Colonization of distant organs by tumor cells generating circulating homotypic clusters adaptive to fluid shear stress

Manabu Maeshiro,Kazuya Iwamoto,Rin Liu,Hideki Nakayama,Hirotaka Matsui,Satoru Shinriki,Ryoji Yoshida,Yoshihiro Komohara,Yutaka Nakachi,Kazuaki Ohtsubo,Yukio Fujiwara

doi:10.1038/s41598-021-85743-z

Manabu Maeshiro, Kazuya Iwamoto + Show 9 more

Open Access

https://doi.org/10.1038/s41598-021-85743-z

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Journal: Scientific Reports	Publication Date: Mar 17, 2021
Citations: 13	License type: open-access

Affiliation: Kumamoto University

Abstract

Once disseminated tumor cells (DTCs) arrive at a metastatic organ, they remain there, latent, and become seeds of metastasis. However, the clonal composition of DTCs in a latent state remains unclear. Here, we applied high-resolution DNA barcode tracking to a mouse model that recapitulated the metastatic dormancy of head and neck squamous cell carcinoma (HNSCC). We found that clones abundantly circulated peripheral blood dominated DTCs. Through analyses of multiple barcoded clonal lines, we identified specific subclonal population that preferentially generated homotypic circulating tumor cell (CTC) clusters and dominated DTCs. Despite no notable features under static conditions, this population significantly generated stable cell aggregates that were resistant to anoikis under fluid shear stress (FSS) conditions in an E-cadherin-dependent manner. Our data from various cancer cell lines indicated that the ability of aggregate-constituting cells to regulate cortical actin-myosin dynamics governed the aggregates’ stability in FSS. The CTC cluster-originating cells were characterized by the expression of a subset of E-cadherin binding factors enriched with actin cytoskeleton regulators. Furthermore, this expression signature was associated with locoregional and metastatic recurrence in HNSCC patients. These results reveal a biological selection of tumor cells capable of generating FSS-adaptive CTC clusters, which leads to distant colonization.

Highlights

Once disseminated tumor cells (DTCs) arrive at a metastatic organ, they remain there, latent, and become seeds of metastasis
Thirty days after the injection, we confirmed primary tumors as clear masses by visual inspection and bioluminescent imaging; we found no metastatic mass in other organs (Fig. 1a)
All the bone marrow (BM)-DTCs we visually detected were negative for a cell proliferation marker Ki-67 (Supplementary Fig. S1a), suggesting that BM-DTCs were in a dormant cellular state

Summary

Introduction

Once disseminated tumor cells (DTCs) arrive at a metastatic organ, they remain there, latent, and become seeds of metastasis. Most DTCs in bone marrow (BM) are negative for proliferation markers[3], the abundance of these cells directly correlates with reduced metastasis-free survival in various types of cancer including head and neck squamous cell carcinoma (HNSCC)[3]. Despite such biological and clinical implications, the composition of DTCs surviving in a latent state remains obscure. We identified a stable functionally homotypic CTC clustering by specific tumor cells that regulated actomyosin machinery in response to FSS, as a crucial clonal selection process leading to dominant colonization of distant organs

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