Oligonucleotide Inhibition for cardiac regeneration in Ischemia

项目摘要 缺血性心脏病（IHD）是全球最大的单一死亡原因。心脏病发作或心肌梗塞 心肌梗死（MI）是由于限制了流向心脏的冠状动脉血流，导致局部缺血，并最终导致心脏衰竭。 心肌细胞的不可逆死亡。心肌梗死的大小与恶化程度相关 心脏功能受损，收缩储备受损，以及心力衰竭死亡的可能性 （HF）.经皮冠状动脉介入治疗中溶栓和抗血小板治疗的及时恢复动脉灌注 冠状动脉介入治疗导致MI的急性死亡率下降。然而， 存活者增加，因为不可逆的心肌细胞死亡导致残余的可诱导缺血， 永久性疤痕一个主要的病理问题是人类成年心肌细胞不能再生 他们自己内在地遵循MI。这是由于缺乏有效的治疗，药理学 或细胞的，其可以与再灌注结合施用，或在刺激心脏再生之后施用。 肌肉.有效促进缺血心脏的内源性心肌细胞再生， 瘢痕大小的减小可能为MI及其不良病理生理学改变提供一种有效的新治疗方法。 后果抑制四种MicroRNA（miR）的特异性组合; miR-99、miR-100、let-7a和let-7 c， 是斑马鱼心肌细胞去分化和心脏再生的关键调节因子。的序列和 这些miR的靶蛋白在人类中是保守的。在体内，腺相关病毒（AAV）递送 这些miR的抑制剂进入左冠状动脉结扎小鼠的心脏， 法尼基转移酶β亚基和SWI/SNF相关基质相关肌动蛋白依赖性调节因子 染色质亚家族A，成员5。心脏再生通过增殖和凋亡的表达来证实。 胞质分裂标记物，标记尿苷掺入DNA，以及瘢痕组织消退和心脏 功能改善。病毒递送的缺点是抗AAV抗体在人体中的高流行率 可以中和它们的效果作为替代方案，JAAN Biotherapeutics将研究两种合成的， 针对miR-99/100和let-7a/c的特异性、有效和核酸酶抗性的核酸miR抑制剂（抗miR）可以 在实验性小鼠缺血再灌注模型中减少缺血性损伤， 方法来再生人类心肌细胞。[The第一阶段的研究将确定抗miR 增殖正常心肌，提供有效剂量和效应持续性的信息，以及 在远端组织中建立任何增殖或脱靶副作用（目的1）。目标2将研究抗- 在再灌注时或再灌注后2天，在小鼠心脏缺血性损伤60分钟后施用miR。 局部缺血可以减小疤痕大小、增加心脏功能和再生心肌。目标1将提供 安全性信息和目的2对于确认抗miR在鼠缺血性心脏中的功效至关重要。这些 研究是未来临床前和临床研究设计的关键。]

PROJECT SUMMARY Ischemic heart disease (IHD) is the single largest cause of death worldwide. A heart attack or myocardial infarction (MI) results from limitation of coronary blood flow to the heart, causing ischemia and ultimately irreversible death of cardiomyocytes. The size of a myocardial infarct correlates with the degree of deterioration of heart function, compromise of contractile reserve, and overtime the likelihood of mortality from heart failure (HF). Prompt restoration of arterial perfusion with thrombolytic and antiplatelet therapy during percutaneous coronary intervention has led to a decline in acute mortality from MI. However, the prevalence of HF among survivors has augmented, because irreversible cardiomyocyte death results in a residual inducible ischemia and permanent scarring. A major pathologic problem is the failure of human adult cardiomyocytes to regenerate themselves endogenously following a MI. This is compounded by a lack of adjunctive treatments, pharmacologic or cellular, that can be administered in conjunction with reperfusion, or after to stimulate regeneration of heart muscle. Effective promotion of endogenous cardiomyocyte regeneration in the ischemic heart with concomitant reduction of scar size would potentially offer a powerful new treatment of MI and its adverse pathophysiologic consequences. Inhibition of a specific combination of four MicroRNAs (miR); miR-99, miR-100, let-7a and let-7c, is a critical regulator of cardiomyocyte dedifferentiation and heart regeneration in zebrafish. The sequences and target proteins of these miRs are conserved in humans. In vivo, adeno-associated virus (AAV) delivery of inhibitors of these miRs into the hearts of mice with left coronary artery ligation increases the expression of the beta subunit of farnesyl-transferase and SWI/SNF-related matrix associated actin-dependent regulator of chromatin subfamily a, member 5. Cardiac regeneration was confirmed by the expression of proliferation and cytokinesis markers, labeled uridine incorporation into DNA, together with scar tissue regression and heart functional improvement. A disadvantage of viral delivery is the high prevalence of anti-AAV antibodies in humans that could neutralize their effect. As an alternative, JAAN Biotherapeutics will investigate whether two synthetic, specific, potent and nuclease resistant nucleic acid miR inhibitors (anti-miRs) to miR-99/100 and let-7a/c can reduce ischemic injury in an experimental murine model of ischemic reperfusion and constitute an innovative approach to regenerate human cardiomyocytes. [The research in Phase I will define whether the anti-miRs proliferate normal heart muscle, provide information on an effective dose and sustainability of effect, and establish any proliferative or off-target side effects in remote tissues (Aim 1). Aim 2 will study whether the anti- miRs administered after a 60 min cardiac ischemic injury in mice either at the time of reperfusion or 2 days after ischemia can reduce scar size, increase cardiac function and regenerate cardiac muscle. Aim 1 will provide safety information and Aim 2 is critical to confirm efficacy of the anti-miRs in the murine ischemic heart. These studies are pivotal for future preclinical and clinical study design.]