Role of novel RNA binding protein LARP6 in alcoholic cardiomyopathy

新型RNA结合蛋白LARP6在酒精性心肌病中的作用

• 批准号: 10593688
• 负责人: Chandrasekar Bysani
• 金额: $ 23.09万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10593688
• 关键词: 5' Untranslated Regions; Adrenergic Agents; Adrenergic beta-Agonists; Alcohol abuse; Alcoholic Cardiomyopathy; Alcohols; Animal Model; Apoptosis; Attenuated; Binding; Cardiac; Cardiac Myocytes; Cardiomyopathies; Chronic; Clinical; Clinical Treatment; Collagen; Data; Deposition; Development; Dilated Cardiomyopathy; Dimensions; Ethanol; Fibrillar Collagen; Fibroblasts; Fibrosis; Functional disorder; Genetic; Genetic Models; Genetic study; Goals; Heart Injuries; Heart failure; Heavy Drinking; Hepatic Stellate Cell; Hypertension; Hypertrophy; Impairment; In Vitro; Inflammation; Infusion procedures; Intervention; Intervention Studies; Isoproterenol; Knock-in; Knock-in Mouse; Left; Left Ventricular Ejection Fraction; Ligation; Liver Fibrosis; Messenger RNA; Modeling; Mus; Mutant Strains Mice; Mutate; Myocardial dysfunction; National Institute on Alcohol Abuse and Alcoholism; Outcome Measure; Oxidative Stress; Pre-Clinical Model; Prevention strategy; Publishing; RNA-Binding Proteins; Rattus; Regulation; Resistance; Ribonucleoproteins; Risk; Role; Stress; Structure; Testing; Therapeutic; Therapeutic Uses; Ventricular; Wild Type Mouse; bile duct; cardioprotection; coronary fibrosis; crosslink; drug-like compound; heart damage; high throughput screening; in vivo; interstitial; mouse model; new therapeutic target; novel; pharmacologic; posttranscriptional; prevent; prophylactic; protein expression; response; small molecule inhibitor; stem; targeted treatment; treatment strategy; vapor

Chronic excessive alcohol consumption can cause cardiac dysfunction and dilation, clinically termed alcoholic cardiomyopathy (AC). While chronic alcohol abuse alone causes AC, clinical evidence indicates that alcohol can exacerbate cardiac dysfunction resulting from other causes of cardiac injury or stress. Alcohol abuse also increases the risk of hypertension and heart failure, with as many as one third of all dilated cardiomyopathies attributed to AC. The development of cardiac fibrosis appears to be a key mechanism of AC dysfunction, as fibrosis impairs ventricular filling and impedes cardiomyocyte contraction. LARP6 is an RNA binding protein that upregulates collagen I expression post-transcriptionally by binding to the 5’SL structure and stabilizing the mRNA. Using a genetic knock-in mouse with modified 5’SL that prevents LARP6 binding, others have shown significantly decreased overall collagen expression and deposition in bile duct ligation-induced hepatic fibrosis. Stefanovic and colleagues discovered a drug-like compound, C9, that blocks the interaction between LARP6 and collagen I. They demonstrated that C9 inhibits collagen I expression in cultured hepatic stellate cells and prevented the development of fibrosis in animal models of hepatic fibrosis. Our preliminary studies indicate that both the knock-in 5’SL mouse and C9-treated wildtype mice are resistant to the development of cardiac fibrosis and dysfunction in response to chronic isoproterenol infusion. In the current proposal, using this unique genetic model and C9 compound, we will determine the role of LARP6 in AC. Our long-term goal is to identify the mechanisms responsible for alcohol-induced cardiac injury, which can then be targeted for clinical benefit. We hypothesize that blocking the interaction between LARP6 and collagen I mRNA significantly blunts cardiac remodeling (fibrosis and hypertrophy) and contractile dysfunction in a preclinical model of AC. Using the well- established NIAAA chronic+binge alcohol mouse model, we propose the following specific aims: Aim 1 determines whether blocking the interaction between LARP6 and collagen I in the 5’SL knock-in mice blunts alcohol abuse-induced adverse cardiac remodeling and dysfunction. We utilize integrated and systematic in vivo and in vitro approaches focused on ethanol-induced cardiac dysfunction, inflammation, oxidative stress, apoptosis, fibroblast activation, and fibrosis. Aim 2 investigates whether pharmacological blockade of LARP6 and collagen I interaction by the small molecule inhibitor C9 inhibits and/or reverses chronic alcohol abuse- induced cardiomyopathy. In this interventional approach, we will determine the therapeutic potential of C9 both prophylactically and therapeutically using the NIAAA model of AC. Outcome measures include cardiac diastolic and systolic function, ventricular chamber dimensions, collagen typing and cross-linking, inflammation, oxidative stress, and interstitial fibrosis. Our proposed genetic and interventional studies will determine if blocking LARP6 and collagen I interaction can blunt chronic alcohol-induced cardiomyopathy and serve as a potential novel therapeutic target for the treatment of clinical AC.

长期过量饮酒可导致心脏功能障碍和扩张，临床上称为酒精性心脏病。 心肌病（AC）。虽然慢性酒精滥用单独导致AC，临床证据表明，酒精 可加重由心脏损伤或压力的其他原因引起的心脏功能障碍。酗酒也 增加高血压和心力衰竭的风险，多达三分之一的扩张型心肌病 因为AC。心脏纤维化的发展似乎是AC功能障碍的关键机制， 纤维化损害心室充盈并阻碍心肌细胞收缩。LARP 6是一种RNA结合蛋白 其通过与5 'SL结构结合并稳定转录后的胶原蛋白I表达， mRNA。使用具有阻止LARP 6结合的修饰的5 'SL的遗传敲入小鼠，其他人已经显示 显著降低胆管结扎诱导的肝纤维化中的总胶原蛋白表达和沉积。 Stefanovic及其同事发现了一种类似药物的化合物C9，它可以阻断LARP 6与 和胶原I。他们证明C9抑制培养的肝星状细胞中胶原I的表达， 在肝纤维化动物模型中阻止纤维化的发展。我们的初步研究表明， 基因敲入的5’SL小鼠和C9处理的野生型小鼠都对心脏纤维化的发展具有抗性 以及对慢性异丙肾上腺素输注的功能障碍。在目前的提案中，利用这种独特的基因， 模型和C9化合物，我们将确定LARP 6在AC中的作用。我们的长期目标是确定 酒精诱导的心脏损伤的机制，然后可以有针对性的临床效益。我们 假设阻断LARP 6和胶原蛋白I mRNA之间的相互作用显著地钝化心脏 在AC的临床前模型中的重构（纤维化和肥大）和收缩功能障碍。利用这口井- 建立NIAAA慢性+酗酒小鼠模型，我们提出以下具体目的：目的1 确定在5 'SL基因敲入小鼠中阻断LARP 6和胶原蛋白I之间的相互作用是否会减弱 酒精滥用引起的不良心脏重塑和功能障碍。我们利用集成和系统化， 体内和体外方法集中在乙醇诱导的心脏功能障碍，炎症，氧化应激， 细胞凋亡、成纤维细胞活化和纤维化。目的2研究LARP 6的药理学阻断是否 和胶原蛋白I的相互作用抑制和/或逆转慢性酒精滥用- 诱发心肌病。在这种介入方法中，我们将确定C9的治疗潜力， 使用AC的NIAAA模型进行药理学和治疗。结果指标包括心脏舒张压 和收缩功能，心室腔尺寸，胶原分型和交联，炎症， 氧化应激和间质纤维化。我们建议的遗传和干预研究将确定， 阻断LARP 6和I型胶原的相互作用可以减弱慢性酒精诱导的心肌病， 为临床AC的治疗提供了潜在的新靶点。