YAP and IRF2BP2 regulation of cardiomyocyte innate immune responses

心肌细胞（Cardiomyocytes，CMs）是心脏的基本组成部分和功能单位，其功能障碍或丧失 是心力衰竭的根源与其他专门的先天免疫细胞类似，CM有自己的 先天免疫分子机制缺血性或非缺血性病理性应激激活CM 先天免疫信号通路，刺激促炎细胞因子释放和反应性 氧物种（ROS）的产生。这些先天性免疫反应有利于防御CM 抵抗病原体入侵和用于组织修复，但也可导致心肌损伤。最终 本项目的目标是确定调节CM先天免疫的新分子机制 反应，这将阐明病原体或非病原体相关的发病机制 心肌病损伤/损伤相关分子模式（DAMP）启动先天免疫 通过与模式识别受体结合的反应，以及最具特征的信号轴之一 是TLR 4/NF-κ B通路。Hippo-YAP通路对心脏发育和心脏功能至关重要。 Hippo-YAP的再生和紊乱与一系列心脏病有关。 IRF 2BP 2（干扰素调节因子2结合蛋白2）是一种转录辅因子，在肿瘤细胞中研究很少。 心我们最近发现雅普通过钝化TLR 4/NF-kB来调节CM先天性免疫应答 IRF 2BP 2是抑制心脏中TLR 4表达的关键。本课题 将检验雅普和IRF 2BP 2是CM先天免疫应答的抑制因子的假设。我们 提出以下两个目标。目标1.定义雅普在CM先天免疫反应中的作用。在这 目的，我们将确定雅普对CM先天免疫应答的调节是否依赖于其 转录活性此外，我们将开发一种新型的雅普modRNA递送系统。我们 先前的研究表明，用心肌内递送的雅普modRNA瞬时激活雅普 减少心脏损伤。然而，原位递送方法限制了aYAP modRNA的临床应用。 在这里，我们将测试CM靶向的和纳米颗粒包装的aYAP modRNA在鼠中的功效。 心肌缺血/再灌注模型。目标2.定义IRF 2BP 2在CM先天免疫中的作用 应答在这个目标中，我们将进行IRF 2BP 2功能获得和丧失的研究，以确定其在 CM先天免疫反应。此外，我们将研究如何在分子机制 IRF 2BP 2调节CM先天免疫应答。使用新的Irf 2bp 2敲入小鼠系， 最近产生的，我们将进行IRF 2BP 2 ChIP-Seq，以确定其直接目标。这项建议 是创新和重要的，因为它侧重于解剖基本的分子机制， CM的先天免疫反应，它的目的是开发新的治疗策略，以减少 心肌梗死损伤

Cardiomyocytes (CMs) are building blocks and function units of the heart, and their dysfunction or loss is the root of heart failure. Similar with other specialized innate immune cells, CMs have their own innate immune molecular machinery. Ischemic or non-ischemic pathological stress activates CM innate immune signaling pathways, which stimulate pro-inflammatory cytokine release and reactive oxygen species (ROS) production. These innate immune responses are beneficial for defending CMs against pathogen invasion and for tissue repair, but can also cause myocardial damage. The ultimate goal of this project is to define new molecular mechanisms that regulate CM innate immune responses, which will shed light on the pathogenesis of pathogen or non-pathogen related cardiomyopathy. Damage/danger-associated molecular patterns (DAMPs) initiate innate immune responses by binding to pattern recognition receptors, and one of the best-characterized signaling axis is TLR4/NF-kB pathway. The Hippo-YAP pathway is crucial for heart development and cardiac regeneration, and disturbances in Hippo-YAP have been implicated in a range of heart diseases. IRF2BP2 (interferon regulatory factor 2 binding protein 2) is a transcription co-factor little studied in the heart. We recently found that YAP regulates CM innate immune responses by blunting TLR4/NF-kB signaling, and that IRF2BP2 is crucial for restraining TLR4 expression in the heart. In this project, we will test the hypothesis that YAP and IRF2BP2 are suppressors of CM innate immune responses. We propose the following two aims. Aim 1. Define the role of YAP in CM innate immune responses. In this aim, we will determine whether YAP regulation of CM innate immune responses depends on its transcriptional activity. Additionally, we will develop a novel YAP modRNA delivery system. We previously showed that transiently activating YAP with intra-myocardial delivered aYAP modRNA reduced heart injury. However, the in situ delivery method limits aYAP modRNA's clinical application. Here, we will test the efficacy of CM targeted and nano-particle packaged aYAP modRNA in a murine cardiac ischemia/reperfusion model. Aim 2. Define the role of IRF2BP2 in CM innate immune responses. In this aim, we will perform IRF2BP2 gain- and loss-of-function studies to define its role in CM innate immune responses. Additionally, we will investigate the molecular mechanism of how IRF2BP2 regulates CM innate immune responses. Using the new Irf2bp2 knock-in mouse line we have recently generated, we will perform IRF2BP2 ChIP-Seq to identify its direct targets. This proposal is innovative and significant, as it focuses on dissecting the basic molecular mechanisms that underlie CMs innate immune responses, and it aims to develop new therapeutic strategies for reducing myocardial infarction injury.