Unraveling mitotic protein networks by 3D multiplexed epitope drug screening

• Verfasst von: Maier, Lorenz Josef [VerfasserIn]
• Verfasst von: Kallenberger, Stefan M. [VerfasserIn]
• Verfasst von: Jechow, Katharina [VerfasserIn]
• Verfasst von: Waschow, Marcel [VerfasserIn]
• Verfasst von: Eils, Roland [VerfasserIn]
• Verfasst von: Conrad, Christian [VerfasserIn]
• Titel: Unraveling mitotic protein networks by 3D multiplexed epitope drug screening
• Verf.angabe: Lorenz J Maier, Stefan M Kallenberger, Katharina Jechow, Marcel Waschow, Roland Eils & Christian Conrad
• E-Jahr: 2018
• Jahr: [1 August 2018]
• Illustrationen: Illustrationen
• Fussnoten: Gesehen am 17.09.2019
• Titel Quelle: Enthalten in: Molecular systems biology
• Ort Quelle: [London] : Nature Publishing Group UK, 2005
• Jahr Quelle: 2018
• Band/Heft Quelle: 14 (2018,8) Artikel-Nummer e8238, Seite 1-15, 15 Seiten
• ISSN Quelle: 1744-4292
• DOI: doi:10.15252/msb.20188238
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: cell profiling
• Sach-SW: mitosis modeling
• Sach-SW: multiplexed immunostaining
• Sach-SW: protein-protein interactions
• K10plus-PPN: 1677147687

Abstract

Abstract Three-dimensional protein localization intricately determines the functional coordination of cellular processes. The complex spatial context of protein landscape has been assessed by multiplexed immunofluorescent staining or mass spectrometry, applied to 2D cell culture with limited physiological relevance or tissue sections. Here, we present 3D SPECS, an automated technology for 3D Spatial characterization of Protein Expression Changes by microscopic Screening. This workflow comprises iterative antibody staining, high-content 3D imaging, and machine learning for detection of mitoses. This is followed by mapping of spatial protein localization into a spherical, cellular coordinate system, a basis for model-based prediction of spatially resolved affinities of proteins. As a proof-of-concept, we mapped twelve epitopes in 3D-cultured spheroids and investigated the network effects of twelve mitotic cancer drugs. Our approach reveals novel insights into spindle fragility and chromatin stress, and predicts unknown interactions between proteins in specific mitotic pathways. 3D SPECS's ability to map potential drug targets by multiplexed immunofluorescence in 3D cell culture combined with our automated high-content assay will inspire future functional protein expression and drug assays.