Image-guided genomics of phenotypically heterogeneous populations reveals vascular signalling during symbiotic collective cancer invasion

Abstract

Phenotypic heterogeneity is widely observed in cancer cell populations. Here, to probe this heterogeneity, we developed an image-guided genomics technique termed spatiotemporal genomic and cellular analysis (SaGA) that allows for precise selection and amplification of living and rare cells. SaGA was used on collectively invading 3D cancer cell packs to create purified leader and follower cell lines. The leader cell cultures are phenotypically stable and highly invasive in contrast to follower cultures, which show phenotypic plasticity over time and minimally invade in a sheet-like pattern. Genomic and molecular interrogation reveals an atypical VEGF-based vasculogenesis signalling that facilitates recruitment of follower cells but not for leader cell motility itself, which instead utilizes focal adhesion kinase-fibronectin signalling. While leader cells provide an escape mechanism for followers, follower cells in turn provide leaders with increased growth and survival. These data support a symbiotic model of collective invasion where phenotypically distinct cell types cooperate to promote their escape.

EACR-MRS conference on Seed and Soil: In Vivo Models of Metastasis

“…The same collective invasion phenomenon was also observed in 3D spheroids [17]. In a video of an invading spheroid, Dr. Marcus showed that some cells initiate invasion by breaking through the matrix and seem to be followed by the other cells, forming cellular streams.…”

“…While spheroids made of leader cells are highly invasive, follower cells remain packed and hardly invaded. When mixing both cell populations, leader cells remain leaders and rescue follower cells promoting their migration again [17]. Leader cells also appear to be more drug resistant.…”

“…In a video of an invading spheroid, Dr. Marcus showed that some cells initiate invasion by breaking through the matrix and seem to be followed by the other cells, forming cellular streams. When these starting cells move too far and lose contact with the spheroid body, the adjacent cells instantly stop following in 84% of the cases [17]. Therefore, invasion seems to be regulated by a specific class of cells that Dr. Marcus termed "leaders".…”

“…Therefore, invasion seems to be regulated by a specific class of cells that Dr. Marcus termed "leaders". These leader cells can move back to the spheroid, allowing the follower cells to re-attach and follow them again [17]. To further investigate this hypothesis, Dr. Marcus and his team applied SaGA and after dissociating leader and follower cells by FACS, they observed that both populations retain their morphology and properties.…”

Molecular Tension Probes to Investigate the Mechanopharmacology of Single Cells: A Step toward Personalized Mechanomedicine

“…[1] Accordingly, mechanical dysregulation contributes to a range of human diseases, especially in processes that involve mechanically active cells such as smooth muscle cells, cardiomyocytes, and cancer cells. [2] The development of drugs that directly target the underlying mechanically-mediated pathology represents a nascent and growing focus of drug development. Underscoring this growing interest, the term mechanopharmacology was recently introduced to describe the study of drugs that target cell mechanics or the study of how a cell’s mechanical state influences its sensitivity to a particular drug molecule.…”