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Immune cells adapt to confined environments in vivo to optimise nuclear plasticity for migration

ABSTRACT: Cells navigating in complex 3D microenvironments frequently encounter narrow spaces that physically challenge migration. Whilst in vitro studies identified nuclear stiffness as a key rate-limiting factor governing the movement of many cell types through artificial constraints, how cells migrating in vivo respond dynamically to confinement imposed by local tissue architecture, and whether these encounters trigger molecular adaptations, is unclear. Here, we establish an innovative in vivo model for mechanistic analysis of nuclear plasticity as Drosophila immune cells transition into increasingly confined microenvironments. Integrating live in vivo imaging with molecular genetic analyses, we demonstrate how rapid molecular adaptation upon environmental confinement (including fine-tuning of the nuclear lamina) primes leukocytes for enhanced nuclear deformation whilst curbing damage (including rupture and micronucleation), ultimately accelerating movement through complex tissues. We find nuclear dynamics in vivo are further impacted by large organelles (phagosomes) and the plasticity of neighbouring cells, which themselves deform during leukocyte passage. The biomechanics of cell migration in vivo are thus shaped both by factors intrinsic to individual immune cells and the malleability of the surrounding microenvironment.

SUBMITTER: Helen Weavers

PROVIDER: S-SCDT-10_1038-S44319-025-00381-0 | biostudies-other |

REPOSITORIES: biostudies-other

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Project description:Recent in vitro and in vivo studies have highlighted the importance of the cell nucleus in governing migration through confined environments. Microfluidic devices that mimic the narrow interstitial spaces of tissues have emerged as important tools to study cellular dynamics during confined migration, including the consequences of nuclear deformation and nuclear envelope rupture. However, while image acquisition can be automated on motorized microscopes, the analysis of the corresponding time-lapse sequences for nuclear transit through the pores and events such as nuclear envelope rupture currently requires manual analysis. In addition to being highly time-consuming, such manual analysis is susceptible to person-to-person variability. Studies that compare large numbers of cell types and conditions therefore require automated image analysis to achieve sufficiently high throughput. Here, we present an automated image analysis program to register microfluidic constrictions and perform image segmentation to detect individual cell nuclei. The MATLAB program tracks nuclear migration over time and records constriction-transit events, transit times, transit success rates, and nuclear envelope rupture. Such automation reduces the time required to analyze migration experiments from weeks to hours, and removes the variability that arises from different human analysts. Comparison with manual analysis confirmed that both constriction transit and nuclear envelope rupture were detected correctly and reliably, and the automated analysis results closely matched a manual analysis gold standard. Applying the program to specific biological examples, we demonstrate its ability to detect differences in nuclear transit time between cells with different levels of the nuclear envelope proteins lamin A/C, which govern nuclear deformability, and to detect an increase in nuclear envelope rupture duration in cells in which CHMP7, a protein involved in nuclear envelope repair, had been depleted. The program thus presents a versatile tool for the study of confined migration and its effect on the cell nucleus.

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Immune cells adapt to confined environments in vivo to optimise nuclear plasticity for migration

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