Synergistically Target Mitochondria for Heart Failure Treatment

总结/摘要 心力衰竭（HF）是全球范围内的主要死亡原因。虽然HF的病理生理学是复杂的， 仍然不完全了解，线粒体功能的缺陷已经涉及进展， 结果，并成为HF治疗的重要目标。一个众所周知的促成因素， 衰竭心脏中的不良心脏重构是过量的血管源性活性氧 （mtROS）。因此，已经开发了多种基于抗氧化剂的疗法用于HF治疗。 过去几十年。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 迄今为止，临床尚未成功，这表明替代或补充线粒体治疗靶点 是必要的。除了过量的线粒体活性氧，线粒体膜电位（线粒体膜电位）的严重丧失是另一个原因。 HF的主要标志。在心肌细胞（CM）中，线粒体破坏不仅影响能量的产生，而且还影响心肌细胞（CM）的功能。 多种对细胞功能和存活至关重要的信号传导途径，如氧化还原平衡、钙稳态 和线粒体质量控制。我们假设协同靶向线粒体，即，同时 保护线粒体线粒体膜和清除过量的mtROS是HF治疗的可行治疗策略。 然而，由于缺乏工具，评估骨小梁保存的治疗潜力是具有挑战性的。 在活体动物中动态和特异性地控制CM真菌。为了克服这一技术障碍，我们开发了一种 通过整合荧光素酶的创新的靶向肿瘤的发光遗传学（命名为mLumOpto）技术- 我们最近发表的线粒体光遗传学研究中，荧光蛋白发出的内源性生物发光。我们 初步数据表明，mLumOpto可以在没有外部光的情况下诱导动态CM光控制 照明。该项目的主要目标是利用这种先进的mLumOpto技术来操纵 CM生物膜在体内的应用，以评估CM生物膜单独保存或与mtROS清除剂协同保存的功效 (i.e.,组织特异性抗氧化剂），在临床前小鼠模型的HF治疗中。平移势 还将在类似人类的大型动物中评估所提出的协同脑靶向HF治疗 (i.e.,猪）。我们提出了三个具体目标来实现我们的目标。目标1将决定 在小鼠病理性心脏重构和HF发展中持续CM介导的去极化， 潜在的分子机制。目的2将评估协同防腐剂的效果， 线粒体ROS清除在改善两个已建立的心脏重构和收缩功能障碍中的作用 小鼠HF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HF治疗在猪中的翻译潜力。该项目的成功完成将导致 不仅是一种能够特异性和动态操纵CM线粒体的创新技术， 体内，而且也是一种新的翻译的神经靶向治疗HF治疗。

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.