Abstract 3150: A characterization of cancer vasculogenic mimicry: Extracellular matrix induced cellular signaling to lumen formation

Abstract

Introduction: Vasculogenic mimicry (VM) describes a process by which cancer cells establish an alternative perfusion pathway in an endothelial cell-free manner. Despite the strong correlation with reduced patient survival, the mechanisms by which a tumor can create this self-generated irrigation system are still not fully understood. The process of VM in vitro can only occur in Matrigel; however, the protein component and signaling pathways involved in this process are unknown. Methods: Using an established in vitro model of VM, of ovarian and breast cancer cells (HEY and MDA-MB-231, respectively) on Matrigel coating, we utilized pharmacological inhibitors, gene silencing and blocking antibodies to elucidate the signaling pathways involved in the process of VM. Immunofluorescence and sirius red staining were used to determine the glycoprotein-rich component lining the lumen of the tubular structures. Results: Differently to what observed in the presence of Matrigel, VM did not occur when cancer cells were cultivated on plastic, glass or heat denatured Matrigel (10 mins at 65°C). Using exclusively Collagen I or Laminin 111 to mimic the extracellular matrix we observed than only in the presence of Laminin 111 could VM formation occur. Laminin is secreted and deposited by HEY cells and constitutes a part of the luminal lining. Silencing of integrin β1, but not β3, by siRNA and antibody blocking prevents this process. Chemical inhibition of PI3K pathway and metalloproteases (MMP) activation demonstrate that these pathways are also essential. RNAseq analysis suggests that this process has minimal dependence on de novo transcriptional activity. Discussion and conclusion: We have shown that VM only occurs when cells are seeded on Matrigel but not on plastic, glass or heat denatured Matrigel, suggesting that this phenomenon is susceptible to substrate/matrix rigidity. Furthermore, we identified Laminin as the essential matrix protein secreted and deposited by cancer cells to allow for VM assembly. Its interaction with integrin β1, and the consequent regulation of the activity of MMP leads to the remodeling of the ECM to favor the connection of the VM channels to the microcirculation system. This pathway is not heavily dependent on transcription but requires the PI3K pathway. As VM is strongly associated with poor patient survival, understanding the formation of this alternative irrigation system may deliver new druggable targets. Citation Format: Gabriel Mingo, Andres Valdivia, Varina Aldana, Javiera Pradenas, Nicole Babbitt, Pamela Gonzalez, Francisco Nualart, Jorge Díaz, Lisette Leyton, Cristina Bertocchi, Gareth Owen. A characterization of cancer vasculogenic mimicry: Extracellular matrix induced cellular signaling to lumen formation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3150.