Synergistically Target Mitochondria for Heart Failure Treatment

摘要/摘要 心力衰竭(HF)是世界范围内主要的死亡原因。尽管心衰的病理生理机制复杂， 目前仍不完全清楚，线粒体功能缺陷与疾病的进展和发展有关 心衰的转归，并成为心力衰竭治疗的重要靶点。一个众所周知的促成因素 衰竭心脏的不良心脏重塑是线粒体衍生的过量的活性氧 (MtROS)。因此，已经开发了各种基于抗氧化剂的疗法来治疗心力衰竭。 过去的几十年里。然而，尽管在临床前研究中取得了有希望的结果，但将这些疗法转化为 到目前为止，临床还没有成功，这表明替代或互补的线粒体治疗靶点 都是需要的。除了mtros过多外，线粒体膜电位的严重丧失(m)是另一个原因。 HF的关键标志。在心肌细胞(CM)中，m的中断不仅影响能量的产生，而且还影响 对细胞功能和生存至关重要的各种信号通路，如氧化还原平衡、钙稳态 和线粒体质量控制。我们假设协同作用靶向线粒体，即同时 保护线粒体m，清除多余的mtROS，是治疗心衰的一种可行的治疗策略。 然而，由于缺乏工具，评估m保存的治疗潜力是具有挑战性的。 活体动物体内CMm的动态和特异性控制。为了克服这一技术障碍，我们开发了一种 整合荧光素酶的线粒体靶向发光遗传(简称mLumOpto)技术 利用我们最近发表的线粒体光遗传学，荧光素发出内源生物发光。我们的 初步数据表明，mLumopto在没有外部光的情况下可以诱导动态的CMm控制 照明。这个项目的主要目标是利用这种先进的mLumOpto技术来操纵 在活体中评估CMm单独保存或与mtROS清除剂协同保存的效果 (即线粒体特异性抗氧化剂)，在临床前小鼠模型的HF治疗中。翻译潜力 拟议的协同线粒体靶向HF治疗的有效性也将在类似人类的大型动物中进行评估。 (即，猪)。为了实现我们的目标，我们提出了三个具体目标。目标1将决定 持续的CMm去极化在小鼠病理性心脏重构和心力衰竭发展中的作用 潜在的分子机制。目的2将评估协同m保存的效果和 MtROS清除在改善两种已知心脏重构和收缩功能障碍中的作用 小鼠心力衰竭模型(即压力超负荷和心肌缺血-再灌注)。目标3将评估 MLumopto介导的猪心衰治疗的翻译潜力。这个项目的成功完成将引领 不仅是一项能够特定和动态地操纵心肌线粒体的创新技术 活体，但也是一种新的翻译线粒体靶向治疗心力衰竭。

SUMMARY/ABSTRACT Heart failure (HF) is a leading cause of death worldwide. Although the pathophysiology of HF is complex and remains incompletely understood, defects in mitochondrial function have been implicated in the progression and outcomes of HF, and emerged as an important target for HF therapy. One well-known contributing factor to adverse cardiac remodeling in the failing hearts is excess mitochondrial-derived reactive oxygen species (mtROS). Accordingly, a variety of antioxidant-based therapies have been developed for HF treatment over the last decades. However, despite promising outcomes in preclinical studies, translation of these therapies to the clinic has not succeeded to date, suggesting that alternative or complementary mitochondrial therapeutic targets are needed. In addition to excess mtROS, profound loss of mitochondrial membrane potential (m) is another key hallmark of HF. In the cardiomyocyte (CM), m disruption affects not only energy production, but also a variety of signaling pathways crucial for cell function and survival, such as redox balance, calcium homeostasis and mitochondrial quality control. We hypothesize that synergistically targeting mitochondria, i.e., concurrently preserving mitochondrial m and scavenging excess mtROS, is a viable therapeutic strategy for HF treatment. However, assessing the therapeutic potential of m preservation is challenging, due to a lack of tools for dynamic and specific control of CM m in live animals. To overcome this technical barrier, we developed an innovative mitochondrial-targeted luminoptogenetic (named mLumOpto) technology by integrating luciferase- luciferin-emitted endogenous bioluminescence with the mitochondrial optogenetics we recently published. Our preliminary data indicate that mLumOpto can induce dynamic CM m control in the absence of external light illumination. The primary goal of this project is to employ this advanced mLumOpto technology to manipulate CM m in vivo to evaluate the efficacy of CM m preservation alone, or in synergism with a mtROS scavenger (i.e., mitochondrial-specific antioxidant), in HF treatment in preclinical mouse models. The translational potential of the proposed synergistic mitochondrial-targeted HF therapy will also be assessed in human-like large animals (i.e., pigs). Three Specific Aims are proposed to accomplish our objectives. Aim 1 will determine the role of sustained CM m depolarization in pathological cardiac remodeling and HF development in mice, and dissect the underlying molecular mechanisms. Aim 2 will evaluate the efficacy of synergistic m preservation and mtROS scavenging in improving adverse cardiac remodeling and contractile dysfunction in two well-established mouse HF models (i.e., pressure overload and myocardial ischemia-reperfusion). Aim 3 will assess the translational potential of mLumOpto-mediated HF therapy in pigs. Successful completion of this project will lead to not only an innovative technology capable of specifically and dynamically manipulating CM mitochondria in vivo, but also a novel translational mitochondrial-targeted therapy for HF treatment.