Development and Application of Optimized Cardio-Specific AdVector-based Systems for In Situ Establishment of Biological Pacemakers

我们的项目致力于建立一个“生物心脏起搏器”，用于未来治疗心脏疾病，总结在术语“病窦综合征”。我们打算通过基于引入优化的编程因子组合的工作心肌细胞的直接重编程来创建生物起搏器。它们将通过我们的SMART腺病毒技术引入靶细胞，该技术通过将高容量Ad载体定制为双特异性衔接蛋白、假型化和磁性纳米颗粒，结合了最高的特异性、有效性和安全性。设想的方法允许在体内原位改变细胞。原理证明最初将依赖于培养的鼠PSC衍生的心肌细胞在体外实现。因此，通过我们建立的正向编程方法产生的鼠PSC衍生的起搏细胞将用作生理和功能参数的参考。然后将该概念转化为临床前模型，该模型依赖于携带实验诱导的病窦综合征的转基因小鼠以及使用猪心切片培养物的离体情况。最近通过正向编程小鼠起搏细胞的RNAseq新鉴定的因子将被纳入其在直接重编程中的适用性。所描述的概念在科学上是独特的，具有最高的临床相关性。

Our project addresses the establishment of a "Biological Cardiac Pacemaker" for future therapy of cardiac diseases summarized under the term "Sick-Sinus-Syndrome". We intend to create a biological pacemaker via direct reprogramming of working cardiomyocytes based on the introduction of optimized programming factor combinations. They will be introduced into the target cells via our SMART adenovirus technology, which combines highest specificity, efficacy and safety by tailoring high capacity Ad vectors to bi-specific adaptor proteins, pseudotyping and magnetic nanoparticles. The envisaged approaches allow in situ alterations of cells in vivo. Proof-of-principle will be achieved initially in vitro relying on cultured murine PSC derived cardiomyocytes. Thereby, murine PSC derived pacemaker cells generated via our established forward programming approach will serve as a reference for physiological and functional parameters. The concept will then be translated to a pre-clinical model relying on transgenic mice bearing an experimentally induced Sick-Sinus-Syndrome as well as an ex vivo situation using pig heart slice cultures. Factors, recently newly identified via RNAseq of forward programmed murine pacemaker cells will be included with respect to their applicability in direct reprogramming. The described concept is scientifically unique and of highest clinical relevance.