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Isolating and targeting the real-time plasticity and malignant properties of epithelial-mesenchymal transition in cancer

Oncogene volume 40, pages 2884–2897 (2021)Cite this article

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Abstract

Epithelial-mesenchymal transition (EMT) is a driving force in promoting malignant cancer, including initiation, growth, and metastasis. EMT is a dynamic process that can undergo a mesenchymal-epithelial transition (MET) and partial transitions between both phenotypes, termed epithelial-mesenchymal plasticity (EMP). In cancer, the acquisition of EMP results in a spectrum of phenotypes, promoting tumor cell heterogeneity and resistance to standard of care therapy. Here we describe a real-time fluorescent dual-reporter for vimentin and E-cadherin, biomarkers of the mesenchymal and epithelial cell phenotypes, respectively. Stable dual-reporter cell lines generated from colorectal (SW620), lung (A549), and breast (MDA-MB-231) cancer demonstrate a spectrum of EMT cell phenotypes. We used the dual-reporter to isolate the quasi epithelial, epithelial/mesenchymal, and mesenchymal phenotypes. Although EMT is a dynamic process, these isolated quasi-EMT-phenotypes remain stable to spontaneous EMP in the absence of stimuli and during prolonged cell culture. However, the quasi-EMT phenotypes can readily be induced to undergo EMT or MET with growth factors or small molecules. Moreover, isolated EMT phenotypes display different tumorigenic properties and are morphologically and metabolically distinct. 3D high-content screening of ~23,000 compounds using dual-reporter mesenchymal SW620 tumor organoids identified small molecule probes that modulate EMT, and a subset of probes that effectively induced MET. The tools, probes, and models described herein provide a coherent mechanistic understanding of mesenchymal cell plasticity. Future applications utilizing this technology and probes are expected to advance our understanding of EMT and studies aimed at therapeutic strategies targeting EMT.

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Fig. 1: The dual-reporter allows for the identification and isolation of distinct EMT subtypes.
Fig. 2: The mesenchymal phenotype has higher stemness and invasive potential.
Fig. 3: Isolated EMT cell populations have distinct metabotypes.
Fig. 4: Treatment of M-phenotype CRC tumor organoids with 7 induces MET by metabolomics.
Fig. 5: A diverse set of probes reverse EMT in a dose-dependent manner.
Fig. 6: Mechanisms regulating mesenchymal cell EMP in CRC.

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Acknowledgements

This research was supported in part by grants awarded to DVL from the Department of Defense Peer Reviewed Cancer Research Program (W81XWH-18-1-0142), the NIH NCI (1R01CA251361-01), and The ALSAM Foundation Therapeutic Innovation Grant. TN was supported by a NIH Training Grant (5T32HL7171-42). SR was supported by the Cancer Foundation of Luxembourg. We thank the Eli Lilly and Company Open Innovation Drug Discovery Program, particularly Evan Castetter and Maria Jose Lorite, for providing the BIC library. We also thank CU AMC shared resource laboratories, including the Drug Discovery and Development Shared Resource (HTS/HCS Discovery Arm), MS Metabolomics, Flow Cytometry, and NMR. These shared resources are supported in part by the UCCC NIH NCI designated cancer center (P30CA046934). We are grateful to Claudia Esquer for graphic design input of Fig. 6.

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  1. These authors contributed equally: Hector Esquer, Qiong Zhou

Authors and Affiliations

  1. The Skaggs School of Pharmacy and Pharmaceutical Sciences, Department of Pharmaceutical Sciences, The University of Colorado Anschutz Medical Campus (CU AMC), Aurora, Colorado, USA

    Hector Esquer, Qiong Zhou, Adedoyin D. Abraham, Sébastien Rinaldetti, Michael V. Orman & Daniel V. LaBarbera

  2. Department of Biochemistry and Molecular Genetics, The University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA

    Travis Nemkov & Angelo D’Alessandro

  3. Early Translation Branch, National Center for Advancing Translational Sciences (NCATS), National Institutes of Health (NIH), Rockville, Maryland, USA

    Yu-Chi Chen, Xiaohu Zhang & Marc Ferrer

  4. The University of Colorado Cancer Center (UCCC); The University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA

    Angelo D’Alessandro, Wells A. Messersmith & Daniel V. LaBarbera

  5. The School of Medicine, Division of Medical Oncology, The University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA

    Wells A. Messersmith

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Esquer, H., Zhou, Q., Nemkov, T. et al. Isolating and targeting the real-time plasticity and malignant properties of epithelial-mesenchymal transition in cancer. Oncogene 40, 2884–2897 (2021). https://doi.org/10.1038/s41388-021-01728-2

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