Developing osteoclasts and aortic valve organoids for the potential treatment of valve calcification

Abstract

Calcific aortic valve disease (CAVD) is the most frequent valvulopathy in the Western world, butthere is currently no pharmacological treatment to halt or delay the progression of the disease. Since the only treatment for severe CAVD is valve replacement, there is a need for alternative therapies. Valve interstitial cells (VICs), especially those in the collagen I rich fibrosa layer, have been identified as key mediators of valve calcification. Previous studies have developed a 2D culture model, but this model fails to mimic cell-extracellular matrix and mechanical interactions known to occur in CAVD. This project addresses the need for a simple 3D model for studying CAVD and begins to investigate a potential treatment that can be integrated into the model in the future. Physiological mineral resorbing cells may potentially be used to reduce or eliminate valve mineralization. The project is divided into two aims: (1) developing and characterizing a 3D fibrosa model and (2) optimizing and documenting a protocol for generating osteoclasts (OCs) from human induced pluripotent stem cells (iPSCs). This dissertation outlines the method and initial characterization of a 3D fibrosa model with confocal microscopy. The differentiation of human iPSCs to OCs is also described as a potential treatment for valve calcification. Generated OCs or their exosomes could be combined with the 3D fibrosa model to study novel treatments for valve calcification.