In vitro Assays and Imaging Methods for Drug Discovery for Cardiac Fibrosis

Abstract

As a result of stress, injury, or aging, cardiac fibrosis is characterized by excessive deposition of extracellular matrix (ECM) components resulting in pathological remodeling, tissue stiffening, ventricular dilatation, and cardiac dysfunction that contribute to heart failure (HF) and eventually death. Currently, there are no effective therapies specifically targeting cardiac fibrosis, partially due to limited understanding of the pathological mechanisms and the lack of predictive in vitro models for high-throughput screening of antifibrotic compounds. The use of more relevant cell models, three-dimensional (3D) models, and coculture systems, together with high-content imaging (HCI) and machine learning (ML)-based image analysis, is expected to improve predictivity and throughput of in vitro models for cardiac fibrosis. In this review, we present an overview of available in vitro assays for cardiac fibrosis. We highlight the potential of more physiological 3D cardiac organoids and coculture systems and discuss HCI and automated artificial intelligence (AI)-based image analysis as key methods able to capture the complexity of cardiac fibrosis in vitro. As 3D and coculture models will soon be sufficiently mature for application in large-scale preclinical drug discovery, we expect the combination of more relevant models and high-content analysis to greatly increase translation from in vitro to in vivo models and facilitate the discovery of novel targets and drugs against cardiac fibrosis.

Bioactivity and miRNome Profiling of Native Extracellular Vesicles in Human Induced Pluripotent Stem Cell‐Cardiomyocyte Differentiation

“…[ 89 ] However, evidence for the role of hiPSC‐EV in these processes is still lacking [ 9 , 90 ] and should be addressed in future in vivo studies, as reliable and predictive in vitro models of cardiac fibrosis able to respond to fibrotic and anti‐fibrotic stimulation are yet to be standardized and currently do not reflect the interaction between fibroblasts, macrophages and endothelial cells, major players driving myocardial fibrosis. [ 91 , 92 ]…”

Organoids revealed: morphological analysis of the profound next generation in-vitro model with artificial intelligence

“…The excessive influence of human factors in the organoid culture process and the low degree of automation have led to inevitable human error. Therefore, development of more automated, intelligent, and integrated systems is the future development trend [ 133 , 134 ]. Using DL-based object detection to obtain useful information, the system or equipment can make intelligent judgments and carry out operations according to the recognition results, which is an important application of technology to avoid human interference.…”

Transcriptome and proteome profiling of activated cardiac fibroblasts supports target prioritization in cardiac fibrosis

“…Since activated CF are one of the major cellular drivers of cardiac fibrosis development, finding novel targets to modulate CF state represents an opportunity to treat these patients. Reasons for the current shortage of actionable targets in cardiac fibrosis include the lack of sustainable tissuespecific in vitro models of CF and reliable cell culture systems (14)(15)(16). Here, the use of primary cells for instance as led to some controversy in the field, namely due to the challenges in defining cell identity markers (11).…”

“…Despite the substantial progress in our understanding of CF activation mechanisms, the number of actionable targets in cardiac fibrosis remains limited. The lack of sustainable sources of CF and reliable cell culture methods have been hampering the identification of putative targets and drug development studies (14)(15)(16)(17).…”