A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma

Tugba Keskin; Beatrice Rucci; Sandrine Cornaz-Buros; Patricia Martin; Carlo Fusco; Liliane Broye; Katarina Cisarova; Elizabeth M Perez; Igor Letovanec; Stefano La Rosa; Stephane Cherix; Manuel Diezi; Raffaele Renella; Paolo Provero; Mario L Suvà; Ivan Stamenkovic; Nicolò Riggi

doi:10.1126/sciadv.abf9394

A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma

Sci Adv. 2021 Jul 2;7(27):eabf9394. doi: 10.1126/sciadv.abf9394. Print 2021 Jul.

Authors

Tugba Keskin¹, Beatrice Rucci¹, Sandrine Cornaz-Buros¹, Patricia Martin¹, Carlo Fusco¹, Liliane Broye¹, Katarina Cisarova², Elizabeth M Perez^{3

4

5}, Igor Letovanec^{6

7}, Stefano La Rosa⁷, Stephane Cherix⁸, Manuel Diezi⁹, Raffaele Renella⁹, Paolo Provero¹⁰, Mario L Suvà^{3

4}, Ivan Stamenkovic¹¹, Nicolò Riggi¹

Affiliations

¹ Experimental Pathology Service, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland.
² Department of Computational Biology, University of Lausanne, Lausanne, Switzerland.
³ Department of Pathology and Center for Cancer Research, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA.
⁴ Broad Institute of MIT and Harvard, Boston, MA, USA.
⁵ Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
⁶ Department of Histopathology, Central Institute, Valais Hospital, Sion, Switzerland.
⁷ Institute of Pathology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland.
⁸ Department of Orthopedics, Faculty of Biology and Medicine, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
⁹ Department Woman-Mother-Child, Division of Pediatrics, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland.
¹⁰ Department of Neurosciences Rita Levi Montalcini, University of Turin, Center of Omics Sciences, Ospedale San Raffaele IRCSS, Milan, Italy.
¹¹ Experimental Pathology Service, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland. [email protected].

Abstract

Targeting of the most aggressive tumor cell subpopulations is key for effective management of most solid malignancies. However, the metastable nature of tumor heterogeneity, which allows cells to transition between strong and weak tumorigenic phenotypes, and the lack of reliable markers of tumor-promoting properties hamper identification of the most relevant cells. To overcome these obstacles, we designed a functional microRNA (miR)-based live-cell reporter assay to identify highly tumorigenic cells in xenotransplants of primary Ewing sarcoma (EwS) 3D cultures. Leveraging the inverse relationship between cell pluripotency and miR-145 expression, we successfully separated highly tumorigenic, metastasis-prone (miR-145^low) cells from poorly tumorigenic, nonmetastatic (miR-145^high) counterparts. Gene expression and functional studies of the two cell populations identified the EPHB2 receptor as a prognostic biomarker in patients with EwS and a major promoter of metastasis. Our study provides a simple and powerful means to identify and isolate tumor cells that display aggressive behavior.

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