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Mechanobiology of Disease
Poster Abstracts
65
64-POS
Board 64
Ligand-Activated Epha2 Transport as Marker for Cancer Progression and Population
Heterogeneity
Andrea Ravasio
1
, Bakya Arasi
1
, Myint Z. Myaing
1
, Shumei Chia
2
, Vin Y. Chung
3
, Hui Ting
Ong
1
, Ruby Yun-Ju Huang
3,4
, Ramanuj DasGupta
2
, Jay T. Groves
1,5,6
, Virgile Viasnoff
1,7,8
.
1
National University of Singapore, Singapore, Singapore,
2
Genome Institute of Singapore, A-
STAR, Singapore, Singapore,
3
National University of Singapore, Singapore,
Singapore,
4
National University Health System, Singapore, Singapore,
5
University of California,
Berkeley, CA, USA,
6
University of California, Berkeley, CA, USA,
7
Centre National de la
Recherche Scientifique, Singapore, Singapore,
8
National University of Singapore, Singapore,
Singapore.
Evolution of cancer into heterogeneous subclones is due to somatic mutations, epigenetic
adaptations and selective pressure, leading to phenotypic differences, that result in different EMT
states, migration capacity and, thus metastatic potential. Resistant subclones survive therapeutic
intervention giving rise to recurrence of the disease. Here, we employ an assay to access the
metastatic potential of cancer cells by their phenotypic ability to cluster EphA2 receptor upon
ligand activation. Carcinoma cell lines from different organs, different EMT state and migratory
capacity were seeded onto fluid lipid bilayers presenting the ligand - ephrinA1. Following
activation of the receptor by the ligand, signalling cascades trigger a cytoskeleton-dependent
clustering of the receptor. Using this assay, we could discriminate the degree of receptor
clustering in single cancer cells and we found that it correlates with the EMT state and migration
potential of the cells. Analysis of population distribution showed that fast migratory
mesenchymal-transformed cell lines had a larger spread of the clustering values, suggesting for a
greater heterogeneity within their population. To further test the correlation between migration
speed and EphA2 clustering, we devised an assay to separate subpopulations of cells for their
migration speed and persistence. Fast migratory cells - potentially more invasive – proved to
have higher EphA2 clustering as compared to slower ones. Finally, to test the possible
application of our assay as screening tool for the invasion potential of cancer cells and their
population heterogeneity, we compared cell derived from biopsies from primary and secondary
tumours. Cells from secondary tumour had higher EphA2 clustering as compared to primary
ones. In summary, we showed that EphA2 receptor clustering proved to be a potential tool to
rapidly assess metastatic potential of cancer cells and phenotypic subclonal differences in their
population.