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Relatively Rare Populations of Invasive Cells Drive Progression of Heterogeneous Tumors

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Abstract

Introduction

Breast tumors often display an astonishing degree of spatial and temporal heterogeneity, which are associated with cancer progression, drug resistance, and relapse. Triple-negative breast cancer (TNBC) is a particularly aggressive and heterogeneous subtype for which targeted therapies are scarce. Consequently, patients with TNBC have a poorer overall prognosis compared to other breast cancer patients. Within heterogeneous tumors, individual clonal subpopulations may exhibit differences in their rates of growth and degrees of invasiveness. We hypothesized that such phenotypic heterogeneity at the single-cell level may accelerate tumor progression by enhancing the overall growth and invasion of the entire tumor.

Methods

To test this hypothesis, we isolated and characterized clonal subpopulations with distinct morphologies and biomarker expression from the inherently heterogeneous 4T1 mouse mammary carcinoma cell line. We then leveraged a 3D microfluidic tumor model to reverse-engineer intratumoral heterogeneity and thus investigate how interactions between phenotypically distinct subpopulations affect tumor growth and invasion.

Results

We found that the growth and invasion of multiclonal tumors were largely dictated by the presence of cells with epithelial and mesenchymal traits, respectively. The latter accelerated overall tumor invasion, even when these cells comprised less than 1% of the initial population. Consistently, tumor progression was delayed by selectively targeting the mesenchymal subpopulation.

Discussion

This work reveals that highly invasive cells can dominate tumor phenotype and that specifically targeting these cells can slow the progression of heterogeneous tumors, which may help inform therapeutic approaches.

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Data availability

RNA-seq data have been deposited at GEO under accession number GSE252177 and are available as of the date of publication.

Abbreviations

CAM:

Chorioallantoic membrane

EMT:

Epithelial–mesenchymal transition

TNBC:

Triple-negative breast cancer

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Acknowledgements

This work was supported in part by grants from the National Institutes of Health (CA187692, CA214292), the New Jersey Health Foundation, and a Faculty Scholars Award from the Howard Hughes Medical Institute. SEL was supported in part by the National Center for Advancing Translational Sciences of the National Institutes of Health (TL1TR003019, UL1TR003017). MCB was supported in part by the Ludwig Princeton Branch and by a New Jersey Commission on Cancer Research Predoctoral Fellowship (COCR22PRF009).

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Author notes

  1. Susan E. Leggett and Molly C. Brennan have contributed equally.

Authors and Affiliations

  1. Department of Chemical & Biological Engineering, Princeton University, 303 Hoyt Laboratory, 25 William Street, Princeton, NJ, 08544, USA

    Susan E. Leggett, Molly C. Brennan, Sophia Martinez & Celeste M. Nelson

  2. Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA

    Susan E. Leggett

  3. Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA

    Joe Tien

  4. Department of Molecular Biology, Princeton University, Princeton, NJ, 08544, USA

    Celeste M. Nelson

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Correspondence to Celeste M. Nelson.

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Leggett, S.E., Brennan, M.C., Martinez, S. et al. Relatively Rare Populations of Invasive Cells Drive Progression of Heterogeneous Tumors. Cel. Mol. Bioeng. 17, 7–24 (2024). https://doi.org/10.1007/s12195-023-00792-w

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