High-Throughput Imaging for the Discovery of Cellular Mechanisms of Disease

HTI is a powerful method in basic research, drug discovery, and identification of disease mechanisms. HTI uses automated microcopy and image analysis to extract numerical cellular features that can be used to measure cellular pathways activity and/or morphological phenotypes. HTI assays can be used to screen and profile large collections of perturbing reagents (compounds, RNAi, CRISPR/Cas9), or to precisely quantify rare biological events. Since HTI assays can be adapted to study a wide variety of cellular phenotypes, this technique has been adopted to identify cellular pathways and genes altered in pathogen infection, cancer, and monogenic diseases, among others. HTI has utility in chemical screens to discover novel cancer vulnerabilities for potential pharmacological intervention. High-throughput imaging (HTI) is a powerful tool in the discovery of cellular disease mechanisms. While traditional approaches to identify disease pathways often rely on knowledge of the causative genetic defect, HTI-based screens offer an unbiased discovery approach based on any morphological or functional defects of disease cells or tissues. In this review, we provide an overview of the use of HTI for the study of human disease mechanisms. We discuss key technical aspects of HTI and highlight representative examples of its practical applications for the discovery of molecular mechanisms of disease, focusing on infectious diseases and host–pathogen interactions, cancer, and rare genetic diseases. We also present some of the current challenges and possible solutions offered by novel cell culture systems and genome engineering approaches. High-throughput imaging (HTI) is a powerful tool in the discovery of cellular disease mechanisms. While traditional approaches to identify disease pathways often rely on knowledge of the causative genetic defect, HTI-based screens offer an unbiased discovery approach based on any morphological or functional defects of disease cells or tissues. In this review, we provide an overview of the use of HTI for the study of human disease mechanisms. We discuss key technical aspects of HTI and highlight representative examples of its practical applications for the discovery of molecular mechanisms of disease, focusing on infectious diseases and host–pathogen interactions, cancer, and rare genetic diseases. We also present some of the current challenges and possible solutions offered by novel cell culture systems and genome engineering approaches. unattended image processing workflow aimed at the identification of cellular and subcellular objects from microscopy images, and at the extraction of cellular features that can be used to quantify or classify these objects. a set of experimental steps to simultaneously move large numbers of liquid samples from one or more source vessels, usually multiwell plates, to destination vessels using programmable, multichannel robotic dispensing devices. method based on Cas9, a programmable bacterial RNA-binding DNA nuclease that can be targeted to bind specific regions in the genome. CRISPR/Cas9 can be used to knockout genes and noncoding DNA regions via DNA double-strand breaks generation, edit genomic regions via DNA homologous recombination or deamination, or to target chromatin modifiers to change the epigenetic environment in a DNA sequence-specific fashion. suppression of cell growth. an HTI approach that uses quantitation of large data sets of multiparametric morphological features as a readout. a technique combining automated liquid handling, high-throughput microscopy, and automated image analysis in a single workflow. a workflow that uses HTI to functionally cluster and/or characterize experimental treatments based on the analysis of large datasets of morphological cellular features. cell lines or organisms sharing the same genetic background. the complete set of genes encoding kinases in a genome. double-stranded RNA-based method to silence the expression of a gene via the sequence-specific targeting of its mRNA for degradation.