Characterization and Validation of a Human 3D Cardiac Microtissue for the Assessment of Changes in Cardiac Pathology

Abstract

Pharmaceutical agents despite their efficacy to treat disease can cause additional unwanted cardiovascular side effects. Cardiotoxicity is characterized by changes in either the function and/or structure of the myocardium. Over recent years, functional cardiotoxicity has received much attention, however morphological damage to the myocardium and/or loss of viability still requires improved detection and mechanistic insights. A human 3D cardiac microtissue containing human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs), cardiac endothelial cells and cardiac fibroblasts was used to assess their suitability to detect drug induced changes in cardiac structure. Histology and clinical pathology confirmed these cardiac microtissues were morphologically intact, lacked a necrotic/apoptotic core and contained all relevant cell constituents. High-throughput methods to assess mitochondrial membrane potential, endoplasmic reticulum integrity and cellular viability were developed and 15 FDA approved structural cardiotoxins and 14 FDA approved non-structural cardiotoxins were evaluated. We report that cardiac microtissues provide a high-throughput experimental model that is both able to detect changes in cardiac structure at clinically relevant concentrations and provide insights into the phenotypic mechanisms of this liability.

Human Cardiac Organoids for Disease Modeling

“…© The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY‐NC) http://creativecommons.org/licenses/by-nc/4.0/; and Archer et al, licensed under CC BY 4.0. © 2018 The Authors.…”

“…In addition to disease modeling, human cardiac organoids (hCOs) can also be produced in high‐throughput as a model to detect changes in cardiac structure and provide insights into the phenotypic mechanisms at clinically relevant drug concentrations ( Figure 3 ). In a study performed by Archer et al .…”

“…In addition to disease modeling, human cardiac organoids (hCOs) can also be produced in high-throughput as a model to detect changes in cardiac structure and provide insights into the phenotypic mechanisms at clinically relevant drug concentrations 34 (Figure 3). In a study performed by Archer et al 34 in 2018, endoplasmic reticulum (ER) membrane integrity, mitochondrial membrane potential (ΔΨm), and cellular viability (ATP depletion) could be used as measurements of primary and secondary cardiotoxic structural effects upon drug administration. It is known that both mitochondria and ER are affect either directly or indirectly in cardiotoxicity.…”

3D Co-culture of hiPSC-Derived Cardiomyocytes With Cardiac Fibroblasts Improves Tissue-Like Features of Cardiac Spheroids

“…A number of studies have made use of cardiac scaffold-free microtissues, also called spheroids, for drug testing and toxicology, using a mix of several cell types such as rodent or human, primary-or hiPSC-derived cardiomyocytes, fibroblasts, stem cells, and endothelial cells (Garzoni et al, 2009). The influence of non-myocytes has been neglected in simple cardiomyocyte in vitro models (Archer et al, 2018). The outcome of co-culturing several cardiac cell types in 3D cultures is incompletely understood and adds complexity and practical challenges.…”

Modeling Hypertrophic Cardiomyopathy: Mechanistic Insights and Pharmacological Intervention

“…Moreover, hPSC-CM disease models fail to recapitulate the physiological complexity of a multicellular intact heart system that encompasses fluid dynamics inherent to the circulatory system as well as neurohormonal control (i.e., metabolic changes) and extracellular matrix alterations (e.g., fibrosis). Recent efforts have included additional cell types such as cardiac fibroblasts or endothelial cells, although these were derived from different sources with multiple genetic backgrounds [68]. Thus, HCM modeling studies using hPSC-CMs should also be interpreted carefully because hallmarks of disease may be under-or overestimated [16].…”