Inhibition of cell–cell adhesion impairs directional epithelial migration on micropatterned surfaces

Abstract

The development of the vertebrate body plan with left-right (LR) asymmetry (also known as handedness and chirality) requires the emerging chiral morphogenesis of epithelial cells at specific embryonic stages. In this process, cell-cell adhesions coordinate cellular organization and collective cell migration, and are critical for the directional looping of developing embryonic organs. However, the underlying biophysical mechanism is not yet well understood. Here we modeled normal and delayed epithelial LR symmetry breaking with patterned epithelial chiral morphogenesis on microscale lines with various widths. The patterned cells exhibited biased migration wherein those on opposing boundaries migrated in different directions. Disrupting adherens junctions with ethylene glycol tetraacetic acid (EGTA) resulted in a decrease in velocity difference in opposing boundaries as well as the associated biased cell alignment, along with an increase in the overall random motion. Altering the distance between the opposing boundaries did not significantly alter alignment, but significantly disturbed the velocity profile of the cell migration field. Further examination of cell polarity indicated that disruption of adherens junctions did not affect cell polarization on the boundaries, but decreased the transmission of chiral bias into the interior region of the epithelial cell sheet. Overall, our results demonstrated the dependence of the scale of collective cell migration on the strength of cell-cell adhesion, and its effects on the chirality of a multicellular structure through mediating cell polarity in the vicinity of geometric boundaries. This study demonstrated that our 2D microscale system provides a simple yet effective tool for studying the influence of collective cell migration on LR symmetry breaking, and possibly for fetal drug screening to prevent birth defects related to alteration in cell-cell adhesion.

Cellular and Nuclear Alignment Analysis for Determining Epithelial Cell Chirality

“…Results were similar to those generated by the intensity gradient based analysis, which have been used in several studies 18, 25, 28 , and the only slight deviation was in the final count of CCW against NC (non-chiral) rings.…”

“…Cell alignment has been identified as a chiral measure and used for several studies 18, 25, 28 . As the nucleus tends to align themselves with the cells 4, 30 , the chiral nuclear alignment is partially expected.…”

“…used a cell micropatterning technique, analyzed still phase contrast images of patterned cells, and found that cells have an innate, distinct chiral bias, which is dependent of cell phenotype and mediated by actin 24–26 . The scale of chiral bias of epithelial cells was shown to depend on cell-cell adhesion 28 . Cells lost their distinct chirality when exposed to carbon nanotubes due to oxidative stress, suggesting a potential application of cell chirality as a biomarker for detecting cell toxicity 18 .…”

A patient with chronic myeloid leukemia and situs inversus totalis: A case report

“…Although the occurrence of malignancy in patients with SIT may be coincidental, certain studies have suggested a possible association between malignancy and SIT (16)(17)(18). Previous studies have proposed that unidentified genes affecting the left-right axis arrangement may be associated with cancer susceptibility (19,20). Although this may potentially explain the occurrence of cancer in the patient with SIT assessed in the present study, no genes have been conclusively identified as candidates for this dual role to the best of our knowledge (16).…”

Cell chirality: its origin and roles in left–right asymmetric development

“…Whether the handedness is dextral or sinistral depends on the cell line [49]. Chirality in the nuclear shape and the involvement of E-cadherin in transmitting a chiral bias to neighbouring cells were shown using Madin–Darby canine kidney epithelial cells [51,52]. …”