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

Abstract

High-throughput imaging (HTI) is a promising 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. Here 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 a rare genetic disease. We also present some of the current challenges and possible solutions offered by recent novel cell culture systems and genome engineering approaches.

Thinking Outside the Box—Novel Antibacterials To Tackle the Resistance Crisis

“…Depending on the global distribution of the target of interest, these efforts aid, in particular,i nt he identification of potent antibiotic and antivirulence (see also Section 4) candidates which spare commensal bacteria and are likely less toxic to eukaryotic cells. [101][102][103][104] Additionally,c omputational methods have been av ery valuable tool for the screening and analysis of potential antibiotics.T he substantial advances in the "-omics" fields including genomics,p roteomics,a nd metabolomics have resulted in av eritable explosion of (phenotypic) data. [88] Reporter strains are genetically engineered bacterial strains that allow the readout of aspecific physiological perturbation by the altered expression of ar eporter gene,s uch as GFP.…”

“…In particular,t he automation of microscopic imaging nowadays allows not only the parallel monitoring of various cellular characteristics and functions in bacterial cells,but also cellular bacterial infection models.B CP can not only be applied to phenotypic drug discovery screening,b ut also to cytological profiling experiments that allow the study of adrugsmode of action. [101][102][103][104] Additionally,c omputational methods have been av ery valuable tool for the screening and analysis of potential antibiotics.T he substantial advances in the "-omics" fields including genomics,p roteomics,a nd metabolomics have resulted in av eritable explosion of (phenotypic) data. Bioinformatic approaches aim to take advantage of these huge data sets to identify new antibacterial compounds and potential new targets or to help clarify the mode of action of new compounds.…”

Über bisherige Denkweisen hinaus – neue Wirkstoffe zur Überwindung der Antibiotika‐Krise

“…[101][102][103][104] Zusätzlich sind computerbasierte Te chniken sehr wertvoll fürdas Screening und die Analyse von potentiellen Antibiotika. Insbesondere die Automatisierung der mikroskopischen Bildgebung erlaubt heute nicht nur die parallele Beobachtung verschiedener zellulärer Charakteristika und Funktionen in Bakterien, sondern auch in Infektionsmodellen.…”

“…Um den unterschiedlichen physiologischen Parametern, wie der Zellwandstruktur oder dem Virulenzmechanismus,v erschiedener bakterieller Stämme gerecht zu werden, kçnnen reporterbasierte Assays direkt im gewünschten Bakterium durchgeführt werden. Screenings in wildtypischen oder multiresistenten Bakterien wurden beispielsweise in S. aureus, [95,96] V. cholerae, [97] K. pneumoniae [98] und P. aeruginosa [99,100] [101][102][103][104] Zusätzlich sind computerbasierte Te chniken sehr wertvoll fürdas Screening und die Analyse von potentiellen Antibiotika. Die wesentlichen Fortschritte in den "-omik"-Feldern wie Genomik, Proteomik oder Metabolomik haben zu einer regelrechten Explosion von (phänotypischen) Daten geführt.…”

Quantitative Phase Imaging Flow Cytometry for Ultra‐Large‐Scale Single‐Cell Biophysical Phenotyping

“…However, continuing development of such imaging assay is fundamentally hampered by two challenges. First, it lacks the imaging throughput to screen a large, heterogeneous cell population at single‐cell precision—a critical attribute demanded in a wide range of biomedical and biotechnology applications, from cancer screening , drug discovery processes , environmental, and biofuel production monitoring to the intense undertaking of single‐cell analysis (SCA) to gain new understanding of complex mechanisms of health and disease . Second, current cellular assay overwhelmingly relies on biochemical markers and biomolecular analyses, which are not always effective, especially when there is poor prior knowledge of the markers of a yet‐to‐be‐identified population.…”