High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells

Dmitry Kuchenov, Vibor Laketa, Frank Stein, Florian Salopiata, Ursula Klingmüller, Carsten Schultz

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Essential characteristics of cellular signaling networks include a complex interconnected architecture and temporal dynamics of protein activity. The latter can be monitored by Förster resonance energy transfer (FRET) biosensors at a single-live-cell level with high temporal resolution. However, these experiments are typically limited to the use of a couple of FRET biosensors. Here, we describe a FRET-based multi-parameter imaging platform (FMIP) that allows simultaneous high-throughput monitoring of multiple signaling pathways. We apply FMIP to monitor the crosstalk between epidermal growth factor receptor (EGFR) and insulin-like growth factor-1 receptor signaling, signaling perturbations caused by pathophysiologically relevant EGFR mutations, and the effects of a clinically important MEK inhibitor (selumetinib) on the EGFR network. We expect that in the future the platform will be applied to develop comprehensive models of signaling networks and will help to investigate the mechanism of action as well as side effects of therapeutic treatments.

Original languageEnglish (US)
Pages (from-to)1550-1559
Number of pages10
JournalCell Chemical Biology
Volume23
Issue number12
DOIs
StatePublished - Dec 22 2016

Fingerprint

Energy Transfer
Epidermal Growth Factor Receptor
Cells
Biosensing Techniques
Energy transfer
Imaging techniques
Biosensors
Somatomedin Receptors
Mitogen-Activated Protein Kinase Kinases
Therapeutic Uses
Cell signaling
Somatomedins
Crosstalk
Mutation
Throughput
Monitoring
Proteins
Experiments

Keywords

  • fluorescence microscopy
  • FRET
  • growth factors
  • kinases
  • principal component analysis
  • sensors
  • signaling cross-talk

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Molecular Biology
  • Molecular Medicine
  • Drug Discovery
  • Pharmacology

Cite this

High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells. / Kuchenov, Dmitry; Laketa, Vibor; Stein, Frank; Salopiata, Florian; Klingmüller, Ursula; Schultz, Carsten.

In: Cell Chemical Biology, Vol. 23, No. 12, 22.12.2016, p. 1550-1559.

Research output: Contribution to journalArticle

Kuchenov, Dmitry ; Laketa, Vibor ; Stein, Frank ; Salopiata, Florian ; Klingmüller, Ursula ; Schultz, Carsten. / High-Content Imaging Platform for Profiling Intracellular Signaling Network Activity in Living Cells. In: Cell Chemical Biology. 2016 ; Vol. 23, No. 12. pp. 1550-1559.
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