Geração de um modelo cardióide humano baseado em PSC para cardiomiopatia não compactada para novas terapias baseadas em EVs.

In its congenital form, Non-Compaction Cardiomyopathy (NCCM) is a congenital heart disease that originates from an interruption in the normal compaction process of the myocardium during embryonic development. The clinical implications of this condition are profound and potentially life-threatening. NCCM is associated with a spectrum of severe cardiac complications including heart failure, arrhythmias, and systemic embolization. These complications not only compromise the quality of life of the affected individuals but also pose a significant risk to their survival. The current treatment approaches primarily focus on managing the symptoms of heart failure and preventing sudden cardiac death, underscoring the need to further fill the critical gap in our understanding of the disease and its underlying pathophysiology. Our proposed research is strategically designed to bridge this existing gap and the development of targeted therapies that can address the fundamental causes of NCCM. The main objectives are: 1. Enhance our understanding of the molecular and cellular factors underlying NCCM, necessary for better tackling the disease. The aim is to identify new NCCM determinants exploring the role of CCBE1 signaling in cardiovascular development. Our previous studies have demonstrated that mutations or defects in CCBE1 expression are associated with impaired heart development (see Fig.1), particularly affecting the formation and compaction of myocardial tissue, which are hallmark features of NCCM. This novel knowledge is pivotal for the design of novel therapies. 2. To establish a cardiac organoid model that recapitulates the phenotypic characteristics of NCCM utilizing human pluripotent stem cells. By using hiPSCs CCBE1-KO (see Fig.2), we aim to fabricate an in vitro model of NCCM that accurately reflects the transmural patterning of ventricles as observed in vivo. The goal is to engineer cardioids with a more densely packed subepicardial cardiomyocyte region in comparison to the inner cardiomyocyte layer. This model will serve as a valuable tool for elucidating the underlying mechanisms of the disease, both at cellular and electrophysiological level, and for evaluating potential therapeutic strategies in a controlled setting. 3. Transform the therapeutic approach to NCCM utilizing Extracellular Vesicles (EVs) as a drug delivery system. By integrating the data from our project, our goal is to identify and test novel biomarkers and potential therapeutic targets, catalyzing groundbreaking advancements in NCCM treatment. The key aim is to a test EV-based strategies designed to specifically correct the functionality of the defective myocardium compaction using our cardioid model. Through this endeavor, we also seek to establish cardioids as a robust preclinical instrument for the investigation of cardiovascular disorders and the enhancement of regenerative medical practices. The proposed research addresses crucial challenges in understanding and treating NCCM by introducing a groundbreaking cardioid model that enables detailed study of the disease’s pathogenesis. Furthermore, the research aims to identify potential therapeutic targets and test EV-based therapy, which could lead to breakthroughs in clinical applications. This is indicative of an ambitious endeavor to transition from discovery to translational and potentially clinical impacts, ultimately improving the prognosis and quality of life for individuals affected by NCCM.