Project 455131

Patients with viral infections, especially COVID 19, frequently developed myocarditis, an inflammation of the myocardium that can result in severe morbidity and mortality. However, the underlying mechanisms are still a mystery. Heart-on-a-chip combined with stem cell technology presents a unique opportunity to tackle the issue, as human tissues can more precisely recapitulate human diseases. However, cardiac muscle contains extensive vascular networks and immune cells that are challenging to recreate in vitro. To overcome this obstacle, I propose to design a culture system with opposing hollow pillars, serving as tissue forming templates, vascular perfusion inlet/outlet and force sensors. Perfusable tissue will be created around the hollow pillars with sequential formation of the vascular layer, heart muscle tissues with resident macrophages and circulating immune cells. The tissues will be electrically conditioned to reach advanced maturation approaching adult-like phenotypes. The cardiac and vascular functional assessments will be benchmarked with healthy adult myocardium. All cells in the matured tissue will go through single-cell sequencing to compare with reported genetic profiles of cells in adult myocardium. Finally, patients with and without viral myocarditis will be added to induce myocarditis. The sars-Cov2 vascular infection will be an alternative approach. The cardiac and vascular injuries will be evaluated to identify the consistency with the clinical results of the donor patients. Moreover, EV, cytokines and antibodies will be compared between groups through RT-PCR, ELISA and mass spectrometry to identify the potential target responsible for myocarditis. The final candidates will be tested on the heart-on-a-chip to confirm the findings. The proposed tissue model will be the first to precisely recreate adult-like cardiac tissues and will be a valuable asset to investigate cardiovascular-related disease mechanisms and therapeutic interventions.