Optogenetic silencing to achieve antiarrhythmic effect of renal denervation in chronic heart failure

光遗传学沉默实现肾去神经支配慢性心力衰竭的抗心律失常作用

• 批准号: 10714486
• 负责人: Yu-Long Li
• 金额: $ 59.44万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-28 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714486
• 关键词: Animals; Anti-Arrhythmia Agents; Aorta; Arrhythmia; Attenuated; Blood Vessels; Calcium; Calmodulin; Cardiac; Cause of Death; Cells; Chronic; Clinic; Clinical; Congestive Heart Failure; Conscious; Data; Denervation; Dependovirus; Ganglia; Genes; Genetic; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor Receptors; In Vitro; Infection; Inflammatory; Infusion procedures; Interleukin-1 beta; Intervention; Kidney; Light; Link; Macrophage; Macrophage Activation; Measures; Mediating; Mediator; Microglia; Molecular; Nerve; Neuroglia; Neurons; Operative Surgical Procedures; Opsin; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Population; Predisposition; Production; Prognosis; Quality of life; Rattus; Renal Artery Stenosis; Reporting; Role; Safety; Serum; Severity of illness; Signal Pathway; Signal Transduction; Specificity; Structure of stellate ganglion; TNF gene; Techniques; Testing; Therapeutic; Tissues; Transfection; Ventricular; Ventricular Arrhythmia; design; excitatory neuron; improved; improved outcome; in vivo; kidney dysfunction; miniaturize; monocyte; mortality; nerve supply; neuroinflammation; neuronal excitability; novel; novel therapeutic intervention; optogenetics; promoter; reinnervation; release factor; wireless

Project Summary Ventricular arrhythmia is the leading cause of death for chronic heart failure (CHF) patients. Although the therapeutic potential of renal denervation (RDN) for ventricular arrhythmias has been reported extensively, RDN- induced adverse complications severely limit its use in the clinic. Our recent study revealed that macrophage expansion and neuroinflammation in the stellate ganglion (SG) contribute to CHF-increased cell excitability of cardiac sympathetic postganglionic (CSP) neurons, which subsequently promotes cardiac sympathetic overactivation and ventricular arrhythmogenesis in CHF rats. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a crucial mediator in macrophage activation. Our pilot data showed that RDN attenuates cardiac sympathetic overactivation and ventricular arrhythmias, which are accompanied by the marked reduction of GM- CSF level and macrophage activation in SGs in CHF rats. However, it remains unclear if the antiarrhythmic effect of RDN is achieved via attenuating GM-CSF-mediated inflammatory pathways in SGs in CHF. Following the discovery of the antiarrhythmic mechanisms of RDN, this proposal aims to develop a novel clinical intervention to achieve the therapeutic role of RDN and avoid its limitations. Since sympathetic innervation of the kidney originates primarily from neurons in the aorticorenal ganglion (ARG), targeting ARG neurons could be a logical therapeutic strategy for achieving the antiarrhythmic role of RDN. Considering the advantages of optogenetics, including rapid, specific control of neuronal activities by light-sensitive opsins, adeno-associated-virus- Archaerhodopsin (ArchT, an inhibitory light-sensitive opsin) gene will be transfected into ARG neurons in CHF rats. Specificity of neuronal expression of ArchT in ARG neurons will be achieved by linking a neuron-specific promoter to the ArchT gene. Continual optogenetic silencing in ARG neurons will be achieved by illuminating a LED probe that is controlled and powered wirelessly in freely moving animals. We hypothesize that optogenetic inhibition of ARG neurons would reduce CHF-elevated GM-CSF level in SGs, which subsequently alleviates macrophage activation and neuroinflammation in SGs, thereby attenuating CSP neuronal excitability, cardiac sympathetic overactivation, and ventricular arrhythmogenesis in CHF. Using multi-faceted technical approaches ranging from whole-animals to cellular-molecular levels, we will design in vivo and in vitro studies to assess these questions. Specific Aim 1, we will test if GM-CSF signaling axis contributes to macrophage activation and neuroinflammation in SGs from CHF animals. Specific Aim 2, we will address if GM-CSF signaling pathway contributes to CHF-increased cell excitability of CSP neurons, cardiac sympathetic overactivation, and ventricular arrhythmogenesis. Specific Aim 3, we will determine if optogenetic silencing in ARGs can achieve the antiarrhythmic effect of RDN by attenuating GM-CSF-induced macrophage activation and neuroinflammation in SGs in CHF. These studies will open a new avenue in therapeutics against lethal ventricular arrhythmia and provide a novel clinical intervention to reduce mortality and improve outcomes and quality of life in CHF patients.

项目摘要 心室心律不齐是慢性心力衰竭（CHF）患者死亡的主要原因。虽然 肾脏神经（RDN）对心室心律不齐的治疗潜力已广泛报道 诱导的不良并发症严重限制了其在诊所中的使用。我们最近的研究表明巨噬细胞 星状神经节（SG）中的膨胀和神经炎症有助于CHF增强的细胞兴奋性 心脏交感神经后（CSP）神经元，随后促进心脏交感神经 CHF大鼠的过度活化和心室心律失常。粒细胞巨噬细胞刺激因子 （GM-CSF）是巨噬细胞激活中的关键介体。我们的飞行员数据显示，RDN减弱了心脏 交感的过度活化和心室心律不齐，伴随着GM-明显减少 CSF水平和CHF大鼠SGS的巨噬细胞激活。但是，尚不清楚抗心律失常效应是否 通过减弱CHF中SGS中的GM-CSF介导的炎症途径来实现RDN的of。跟随 发现RDN的抗心律失常机制，该提案旨在开发新的临床干预措施 为了达到RDN的治疗作用并避免其局限性。由于肾脏的同情神经 主要来自主动脉神经节（ARG）中的神经元，靶向ARG神经元可能是逻辑 实现RDN的抗心律失常作用的治疗策略。考虑到光遗传学的优势， 包括通过光敏敏感蛋白对神经元活性的快速，特定的控制，腺相关病毒 - Archaerhopopsin（Archt，抑制光敏的Opsin）基因将被转染为CHF中的ARG神经元 老鼠。通过链接神经元特异性，将实现ARGT神经元中ARCT神经元表达的特异性 Archt基因的启动子。 ARG神经元中的持续光遗传沉默将通过照亮A 在自由移动的动物中无线控制和无线驱动的LED探针。我们假设该光学遗传学 抑制ARG神经元将降低SGS中CHF的GM-CSF水平，随后减轻 SGS中的巨噬细胞激活和神经炎症，从而衰减CSP神经元兴奋性，心脏 CHF中的交感神经过度活化和心室心律失常。使用多方面的技术方法 从全动物到细胞分子水平，我们将在体内和体外研究中进行设计以评估这些水平 问题。特定目标1，我们将测试GM-CSF信号轴是否有助于巨噬细胞激活和 CHF动物的SGS中神经炎症。特定目标2，我们将解决GM-CSF信号通路 有助于CSP神经元，心脏交感神经过度激活和 心律失常发生。特定目标3，我们将确定ARG中的光遗传学沉默是否可以实现 通过衰减GM-CSF诱导的巨噬细胞激活和神经炎症，RDN的抗心律失常效应 在瑞士法郎的SGS中。这些研究将为致命性心律不齐和 提供一种新型的临床干预措施，以降低CHF患者的死亡率并改善预后和生活质量。