University of Twente Student Theses


Modeling Familial Hypertrophic Cardiomyopathy using Human Engineered Heart Tissues

Iddekinge, T.R. van (2021) Modeling Familial Hypertrophic Cardiomyopathy using Human Engineered Heart Tissues.

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Abstract:Familial hypertrophic cardiomyopathy (FHCM) is a prevalent and possibly fatal heart condition that strikes about 1 in 500 individuals. It can be caused by mutations in several genes, mutations in the MYBPC3 gene account for 20-25% of FHCM cases. These mutations can cause sarcomeric disarray and disorganization which can eventually cause hypertrophy of the intraventricular septum, defined as FHCM. As diagnosis and treatment of this condition remain a challenge, disease modeling of FHCM can give insight into the characteristics of the disease. It can give rise to new therapies and patient specific drug testing. human induced pluripotent stem cells (hiPSCs) are a good candidate for disease modeling. However, most disease models rely on 2D monolayer cultures that are generally immature and unable to efficiently recapitulate the contractile deficits present in a patient carrying the MYBPC3 mutation. A way to overcome these limitations is the use of 3D engineered heart tissues (EHTs). In this research, we developed an EHT disease model for FHCM using hIPSC derived cardiomyocytes (CMs), un-purified and lactate purified, with a CRISPR-CAS9 induced mutation in the MYBPC3 gene to understand the contractile parameters that are present in a patient carrying this mutation. We also attempted to develop an EHT based disease model treating healthy cells with hypertrophic stimulus phenylephrine (PE). We attempted to unveil a similar response in contraction force and velocity as in the MYBPC3 mutated disease model, due to the cells becoming hypertrophic. We found a significant decrease in contraction force and velocity between our control group and the mutated cell line, in experiments with and without lactate purified cells, more apparent than previous studies using 2D models, animal models, or 3D models. Stimulation with PE did not affect the contraction of our EHTs, thus having failed making a disease model with PE stimulation. Furthermore, we succeeded in making EHTs in a microfluidic chip co-cultured with endothelial cells (ECs), which is a very stable, reproducible and relatively easy method of EHT generation and can potentially be used in the future for making 3D hIPSC derived cardiac disease models, with the option of a constant medium perfusion making the EHT culture dynamic.
Item Type:Essay (Master)
Faculty:TNW: Science and Technology
Subject:42 biology
Programme:Biomedical Engineering MSc (66226)
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