Microscopy-based Single-cell Proteomic Profiling Reveals Heterogeneity in DNA Damage Response Dynamics

Overview

Journal Cell Rep Methods

Specialty Cell Biology

Date 2022 Jul 5

PMID 35784653

Authors

Pin-Rui Su

Li You

Cecile Beerens

Karel Bezstarosti

Jeroen Demmers

Martin Pabst

Roland Kanaar

Cheng-Chih Hsu

Miao-Ping Chien

Affiliations

Soon will be listed here.

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.

Citing Articles

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Research progress and the prospect of using single-cell sequencing technology to explore the characteristics of the tumor microenvironment.

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Chen T, You L, Hardillo J, Chien M Cells. 2023; 12(16).

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