bims-ecemfi 2024-06-09 papers

bims-ecemfi

Biomed News

on ECM and fibroblasts

Issue of 2024‒06‒09
twenty papers selected by
Badri Narayanan Narasimhan, University of California, San Diego

Rationally designed β-cyclodextrin-crosslinked polyacrylamide hydrogels for cell spheroid formation and 3D tumor model construction.
Assessing the impact of extracellular matrix fiber orientation on breast cancer cellular metabolism.
How cytoskeletal crosstalk makes cells move: Bridging cell-free and cell studies.
Elucidating the interaction between stretch and stiffness using an agent-based spring network model of progressive pulmonary fibrosis.
Cell-size-dependent regulation of Ezrin dictates epithelial resilience to stretch by countering myosin-II-mediated contractility.
Rear cortex contraction aids in nuclear transit during confined migration by increasing pressure in the cell posterior.
Remodeling of tumor microenvironment by extracellular matrix protein 1a differentially regulates ovarian cancer metastasis.
Photoactivatable substrates show diverse phenotypes of leader cells in collective migration when moving along different extracellular matrix proteins.
Macrophages enhance contractile force in iPSC-derived human engineered cardiac tissue.
Photoacoustic Processing of Decellularized Extracellular Matrix for Biofabricating Living Constructs.
Multiplex, high-throughput method to study cancer and immune cell mechanotransduction.
Cellular memory function from 3D to 2D: Three-dimensional high density collagen microfiber cultures induce their resistance to reactive oxygen species.
Agent-based model demonstrates the impact of nonlinear, complex interactions between cytokinces on muscle regeneration.
Spatiotemporal heterogeneity of LMOD1 expression summarizes two modes of cell communication in colorectal cancer.
Vimentin supports cell polarization by enhancing centrosome function and microtubule acetylation.
Mapping the unicellular transcriptome of the ascending thoracic aorta to changes in mechanosensing and mechanoadaptation during aging.
FAK regulates tension transmission to the nucleus and endothelial transcriptome independent of kinase activity.
Viscoelasticity of diverse biological samples quantified by Acoustic Force Microrheology (AFMR).
A YAP-centered mechanotransduction loop drives collective breast cancer cell invasion.
Beyond stiffness: multiscale viscoelastic features as biomechanical markers for assessing cell types and states.

Carbohydr Polym. 2024 Sep 01. pii: S0144-8617(24)00479-X. [Epub ahead of print]339 122253

Rationally designed β-cyclodextrin-crosslinked polyacrylamide hydrogels for cell spheroid formation and 3D tumor model construction.

Taili Chen, Yuting Wen, Xia Song, Zhongxing Zhang, Jingling Zhu, Xuehao Tian, Shan Zeng, Jun Li.

  In vitro tumor models are essential for understanding tumor behavior and evaluating tumor biological properties. Hydrogels that can mimic the tumor extracellular matrix have become popular for creating 3D in vitro tumor models. However, designing biocompatible hydrogels with appropriate chemical and physical properties for constructing tumor models is still a challenge. In this study, we synthesized a series of β-cyclodextrin (β-CD)-crosslinked polyacrylamide hydrogels with different β-CD densities and mechanical properties and evaluated their potential for use in 3D in vitro tumor model construction, including cell capture and spheroid formation. By utilizing a combination of β-CD-methacrylate (CD-MA) and a small amount of N,N'-methylene bisacrylamide (BIS) as hydrogel crosslinkers and optimizing the CD-MA/BIS ratio, the hydrogels performed excellently for tumor cell 3D culture and spheroid formation. Notably, when we co-cultured L929 fibroblasts with HeLa tumor cells on the hydrogel surface, co-cultured spheroids were formed, showing that the hydrogel can mimic the complexity of the tumor extracellular matrix. This comprehensive investigation of the relationship between hydrogel mechanical properties and biocompatibility provides important insights for hydrogel-based in vitro tumor modeling and advances our understanding of the mechanisms underlying tumor growth and progression.

Keywords:  3D tumor model; Cell spheroid; HeLa cell; Hydrogel; Polyacrylamide; β-Cyclodextrin

DOI:  https://doi.org/10.1016/j.carbpol.2024.122253
Cancer Cell Int. 2024 Jun 05. 24(1): 199

Assessing the impact of extracellular matrix fiber orientation on breast cancer cellular metabolism.

Madison R Pickett, Yuan-I Chen, Mohini Kamra, Sachin Kumar, Nikhith Kalkunte, Gabriella P Sugerman, Kelsey Varodom, Manuel K Rausch, Janet Zoldan, Hsin-Chin Yeh, Sapun H Parekh.

  The extracellular matrix (ECM) is a dynamic and complex microenvironment that modulates cell behavior and cell fate. Changes in ECM composition and architecture have been correlated with development, differentiation, and disease progression in various pathologies, including breast cancer [1]. Studies have shown that aligned fibers drive a pro-metastatic microenvironment, promoting the transformation of mammary epithelial cells into invasive ductal carcinoma via the epithelial-to-mesenchymal transition (EMT) [2]. The impact of ECM orientation on breast cancer metabolism, however, is largely unknown. Here, we employ two non-invasive imaging techniques, fluorescence-lifetime imaging microscopy (FLIM) and intensity-based multiphoton microscopy, to assess the metabolic states of cancer cells cultured on ECM-mimicking nanofibers in a random and aligned orientation. By tracking the changes in the intrinsic fluorescence of nicotinamide adenine dinucleotide and flavin adenine dinucleotide, as well as expression levels of metastatic markers, we reveal how ECM fiber orientation alters cancer metabolism and EMT progression. Our study indicates that aligned cellular microenvironments play a key role in promoting metastatic phenotypes of breast cancer as evidenced by a more glycolytic metabolic signature on nanofiber scaffolds of aligned orientation compared to scaffolds of random orientation. This finding is particularly relevant for subsets of breast cancer marked by high levels of collagen remodeling (e.g. pregnancy associated breast cancer), and may serve as a platform for predicting clinical outcomes within these subsets [3-6].

Keywords:  Breast cancer microenvironment; Cellular metabolism; Epithelial-to-mesenchymal transition (EMT); Extracellular matrix (ECM); Fluorescence lifetime imaging microscopy (FLIM); Optical redox ratio (ORR)

DOI:  https://doi.org/10.1186/s12935-024-03385-3
Biophys Rev (Melville). 2024 Jun;5(2): 021307

How cytoskeletal crosstalk makes cells move: Bridging cell-free and cell studies.

James P Conboy, Irene Istúriz Petitjean, Anouk van der Net, Gijsje H Koenderink.

Cell migration is a fundamental process for life and is highly dependent on the dynamical and mechanical properties of the cytoskeleton. Intensive physical and biochemical crosstalk among actin, microtubules, and intermediate filaments ensures their coordination to facilitate and enable migration. In this review, we discuss the different mechanical aspects that govern cell migration and provide, for each mechanical aspect, a novel perspective by juxtaposing two complementary approaches to the biophysical study of cytoskeletal crosstalk: live-cell studies (often referred to as top-down studies) and cell-free studies (often referred to as bottom-up studies). We summarize the main findings from both experimental approaches, and we provide our perspective on bridging the two perspectives to address the open questions of how cytoskeletal crosstalk governs cell migration and makes cells move.

DOI: https://doi.org/10.1063/5.0198119
Front Netw Physiol. 2024 ;4 1396383

Elucidating the interaction between stretch and stiffness using an agent-based spring network model of progressive pulmonary fibrosis.

Joseph K Hall, Jason H T Bates, Ramaswamy Krishnan, Jae Hun Kim, Yuqing Deng, Kenneth R Lutchen, Béla Suki.

  Pulmonary fibrosis is a deadly disease that involves the dysregulation of fibroblasts and myofibroblasts, which are mechanosensitive. Previous computational models have succeeded in modeling stiffness-mediated fibroblasts behaviors; however, these models have neglected to consider stretch-mediated behaviors, especially stretch-sensitive channels and the stretch-mediated release of latent TGF-β. Here, we develop and explore an agent-based model and spring network model hybrid that is capable of recapitulating both stiffness and stretch. Using the model, we evaluate the role of mechanical signaling in homeostasis and disease progression during self-healing and fibrosis, respectively. We develop the model such that there is a fibrotic threshold near which the network tends towards instability and fibrosis or below which the network tends to heal. The healing response is due to the stretch signal, whereas the fibrotic response occurs when the stiffness signal overpowers the stretch signal, creating a positive feedback loop. We also find that by changing the proportional weights of the stretch and stiffness signals, we observe heterogeneity in pathological network structure similar to that seen in human IPF tissue. The system also shows emergent behavior and bifurcations: whether the network will heal or turn fibrotic depends on the initial network organization of the damage, clearly demonstrating structure's pivotal role in healing or fibrosis of the overall network. In summary, these results strongly suggest that the mechanical signaling present in the lungs combined with network effects contribute to both homeostasis and disease progression.

Keywords:  bifurcation (mathematics); extracellar matrix; fibroblasts; mechanosensitive; mechanotransduction; network effects

DOI:  https://doi.org/10.3389/fnetp.2024.1396383
Cell Rep. 2024 May 31. pii: S2211-1247(24)00599-0. [Epub ahead of print]43(6): 114271

Cell-size-dependent regulation of Ezrin dictates epithelial resilience to stretch by countering myosin-II-mediated contractility.

Geetika Chouhan, Natasha Steffi Lewis, Vallari Ghanekar, Sri Rama Koti Ainavarapu, Mandar M Inamdar, Mahendra Sonawane.

  The epithelial adaptations to mechanical stress are facilitated by molecular and tissue-scale changes that include the strengthening of junctions, cytoskeletal reorganization, and cell-proliferation-mediated changes in tissue rheology. However, the role of cell size in controlling these properties remains underexplored. Our experiments in the zebrafish embryonic epidermis, guided by theoretical estimations, reveal a link between epithelial mechanics and cell size, demonstrating that an increase in cell size compromises the tissue fracture strength and compliance. We show that an increase in E-cadherin levels in the proliferation-deficient epidermis restores epidermal compliance but not the fracture strength, which is largely regulated by Ezrin-an apical membrane-cytoskeleton crosslinker. We show that Ezrin fortifies the epithelium in a cell-size-dependent manner by countering non-muscle myosin-II-mediated contractility. This work uncovers the importance of cell size maintenance in regulating the mechanical properties of the epithelium and fostering protection against future mechanical stresses.

Keywords:  2D vertex model; CP: Cell biology; CP: Developmental biology; Ezrin; cell adhesion; cell proliferation; cell size; epithelial resilience; epithelium; myosin contractility

DOI:  https://doi.org/10.1016/j.celrep.2024.114271
J Cell Sci. 2024 Jun 04. pii: jcs.260623. [Epub ahead of print]

Rear cortex contraction aids in nuclear transit during confined migration by increasing pressure in the cell posterior.

Jeremy Keys, Brian C H Cheung, Margaret A Elpers, Mingming Wu, Jan Lammerding.

  As cells migrate through biological tissues, they must frequently squeeze through micron-sized constrictions in the form of interstitial pores between extracellular matrix fibers and/or other cells. Although it is now well recognized that such confined migration is limited by the nucleus, which is the largest and stiffest organelle, it remains incompletely understood how cells apply sufficient force to move their nucleus through small constrictions. Here, we report a mechanism by which contraction of the cell rear cortex pushes the nucleus forward to mediate nuclear transit through constrictions. Laser ablation of the rear cortex reveals that pushing forces behind the nucleus are the result of increased intracellular pressure in the rear compartment of the cell. The pushing forces behind the nucleus depend on accumulation of actomyosin in the rear cortex and require Rho-kinase (ROCK) activity. Collectively, our results suggest a mechanism by which cells generate elevated, intracellular pressure in the posterior compartment to facilitate nuclear transit through 3D constrictions. This mechanism may supplement or even substitute for other mechanisms supporting nuclear transit, ensuring robust cell migrations in confined 3D environments.

Keywords:  Confined Migration; Cortex; Invasion; Mechanobiology; Nucleus; Pressure

DOI:  https://doi.org/10.1242/jcs.260623
Cancer Lett. 2024 Jun 05. pii: S0304-3835(24)00416-6. [Epub ahead of print] 217022

Remodeling of tumor microenvironment by extracellular matrix protein 1a differentially regulates ovarian cancer metastasis.

Yinjue Yu, Cuiting Lyu, Xiaojing Li, Lina Yang, Jingshu Wang, Hui Li, Zhaochen Xin, Xinyi Xu, Chunxia Ren, Gong Yang.

  We previously reported that extracellular matrix protein 1 isoform a (ECM1a) promotes epithelial ovarian cancer (EOC) through autocrine signaling through binding to cell surface receptors αXβ2. However, the role of ECM1a as a secretory molecule in the tumor microenvironment is rarely reported. In this study, we constructed murine Ecm1-knockout mice and human ECM1a-knockin mice and further generated orthotopic or peritoneal xenograft tumor models to mimic the different metastatic stages of EOC. We show that ECM1a induces oncogenic metastasis of orthotopic xenograft tumors, but inhibits early-metastasis of peritoneal xenograft tumors. ECM1a remodels extracellular matrices (ECM) and promotes remote metastases by recruiting and transforming bone marrow mesenchymal stem cells (BMSCs) into platelet-derived growth factor receptor beta (PDGFRβ+) cancer-associated fibroblasts (CAFs) and facilitating the secretion of angiopoietin-like protein 2 (ANGPTL2). Competing with ECM1a, ANGPTL2 also binds to integrin αX through the P1/P2 peptides, resulting in negative effects on BMSC differentiation. Collectively, this study reveals the dual functions of ECM1a in remodeling of TME during tumor progression, emphasizing the complexity of EOC phenotypic heterogeneity and metastasis.

Keywords:  Angiopoietin-like protein 2 (ANGPTL2); Cancer associated fibroblasts (CAFs); Epithelial ovarian cancer (EOC); Extracellular matrix protein 1 (ECM1); Extracellular matrix remodeling (ECM remodeling)

DOI:  https://doi.org/10.1016/j.canlet.2024.217022
Biomater Sci. 2024 Jun 04.

Photoactivatable substrates show diverse phenotypes of leader cells in collective migration when moving along different extracellular matrix proteins.

Shimaa A Abdellatef, Francesca Bard, Jun Nakanishi.

In cancer metastasis, collectively migrating clusters are discriminated into leader and follower cells that move through extracellular matrices (ECMs) with different characteristics. The impact of changes in ECM protein types on leader cells and migrating clusters is unknown. To address this, we investigated the response of leader cells and migrating clusters upon moving from one ECM protein to another using a photoactivatable substrate bearing photocleavable PEG (PCP), whose surface changes from protein-repellent to protein-adhesive in response to light. We chose laminin and collagen I for our study since they are abundant in two distinct regions in living tissues, namely basement membrane and connective tissue. Using the photoactivatable substrates, the precise deposition of the first ECM protein in the irradiated areas was achieved, followed by creating well-defined cellular confinements. Secondary irradiation enabled the deposition of the second ECM protein in the new irradiated regions, resulting in region-selective heterogeneous and homogenous ECM protein-coated surfaces. Different tendencies in leader cell formation from laminin into laminin compared to those migrating from laminin into collagen were observed. The formation of focal adhesion and actin structures for cells within the same cluster in the ECM proteins responded according to the underlying ECM protein type. Finally, integrin β1 was crucial for the appearance of leader cells for clusters migrating from laminin into collagen. However, when it came to laminin into laminin, integrin β1 was not responsible. This highlights the correlation between leader cells in collective migration and the biochemical signals that arise from underlying extracellular matrix proteins.

DOI: https://doi.org/10.1039/d4bm00225c
Cell Rep. 2024 May 31. pii: S2211-1247(24)00630-2. [Epub ahead of print]43(6): 114302

Macrophages enhance contractile force in iPSC-derived human engineered cardiac tissue.

Roberta I Lock, Pamela L Graney, Daniel Naveed Tavakol, Trevor R Nash, Youngbin Kim, Eloy Sanchez, Margaretha Morsink, Derek Ning, Connie Chen, Sharon Fleischer, Ilaria Baldassarri, Gordana Vunjak-Novakovic.

  Resident cardiac macrophages are critical mediators of cardiac function. Despite their known importance to cardiac electrophysiology and tissue maintenance, there are currently no stem-cell-derived models of human engineered cardiac tissues (hECTs) that include resident macrophages. In this study, we made an induced pluripotent stem cell (iPSC)-derived hECT model with a resident population of macrophages (iM0) to better recapitulate the native myocardium and characterized their impact on tissue function. Macrophage retention within the hECTs was confirmed via immunofluorescence after 28 days of cultivation. The inclusion of iM0s significantly impacted hECT function, increasing contractile force production. A potential mechanism underlying these changes was revealed by the interrogation of calcium signaling, which demonstrated the modulation of β-adrenergic signaling in +iM0 hECTs. Collectively, these findings demonstrate that macrophages significantly enhance cardiac function in iPSC-derived hECT models, emphasizing the need to further explore their contributions not only in healthy hECT models but also in the contexts of disease and injury.

Keywords:  CP: Developmental biology; cardiac; iPSC; immune; macrophage; myocardium; stem cell; tissue engineering

DOI:  https://doi.org/10.1016/j.celrep.2024.114302
Acta Biomater. 2024 Jun 03. pii: S1742-7061(24)00304-0. [Epub ahead of print]

Photoacoustic Processing of Decellularized Extracellular Matrix for Biofabricating Living Constructs.

Luís P Ferreira, Carole Jorge, Matilde R Lagarto, Maria V Monteiro, Iola F Duarte, Vítor M Gaspar, João F Mano.

  The diverse biomolecular landscape of tissue-specific decellularized extracellular matrix (dECM) biomaterials provides a multiplicity of bioinstructive cues to target cells, rendering them highly valuable for various biomedical applications. However, the isolation of dECM biomaterials entails cumbersome xenogeneic enzymatic digestions and also additional inactivation procedures. Such, increases processing time, increments costs and introduces residues of non-naturally present proteins in dECM formulations that remain present even after inactivation. To overcome these limitations, herein we report an innovative conjugation of light and ultrasound-mediated dECM biomaterial processing for fabricating dECM biomaterials. Such approach gathers on ultrasound waves to facilitate dECM-in-liquid processing and visible light photocrosslinking of tyrosine residues naturally present in dECM biomaterials. This dual step methodology unlocked the in-air production of cell laden dECM hydrogels or programmable dECM hydrogel spherical-like beads by using superhydrophobic surfaces. These in-air produced units do not require any additional solvents and successfully supported both fibroblasts and breast cancer cells viability upon encapsulation or surface seeding. In addition, the optimized photoacoustic methodology also enabled a rapid formulation of dECM biomaterial inks with suitable features for biofabricating volumetrically defined living constructs through embedded 3D bioprinting. The biofabricated SdECM hydrogel constructs supported cell adhesion, spreading and viability for 7 days. Overall, the implemented photoacoustic processing methodology of dECM biomaterials offers a rapid and universal strategy for upgrading the processing of dECM biomaterials from virtually any tissue. STATEMENT OF SIGNIFICANCE: Leveraging decellularized matrix biomaterials as cell instructive has potential to open new avenues for tissue engineering and in vitro disease modelling. The processing of dECM remains however, lengthy, costly and introduces non-naturally present proteins in the final biomaterials formulations. In this regard, here we report an innovative light and ultrasound mediated dECM hydrogel crosslinking methodology that enables rapid dECM-in-liquid processing and downstream photocrosslinking of hydrogel beads and 3D bioprinted constructs. Such acoustic-light based processing constitutes a universally applicable method for processing any type of tissue-derived dECM biomaterials.

Keywords:  3D Bioprinting; Decellularized Extracellular Matrix; In Vitro Models; Photocrosslinking; Ultrasound

DOI:  https://doi.org/10.1016/j.actbio.2024.05.054
Commun Biol. 2024 Jun 01. 7(1): 674

Multiplex, high-throughput method to study cancer and immune cell mechanotransduction.

Abigail R Fabiano, Spencer C Robbins, Samantha V Knoblauch, Schyler J Rowland, Jenna A Dombroski, Michael R King.

Studying cellular mechanoresponses during cancer metastasis is limited by sample variation or complex protocols that current techniques require. Metastasis is governed by mechanotransduction, whereby cells translate external stimuli, such as circulatory fluid shear stress (FSS), into biochemical cues. We present high-throughput, semi-automated methods to expose cells to FSS using the VIAFLO96 multichannel pipetting device custom-fitted with 22 G needles, increasing the maximum FSS 94-fold from the unmodified tips. Specifically, we develop protocols to semi-automatically stain live samples and to fix, permeabilize, and intracellularly process cells for flow cytometry analysis. Our first model system confirmed that the pro-apoptotic effects of TRAIL therapeutics in prostate cancer cells can be enhanced via FSS-induced Piezo1 activation. Our second system implements this multiplex methodology to show that FSS exposure (290 dyn cm-2) increases activation of murine bone marrow-derived dendritic cells. These methodologies greatly improve the mechanobiology workflow, offering a high-throughput, multiplex approach.

DOI: https://doi.org/10.1038/s42003-024-06327-x
Mater Today Bio. 2024 Jun;26 101097

Cellular memory function from 3D to 2D: Three-dimensional high density collagen microfiber cultures induce their resistance to reactive oxygen species.

Asuka Yamada, Shiro Kitano, Michiya Matsusaki.

  Cell properties generally change when the culture condition is changed. However, mesenchymal stem cells cultured on a hard material surface maintain their differentiation characteristics even after being cultured on a soft material surface. This phenomenon suggests the possibility of a cell culture material to memorize stem cell function even in changing cell culture conditions. However, there are no reports about cell memory function in three-dimensional (3D) culture. In this study, colon cancer cells were cultured with collagen microfibers (CMF) in 3D to evaluate their resistance to reactive oxygen species (ROS) in comparison with a monolayer (2D) culture condition and to understand the effect of 3D-culture on cell memory function. The ratio of ROS-negative cancer cells in 3D culture increased with increasing amounts of CMF and the highest amount of CMF was revealed to be 35-fold higher than that of the 2D condition. The ROS-negative cells ratio was maintained for 7 days after re-seeding in a 2D culture condition, suggesting a 3D-memory function of ROS resistance. The findings of this study will open up new opportunities for 3D culture to induce cell memory function.

Keywords:  3D culture; Cancer tissue; Cellular memory function; Collagen microfibers; Reactive oxygen species

DOI:  https://doi.org/10.1016/j.mtbio.2024.101097
Elife. 2024 Jun 03. pii: RP91924. [Epub ahead of print]13

Agent-based model demonstrates the impact of nonlinear, complex interactions between cytokinces on muscle regeneration.

Megan Haase, Tien Comlekoglu, Alexa Petrucciani, Shayn M Peirce, Silvia S Blemker.

  Muscle regeneration is a complex process due to dynamic and multiscale biochemical and cellular interactions, making it difficult to identify microenvironmental conditions that are beneficial to muscle recovery from injury using experimental approaches alone. To understand the degree to which individual cellular behaviors impact endogenous mechanisms of muscle recovery, we developed an agent-based model (ABM) using the Cellular-Potts framework to simulate the dynamic microenvironment of a cross-section of murine skeletal muscle tissue. We referenced more than 100 published studies to define over 100 parameters and rules that dictate the behavior of muscle fibers, satellite stem cells (SSCs), fibroblasts, neutrophils, macrophages, microvessels, and lymphatic vessels, as well as their interactions with each other and the microenvironment. We utilized parameter density estimation to calibrate the model to temporal biological datasets describing cross-sectional area (CSA) recovery, SSC, and fibroblast cell counts at multiple timepoints following injury. The calibrated model was validated by comparison of other model outputs (macrophage, neutrophil, and capillaries counts) to experimental observations. Predictions for eight model perturbations that varied cell or cytokine input conditions were compared to published experimental studies to validate model predictive capabilities. We used Latin hypercube sampling and partial rank correlation coefficient to identify in silico perturbations of cytokine diffusion coefficients and decay rates to enhance CSA recovery. This analysis suggests that combined alterations of specific cytokine decay and diffusion parameters result in greater fibroblast and SSC proliferation compared to individual perturbations with a 13% increase in CSA recovery compared to unaltered regeneration at 28 days. These results enable guided development of therapeutic strategies that similarly alter muscle physiology (i.e. converting extracellular matrix [ECM]-bound cytokines into freely diffusible forms as studied in cancer therapeutics or delivery of exogenous cytokines) during regeneration to enhance muscle recovery after injury.

Keywords:  agent-based model; cell biology; computational biology; cytokine dynamics; mouse; muscle regeneration; skeletal muscle; systems biology

DOI:  https://doi.org/10.7554/eLife.91924
J Transl Med. 2024 Jun 07. 22(1): 549

Spatiotemporal heterogeneity of LMOD1 expression summarizes two modes of cell communication in colorectal cancer.

Jie-Pin Li, Yuan-Jie Liu, Yang Li, Yi Yin, Qian-Wen Ye, Zhi-Hua Lu, Yu-Wei Dong, Jin-Yong Zhou, Xi Zou, Yu-Gen Chen.

  Cellular communication (CC) influences tumor development by mediating intercellular junctions between cells. However, the role and underlying mechanisms of CC in malignant transformation remain unknown. Here, we investigated the spatiotemporal heterogeneity of CC molecular expression during malignant transformation. It was found that although both tight junctions (TJs) and gap junctions (GJs) were involved in maintaining the tumor microenvironment (TME), they exhibited opposite characteristics. Mechanistically, for epithelial cells (parenchymal component), the expression of TJ molecules consistently decreased during normal-cancer transformation and is a potential oncogenic factor. For fibroblasts (mesenchymal component), the expression of GJs consistently increased during normal-cancer transformation and is a potential oncogenic factor. In addition, the molecular profiles of TJs and GJs were used to stratify colorectal cancer (CRC) patients, where subtypes characterized by high GJ levels and low TJ levels exhibited enhanced mesenchymal signals. Importantly, we propose that leiomodin 1 (LMOD1) is biphasic, with features of both TJs and GJs. LMOD1 not only promotes the activation of cancer-associated fibroblasts (CAFs) but also inhibits the Epithelial-mesenchymal transition (EMT) program in cancer cells. In conclusion, these findings demonstrate the molecular heterogeneity of CC and provide new insights into further understanding of TME heterogeneity.

Keywords:  Colorectal cancer; Epithelial cells; Fibroblasts; Gap junctions; LMOD1; Tight junctions

DOI:  https://doi.org/10.1186/s12967-024-05369-3
J R Soc Interface. 2024 Jun;21(215): 20230641

Vimentin supports cell polarization by enhancing centrosome function and microtubule acetylation.

Renita Saldanha, Minh Tri Ho Thanh, Nikhila Krishnan, Heidi Hehnly, Alison Patteson.

  Cell polarity is important for controlling cell shape, motility and cell division processes. Vimentin intermediate filaments are important for cell migration and cell polarization in mesenchymal cells and assembly of vimentin and microtubule networks is dynamically coordinated, but the precise details of how vimentin mediates cell polarity remain unclear. Here, we characterize the effects of vimentin on the structure and function of the centrosome and the stability of microtubule filaments in wild-type and vimentin-null mouse embryonic fibroblasts. We find that vimentin mediates the structure of the pericentriolar material, promotes centrosome-mediated microtubule regrowth and increases the level of stable acetylated microtubules in the cell. Loss of vimentin also impairs centrosome repositioning during cell polarization and migration processes that occur during wound closure. Our results suggest that vimentin modulates centrosome structure and function as well as microtubule network stability, which has important implications for how cells establish proper cell polarization and persistent migration.

Keywords:  cell polarization; centrosome; cytoskeleton; microtubules; vimentin; wound healing

DOI:  https://doi.org/10.1098/rsif.2023.0641
Aging Cell. 2024 Jun 02. e14197

Mapping the unicellular transcriptome of the ascending thoracic aorta to changes in mechanosensing and mechanoadaptation during aging.

Cristobal F Rivera, Yasmeen M Farra, Michele Silvestro, Steven Medvedovsky, Jacqueline Matz, Muhammad Yogi Pratama, John Vlahos, Bhama Ramkhelawon, Chiara Bellini.

  Aortic stiffening is an inevitable manifestation of chronological aging, yet the mechano-molecular programs that orchestrate region- and layer-specific adaptations along the length and through the wall of the aorta are incompletely defined. Here, we show that the decline in passive cyclic distensibility is more pronounced in the ascending thoracic aorta (ATA) compared to distal segments of the aorta and that collagen content increases in both the medial and adventitial compartments of the ATA during aging. The single-cell RNA sequencing of aged ATA tissues reveals altered cellular senescence, remodeling, and inflammatory responses accompanied by enrichment of T-lymphocytes and rarefaction of vascular smooth muscle cells, compared to young samples. T lymphocyte clusters accumulate in the adventitia, while the activation of mechanosensitive Piezo-1 enhances vasoconstriction and contributes to the overall functional decline of ATA tissues. These results portray the immuno-mechanical aging of the ATA as a process that culminates in a stiffer conduit permissive to the accrual of multi-gerogenic signals priming to disease development.

Keywords:  Piezo‐1; collagen remodeling; distensibility; elastic energy; inflammation; tissue stiffness; vasoconstriction

DOI:  https://doi.org/10.1111/acel.14197
Cell Rep. 2024 May 31. pii: S2211-1247(24)00625-9. [Epub ahead of print]43(6): 114297

FAK regulates tension transmission to the nucleus and endothelial transcriptome independent of kinase activity.

Md Zahid Akhter, Pascal Yazbeck, Mohammad Tauseef, Mumtaz Anwar, Faruk Hossen, Sayanti Datta, Vigneshwaran Vellingiri, Jagdish Chandra Joshi, Peter T Toth, Nityanand Srivastava, Stephen Lenzini, Guangjin Zhou, James Lee, Mukesh K Jain, Jae-Won Shin, Dolly Mehta.

  The mechanical environment generated through the adhesive interaction of endothelial cells (ECs) with the matrix controls nuclear tension, preventing aberrant gene synthesis and the transition from restrictive to leaky endothelium, a hallmark of acute lung injury (ALI). However, the mechanisms controlling tension transmission to the nucleus and EC-restrictive fate remain elusive. Here, we demonstrate that, in a kinase-independent manner, focal adhesion kinase (FAK) safeguards tension transmission to the nucleus to maintain EC-restrictive fate. In FAK-depleted ECs, robust activation of the RhoA-Rho-kinase pathway increased EC tension and phosphorylation of the nuclear envelope protein, emerin, activating DNMT3a. Activated DNMT3a methylates the KLF2 promoter, impairing the synthesis of KLF2 and its target S1PR1 to induce the leaky EC transcriptome. Repleting FAK (wild type or kinase dead) or inhibiting RhoA-emerin-DNMT3a activities in damaged lung ECs restored KLF2 transcription of the restrictive EC transcriptome. Thus, FAK sensing and control of tension transmission to the nucleus govern restrictive endothelium to maintain lung homeostasis.

Keywords:  CP: Cell biology; DNA methylation; DNMT3a; FAK; KLF2; S1PR1; emerin; endothelial cell; intracellular tension; stiffness

DOI:  https://doi.org/10.1016/j.celrep.2024.114297
Commun Biol. 2024 Jun 04. 7(1): 683

Viscoelasticity of diverse biological samples quantified by Acoustic Force Microrheology (AFMR).

Giulia Bergamaschi, Kees-Karel H Taris, Andreas S Biebricher, Xamanie M R Seymonson, Hannes Witt, Erwin J G Peterman, Gijs J L Wuite.

In the context of soft matter and cellular mechanics, microrheology - the use of micron-sized particles to probe the frequency-dependent viscoelastic response of materials - is widely used to shed light onto the mechanics and dynamics of molecular structures. Here we present the implementation of active microrheology in an Acoustic Force Spectroscopy setup (AFMR), which combines multiplexing with the possibility of probing a wide range of forces ( ~ pN to ~nN) and frequencies (0.01-100 Hz). To demonstrate the potential of this approach, we perform active microrheology on biological samples of increasing complexity and stiffness: collagen gels, red blood cells (RBCs), and human fibroblasts, spanning a viscoelastic modulus range of five orders of magnitude. We show that AFMR can successfully quantify viscoelastic properties by probing many beads with high single-particle precision and reproducibility. Finally, we demonstrate that AFMR to map local sample heterogeneities as well as detect cellular responses to drugs.

DOI: https://doi.org/10.1038/s42003-024-06367-3
Nat Commun. 2024 Jun 07. 15(1): 4866

A YAP-centered mechanotransduction loop drives collective breast cancer cell invasion.

Antoine A Khalil, Daan Smits, Peter D Haughton, Thijs Koorman, Karin A Jansen, Mathijs P Verhagen, Mirjam van der Net, Kitty van Zwieten, Lotte Enserink, Lisa Jansen, Abdelrahman G El-Gammal, Daan Visser, Milena Pasolli, Max Tak, Denise Westland, Paul J van Diest, Cathy B Moelans, M Guy Roukens, Sandra Tavares, Anne-Marie Fortier, Morag Park, Riccardo Fodde, Martijn Gloerich, Fried J T Zwartkruis, Patrick Wb Derksen, Johan de Rooij.

Dense and aligned Collagen I fibers are associated with collective cancer invasion led by protrusive tumor cells, leader cells. In some breast tumors, a population of cancer cells (basal-like cells) maintain several epithelial characteristics and express the myoepithelial/basal cell marker Keratin 14 (K14). Emergence of leader cells and K14 expression are regarded as interconnected events triggered by Collagen I, however the underlying mechanisms remain unknown. Using breast carcinoma organoids, we show that Collagen I drives a force-dependent loop, specifically in basal-like cancer cells. The feed-forward loop is centered around the mechanotransducer Yap and independent of K14 expression. Yap promotes a transcriptional program that enhances Collagen I alignment and tension, which further activates Yap. Active Yap is detected in invading breast cancer cells in patients and required for collective invasion in 3D Collagen I and in the mammary fat pad of mice. Our work uncovers an essential function for Yap in leader cell selection during collective cancer invasion.

DOI: https://doi.org/10.1038/s41467-024-49230-z
Biophys J. 2024 Jun 03. pii: S0006-3495(24)00382-5. [Epub ahead of print]

Beyond stiffness: multiscale viscoelastic features as biomechanical markers for assessing cell types and states.

Zhuo Chang, Li-Ya Li, Zhi-Jun Shi, Wenjia Liu, Guang-Kui Xu.

Cell mechanics are pivotal in regulating cellular activities, diseases progression, and cancer development. However, the understanding of how cellular viscoelastic properties varying in physiological and pathological stimuli remains scarce. Here, we develop a hybrid self-similar hierarchical theory-microrheology approach to accurately and efficiently characterize cellular viscoelasticity. Focusing on two key cell types associated with livers fibrosis - the capillarized liver sinusoidal endothelial cells (cLSEC) and activated hepatic stellate cells (aLX-2) - we uncover a universal two-stage power-law rheology characterized by two distinct exponents αshort and αlong. The mechanical profiles derived from both exponents exhibit significant potential for discriminating among diverse cells. This finding suggests a potential common dynamic creep characteristic across biological systems, extending our earlier observations in soft tissues. Using a tailored hierarchical model for cellular mechanical structures, we discern significant variations in the viscoelastic properties and their distribution profiles across different cell types and states from the cytoplasm (elastic stiffness E1 and viscosity η), to a single cytoskeleton fiber (elastic stiffness E2), and then to the cell level (transverse expansion stiffness E3). Importantly, we construct a logistic regression based-machine learning (ML) model using the dynamic parameters outperforms conventional cell stiffness-based classifiers in assessing cell states, achieving an area under the curve (AUC) of 97% vs. 78%. Our findings not only advance a robust framework for monitoring intricate cell dynamics but also highlight the crucial role of cellular viscoelasticity in discerning cell states across a spectrum of liver diseases and prognosis, offering new avenues for developing diagnostic and therapeutic strategies based on cellular viscoelasticity.

DOI: https://doi.org/10.1016/j.bpj.2024.05.033