Contractile forces direct the chiral swirling of minimal cell collectives

Abstract Chirality is a conserved biological feature with critical implications in tissue morphogenesis and embryonic development. In culture, large multicellular groups exhibit spontaneous chiral symmetry break when moving collectively on micropatterned surfaces. Although several studies have shown that actin network integrity and acto-myosin network contractility participate to the establishment of the chirality of the movement, the exact contribution of contractile forces to the directionality of the chiral bias in collectives remains to be elucidated. Here we studied the contractile forces produced by a minimal collective constituted of a pair of endothelial cells. We first show that cell doublets confined on disk-shaped micropatterns undergo spontaneous and persistent chiral swirling, displaying a mild but robust clockwise (CW) bias, as the one observed in bigger collectives. This bias could be amplified or reversed by modulating contractile forces. Traction force measurements revealed that large forces tend to drive counter-clockwise (CCW) rotation whereas low forces rather favor a CW rotation. Furthermore, the study of heterotypic doublets indicates that the speed and direction of the rotation is determined by the more contractile cells within the doublets. These results thus revealed that contractile leader cells could drive the chiral motion of minimal collectives. Significance Statement Chirality, which represents a fundamental property of living systems, manifests in cell collectives by their persistent biased directional swirling. Despite the clear identification of the implication of actomyosin cytoskeleton in driving the internal chiral symmetry break occurring in cells, little is known about the actual role of cellular forces produced by this network in the development of handedness in collectives. Our findings establish that the level of mechanical energy developed by pairs of confined endothelial cells regulates the strength and direction of their rotation. Our results also identify the more contractile cell of the doublet as the cell driving the direction and speed of rotation of the pair. This study thus sheds new light on the importance of the generation and integration of mechanical forces within a small collective in the determination of its chiral rotation.