane Potentials Mesenteric Arteries Mesentery Methods MicroRNAs Microelectrodes Minority Mus Myosin Heavy Chains Norepinephrine Patients Pharmaceutical Preparations Phenotype Potassium Channel Proteins Regimen Rest Site Small RNA Smooth Muscle Smooth Muscle Myocytes Splanchnic Circulation Technology Testing Therapeutic Time Transcript Vascular Smooth Muscle Vasoconstrictor Agents Viral Voltage-Gated Potassium Channel Western Blotting Western World adeno-associated viral vector base cell type channel blockers compliance behavior design in vivo mouse model normotensive novel patch clamp pressure promoter public health relevance restoration vasoconstriction

项目摘要

DESCRIPTION (provided by applicant): There are many short-acting drugs available to treat hypertension. However, most patients fail to adhere to a daily, multi-drug antihypertensive regimen with frequent side effects. Thus, normal blood pressure is restored in only a minority of hypertensive patients, and only long-term, targeted antihypertensive therapies with fewer side effects will fundamentally impact the epidemic levels of hypertension in the Western world. The studies in this proposal seek to develop a novel, long-term therapy for hypertension by using an adeno-associated viral (AAV) vector containing an enhanced smooth muscle specific promoter (EnSM221) to deliver exogenous microRNA directed against vascular L-type (CaV1.2) calcium channels. Notably, CaV1.2 channels already are the target of clinical calcium channel blocking drugs used to lower blood pressure. However, our strategy will use AAV-mediated delivery of CaV1.2 channel microRNA to enable long- term and vascular-specific knockdown of calcium channel expression. Our aims include: 1) to demonstrate the ability of CaV1.2 channel microRNA to reduce blood pressure for at least 8 weeks without serious side effects in hypertensive mice, 2) confirm in vivo and in vitro that the vasoconstrictor function of CaV1.2 channels is suppressed in small mesenteric arteries, and 3) demonstrate that decreases in vascular CaV1.2 channel expression, whole-cell Ca2+ current, and restoration of K+ channel expression and resting membrane potential in mesenteric arterial cells correlate with the antihypertensive effect of CaV1.2 channel microRNA delivered by AAV to the vasculature. The results of this study will help to establish a foundation for using targeted AAV and microRNA technology to treat systemic hypertension and other chronic vascular abnormalities that are poorly controlled by short-term therapies. PUBLIC HEALTH RELEVANCE: High blood pressure known as hypertension afflicts over 60 million Americans and can lead to even more debilitating conditions. While there are numerous short-acting medications (including calcium channel blockers) to treat hypertension, most patients do not take their medications faithfully and consequently less than one third of patients have their blood pressure adequately controlled. We have developed a small RNA-based therapeutic (microRNA) which decreases expression of a calcium channel found in blood vessels that is also the site of action of all antihypertensive calcium channel blockers. When incorporated into a safe adeno associated virus, our therapeutic can continuously produce more microRNA over a period of months. In addition our therapeutic is coupled to a DNA sequence that will selectively generate the desired microRNA only in vascular smooth muscle cells that make up the wall of blood vessels. This is likely to reduce side effects encountered with many orally administered antihypertensive medications, which spread throughout the body. A longer lasting therapy with fewer side effects may be extremely beneficial for the >40 million Americans whose hypertension is not properly managed.

描述（由申请人提供）：有许多短效药物可用于治疗高血压。然而，大多数患者未能坚持每日的多种药物抗高血压治疗方案，并且经常出现副作用。因此，只有少数高血压患者的血压能够恢复正常，只有长期、副作用较少的针对性降压治疗才能从根本上影响西方世界高血压的流行水平。该提案中的研究旨在开发一种新的、长期的高血压疗法，通过使用含有增强型平滑肌特异性启动子（EnSM221）的腺相关病毒（AAV）载体来递送针对血管L型（CaV1）的外源microRNA .2) 钙通道。值得注意的是，CaV1.2通道已经成为临床用于降低血压的钙通道阻断药物的靶标。然而，我们的策略将使用 AAV 介导的 CaV1.2 通道 microRNA 的传递，以实现钙通道表达的长期和血管特异性敲低。我们的目标包括：1) 证明 CaV1.2 通道 microRNA 能够在高血压小鼠中降低血压至少 8 周而不产生严重副作用，2) 在体内和体外证实 CaV1.2 通道的血管收缩功能在小肠系膜动脉中受到抑制，并且 3) 表明血管 CaV1.2 通道表达、全细胞 Ca2+ 电流减少，K+ 通道表达和静息膜电位恢复肠系膜动脉细胞与 AAV 递送至脉管系统的 CaV1.2 通道 microRNA 的抗高血压作用相关。这项研究的结果将有助于为使用靶向 AAV 和 microRNA 技术治疗短期治疗效果不佳的全身性高血压和其他慢性血管异常奠定基础。公共卫生相关性：高血压（又称高血压）困扰着超过 6000 万美国人，并可能导致更严重的疾病。虽然有许多短效药物（包括钙通道阻滞剂）可以治疗高血压，但大多数患者并没有忠实地服药，因此只有不到三分之一的患者血压得到充分控制。我们开发了一种基于小 RNA 的治疗药物 (microRNA)，它可以降低血管中钙通道的表达，而血管中钙通道也是所有抗高血压钙通道阻滞剂的作用部位。当掺入安全的腺相关病毒时，我们的治疗方法可以在几个月内持续产生更多的 microRNA。此外，我们的治疗方法与 DNA 序列相结合，该序列将仅在构成血管壁的血管平滑肌细胞中选择性地产生所需的 microRNA。这可能会减少许多口服抗高血压药物所遇到的副作用，这些副作用会扩散到全身。对于超过 4000 万高血压未得到妥善管理的美国人来说，一种副作用更少、更持久的治疗方法可能极其有益。