Oligonucleotide Inhibition for cardiac regeneration in Ischemia

项目总结 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)是由于冠脉流向心脏的血流受限，导致心肌缺血，最终 心肌细胞不可逆转的死亡。心肌梗死的大小与恶化的程度有关。 心功能受损，收缩储备受损，心力衰竭死亡的可能性增加 (Hf)。经皮穿刺术中溶栓和抗血小板治疗可迅速恢复动脉灌注 冠状动脉介入治疗降低了急性心肌梗塞的死亡率。然而，心力衰竭的患病率在 存活的人数增加了，因为不可逆转的心肌细胞死亡会导致残余的诱导性缺血和 永久的疤痕。一个主要的病理问题是成人心肌细胞不能再生。 他们内在地跟随着MI。缺乏辅助治疗、药物治疗使情况更加严重。 或细胞，可与再灌流一起使用，或在刺激心脏再生后使用 肌肉。伴发心肌缺血对内源性心肌细胞再生的促进作用 减少瘢痕大小可能为治疗心肌梗塞及其不利的病理生理提供一种有效的新方法 后果。抑制四种microRNAs(MiR)的特定组合；miR-99、miR-100、let-7a和let-7c， 是斑马鱼心肌细胞去分化和心脏再生的关键调节因子。序列和 这些miRs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增加了 法尼基转移酶β亚基和SWI/SNF相关的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。心脏再生由增殖和 胞质分裂标记物，标记的尿苷掺入DNA，与瘢痕组织退变和心脏 功能完善。病毒传播的一个缺点是抗AAV抗体在人类中的高流行率 这可能会抵消它们的影响。作为替代方案，Jaan BioTreatetics将调查两种合成的， 针对miR-99/100和let-7a/c的特异性、强效和抗核酸酶的miR抑制剂(抗miRs)可以 减少实验性小鼠缺血再灌注模型的缺血损伤并构成创新的 再生人心肌细胞的方法。[第一阶段的研究将定义抗MIR 使正常心肌增殖，提供有效剂量和效果可持续性的信息，以及 在远程组织中确定任何增殖性或非靶点副作用(目标1)。目标2将研究是否反- MIRS在小鼠心肌缺血损伤60min后再灌流时和再灌流后2天给药 缺血可以缩小瘢痕大小，增加心功能，再生心肌。AIM 1将提供 安全性信息和目标2对于证实抗MIR在小鼠缺血心脏中的有效性至关重要。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.]