Tubulin-Based Microtentacles Aid in Heterotypic Clustering of Neutrophil-Differentiated HL-60 Cells and Breast Tumor Cells.

Abstract

Circulating tumor cells (CTCs) travel through the vasculature to seed secondary sites and serve as direct precursors of metastatic outgrowth for many solid tumors. Heterotypic cell clusters form between CTCs and white blood cells (WBCs) and recent studies report that a majority of these WBCs are neutrophils in patient and mouse models. The lab discovered that CTCs produce tubulin-based protrusions, microtentacles (McTNs), which promote reattachment, retention in distant sites during metastasis and formation of tumor cell clusters. Neutrophil-CTC clusters help CTCs survive the harsh vascular environment to promote successful metastasis, however, the specific mechanism of this interaction is not fully understood. Utilizing TetherChip technology, it is found that primary and differentiated neutrophils produce McTNs composed of detyrosinated and acetylated α-tubulin and vimentin. Neutrophil McTNs aid in cluster formation, migration, and reattachment, which are suppressed with the tubulin-depolymerizing agent, Vinorelbine. Co-culturing differentiated neutrophils and tumor cells formed heterotypic clusters that enhanced migration. CTC-neutrophil clusters have higher metastatic efficiency, and by demonstrating that neutrophils form McTNs, a new possible mechanism for how neutrophils interact with tumor cells is revealed. These findings further support the idea that developing cluster-disrupting therapies can provide a new targeted strategy to reduce the metastatic potential of cancer cells.