Unraveling mitotic protein networks by 3D multiplexed epitope drug screening
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 physiol...
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| Format: | Article |
| Language: | English |
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Springer Nature
2018-08-01
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| Series: | Molecular Systems Biology |
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| Online Access: | https://doi.org/10.15252/msb.20188238 |
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| author | Lorenz J Maier Stefan M Kallenberger Katharina Jechow Marcel Waschow Roland Eils Christian Conrad |
| author_facet | Lorenz J Maier Stefan M Kallenberger Katharina Jechow Marcel Waschow Roland Eils Christian Conrad |
| author_sort | Lorenz J Maier |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-cda07af543e74cdd9c475a283af419cc |
| institution | DOAJ |
| issn | 1744-4292 |
| language | English |
| publishDate | 2018-08-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-cda07af543e74cdd9c475a283af419cc2025-08-20T03:06:27ZengSpringer NatureMolecular Systems Biology1744-42922018-08-0114811510.15252/msb.20188238Unraveling mitotic protein networks by 3D multiplexed epitope drug screeningLorenz J Maier0Stefan M Kallenberger1Katharina Jechow2Marcel Waschow3Roland Eils4Christian Conrad5Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), Heidelberg UniversityCenter for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), Heidelberg UniversityDivision of Theoretical Bioinformatics, German Cancer Research Center (DKFZ)Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ)Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), Heidelberg UniversityCenter for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), Heidelberg UniversityAbstract 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.https://doi.org/10.15252/msb.20188238cell profilingmitosis modelingmultiplexed immunostainingprotein–protein interactions |
| spellingShingle | Lorenz J Maier Stefan M Kallenberger Katharina Jechow Marcel Waschow Roland Eils Christian Conrad Unraveling mitotic protein networks by 3D multiplexed epitope drug screening Molecular Systems Biology cell profiling mitosis modeling multiplexed immunostaining protein–protein interactions |
| title | Unraveling mitotic protein networks by 3D multiplexed epitope drug screening |
| title_full | Unraveling mitotic protein networks by 3D multiplexed epitope drug screening |
| title_fullStr | Unraveling mitotic protein networks by 3D multiplexed epitope drug screening |
| title_full_unstemmed | Unraveling mitotic protein networks by 3D multiplexed epitope drug screening |
| title_short | Unraveling mitotic protein networks by 3D multiplexed epitope drug screening |
| title_sort | unraveling mitotic protein networks by 3d multiplexed epitope drug screening |
| topic | cell profiling mitosis modeling multiplexed immunostaining protein–protein interactions |
| url | https://doi.org/10.15252/msb.20188238 |
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