Biologie cellulaire systémique de la polarité et de la division

Liste des publications

Année de publication : 2021

Guilherme Pedreira de Freitas Nader, Sonia Agüera-Gonzalez, Fiona Routet, Matthieu Gratia, Mathieu Maurin, Valeria Cancila, Clotilde Cadart, Andrea Palamidessi, Rodrigo Nalio Ramos, Mabel San Roman, Matteo Gentili, Ayako Yamada, Alice Williart, Catalina Lodillinsky, Emilie Lagoutte, Catherine Villard, Jean-Louis Viovy, Claudio Tripodo, Jérôme Galon, Giorgio Scita, Nicolas Manel, Philippe Chavrier, Matthieu Piel (2021 Sep 22)

Compromised nuclear envelope integrity drives TREX1-dependent DNA damage and tumor cell invasion.

Cell : DOI : S0092-8674(21)01046-1 En savoir plus
Résumé

Although mutations leading to a compromised nuclear envelope cause diseases such as muscular dystrophies or accelerated aging, the consequences of mechanically induced nuclear envelope ruptures are less known. Here, we show that nuclear envelope ruptures induce DNA damage that promotes senescence in non-transformed cells and induces an invasive phenotype in human breast cancer cells. We find that the endoplasmic reticulum (ER)-associated exonuclease TREX1 translocates into the nucleus after nuclear envelope rupture and is required to induce DNA damage. Inside the mammary duct, cellular crowding leads to nuclear envelope ruptures that generate TREX1-dependent DNA damage, thereby driving the progression of in situ carcinoma to the invasive stage. DNA damage and nuclear envelope rupture markers were also enriched at the invasive edge of human tumors. We propose that DNA damage in mechanically challenged nuclei could affect the pathophysiology of crowded tissues by modulating proliferation and extracellular matrix degradation of normal and transformed cells.

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Valentin Laplaud, Nicolas Levernier, Judith Pineau, Mabel San Roman, Lucie Barbier, Pablo J Sáez, Ana-Maria Lennon-Duménil, Pablo Vargas, Karsten Kruse, Olivia du Roure, Matthieu Piel, Julien Heuvingh (2021 Jul 3)

Pinching the cortex of live cells reveals thickness instabilities caused by myosin II motors.

Science advances : DOI : eabe3640 En savoir plus
Résumé

The cell cortex is a contractile actin meshwork, which determines cell shape and is essential for cell mechanics, migration, and division. Because its thickness is below optical resolution, there is a tendency to consider the cortex as a thin uniform two-dimensional layer. Using two mutually attracted magnetic beads, one inside the cell and the other in the extracellular medium, we pinch the cortex of dendritic cells and provide an accurate and time-resolved measure of its thickness. Our observations draw a new picture of the cell cortex as a highly dynamic layer, harboring large fluctuations in its third dimension because of actomyosin contractility. We propose that the cortex dynamics might be responsible for the fast shape-changing capacity of highly contractile cells that use amoeboid-like migration.

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Année de publication : 2020

A J Lomakin, C J Cattin, D Cuvelier, Z Alraies, M Molina, G P F Nader, N Srivastava, P J Sáez, J M Garcia-Arcos, I Y Zhitnyak, A Bhargava, M K Driscoll, E S Welf, R Fiolka, R J Petrie, N S De Silva, J M González-Granado, N Manel, A M Lennon-Duménil, D J Müller, M Piel (2020 Oct 16)

The nucleus acts as a ruler tailoring cell responses to spatial constraints.

Science (New York, N.Y.) : DOI : eaba2894 En savoir plus
Résumé

The microscopic environment inside a metazoan organism is highly crowded. Whether individual cells can tailor their behavior to the limited space remains unclear. In this study, we found that cells measure the degree of spatial confinement by using their largest and stiffest organelle, the nucleus. Cell confinement below a resting nucleus size deforms the nucleus, which expands and stretches its envelope. This activates signaling to the actomyosin cortex via nuclear envelope stretch-sensitive proteins, up-regulating cell contractility. We established that the tailored contractile response constitutes a nuclear ruler-based signaling pathway involved in migratory cell behaviors. Cells rely on the nuclear ruler to modulate the motive force that enables their passage through restrictive pores in complex three-dimensional environments, a process relevant to cancer cell invasion, immune responses, and embryonic development.

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Gururaj Rao Kidiyoor, Qingsen Li, Giulia Bastianello, Christopher Bruhn, Irene Giovannetti, Adhil Mohamood, Galina V Beznoussenko, Alexandre Mironov, Matthew Raab, Matthieu Piel, Umberto Restuccia, Vittoria Matafora, Angela Bachi, Sara Barozzi, Dario Parazzoli, Emanuela Frittoli, Andrea Palamidessi, Tito Panciera, Stefano Piccolo, Giorgio Scita, Paolo Maiuri, Kristina M Havas, Zhong-Wei Zhou, Amit Kumar, Jiri Bartek, Zhao-Qi Wang, Marco Foiani (2020 Sep 25)

ATR is essential for preservation of cell mechanics and nuclear integrity during interstitial migration.

Nature communications : 4828 : DOI : 10.1038/s41467-020-18580-9 En savoir plus
Résumé

ATR responds to mechanical stress at the nuclear envelope and mediates envelope-associated repair of aberrant topological DNA states. By combining microscopy, electron microscopic analysis, biophysical and in vivo models, we report that ATR-defective cells exhibit altered nuclear plasticity and YAP delocalization. When subjected to mechanical stress or undergoing interstitial migration, ATR-defective nuclei collapse accumulating nuclear envelope ruptures and perinuclear cGAS, which indicate loss of nuclear envelope integrity, and aberrant perinuclear chromatin status. ATR-defective cells also are defective in neuronal migration during development and in metastatic dissemination from circulating tumor cells. Our findings indicate that ATR ensures mechanical coupling of the cytoskeleton to the nuclear envelope and accompanying regulation of envelope-chromosome association. Thus the repertoire of ATR-regulated biological processes extends well beyond its canonical role in triggering biochemical implementation of the DNA damage response.

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Andrea Dimitracopoulos, Pragya Srivastava, Agathe Chaigne, Zaw Win, Roie Shlomovitz, Oscar M Lancaster, Maël Le Berre, Matthieu Piel, Kristian Franze, Guillaume Salbreux, Buzz Baum (2020 Aug 1)

Mechanochemical Crosstalk Produces Cell-Intrinsic Patterning of the Cortex to Orient the Mitotic Spindle.

Current biology : CB : DOI : S0960-9822(20)30984-2 En savoir plus
Résumé

Proliferating animal cells are able to orient their mitotic spindles along their interphase cell axis, setting up the axis of cell division, despite rounding up as they enter mitosis. This has previously been attributed to molecular memory and, more specifically, to the maintenance of adhesions and retraction fibers in mitosis [1-6], which are thought to act as local cues that pattern cortical Gαi, LGN, and nuclear mitotic apparatus protein (NuMA) [3, 7-18]. This cortical machinery then recruits and activates Dynein motors, which pull on astral microtubules to position the mitotic spindle. Here, we reveal a dynamic two-way crosstalk between the spindle and cortical motor complexes that depends on a Ran-guanosine triphosphate (GTP) signal [12], which is sufficient to drive continuous monopolar spindle motion independently of adhesive cues in flattened human cells in culture. Building on previous work [1, 12, 19-23], we implemented a physical model of the system that recapitulates the observed spindle-cortex interactions. Strikingly, when this model was used to study spindle dynamics in cells entering mitosis, the chromatin-based signal was found to preferentially clear force generators from the short cell axis, so that cortical motors pulling on astral microtubules align bipolar spindles with the interphase long cell axis, without requiring a fixed cue or a physical memory of interphase shape. Thus, our analysis shows that the ability of chromatin to pattern the cortex during the process of mitotic rounding is sufficient to translate interphase shape into a cortical pattern that can be read by the spindle, which then guides the axis of cell division.

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Martina Bonucci, Nicolas Kuperwasser, Serena Barbe, Vonda Koka, Delphine de Villeneuve, Chi Zhang, Nishit Srivastava, Xiaoying Jia, Matthew P Stokes, Frank Bienaimé, Virginie Verkarre, Jean Baptiste Lopez, Fanny Jaulin, Marco Pontoglio, Fabiola Terzi, Benedicte Delaval, Matthieu Piel, Mario Pende (2020 Jun 26)

mTOR and S6K1 drive polycystic kidney by the control of Afadin-dependent oriented cell division.

Nature communications : 3200 : DOI : 10.1038/s41467-020-16978-z En savoir plus
Résumé

mTOR activation is essential and sufficient to cause polycystic kidneys in Tuberous Sclerosis Complex (TSC) and other genetic disorders. In disease models, a sharp increase of proliferation and cyst formation correlates with a dramatic loss of oriented cell division (OCD). We find that OCD distortion is intrinsically due to S6 kinase 1 (S6K1) activation. The concomitant loss of S6K1 in Tsc1-mutant mice restores OCD but does not decrease hyperproliferation, leading to non-cystic harmonious hyper growth of kidneys. Mass spectrometry-based phosphoproteomics for S6K1 substrates revealed Afadin, a known component of cell-cell junctions required to couple intercellular adhesions and cortical cues to spindle orientation. Afadin is directly phosphorylated by S6K1 and abnormally decorates the apical surface of Tsc1-mutant cells with E-cadherin and α-catenin. Our data reveal that S6K1 hyperactivity alters centrosome positioning in mitotic cells, affecting oriented cell division and promoting kidney cysts in conditions of mTOR hyperactivity.

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Ido Lavi, Nicolas Meunier, Raphael Voituriez, Jaume Casademunt (2020 Mar 15)

Motility and morphodynamics of confined cells.

Physical review. E : 022404 : DOI : 10.1103/PhysRevE.101.022404 En savoir plus
Résumé

We introduce a minimal hydrodynamic model of polarization, migration, and deformation of a biological cell confined between two parallel surfaces. In our model, the cell is driven out of equilibrium by an active cytsokeleton force that acts on the membrane. The cell cytoplasm, described as a viscous droplet in the Darcy flow regime, contains a diffusive solute that actively transduces the applied cytoskeleton force. While fairly simple and analytically tractable, this quasi-two-dimensional model predicts a range of compelling dynamic behaviours. A linear stability analysis of the system reveals that solute activity first destabilizes a global polarization-translation mode, prompting cell motility through spontaneous symmetry breaking. At higher activity, the system crosses a series of Hopf bifurcations leading to coupled oscillations of droplet shape and solute concentration profiles. At the nonlinear level, we find traveling-wave solutions associated with unique polarized shapes that resemble experimental observations. Altogether, this model offers an analytical paradigm of active deformable systems in which viscous hydrodynamics are coupled to diffusive force transducers.

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Nishit Srivastava, David Traynor, Matthieu Piel, Alexandre J Kabla, Robert R Kay (2020 Jan 23)

Pressure sensing through Piezo channels controls whether cells migrate with blebs or pseudopods.

Proceedings of the National Academy of Sciences of the United States of America : DOI : 201905730 En savoir plus
Résumé

Blebs and pseudopods can both power cell migration, with blebs often favored in tissues, where cells encounter increased mechanical resistance. To investigate how migrating cells detect and respond to mechanical forces, we used a « cell squasher » to apply uniaxial pressure to cells chemotaxing under soft agarose. As little as 100 Pa causes a rapid (<10 s), sustained shift to movement with blebs rather than pseudopods. Cells are flattened under load and lose volume; the actin cytoskeleton is reorganized, with myosin II recruited to the cortex, which may pressurize the cytoplasm for blebbing. The transition to bleb-driven motility requires extracellular calcium and is accompanied by increased cytosolic calcium. It is largely abrogated in cells lacking the Piezo stretch-operated channel; under load, these cells persist in using pseudopods and chemotax poorly. We propose that migrating cells sense pressure through Piezo, which mediates calcium influx, directing movement with blebs instead of pseudopods.

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Année de publication : 2019

Ritsuya Niwayama, Prachiti Moghe, Yan-Jun Liu, Dimitri Fabrèges, Frank Buchholz, Matthieu Piel, Takashi Hiiragi (2019 Nov 19)

A Tug-of-War between Cell Shape and Polarity Controls Division Orientation to Ensure Robust Patterning in the Mouse Blastocyst.

Developmental cell : DOI : S1534-5807(19)30856-1 En savoir plus
Résumé

Oriented cell division patterns tissues by modulating cell position and fate. While cell geometry, junctions, cortical tension, and polarity are known to control division orientation, relatively little is known about how these are coordinated to ensure robust patterning. Here, we systematically characterize cell division, volume, and shape changes during mouse pre-implantation development by in toto live imaging. The analysis leads us to a model in which the apical domain competes with cell shape to determine division orientation. Two key predictions of the model are verified experimentally: when outside cells of the 16-cell embryo are released from cell shape asymmetry, the axis of division is guided by the apical domain. Conversely, orientation cues from the apical domain can be overcome by applied shape asymmetry in the 8-cell embryo. We propose that such interplay between cell shape and polarity in controlling division orientation ensures robust patterning of the blastocyst and possibly other tissues.

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Clotilde Cadart, Larisa Venkova, Pierre Recho, Marco Cosentino Lagomarsino, Matthieu Piel (2019 Aug 19)

The physics of cell-size regulation across timescales

Nature Physics : 15 : 993–1004 : DOI : 10.1038/s41567-019-0629-y En savoir plus
Résumé

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Lucie Barbier, Pablo J Sáez, Rafaele Attia, Ana-Maria Lennon-Duménil, Ido Lavi, Matthieu Piel, Pablo Vargas (2019 Apr 30)

Myosin II Activity Is Selectively Needed for Migration in Highly Confined Microenvironments in Mature Dendritic Cells.

Frontiers in immunology : 747 : DOI : 10.3389/fimmu.2019.00747 En savoir plus
Résumé

Upon infection, mature dendritic cells (mDCs) migrate from peripheral tissue to lymph nodes (LNs) to activate T lymphocytes and initiate the adaptive immune response. This fast and tightly regulated process is tuned by different microenvironmental factors, such as the physical properties of the tissue. Mechanistically, mDCs migration mostly relies on acto-myosin flow and contractility that depend on non-muscular Myosin IIA (MyoII) activity. However, the specific contribution of this molecular motor for mDCs navigation in complex microenvironments has yet to be fully established. Here, we identified a specific role of MyoII activity in the regulation of mDCs migration in highly confined microenvironments. Using microfluidic systems, we observed that during mDCs chemotaxis in 3D collagen gels under defined CCL21 gradients, MyoII activity was required to sustain their fast speed but not to orientate them toward the chemokine. Indeed, despite the fact that mDCs speed declined, these cells still migrated through the 3D gels, indicating that this molecular motor has a discrete function during their motility in this irregular microenvironment. Consistently, using microchannels of different sizes, we found that MyoII activity was essential to maintain fast cell speed specifically under strong confinement. Analysis of cell motility through micrometric holes further demonstrated that cell contractility facilitated mDCs passage only over very small gaps. Altogether, this work highlights that high contractility acts as an adaptation mechanism exhibited by mDCs to optimize their motility in restricted landscapes. Hence, MyoII activity ultimately facilitates their navigation in highly confined areas of structurally irregular tissues, contributing to the fine-tuning of their homing to LNs to initiate adaptive immune responses.

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Hélène D Moreau, Carles Blanch-Mercader, Rafaele Attia, Mathieu Maurin, Zahraa Alraies, Doriane Sanséau, Odile Malbec, Maria-Graciela Delgado, Philippe Bousso, Jean-François Joanny, Raphaël Voituriez, Matthieu Piel, Ana-Maria Lennon-Duménil (2019 Apr 16)

Macropinocytosis Overcomes Directional Bias in Dendritic Cells Due to Hydraulic Resistance and Facilitates Space Exploration.

Developmental cell : 171-188.e5 : DOI : S1534-5807(19)30235-7 En savoir plus
Résumé

The migration of immune cells can be guided by physical cues imposed by the environment, such as geometry, rigidity, or hydraulic resistance (HR). Neutrophils preferentially follow paths of least HR in vitro, a phenomenon known as barotaxis. The mechanisms and physiological relevance of barotaxis remain unclear. We show that barotaxis results from the amplification of a small force imbalance by the actomyosin cytoskeleton, resulting in biased directional choices. In immature dendritic cells (DCs), actomyosin is recruited to the cell front to build macropinosomes. These cells are therefore insensitive to HR, as macropinocytosis allows fluid transport across these cells. This may enhance their space exploration capacity in vivo. Conversely, mature DCs down-regulate macropinocytosis and are thus barotactic. Modeling suggests that HR may help guide these cells to lymph nodes where they initiate immune responses. Hence, DCs can either overcome or capitalize on the physical obstacles they encounter, helping their immune-surveillance function.

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Juan Manuel Garcia-Arcos, Renaud Chabrier, Mathieu Deygas, Guilherme Nader, Lucie Barbier, Pablo José Sáez, Aastha Mathur, Pablo Vargas, Matthieu Piel (2019 Feb 13)

Reconstitution of cell migration at a glance.

Journal of cell science : DOI : jcs225565 En savoir plus
Résumé

Single cells migrate in a myriad of physiological contexts, such as tissue patrolling by immune cells, and during neurogenesis and tissue remodeling, as well as in metastasis, the spread of cancer cells. To understand the basic principles of single-cell migration, a reductionist approach can be taken. This aims to control and deconstruct the complexity of different cellular microenvironments into simpler elementary constrains that can be recombined together. This approach is the cell microenvironment equivalent of reconstituted systems that combine elementary molecular players to understand cellular functions. In this Cell Science at a Glance article and accompanying poster, we present selected experimental setups that mimic different events that cells undergo during migration These include polydimethylsiloxane (PDMS) devices to deform whole cells or organelles, micro patterning, nano-fabricated structures like grooves, and compartmentalized collagen chambers with chemical gradients. We also outline the main contribution of each technique to the understanding of different aspects of single-cell migration.

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Maria Duda, Natalie J Kirkland, Nargess Khalilgharibi, Melda Tozluoglu, Alice C Yuen, Nicolas Carpi, Anna Bove, Matthieu Piel, Guillaume Charras, Buzz Baum, Yanlan Mao (2019 Jan 30)

Polarization of Myosin II Refines Tissue Material Properties to Buffer Mechanical Stress.

Developmental cell : 245-260.e7 : DOI : S1534-5807(18)31088-8 En savoir plus
Résumé

As tissues develop, they are subjected to a variety of mechanical forces. Some of these forces are instrumental in the development of tissues, while others can result in tissue damage. Despite our extensive understanding of force-guided morphogenesis, we have only a limited understanding of how tissues prevent further morphogenesis once the shape is determined after development. Here, through the development of a tissue-stretching device, we uncover a mechanosensitive pathway that regulates tissue responses to mechanical stress through the polarization of actomyosin across the tissue. We show that stretch induces the formation of linear multicellular actomyosin cables, which depend on Diaphanous for their nucleation. These stiffen the epithelium, limiting further changes in shape, and prevent fractures from propagating across the tissue. Overall, this mechanism of force-induced changes in tissue mechanical properties provides a general model of force buffering that serves to preserve the shape of tissues under conditions of mechanical stress.

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Année de publication : 2018

Grégory Beaune, Carles Blanch-Mercader, Stéphane Douezan, Julien Dumond, David Gonzalez-Rodriguez, Damien Cuvelier, Thierry Ondarçuhu, Pierre Sens, Sylvie Dufour, Michael P Murrell, Françoise Brochard-Wyart (2018 Dec 4)

Spontaneous migration of cellular aggregates from giant keratocytes to running spheroids.

Proceedings of the National Academy of Sciences of the United States of America : 12926-12931 : DOI : 10.1073/pnas.1811348115 En savoir plus
Résumé

Despite extensive knowledge on the mechanisms that drive single-cell migration, those governing the migration of cell clusters, as occurring during embryonic development and cancer metastasis, remain poorly understood. Here, we investigate the collective migration of cell on adhesive gels with variable rigidity, using 3D cellular aggregates as a model system. After initial adhesion to the substrate, aggregates spread by expanding outward a cell monolayer, whose dynamics is optimal in a narrow range of rigidities. Fast expansion gives rise to the accumulation of mechanical tension that leads to the rupture of cell-cell contacts and the nucleation of holes within the monolayer, which becomes unstable and undergoes dewetting like a liquid film. This leads to a symmetry breaking and causes the entire aggregate to move as a single entity. Varying the substrate rigidity modulates the extent of dewetting and induces different modes of aggregate motion: « giant keratocytes, » where the lamellipodium is a cell monolayer that expands at the front and retracts at the back; « penguins, » characterized by bipedal locomotion; and « running spheroids, » for nonspreading aggregates. We characterize these diverse modes of collective migration by quantifying the flows and forces that drive them, and we unveil the fundamental physical principles that govern these behaviors, which underscore the biological predisposition of living material to migrate, independent of length scale.

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