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Pre-existing Cell States Control Heterogeneity of Both EGFR and CXCR4 Signaling

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Abstract

Introduction

CXCR4 and epidermal growth factor receptor (EGFR) represent two major families of receptors, G-protein coupled receptors and receptor tyrosine kinases, with central functions in cancer. While utilizing different upstream signaling molecules, both CXCR4 and EGFR activate kinases ERK and Akt, although single-cell activation of these kinases is markedly heterogeneous. One hypothesis regarding the origin of signaling heterogeneity proposes that intercellular variations arise from differences in pre-existing intracellular states set by extrinsic noise. While pre-existing cell states vary among cells, each pre-existing state defines deterministic signaling outputs to downstream effectors. Understanding causes of signaling heterogeneity will inform treatment of cancers with drugs targeting drivers of oncogenic signaling.

Methods

We built a single-cell computational model to predict Akt and ERK responses to CXCR4- and EGFR-mediated stimulation. We investigated signaling heterogeneity through these receptors and tested model predictions using quantitative, live-cell time-lapse imaging.

Results

We show that the pre-existing cell state predicts single-cell signaling through both CXCR4 and EGFR. Computational modeling reveals that the same set of pre-existing cell states explains signaling heterogeneity through both EGFR and CXCR4 at multiple doses of ligands and in two different breast cancer cell lines. The model also predicts how phosphatidylinositol-3-kinase (PI3K) targeted therapies potentiate ERK signaling in certain breast cancer cells and that low level, combined inhibition of MEK and PI3K ablates potentiated ERK signaling.

Conclusions

Our data demonstrate that a conserved motif exists for EGFR and CXCR4 signaling and suggest potential clinical utility of the computational model to optimize therapy.

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Acknowledgments

The authors acknowledge funding from United States National Institutes of Health Grants R01CA196018, R01CA238042, R01CA238023, R33CA225549, R37CA222563, R50CA221807, and U01CA210152. P.C.S was supported by the NIH Microfluidics in Biomedical Sciences Training Program NIBIB T32 EB005582. B.H was supported by an American Cancer Society - Michigan Cancer Research Fund Postdoctoral Fellowship, PF-18-236-01-CCG. The authors also thank Annabel Levinson for technical assistance.

Author Contributions

P.C.S, P.C.K, J.J.L, G.D.L, and K.E.L conceptualized the study. B.H and K.E.L provided reagents. P.C.S, P.C.K, and K.E.L performed experiments and analyzed data. P.C.S, P.C.K., J.J.L, G.D.L, and K.E.L wrote the manuscript. All authors edited the manuscript before submission.

Conflict of interest

G.D.L receives research funding and serves on the scientific advisory board for Polyphor.

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Authors and Affiliations

  1. Department of Chemical Engineering, University of Michigan, 2800 Plymouth Road, Ann Arbor, MI, 48109-2800, USA

    Phillip C. Spinosa, Patrick C. Kinnunen & Jennifer J. Linderman

  2. Department of Radiology Center for Molecular Imaging, University of Michigan Medical School, Ann Arbor, MI, 48109, USA

    Brock A. Humphries, Gary D. Luker & Kathryn E. Luker

  3. Department of Biomedical Engineering, University of Michigan Medical School, Ann Arbor, MI, USA, 48109

    Gary D. Luker & Jennifer J. Linderman

  4. Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA, 48109

    Gary D. Luker

  5. Department of Radiology, Center for Molecular Imaging, University of Michigan, 109 Zina Pitcher Place, A526 BSRB, Ann Arbor, MI, 48109-2200, USA

    Kathryn E. Luker

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  1. Phillip C. Spinosa
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Spinosa, P.C., Kinnunen, P.C., Humphries, B.A. et al. Pre-existing Cell States Control Heterogeneity of Both EGFR and CXCR4 Signaling. Cel. Mol. Bioeng. 14, 49–64 (2021). https://doi.org/10.1007/s12195-020-00640-1

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