Basal extrusion drives cell invasion and mechanical stripping of E-cadherin
Preprint posted on November 06, 2018 https://www.biorxiv.org/content/early/2018/11/06/463646
Categories: cancer biology, cell biology
Background
Oncogenesis can start with a mutation to a single cell, resulting in a transformed malignant cell, within a healthy epithelial sheet. It is increasingly apparent that single transformed cells can be detected and squeezed out of the tissue by the surrounding normal neighbours. This process has been observed in flies, cell lines, organoids, zebrafish and in vivo using mouse models (Bielmier et al., 2016; Hogan et al., 2009; Kon et al., 2017; Anton et al., 2018).

However, what is less clear is the fate of extruded cells. It is currently thought that the direction of extrusion can impact an extruded cells’ fate; if a cell is pushed out apically then it dies or is cleared through the lumen. However, it is unclear what happens to cells that extrude basally.
In this preprint, the authors investigate the fate of KRas-transformed cells surrounded by normal neighbours in zebrafish. Expression of different oncogenes was combined with fluorescent markers to track mutant cells in real-time. Oncogenes which promote extrusion are associated with metastatic cancers so the authors hypothesise that basal extrusion could be a novel mechanism of metastasis.
Key findings of the preprint
KRas-transformed cells can be extruded basally and apically and initiate tumourigenesis.
Firstly, the authors injected EGFP-KRasV12 plasmids driven by the krt4 promoter into one-cell embryos to generate mosaic expression of KRas in epidermal cells. They found that this was sufficient to form cell masses in 58% of zebrafish embryos compared to 1% of EGFP-CAAX controls. Time-lapse imaging of labelled cells over time revealed that KRasV12 cells can extrude both apically and basally, although significantly more basal extrusions were observed. They saw that basal extrusion can lead to invasion and internalisation of mutant cells into different parts of the zebrafish. Although KRas-transformed cells could form cell masses, a large number of mutant cells were lost over time, which could be a result of extrusion observed in time-lapse imaging.
Loss of p53 promotes the survival extruded cells.
During the evolution of cancer, transformed cells accumulate additional mutations which drive tumourigenesis. To model this the authors combined expression of KRasV12 with depletion of p53 via a translation-blocking morpholino. This led to a reduction in basal extrusion of aberrant cells. However, loss of p53 increased the survival of basally invading cells and their ability to survive as internalised masses at later time points.
Extruded cells can spread through the bloodstream and may initiate EMT
After establishing the system, the authors began characterising cells that invade basally. They found that all internalised cells lose the epithelial marker E-cadherin and a small subset expressed the mesenchymal marker N-cadherin. Intriguingly, time lapse imaging in combination with a vasculature reporter was used to identify GFP-KRasV12 cells in the bloodstream with 65% of 42 zebrafish having circulating cells. Finally, using different fluorescent markers for the apical and cytoplasm it was observed that the apical membrane is ripped off during extrusion and the remainder of the cell migrates away. The authors suggest that basal extrusion drives invasion of KRasV12 cells and strips off the apical surfaces and associated E-cadherin.
What I like about this preprint
This preprint demonstrates that extrusion can lead to mutant cells in the circulation of zebrafish, which is a possible new way for oncogenic cells to spread around the body. Although lots of work on the mechanism of extrusion has been carried out, this paper begins to characterise the fate of extruded cells.
The authors looked at how the mechanical force of KrasV12 cells being squeezed out constricts the nucleus and can scrape off the E-cadherin apical membrane proteins. This sudden loss of the apical membrane could have important implications for metastasis and EMT.
I also liked the great videos of cells extruding from zebrafish!
Questions to the authors
What do you think could be regulating whether mutant cells either accumulate into masses or extrude in the different parts of the zebrafish?
In the movies do you ever see KRasV12 cells extravasate from the circulation?
Mechanical stretch can promote YAP translocation into the nucleus, have you looked for YAP nuclear localisation in extruding cells?
References:
Anton, K. A., Kajita, M., Narumi, R., Fujita, Y., & Tada, M. (2018). Src-transformed cells hijack mitosis to extrude from the epithelium. Nature communications, 9(1), 4695.
Bielmeier, C., Alt, S., Weichselberger, V., La Fortezza, M., Harz, H., Jülicher, F., … & Classen, A. K. (2016). Interface contractility between differently fated cells drives cell elimination and cyst formation. Current Biology, 26(5), 563-574.
Hogan, C., Dupré-Crochet, S., Norman, M., Kajita, M., Zimmermann, C., Pelling, A. E., … & Hosoya, H. (2009). Characterization of the interface between normal and transformed epithelial cells. Nature cell biology, 11(4), 460.
Kon, S., Ishibashi, K., Katoh, H., Kitamoto, S., Shirai, T., Tanaka, S., … & Kamasaki, T. (2017). Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes. Nature cell biology, 19(5), 530.
Posted on: 2nd January 2019
Read preprintSign up to customise the site to your preferences and to receive alerts
Register hereAlso in the cancer biology category:
Subcellular mRNA localization regulates ribosome biogenesis in migrating cells
Selected by | Nidhi Kanwal |
1
Colibactin DNA damage signature indicates causative role in colorectal cancer
Selected by | Connor Rosen |
Mechanotransduction of strain regulates an invasive phenotype in newly transformed epithelial cells
Selected by | William Hill |
Also in the cell biology category:
Nix induced mitochondrial fission, mitophagy, and myocyte insulin resistance are abrogated by PKA phosphorylation
Selected by | Sandra Franco Iborra |
Apical Constriction Reversal upon Mitotic Entry Underlies Different Morphogenetic Outcomes of Cell Division
Selected by | Grace Lim |
Architectural RNA is required for heterochromatin organization
Selected by | Ramona Jühlen |
1
preListscancer biology category:
in theASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
List by | Madhuja Samaddar, Ramona Jühlen, Amanda Haage, Laura McCormick |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
List by | Rob Hynds |
Biophysical Society Annual Meeting 2019
Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA
List by | Joseph Jose Thottacherry |
Also in the cell biology category:
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
List by | Madhuja Samaddar, Ramona Jühlen, Amanda Haage, Laura McCormick |
EMBL Seeing is Believing – Imaging the Molecular Processes of Life
Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019
List by | Gautam Dey |
Autophagy
Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.
List by | Sandra Malmgren Hill |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
List by | Rob Hynds |
Cellular metabolism
A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.
List by | Pablo Ranea Robles |
BSCB/BSDB Annual Meeting 2019
Preprints presented at the BSCB/BSDB Annual Meeting 2019
List by | Gautam Dey |
MitoList
This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.
List by | Sandra Franco Iborra |
ASCB/EMBO Annual Meeting 2018
This list relates to preprints that were discussed at the recent ASCB conference.
List by | Gautam Dey, Amanda Haage |