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ePoster Display

23P - Transcriptional profiling of circulating tumor and hybrid cells in breast cancer patients


16 Sep 2021


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Breast Cancer


Evgeny Denisov


Annals of Oncology (2021) 32 (suppl_5): S361-S375. 10.1016/annonc/annonc684


E.V. Denisov1, M.E. Menyailo1, S.Y. Zolotareva1, T.S. Gerashchenko1, V.V. Alifanov2, L.A. Tashireva2, O.E. Savelieva2, E.S. Grigoryeva3, N.V. Cherdyntseva3, V.M. Perelmuter2

Author affiliations

  • 1 Laboratory Of Cancer Progression Biology, Tomsk National Research Medical Center, 634009 - Tomsk/RU
  • 2 Department Of General And Molecular Pathology, Tomsk National Research Medical Center, 634009 - Tomsk/RU
  • 3 Laboratory Of Molecular Oncology And Immunology, Tomsk National Research Medical Center, 634009 - Tomsk/RU


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Abstract 23P


Metastasis is initiated by tumor cells detached from the primary tumor and entering the bloodstream. Circulating tumor cells (CTCs) are presented by clusters and individual cells which consist of phenotypically and genetically distinct subpopulations. However, only a few CTCs can survive in the bloodstream and form metastases. Deciphering CTC heterogeneity represents a promising instrument for the identification of the metastasis-initiating cells and developing therapeutic strategies against metastatic disease. In this study, we used single-cell RNA sequencing to assess CTC composition in patients with breast cancer.


CTCs were enriched by RosetteSep Human CD45 Depletion Cocktail (STEMCELL Technologies Inc.) and sequenced using Chromium Single Cell 3' Reagents v3.1 (10x Genomics) on NextSeq 500 instrument (Illumina). The data were analyzed using Cell Ranger and Loupe Browser (10x Genomics).


In total, 100 CD45-negative CTCs expressing different epithelial genes (EpCAM, KRT5, 8, 17-19, and E-cadherin) have been identified. These CTCs were represented by two subpopulations one of which was enriched by NF-kB, Notch, and HIF-1 signaling pathways, whereas another one showed hematopoietic cell lineage features. Surprisingly, despite the CD45-negative selection of CTCs used in the study, we detected 67 circulating hybrid cells (CHCs) positive for CD45 and epithelial genes. CHCs were represented by two subpopulations expressing either CD4 or CD14. Both CTCs and CHCs demonstrated significant heterogeneity in the expression of mesenchymal (VIM and CDH2) and stem (CD44, CD133, and ALDH1) genes. However, only CTCs and CHCs with mesenchymal and stem features were enriched in signaling pathways involved in transcriptional misregulation in cancer and inflammation.


CTCs and CHCs are represented by transcriptionally distinct subpopulations of which mesenchymal stem-like cells are characterized by multiple cancer and inflammation signaling pathways.

Clinical trial identification

Editorial acknowledgement

Legal entity responsible for the study

The authors.


Russian Science Foundation, grant number 19-75-30016.


All authors have declared no conflicts of interest.

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