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.

项目概要 室性心律失常是慢性心力衰竭（CHF）患者死亡的主要原因。虽然 去肾神经术（RDN）治疗室性心律失常的潜力已被广泛报道，RDN- 引起的不良并发症严重限制了其在临床的应用。我们最近的研究表明，巨噬细胞 星状神经节 (SG) 的扩张和神经炎症导致 CHF 增加的细胞兴奋性 心脏交感神经节后（CSP）神经元，随后促进心脏交感神经 CHF 大鼠的过度激活和室性心律失常发生。粒细胞-巨噬细胞集落刺激因子 (GM-CSF) 是巨噬细胞激活的重要介质。我们的试验数据显示 RDN 会减弱心脏 交感神经过度激活和室性心律失常，伴随着 GM- 的显着减少 CHF 大鼠 SG 中的 CSF 水平和巨噬细胞活化。然而，尚不清楚其是否具有抗心律失常作用。 RDN 的治疗是通过减弱 CHF SG 中 GM-CSF 介导的炎症途径来实现的。继 发现 RDN 的抗心律失常机制，该提案旨在开发一种新的临床干预措施 达到RDN的治疗作用并避免其局限性。由于肾脏受交感神经支配 主要起源于主动脉肾神经节 (ARG) 中的神经元，针对 ARG 神经元可能是一个合乎逻辑的选择 实现 RDN 抗心律失常作用的治疗策略。考虑到光遗传学的优点， 包括通过光敏视蛋白、腺相关病毒对神经元活动的快速、特异性控制 Archaerhodopsin（ArchT，一种抑制性光敏视蛋白）基因将被转染到 CHF 的 ARG 神经元中 老鼠。 ARG 神经元中 ArchT 神经元表达的特异性将通过连接神经元特异性来实现 ArchT 基因的启动子。 ARG 神经元的持续光遗传学沉默将通过照射 LED 探头在自由移动的动物中进行无线控制和供电。我们假设光遗传学 抑制 ARG 神经元会降低 SG 中 CHF 升高的 GM-CSF 水平，从而缓解 SG 中的巨噬细胞活化和神经炎症，从而减弱 CSP 神经元兴奋性，心脏 CHF 中的交感神经过度激活和室性心律失常发生。使用多方面的技术方法 从整个动物到细胞分子水平，我们将设计体内和体外研究来评估这些 问题。具体目标 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 SG 中的神经炎症。具体目标 2，我们将解决 GM-CSF 信号通路 有助于 CHF 增加 CSP 神经元的细胞兴奋性、心脏交感神经过度激活，以及 室性心律失常发生。具体目标3，我们将确定ARGs中的光遗传学沉默是否可以实现 RDN 通过减弱 GM-CSF 诱导的巨噬细胞活化和神经炎症来发挥抗心律失常作用 SG 换算 CHF。这些研究将为致命性室性心律失常的治疗开辟一条新途径 提供一种新颖的临床干预措施，以降低慢性心力衰竭患者的死亡率并改善预后和生活质量。