Interplay of the physical microenvironment, contact guidance, and intracellular signaling in cell decision making

Abstract

The peritumoral physical microenvironment consists of complex topographies that influence cell migration. Cell decision making, upon encountering anisotropic, physiologically relevant physical cues, has yet to be elucidated. By integrating microfabrication with cell and molecular biology techniques, we provide a quantitative and mechanistic analysis of cell decision making in a variety of well-defined physical microenvironments. We used MDA-MB-231 breast carcinoma and HT1080 fibrosarcoma as cell models. Cell decision making after lateral confinement in 2-dimensional microcontact printed lines is governed by branch width at bifurcations. Cells confined in narrow feeder microchannels prefer to enter wider branches at bifurcations. In contrast, in feeder channels that are wider than the cell body, cells elongate along one side wall of the channel and are guided by contact with the wall to the contiguous branch channel independent of its width. Knockdown of β1-integrins or inhibition of cellular contractility suppresses contact guidance. Concurrent, but not individual, knockdown of nonmuscle myosin isoforms IIA and IIB also decreases contact guidance, which suggests the existence of a compensatory mechanism between myosin IIA and myosin IIB. Conversely, knockdown or inhibition of cell division control protein 42 homolog promotes contact guidance-mediated decision making. Taken together, the dimensionality, length scales of the physical microenvironment, and intrinsic cell signaling regulate cell decision making at intersections.-Paul, C. D., Shea, D. J., Mahoney, M. R., Chai, A., Laney, V., Hung, W.-C., Konstantopoulos, K. Interplay of the physical microenvironment, contact guidance, and intracellular signaling in cell decision making.

Myosin phosphorylation on stress fibers predicts contact guidance behavior across diverse breast cancer cells

“…Recently, β1 integrin was shown to be important in confined migration [69] and β1 integrin is important in generating traction forces [24]. Both of these lines of evidence indicate a role for β1 integrin during contact guidance.…”

Cancer cell motility: lessons from migration in confined spaces

“…As such, these cells, when they reach an intersection, ‘decide’ to follow the path of least hydraulic resistance, meaning that cells choose the shortest path or the path with wider microchannels. Similarly, MDA-MB-231 cells confined in narrow (30 μm 2 ) channels preferentially enter the wider branches at asymmetrical bifurcations 40 . In wider (200 μm 2 ) 3D tracks, cell elongation and alignment along either the left or right side wall of the feeder channel manifests in persistent cell migration through the phenomenon termed contact guidance 40 .…”

“…Similarly, MDA-MB-231 cells confined in narrow (30 μm 2 ) channels preferentially enter the wider branches at asymmetrical bifurcations 40 . In wider (200 μm 2 ) 3D tracks, cell elongation and alignment along either the left or right side wall of the feeder channel manifests in persistent cell migration through the phenomenon termed contact guidance 40 . As such, when they reach an asymmetrical bifurcation, cells enter both narrow (30 μm 2 ) and wide (200 μm 2 ) branches with an equal probability 40 .…”

“…Engineered microenvironments enable high-throughput mechanistic studies in well-defined models of migration spaces in which the individual factors influencing migrating cells (for example, the cross-sectional area available for migration, substrate stiffness, ligand density and the presence of external gradients) are decoupled 37–39 . These in vitro assays providea simplified view of the in vivo setting and impose well-controlled constraints on cells, thereby enabling fine control of the microenvironment so that cell shape 40 , protein localization and actin polymerization 41–44 , as well as response to chemical stimuli 45 , during migration can be studied.…”

Probing cellular response to topography in three dimensions

“…Cell mechanosensing is a complex phenomenon where topographical cues via cellextracellular matrix (ECM) adhesion molecules act concomitantly with intracellular machinery to drive cellular responses (34). While studies using patterned two-dimensional substrates (35,36) and microfabricated environments (37) have revealed aspects of cell response to topography, reaction to topographic cues in more physiologically relevant three-dimensional environments are not well understood. Importantly, three-dimensional cues are likely vital to recapitulate some aspects of physiological mechanosensing.…”