المؤلفون: Fabrizio A. Pennacchio, Alessandro Poli, Francesca Michela Pramotton, Stefania Lavore, Ilaria Rancati, Mario Cinquanta, Daan Vorselen, Elisabetta Prina, Orso Maria Romano, Aldo Ferrari, Matthieu Piel, Marco Cosentino Lagomarsino, Paolo Maiuri

المصدر: Nature Communications, Vol 15, Iss 1, Pp 1-10 (2024)

مصطلحات موضوعية: Science

الوصف: Abstract In eukaryotes, cytoplasmic and nuclear volumes are tightly regulated to ensure proper cell homeostasis. However, current methods to measure cytoplasmic and nuclear volumes, including confocal 3D reconstruction, have limitations, such as relying on two-dimensional projections or poor vertical resolution. Here, to overcome these limitations, we describe a method, N2FXm, to jointly measure cytoplasmic and nuclear volumes in single cultured adhering human cells, in real time, and across cell cycles. We find that this method accurately provides joint size over dynamic measurements and at different time resolutions. Moreover, by combining several experimental perturbations and analyzing a mathematical model including osmotic effects and tension, we show that N2FXm can give relevant insights on how mechanical forces exerted by the cytoskeleton on the nuclear envelope can affect the growth of nucleus volume by biasing nuclear import. Our method, by allowing for accurate joint nuclear and cytoplasmic volume dynamic measurements at different time resolutions, highlights the non-constancy of the nucleus/cytoplasm ratio along the cell cycle.

وصف الملف: electronic resource

Relation: https://doaj.org/toc/2041-1723

URL الوصول: https://doaj.org/article/28d7b0b6342e4134aecf5e7c110002be

المؤلفون: Sonal Joshi, Lucía López, Luciano Gastón Morosi, Roberto Amadio, Manendra Pachauri, Marco Bestagno, Ironya Paul Ogar, Mauro Giacca, Giulia Maria Piperno, Daan Vorselen, Federica Benvenuti

المصدر: Cell Reports, Vol 43, Iss 4, Pp 114096- (2024)

مصطلحات موضوعية: CP: Immunology, CP: Cancer, Biology (General), QH301-705.5

الوصف: Summary: Receptors controlling the cross-presentation of tumor antigens by macrophage subsets in cancer tissues are poorly explored. Here, we show that TIM4+ large peritoneal macrophages efficiently capture and cross-present tumor-associated antigens at early stages of peritoneal infiltration by ovarian cancer cells. The phosphatidylserine (PS) receptor TIM4 promotes maximal uptake of dead cells or PS-coated artificial targets and triggers inflammatory and metabolic gene programs in combination with cytoskeletal remodeling and upregulation of transcriptional signatures related to antigen processing. At the cellular level, TIM4-mediated engulfment induces nucleation of F-actin around nascent phagosomes, delaying the recruitment of vacuolar ATPase, acidification, and cargo degradation. In vivo, TIM4 deletion blunts induction of early anti-tumoral effector CD8 T cells and accelerates the progression of ovarian tumors. We conclude that TIM4-mediated uptake drives the formation of specialized phagosomes that prolong the integrity of ingested antigens and facilitate cross-presentation, contributing to immune surveillance of the peritoneum.

وصف الملف: electronic resource

Relation: http://www.sciencedirect.com/science/article/pii/S2211124724004248; https://doaj.org/toc/2211-1247

URL الوصول: https://doaj.org/article/c2dde8a380ab4786a23705a24951b4bc

المؤلفون: Daan Vorselen, Yifan Wang, Miguel M. de Jesus, Pavak K. Shah, Matthew J. Footer, Morgan Huse, Wei Cai, Julie A. Theriot

المصدر: Nature Communications, Vol 11, Iss 1, Pp 1-14 (2020)

مصطلحات موضوعية: Science

الوصف: Traction force microscopy is an effective method for measuring cellular forces but it is limited by planar geometry. Here the authors develop a facile method to produce deformable hydrogel particles and a reference-free computational method to resolve surface traction forces from particle shape deformation.

وصف الملف: electronic resource

Relation: https://doaj.org/toc/2041-1723

URL الوصول: https://doaj.org/article/fbc3f6afb17f46f49e6de46d02f86e78

المؤلفون: Daan Vorselen, Sarah R Barger, Yifan Wang, Wei Cai, Julie A Theriot, Nils C Gauthier, Mira Krendel

المصدر: eLife, Vol 10 (2021)

مصطلحات موضوعية: phagocytosis, cytoskeleton, actin, myosin, Medicine, Science, Biology (General), QH301-705.5

الوصف: Phagocytosis requires rapid actin reorganization and spatially controlled force generation to ingest targets ranging from pathogens to apoptotic cells. How actomyosin activity directs membrane extensions to engulf such diverse targets remains unclear. Here, we combine lattice light-sheet microscopy (LLSM) with microparticle traction force microscopy (MP-TFM) to quantify actin dynamics and subcellular forces during macrophage phagocytosis. We show that spatially localized forces leading to target constriction are prominent during phagocytosis of antibody-opsonized targets. This constriction is largely driven by Arp2/3-mediated assembly of discrete actin protrusions containing myosin 1e and 1f (‘teeth’) that appear to be interconnected in a ring-like organization. Contractile myosin-II activity contributes to late-stage phagocytic force generation and progression, supporting a specific role in phagocytic cup closure. Observations of partial target eating attempts and sudden target release via a popping mechanism suggest that constriction may be critical for resolving complex in vivo target encounters. Overall, our findings present a phagocytic cup shaping mechanism that is distinct from cytoskeletal remodeling in 2D cell motility and may contribute to mechanosensing and phagocytic plasticity.

وصف الملف: electronic resource

Relation: https://elifesciences.org/articles/68627; https://doaj.org/toc/2050-084X

URL الوصول: https://doaj.org/article/aef0f4ad09434fbab2364d8bb7e93e64

المؤلفون: Daan Vorselen, Susan M. van Dommelen, Raya Sorkin, Melissa C. Piontek, Jürgen Schiller, Sander T. Döpp, Sander A. A. Kooijmans, Brigitte A. van Oirschot, Birgitta A. Versluijs, Marc B. Bierings, Richard van Wijk, Raymond M. Schiffelers, Gijs J. L. Wuite, Wouter H. Roos

المصدر: Nature Communications, Vol 9, Iss 1, Pp 1-9 (2018)

مصطلحات موضوعية: Science

الوصف: Red blood cell disorders are often accompanied by increased release of extracellular vesicles (EVs), but their structural and mechanical properties are not fully understood. Here, the authors show that red blood cell EVs show liposome-like mechanical features and are softened in blood disorder patients.

وصف الملف: electronic resource

Relation: https://doaj.org/toc/2041-1723

URL الوصول: https://doaj.org/article/ee3d649d089e45a59a0d4b1fccb687f2

المؤلفون: Daan Vorselen, Melissa C. Piontek, Wouter H. Roos, Gijs J. L. Wuite

المصدر: Frontiers in Molecular Biosciences, Vol 7 (2020)

مصطلحات موضوعية: (small) vesicles, liposomes, extracellular vesicles, atomic force microscopy (AFM), nanoindentation, bending modulus, Biology (General), QH301-705.5

الوصف: Both natural as well as artificial vesicles are of tremendous interest in biology and nanomedicine. Small vesicles (

وصف الملف: electronic resource

Relation: https://www.frontiersin.org/article/10.3389/fmolb.2020.00139/full; https://doaj.org/toc/2296-889X

URL الوصول: https://doaj.org/article/2104cf73f43e49fe8f55914d1b407d9f

المؤلفون: Daan Vorselen

المصدر: Biochemical Society Transactions 50 (2022) 5
Biochemical Society Transactions, 50(5), 1281-1291

مصطلحات موضوعية: Cell Biology and Immunology, Phagocytosis, Life Science, Humans, Celbiologie en Immunologie, Neurodegenerative Diseases, Apoptosis, Phosphatidylserines, Biochemistry, Signal Transduction

الوصف: Phagocytosis triggered by the phospholipid phosphatidylserine (PS) is key for the removal of apoptotic cells in development, tissue homeostasis and infection. Modulation of PS-mediated phagocytosis is an attractive target for therapeutic intervention in the context of atherosclerosis, neurodegenerative disease, and cancer. Whereas the mechanisms of target recognition, lipid and protein signalling, and cytoskeletal remodelling in opsonin-driven modes of phagocytosis are increasingly well understood, PS-mediated phagocytosis has remained more elusive. This is partially due to the involvement of a multitude of receptors with at least some redundancy in functioning, which complicates dissecting their contributions and results in complex downstream signalling networks. This review focusses on the receptors involved in PS-recognition, the signalling cascades that connect receptors to cytoskeletal remodelling required for phagocytosis, and recent progress in our understanding of how phagocytic cup formation is coordinated during PS-mediated phagocytosis.

وصف الملف: application/pdf

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3cd042041d1d0a3d2898d48083ef16c1
https://pubmed.ncbi.nlm.nih.gov/36281986

المؤلفون: Daan Vorselen, Roarke A. Kamber, Ramon Lorenzo D. Labitigan, Aaron P. van Loon, Eric Peterman, Melissa K. Delgado, Sijie Lin, Jeffrey P. Rasmussen, Michael C. Bassik, Julie A. Theriot

الوصف: SummaryMacrophages phagocytose and thereby eliminate a wide array of extracellular threats, ranging from antibody-coated bacteria to apoptotic cells. Precision modulation of phagocytosis has emerged as a therapeutic strategy across a range of diseases, but is limited by our incomplete understanding of how macrophages recognize, engulf, and respond to different phagocytic targets. Here, we undertook a systematic investigation of the morphological, biophysical and regulatory differences between two major types of phagocytosis: an immunostimulatory form of phagocytosis triggered by antibody-coated targets and an immunosuppressive form triggered by phosphatidylserine (PS)-coated targets. We confirmed classic observations that antibody-mediated phagocytosis involves the extension of thin actin-rich protrusions around the target, but find that PS-mediated phagocytosis involves an unexpected combination of filopodial probing, piecemeal phagocytosis and a distinct ‘sinking’ mechanism of uptake. Using a genome-wide screening approach, we identified genes specifically required for each form of phagocytosis, including actin regulators, cell surface receptors and intracellular signaling molecules. Three cell surface receptors - TREM2, CD14 and integrin αMβ2- were revealed as essential for PS-mediated uptake. Strikingly, each receptor exhibited a distinct pattern of localization at the plasma membrane and contributed uniquely to the organization of the PS-dependent phagocytic cup. Overall, this work reveals divergent genetic requirements for the morphologically and mechanically distinct forms of PS-mediated and antibody-mediated phagocytosis, thereby informing therapeutic strategies for substrate-specific phagocytosis modulation.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::6bcb06001cc69ece8469904ef0d0fbec
https://doi.org/10.1101/2022.07.30.502145

المؤلفون: Fabrizio A. Pennacchio, Alessandro Poli, Francesca Michela Pramotton, Stefania Lavore, Ilaria Rancati, Mario Cinquanta, Daan Vorselen, Elisabetta Prina, Orso Maria Romano, Aldo Ferrari, Matthieu Piel, Marco Cosentino Lagomarsino, Paolo Maiuri

الوصف: In eukaryotes, cytoplasmic and nuclear volumes are tightly regulated to ensure proper cell homeostasis. However, the detailed mechanisms underlying nucleus-cytoplasm volumetric coupling remain unknown. Recent evidence supports a primary role of osmotic mechanisms in determining a tight link between nuclear and cytoplasmic volume, but this hypothesis remains largely untested in mammalian cells. We approach the question in single cultured adhering human cells, by jointly measuring cytoplasmic and nuclear volumes, in real time and across cell cycles. Surprisingly, we find that cytoplasmic and nuclear volumes follow different average growth laws: while the cytoplasm grows exponentially, the nucleus grows linearly. Moreover, by combining several experimental perturbations and analyzing a mathematical model including osmotic effects and tension, we conclude that the mechanical forces exerted by the cytoskeleton on the nuclear envelope can strongly affect nucleus-cytoplasm volumetric coupling by biasing nuclear import. Our results unveil how osmo-mechanical equilibrium regulates nuclear size in mammalian cells.One-Sentence SummaryCytoskeletal forces exerted on the nuclear envelope impact on nuclear volume through modulation of force-coupled nucleo-cytoplasmic transport, affecting osmosis.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::addfb27103a1431dd1a734cc0244d17b
https://doi.org/10.1101/2022.06.07.494975

المؤلفون: Miguel de Jesus, Alexander Settle, Daan Vorselen, Michael Galiano, Endi Santosa, Taha Merghoub, Joseph Sun, Pavak Shah, Julie Theriot, Morgan Huse

المصدر: Biophysical Journal. 122:534a

مصطلحات موضوعية: Biophysics

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::f35a7111c0c5e00ad968e2904206a4b9
https://doi.org/10.1016/j.bpj.2022.11.2831