Microscopy-based single-cell proteomic profiling reveals heterogeneity in DNA damage response dynamics

Journal article (2022)

Authors

Pin Rui Su Erasmus MC, National Taiwan University
L. You Erasmus MC
Cecile Beerens Erasmus MC
Karel Bezstarosti Erasmus MC
J.A.A. Demmers Erasmus MC
Martin Pabst BT/Environmental Biotechnology - AS
R. Kanaar Education and Student Affairs , Erasmus MC, The Oncode Institute
Cheng Chih Hsu National Taiwan University
M.P. Chien Erasmus MC, Education and Student Affairs , The Oncode Institute
Research Group
BT/Environmental Biotechnology (AS) (TU Delft)
Copyright: © 2022 Pin Rui Su, L. You, Cecile Beerens, Karel Bezstarosti, J.A.A. Demmers, Martin Pabst, R. Kanaar, Cheng Chih Hsu, M.P. Chien
DOI
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https://doi.org/10.1016/j.crmeth.2022.100237
53BP1 DDR foci dynamics DNA damage response Functional single-cell selection PDS5A Phenotype-to-proteome linking Phototagging Single-cell proteomics Tumor heterogeneity Ultrawide field-of-view optical microscope
More Info
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Published Date

2022

Language

English

Faculty

Applied Sciences

Department

BT/Biotechnologie

Research Group

BT/Environmental Biotechnology

Abstract

Single-cell proteomics has the potential to decipher tumor heterogeneity, and a method like single-cell proteomics by mass spectrometry (SCoPE-MS) allows profiling several tens of single cells for >1,000 proteins per cell. This method, however, cannot link the proteome of individual cells with phenotypes of interest. Here, we developed a microscopy-based functional single-cell proteomic-profiling technology, called FUNpro, to address this. FUNpro enables screening, identification, and isolation of single cells of interest in a real-time fashion, even if the phenotypes are dynamic or the cells of interest are rare. We applied FUNpro to proteomically profile a newly identified small subpopulation of U2OS osteosarcoma cells displaying an abnormal, prolonged DNA damage response (DDR) after ionizing radiation (IR). With this, we identified the PDS5A protein contributing to the abnormal DDR dynamics and helping the cells survive after IR.