ShRNA-Based Targeting of mAKAPβ in Heart Failure

病理性心脏重塑是疾病中心力衰竭的常见途径。尽管目前 药物治疗和其他减弱重塑的进展，心力衰竭导致的死亡率仍然存在 高新的，更有效的治疗选择是迫切需要在不断增加的患者群体， 提高心力衰竭患者或易患心力衰竭患者的生存率和生活质量。肌A激酶 锚定蛋白β（mAKAPβ）是多分子信号复合物的组织者，对于诱导 和病理性心脏重塑的进展。mAKAPβ通过与多种信号分子结合， 动态协调多个信号传导模块， （MAPK）、Ca 2+、磷酸肌醇和缺氧应激信号。因此，心肌细胞特异性mAKAPβ 在多种心血管疾病模型中，小鼠中的基因敲除抑制了重构和心力衰竭的发展。 疾病CRI是一家开发新型专利保护疗法的公司，用于预防和/或治疗 心脏衰竭的症状在这个快速道SBIR，CRI将测试一种新的基因治疗载体，旨在抑制mAKAPβ 通过RNA干扰在心肌细胞中选择性表达。AAV9sc.shmAKAP生物制剂是一种自我- 互补的、亲心的、血清型9腺相关病毒（AAV 9），其表达mAKAP特异性的 在心肌细胞特异性启动子控制下的小发夹RNA（shRNA）。在此应用中，CRI将 在临床相关的大型动物模型中测试新生物制剂治疗心肌梗死后（MI）心力衰竭。 I期-特定目的：我们将在约克郡猪中绘制生物制剂的剂量反应曲线，以确定 持续抑制心脏中mAKAPβ表达所需的最小剂量。第二阶段-具体目标 图1：mAKAP RNAi在大型动物模型中对心力衰竭的功效。这个项目的核心是测试 mAKAPβ RNAi是否能减轻猪心肌梗死诱导的病理性重构，预防心力衰竭。 猪将经受缺血-再灌注以诱导MI或假手术，然后用 AAV9sc.shmAKAP生物制剂在确定剂量后立即或1个月后通过冠状动脉内输注 在第一阶段将对猪进行连续超声心动图随访，并在MI后3个月的终点进行研究， 左心室压力-容积环血流动力学的导管插入术。本目标的目标是 证明mAKAP RNAi将在MI的大型动物模型中保护心脏结构和功能 疾病具体目的2：AAV 9 sc. shmAKAP介导的分子和细胞病理学抑制 与心力衰竭有关利用从目标1中使用的相同动物收集的组织， mAKAP RNAi在猪中的益处将通过重量分析、组织学和分子分析来证明， 纤维化和其他心脏重塑和心力衰竭的标志物。此外，初步毒理学筛选将 来保证生物制品的安全性该项目将表明mAKAPβ靶向是一种可行的治疗策略 并证明心力衰竭的首次人体临床试验是合理的。

Pathological cardiac remodeling constitutes a common pathway to heart failure in disease. Despite current pharmacologic therapy and other advances that attenuate remodeling, mortality due to heart failure remains high. New, more effective therapeutic options are desperately needed in an increasing patient population to improve both the survival and quality of life for patients with or susceptible to heart failure. Muscle A-kinase anchoring protein β (mAKAPβ) is the organizer of multimolecular signaling complexes critical for the induction and progression of pathological cardiac remodeling. By binding a diverse set of signaling molecules, mAKAPβ dynamically orchestrates multiple signaling modules that transduce cAMP, mitogen-activated protein kinase (MAPK), Ca2+, phosphoinositide, and hypoxic stress signals. Accordingly, cardiomyocyte-specific mAKAPβ knock-out in mice inhibited remodeling and the development heart failure in multiple models of cardiovascular disease. CRI is a company developing novel, patent protected therapeutics for the prevention and/or treatment of heart failure. In this Fast-Tract SBIR, CRI will test a new gene therapy vector designed to inhibit mAKAPβ expression selectively in the cardiac myocyte by RNA interference. The AAV9sc.shmAKAP biologic is a self- complementary, cardiotropic, serotype 9 adeno-associated virus (AAV9) that expresses an mAKAP-specific small hairpin RNA (shRNA) under the control of a cardiac myocyte-specific promoter. In this application, CRI will test the new biologic in a clinically relevant large animal model for post-myocardial infarction (MI) heart failure. Phase I - Specific Aim: We will perform a dose response curve for the biologic in Yorkshire swine to determine the minimum dose required for consistent inhibition of mAKAPβ expression in the heart. Phase II - Specific Aim 1: Efficacy of mAKAP RNAi for heart failure in a large animal model. The core of this project is to test whether mAKAPβ RNAi will mitigate pathological remodeling induced by MI in swine, preventing heart failure. Swine will be subjected to ischemia-reperfusion to induce MI or sham procedure and then treated with the AAV9sc.shmAKAP biologic by intracoronary infusion immediately after or 1 month later at the dose determined in Phase I. The pigs will be followed by serial echocardiography and studied at endpoint 3 months post-MI by catheterization for left ventricular pressure-volume loop hemodynamics. The goal for this Aim is the demonstration that mAKAP RNAi will preserve cardiac structure and function in a large animal model of MI disease. Specific Aim 2: AAV9sc.shmAKAP-mediated inhibition of the molecular and cellular pathology associated with heart failure. Taking advantage of tissue collected from the same animals used in Aim 1, the benefits of mAKAP RNAi in swine will be demonstrated by gravimetric, histological, and molecular analyses for fibrosis and other markers of cardiac remodeling and heart failure. In addition, initial toxicology screens will be done to support the biologic’s safety. This project will show that mAKAPβ targeting is a viable therapeutic strategy for heart failure post-MI and justify a first-in-human clinical trial for heart failure.