VIRAL DELIVERED CARDIAC REGENERATION

项目摘要 缺血性心脏病（IHD）是工业化国家中最大的单一死亡原因， 成人心力衰竭（HF）的发展1-2。IHD可由心肌梗死（MI）引起， 限制了冠状动脉血液流向心脏，导致局部缺血并最终导致心肌细胞不可逆的死亡。 心肌梗死的大小与心脏功能恶化的程度相关3， 收缩储备，以及随着时间的推移HF 4死亡的可能性。迅速恢复动脉灌注 在经皮冠状动脉介入治疗期间进行溶栓和抗血小板治疗， MI死亡率。然而，存活者中HF的患病率增加，因为不可逆的 心肌细胞死亡导致残余的可诱导局部缺血和永久性瘢痕形成。一个主要的病理 问题是成年人心肌细胞在MI后不能内源性地自我再生。 这是由于缺乏预防性治疗，药理学或细胞，可以管理， 与再灌注结合以刺激心肌再生并防止转变为HF。的 有效促进缺血心脏中的内源性心肌细胞再生将潜在地提供 MI及其不良病理生理后果的有效新治疗。抑制特定 四种微小RNA（miR-99，miR-100，let-7a和let-7 c）的组合是一种关键的调节因子， 哺乳动物心肌细胞的去分化和增殖6. JBT设计并测试了一种腺病毒， 相关病毒称为JBT-miR 2，其允许时间，心脏和非整合表达， 这四种miR的抑制剂。在心脏缺血再灌注（IR）损伤的年轻小鼠的初步研究中， 通过静脉内注射给予单剂量JBT-miR 2的有效性和安全性， 再灌注与对照病毒进行比较。JBT-miR 2增加心脏功能并降低心脏功能。 IR后2周的体积，IR后8周的瘢痕组织相应减少约47%， 控制病毒。为了继续进行临床试验所需的研发，有必要 证实了在临床平移较大年龄动物中的有效性和安全性。第一阶段的具体目标 SBIR资助的目的是：1：在中年人中开展JBT-miR 2的剂量范围发现有效性、安全性和PK研究 具有IR损伤的小型猪，评价多效性作用和作用机制。重要的，可重复的， 在JBT-miR 2中，预期心脏尺寸和功能指标的临床相关终点变化 与对照病毒相比，处理的动物。2.开发和验证PK和体外生物活性测定。评估 用于免疫原性、中和抗体和病毒颗粒滴度水平，以确定猪体内的病毒脱落。一 将起草目标产品概况，了解制剂鉴别、活性和 纯度3.这些结果将允许准备和进行与FDA的C类会议， 就临床翻译所需的药物非临床研究质量管理规范安全性毒理学研究的设计达成一致。

PROJECT SUMMARY Ischemic heart disease (IHD) is the single largest cause of death in the industrialized world and contributor to the development of Heart Failure (HF) in adults 1-2. IHD can be caused by a myocardial infarction (MI), which limits 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 3, compromise of contractile reserve, and over time the likelihood of mortality from HF 4. 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 to stimulate regeneration of heart muscle and prevent the transition to HF. The effective promotion of endogenous cardiomyocyte regeneration in the ischemic heart would potentially offer a powerful new treatment for MI and its adverse pathophysiologic consequences. Inhibition of a specific combination of four MicroRNAs (miRs); miR-99, miR-100, let-7a and let-7c, is a critical regulator of cardiomyocyte dedifferentiation and proliferation in mammals 6. JBT has designed and tested an adeno associated virus known as JBT-miR2, which allows for temporal, cardiac and non-integrative expression of inhibitors to these four miRs. In preliminary studies in young mice with cardiac Ischemic Reperfusion (IR) injury the efficacy and safety of a single dose of JBT-miR2 administered by intravenous injection at the time of reperfusion was compared with Control virus. JBT-miR2 increased heart function and decreased cardiac volumes at 2-weeks post IR with a corresponding ~47% reduction in scar tissue at 8-weeks post-IR compared with control virus. To continue with Research and Development required for clinical trials it is necessary to confirm the efficacy and safety in clinically translational larger aged animals. The Specific Aims of this Phase I SBIR grant are to: 1: Conduct a dose range finding efficacy, safety and PK study of JBT-miR2 in middle aged mini-pigs with IR injury, evaluating pleiotropic effects and mechanism of action. Significant, reproducible, clinically relevant end-point changes in measures of cardiac dimension and function are expected in JBT-miR2 treated animals compared to Control virus. 2. Develop and validate PK and in vitro bioactivity assays. Assess for immunogenicity, neutralizing antibodies, and viral particle titer levels to determine viral shedding in pigs. A Target Product Profile will be drafted, with an understanding of measures of drug product identity, activity and purity. 3. These results will allow preparation for, and conduction of a type C meeting with FDA to provide agreement on the design of a Good Laboratory Practice safety toxicology studies required for clinical translation.