Impact of cell shapes on cell behaviour and fate

Project Information

SHAPINCELLFATE

Grant agreement ID: 101071470

DOI

10.3030/101071470

EC signature date 6 March 2023

Start date 1 April 2023

End date 31 March 2029

Funded under

• European Research Council (ERC)

Total cost

€ 10 834 795,00

EU contribution

€ 10 834 795,00

Coordinated by

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
France

Periodic Reporting for period 2 - SHAPINCELLFATE (Impact of cell shapes on cell behaviour and fate)

Reporting period: 2024-10-01 to 2026-03-31

Summary of the context and overall objectives of the project

Cells are often depicted as irregular spherical objects - the shape they adopt when in suspension. However, the packed environment of tissues alters this simple shape, causing large cell deformations. This occurs during normal tissue growth and is even more pronounced upon tissue overgrowth, as in the case of solid tumors. Cell shape changes frequently occur in migratory cells, such as immune cells that patrol the organism within interstitial tissues, and cancer metastases that escape from the primary tumor to invade healthy tissues. In all cases, cells adapt and survive even to very large deformations. The mechanisms enabling this adaptation and the long-term consequences that these repeated changes in form have on physiology and pathology remain largely unknown.
It has been observed that changes in the shape of cells and organelle(s) induce reversible and irreversible modifications in their behaviour and function(s). The aim of SHAPINCELLFATE is thus to reveal whether cell shape alterations imposed by physical constraints in the tissues have an influence on the cell’s present and future behavior.
We propose to tackle the molecular mechanisms and physical principles accounting for shape-induced memory effects on two cell types that undergo large shape changes in vivo, and communicate to establish cancer immunity: (1) dendritic cells (hereafter referred to as DCs), which initiate adaptive immune responses, and (2) cancer cells that can migrate leading to metastasis.
Our project will reveal whether physical confinement and the physical history of a cell are determinants of cellular behaviours, and may further establish novel clinical paths for a holistic understanding of early malignancies and their recognition by the immune system.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

DCs are immune cells found in a variety of peripheral tissues, where they sample the environment in the lookout for threats such as microbial infections. When they find such threats, they subsequently migrate from the peripheral tissues to the lymph nodes where they transmit this information to T cells. These migration steps are accompanied by changes in DC shape and we set out to quantify such changes, their molecular mechanisms and long-term phenotypic and functional consequences. DC migration from the peripheral tissues to the lymph nodes requires the expression of specific chemokine receptors (CCR7) that recognize chemokine gradients guiding them to the lymphatic vessels. To understand the effects of environmentally-induced shape changes on DCs, we subjected skin DCs to various levels of physical confinement they can find in tissues and we unexpectedly uncovered a precise deformation amplitude at which DCs activate a shape-sensing mechanism that leads to an increased expression of the CCR7 chemokine receptor. This subsequently leads to the steady state migration of DCs from peripheral tissues to the lymph nodes and a global transcriptional reprogramming that is different from the one triggered upon microbial sensing. These results show that DCs have a shape-sensing mechanism that defines their migratory behaviour and immune phenotype in steady state conditions. Rather than sensing molecular biochemical signals, these cells might sense the physical constraints they encounter while patrolling their environment. These unexpected results highlight the importance of physical properties and their subsequent cues to tissue organization and function, and how these can shape immunity (published in 2024 DOI 10.1038/s41590-024-01856-3).

We also uncovered the molecular drivers of migration of cells that do not attach to surfaces, such as the two cell types we focus on in this project. These cells move through the creation of intracellular actomyosin zones of distinct rigidity: a soft front that deforms to squeeze through spaces, a rigid middle to hold its shape, and a rear that acts as the cell’s ‘muscle’ to push forward. This organization allows cells to generate force and move effectively. When physically confined, these cells can also create protrusions that eventually detach as motile cell fragments with capacity to migrate. This finding mirrors recent intravital microscopy studies in live tumors, where scientists observed cancer cells fragmenting in real time, with pieces breaking off and moving independently through tissues (published in 2024 DOI 10.1016/j.devcel.2024.06.023 ).

As stated above, we also set out to analyze the influence of physical constraints in breast cancer cells (BCCs). It has been observed that rounder BCCs are correlated with solid-like and tumor suppressive states. The transition to a malignant state is often accompanied by the acquisition of more fluid-like features required for cell proliferation, migration and dissemination, a process we call unjamming. Here we uncovered that the changes in cell density resulting from the unjamming phenomenon result in mechanical deformations of cells and nuclei that ultimately lead to an alteration of the cell states towards the emergence of malignant features that include epithelial-to-mesenchymal plasticity and chemoresistance (published in 2022 DOI 10.1038/s41563-022-01431-x).
SHAPINCELLFATE teams continue the project, aiming at understanding the mechanisms linked to cell shape and the memory-induced changes.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

Even though the project just arrived to the term of its first 18 months, we consider that some of the obtained results are significant scientific breakthroughs:

Alraies et al. reports an unexpected finding, since it was not our initial hypothesis that the deformation of the nuclei of immune cells, for a specific range of deformation, would be informative for these cells and trigger a very specific state, likely contributing to immune tolerance. It is also a breakthrough, since there was so far no known stimulus to trigger the migration of dendritic cells to lymph nodes in the absence of inflammation or infection.

Frittoli & Palamidessi et al. also represents a major advance in the contribution of tissue and cell mechanical cues on cellular phenotypes and states. Using various tumor modesl, we uncovered that changes in cell density resulting from the unjamming phenomenon result in repeated mechanical deformations of cells and nuclei (increased size and stiffness) that ultimately lead to a transcriptional rewiring altering cell states towards the emergence of malignant features including epithelial-to-mesenchymal plasticity and chemoresistance. These results highlight the importance of tissue mechanical properties triggering the acquisition of intracellular traits leading to changes in cell states.

Garcia et al. reports a very basic finding that clarifies how almost any cell can develop a fast mode of migration, by unleashing a generic property of acto-myosin. It is advancing the field of cell migration beyond the state of the art and represents mostly a conceptual advance, with the notion of advected percolation, which is new not only for biology but even for the physics of materials in general and could serve as a basic principle to produce moving objects.