Dynamic Mechanical Cue Facilitate Collective Responses of Crowded Cell Population

Collective cell behavior is essential for tissue growth, development and function, e.g. heartbeat1, immune responses2 and cerebral consciousness3. In recent years, studies on population cells uncover that collective behavior emerges in both inter- and intra-cellular activities, e.g. synchronized signal cascade4, and collective migration5. As the movement and shape transition of cells within the crowded environment of biological tissue can generate mechanical cues at the cell-cell interface, which may affect the signaling cascade6,7, we suspect that the inter- and intra-cellular collective behavior interplay with one another and cooperatively regulate life machinery. To verify our hypothesis, we study the collective responses of fibroblasts in a confluent cell monolayer (CCM). Our results demonstrate that cells in CCM show distinctive behavior as compared to the stand-alone (SA) cells, suggesting effect of inter-cellular interactions. Upon periodic TNF-α stimulation, collective behavior emerges simultaneously in NF-κB signaling cascade and nuclear shape fluctuations in CCM but not SA cells. We then model the inter-cellular interactions in CCM using a customized microfluidic device, and discover a feedback loop intrinsic to CCM, in which dynamic mechanical cues and mechano-signaling act as link connecting the inter- and intra-cellular collective activities. We found that mechano-signaling triggered by the dynamic mechanical cues causes collective nuclear shape fluctuation (NSF), which subsequently facilitates the collective behavior in NF-κB dynamics. Furthermore, our studies reveal that regardless of the input TNF-α periodicity, cellular responses of single fibroblasts are elevated when the dynamic mechanical cues synergize with the chemical inputs, and inhibited when there is phase-mismatching. We, therefore, postulate that besides the biological significance of mechano-signaling in regulating collective cell responses, the induction of dynamic mechanical cues to human body may be a potential therapeutic approach, allowing us to regulate the action of single cells to achieve optimal tissue performance.

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Modulatory Effects of Osthole on Lipopolysaccharides-Induced Inflammation in Caco-2 Cell Monolayer and Co-Cultures with THP-1 and THP-1-Derived Macrophages

Nutrients ◽

10.3390/nu13010123 ◽

2020 ◽

Vol 13 (1) ◽

pp. 123

Author(s):

Natalia K. Kordulewska ◽

Justyna Topa ◽

Małgorzata Tańska ◽

Anna Cieślińska ◽

Ewa Fiedorowicz ◽

...

Keyword(s):

Gene Expression ◽

Inflammatory Reaction ◽

Wide Spectrum ◽

Cell Monolayer ◽

In Vivo Studies ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Tnf Α

Lipopolysaccharydes (LPS) are responsible for the intestinal inflammatory reaction, as they may disrupt tight junctions and induce cytokines (CKs) secretion. Osthole has a wide spectrum of pharmacological effects, thus its anti-inflammatory potential in the LPS-treated Caco-2 cell line as well as in Caco-2/THP-1 and Caco-2/macrophages co-cultures was investigated. In brief, Caco-2 cells and co-cultures were incubated with LPS to induce an inflammatory reaction, after which osthole (150–450 ng/mL) was applied to reduce this effect. After 24 h, the level of secreted CKs and changes in gene expression were examined. LPS significantly increased the levels of IL-1β, -6, -8, and TNF-α, while osthole reduced this effect in a concentration-dependent manner, with the most significant decrease when a 450 ng/mL dose was applied (p < 0.0001). A similar trend was observed in changes in gene expression, with the significant osthole efficiency at a concentration of 450 ng/μL for IL1R1 and COX-2 (p < 0.01) and 300 ng/μL for NF-κB (p < 0.001). Osthole increased Caco-2 monolayer permeability, thus if it would ever be considered as a potential drug for minimizing intestinal inflammatory symptoms, its safety should be confirmed in extended in vitro and in vivo studies.

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PKCδ–iPLA2–PGE2–PPARγ signaling cascade mediates TNF-α induced Claudin 1 expression in human lung carcinoma cells

Cellular Signalling ◽

10.1016/j.cellsig.2014.12.015 ◽

2015 ◽

Vol 27 (3) ◽

pp. 568-577 ◽

Cited By ~ 19

Author(s):

Daisuke Iitaka ◽

Serisha Moodley ◽

Hiroki Shimizu ◽

Xiao-Hui Bai ◽

Mingyao Liu

Keyword(s):

Lung Carcinoma ◽

Human Lung ◽

Carcinoma Cells ◽

Signaling Cascade ◽

Lung Carcinoma Cells ◽

Tnf Α ◽

Human Lung Carcinoma

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Intestinal Intraepithelial Lymphocytes Sustain the Epithelial Barrier Function against Eimeria vermiformis Infection

Infection and Immunity ◽

10.1128/iai.02024-05 ◽

2006 ◽

Vol 74 (9) ◽

pp. 5292-5301 ◽

Cited By ~ 68

Author(s):

Kyoko Inagaki-Ohara ◽

Fitriya Nurannisa Dewi ◽

Hajime Hisaeda ◽

Adrian L. Smith ◽

Fumiko Jimi ◽

...

Keyword(s):

Transforming Growth Factor ◽

Cell Monolayer ◽

Direct Interaction ◽

Intraepithelial Lymphocytes ◽

Epithelial Barrier ◽

Necrosis Factor Alpha ◽

Intestinal Intraepithelial Lymphocytes ◽

Tnf Α ◽

Ifn Γ ◽

Eimeria Vermiformis

ABSTRACT Eimeria spp. are intracellular protozoa that infect intestinal epithelia of most vertebrates, causing coccidiosis. Intestinal intraepithelial lymphocytes (IEL) that reside at the basolateral site of epithelial cells (EC) have immunoregulatory and immunoprotective roles against Eimeria spp. infection. However, it remains unknown how IEL are involved in the regulation of epithelial barrier during Eimeria sp. infection. Here, we demonstrated two distinct roles of IEL against infection with Eimeria vermiformis, a murine pathogen: production of cytokines to induce protective immunity and expression of junctional molecules to preserve epithelial barrier. The number of IEL markedly increased when oocyst production reached a peak. During infection, IEL increased production of gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) and decreased transforming growth factor β (TGF-β) production. Addition of IFN-γ and TNF-α or supernatants obtained from cultured IEL from E. vermiformis-infected mice reduced transepithelial electrical resistance (TER) in a confluent CMT93 cell monolayer, a murine intestine-derived epithelial line, but antibodies against these cytokines suppressed the decline of TER. Moreover, TGF-β attenuated the damage of epithelial monolayer and changes in TER caused by IFN-γ and TNF-α. The expression of junctional molecules by EC was decreased when IEL produced a high level of IFN-γ and TNF-α and a low level of TGF-β in E. vermiformis-infected mice. Interestingly, IEL constantly expressed junctional molecules and a coculture of EC with IEL increased TER. These results suggest that IEL play important multifunctional roles not only in protection of the epithelium against E. vermiformis-induced change by cytokine production but also in direct interaction with the epithelial barrier when intra-EC junctions are down-regulated.

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A tunable multicellular timer in bacterial consortia

10.1101/2020.10.24.353334 ◽

2020 ◽

Author(s):

Carlos Toscano-Ochoa ◽

Jordi Garcia-Ojalvo

Keyword(s):

Collective Behavior ◽

Bacterial Population ◽

Single Cells ◽

Bacterial Strains ◽

Intracellular Space ◽

Bacterial Consortia ◽

Robust Implementation ◽

Time Encoding ◽

Stationary Conditions ◽

Molecular Components

Processing time-dependent information requires cells to quantify the durations of past regulatory events and program the time span of future signals. Such timer mechanisms are difficult to implement at the level of single cells, however, due to saturation in molecular components and stochasticity in the limited intracellular space. Multicellular implementations, on the other hand, outsource some of the components of information-processing circuits to the extracellular space, and thereby might escape those constraints. Here we develop a theoretical framework, based on a trilinear coordinate representation, to study the collective behavior of a three-strain bacterial population under stationary conditions. This framework reveals that distributing different processes (in our case the production, detection and degradation of a time-encoding signal) across distinct bacterial strains enables the robust implementation of a multicellular timer. Our analysis also shows the circuit to be easily tunable by varying the relative frequencies of the bacterial strains composing the consortium.

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Induction of TNF‐α signaling cascade in neonatal rat brain during propofol anesthesia

International Journal of Developmental Neuroscience ◽

10.1016/j.ijdevneu.2015.05.003 ◽

2015 ◽

Vol 44 (1) ◽

pp. 22-32 ◽

Cited By ~ 11

Author(s):

Jelena Popić ◽

Vesna Pešić ◽

Desanka Milanović ◽

Nataša Lončarević‐Vasiljković ◽

Kosara Smiljanić ◽

...

Keyword(s):

Rat Brain ◽

Neonatal Rat ◽

Signaling Cascade ◽

Tnf Α ◽

Neonatal Rat Brain

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Cerevisterol Alleviates Inflammation via Suppression of MAPK/NF-κB/AP-1 and Activation of the Nrf2/HO-1 Signaling Cascade

Biomolecules ◽

10.3390/biom10020199 ◽

2020 ◽

Vol 10 (2) ◽

pp. 199 ◽

Cited By ~ 2

Author(s):

Md Badrul Alam ◽

Nargis Sultana Chowdhury ◽

Md Hossain Sohrab ◽

Md Sohel Rana ◽

Choudhury Mahmood Hasan ◽

...

Keyword(s):

Endophytic Fungi ◽

Fusarium Solani ◽

Nuclear Translocation ◽

Mapk Signaling ◽

Inflammatory Activity ◽

Signaling Cascade ◽

Nuclear Factor ◽

Tnf Α ◽

Gene Assay ◽

Anti Inflammatory

As part of our continuous effort to find potential anti-inflammatory agents from endophytic fungi, a Fusarium solani strain, isolated from the plant Aponogeton undulatus Roxb., was investigated. Cerevisterol (CRVS) was identified from endophytic fungi, a Fusarium solani strain, and moreover exhibited anti-inflammatory activity. However, the underlying mode of action remains poorly understood. The aim of this study is to reveal the potential mechanisms of CRVS against inflammation on a molecular level in LPS-activated RAW 264.7 peritoneal macrophage cells. CRVS was isolated from F. solani and characterized based on spectral data analysis. The MTT assay was performed to measure cell viability in CRVS-treated macrophages. Anti-inflammatory activity was assessed by measurement of nitric oxide (NO) and prostaglandin E2 (PGE2) levels, as well as the production of various cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and -6 (IL-6) in LPS-stimulated macrophages. RT-PCR and immunoblotting analyses were done to examine the expression of various inflammatory response genes. A reporter gene assay was conducted to measure the level of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein-1 (AP-1) transactivation. CRVS suppresses the LPS-induced production of NO and PGE2, which is a plausible mechanism for this effect is by reducing the expression of iNOS and COX-2. CRVS also decreases the expression of pro-inflammatory cytokines, such as TNF-α, IL-6, and IL-1β. CRVS halted the nuclear translocation of NF-κB by blocking the phosphorylation of inhibitory protein κBα (IκBα) and suppressing NF-κB transactivation. The mitogen-activated protein kinases (MAPK) signaling pathways are also suppressed. CRVS treatment also inhibited the transactivation of AP-1 and the phosphorylation of c-Fos. Furthermore, CRVS could induce the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) by down-regulating Kelch-like ECH-associated protein 1 (Keap-1) and up-regulating hemeoxygenases-1 (HO-1) expression. The results suggest that CRVS acts as a natural agent for treating inflammatory diseases by targeting an MAPK, NF-κB, AP-1, and Nrf2-mediated HO-1 signaling cascade.

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Design, Implementation, and Validation of a Piezoelectric Device to Study the Effects of Dynamic Mechanical Stimulation on Cell Proliferation, Migration and Morphology

Sensors ◽

10.3390/s20072155 ◽

2020 ◽

Vol 20 (7) ◽

pp. 2155 ◽

Cited By ~ 1

Author(s):

Dahiana Mojena-Medina ◽

Marina Martínez-Hernández ◽

Miguel de la Fuente ◽

Guadalupe García-Isla ◽

Julio Posada ◽

...

Keyword(s):

Mechanical Stimulation ◽

Cell Monolayer ◽

Mechanical Stimuli ◽

Mechanical Parameters ◽

Dynamic Stimuli ◽

New Device ◽

Dynamic Mechanical ◽

Cell Functions ◽

Biochemical Signals

Cell functions and behavior are regulated not only by soluble (biochemical) signals but also by biophysical and mechanical cues within the cells’ microenvironment. Thanks to the dynamical and complex cell machinery, cells are genuine and effective mechanotransducers translating mechanical stimuli into biochemical signals, which eventually alter multiple aspects of their own homeostasis. Given the dominant and classic biochemical-based views to explain biological processes, it could be challenging to elucidate the key role that mechanical parameters such as vibration, frequency, and force play in biology. Gaining a better understanding of how mechanical stimuli (and their mechanical parameters associated) affect biological outcomes relies partially on the availability of experimental tools that may allow researchers to alter mechanically the cell’s microenvironment and observe cell responses. Here, we introduce a new device to study in vitro responses of cells to dynamic mechanical stimulation using a piezoelectric membrane. Using this device, we can flexibly change the parameters of the dynamic mechanical stimulation (frequency, amplitude, and duration of the stimuli), which increases the possibility to study the cell behavior under different mechanical excitations. We report on the design and implementation of such device and the characterization of its dynamic mechanical properties. By using this device, we have performed a preliminary study on the effect of dynamic mechanical stimulation in a cell monolayer of an epidermal cell line (HaCaT) studying the effects of 1 Hz and 80 Hz excitation frequencies (in the dynamic stimuli) on HaCaT cell migration, proliferation, and morphology. Our preliminary results indicate that the response of HaCaT is dependent on the frequency of stimulation. The device is economic, easily replicated in other laboratories and can support research for a better understanding of mechanisms mediating cellular mechanotransduction.

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Multiple myeloma increases nerve growth factor and other pain-related markers through interactions with the bone microenvironment

Scientific Reports ◽

10.1038/s41598-019-50591-5 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 3

Author(s):

Sam W. Z. Olechnowicz ◽

Megan M. Weivoda ◽

Seint T. Lwin ◽

Szi K. Leung ◽

Sarah Gooding ◽

...

Keyword(s):

Multiple Myeloma ◽

Nerve Growth Factor ◽

Growth Factor ◽

Cell Activation ◽

Cellular Interactions ◽

Bone Microenvironment ◽

Related Factors ◽

Nerve Growth ◽

Tnf Α

Abstract Interactions between multiple myeloma (MM) and bone marrow (BM) are well documented to support tumour growth, yet the cellular mechanisms underlying pain in MM are poorly understood. We have used in vivo murine models of MM to show significant induction of nerve growth factor (NGF) by the tumour-bearing bone microenvironment, alongside other known pain-related characteristics such as spinal glial cell activation and reduced locomotion. NGF was not expressed by MM cells, yet bone stromal cells such as osteoblasts expressed and upregulated NGF when cultured with MM cells, or MM-related factors such as TNF-α. Adiponectin is a known MM-suppressive BM-derived factor, and we show that TNF-α-mediated NGF induction is suppressed by adiponectin-directed therapeutics such as AdipoRON and L-4F, as well as NF-κB signalling inhibitor BMS-345541. Our study reveals a further mechanism by which cellular interactions within the tumour-bone microenvironment contribute to disease, by promoting pain-related properties, and suggests a novel direction for analgesic development.

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IAP Suppression of Apoptosis Involves Distinct Mechanisms: the TAK1/JNK1 Signaling Cascade and Caspase Inhibition

Molecular and Cellular Biology ◽

10.1128/mcb.22.6.1754-1766.2002 ◽

2002 ◽

Vol 22 (6) ◽

pp. 1754-1766 ◽

Cited By ~ 146

Author(s):

M. Germana Sanna ◽

Jean da Silva Correia ◽

Odile Ducrey ◽

Jongdae Lee ◽

Ken Nomoto ◽

...

Keyword(s):

Signaling Cascade ◽

Inhibitor Of Apoptosis ◽

Alternative Mechanism ◽

Caspase Activity ◽

Selective Activation ◽

Antiapoptotic Effect ◽

Caspase Inhibition ◽

Tnf Α ◽

Antiapoptotic Activity ◽

Induced Apoptosis

ABSTRACT The antiapoptotic properties of the inhibitor of apoptosis (IAP) family of proteins have been linked to caspase inhibition. We have previously described an alternative mechanism of XIAP inhibition of apoptosis that depends on the selective activation of JNK1. Here we report that two other members of the IAP family, NAIP and ML-IAP, both activate JNK1. Expression of catalytically inactive JNK1 blocks NAIP and ML-IAP protection against ICE- and TNF-α-induced apoptosis, indicating that JNK1 activation is necessary for the antiapoptotic effect of these proteins. The MAP3 kinase, TAK1, appears to be an essential component of this antiapoptotic pathway since IAP-mediated activation of JNK1, as well as protection against TNF-α- and ICE-induced apoptosis, is inhibited when catalytically inactive TAK1 is expressed. In addition, XIAP, NAIP, and JNK1 bind to TAK1. Importantly, expression of catalytically inactive TAK1 did not affect XIAP inhibition of caspase activity. These data suggest that XIAP's antiapoptotic activity is achieved by two separate mechanisms: one requiring TAK1-dependent JNK1 activation and the second involving caspase inhibition.

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Construction of 3D Cellular Composites with Stem Cells Derived from Adipose Tissue and Endothelial Cells by Use of Optical Tweezers in a Natural Polymer Solution

Materials ◽

10.3390/ma12111759 ◽

2019 ◽

Vol 12 (11) ◽

pp. 1759 ◽

Cited By ~ 1

Author(s):

Takehiro Yamazaki ◽

Toshifumi Kishimoto ◽

Paweł Leszczyński ◽

Koichiro Sadakane ◽

Takahiro Kenmotsu ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Adipose Tissue ◽

Optical Tweezers ◽

Single Cells ◽

Three Dimensional ◽

Cell Types ◽

Cellular Interactions ◽

Research Fields ◽

Wide Range

To better understand the regulation and function of cellular interactions, three-dimensional (3D) assemblies of single cells and subsequent functional analysis are gaining popularity in many research fields. While we have developed strategies to build stable cellular structures using optical tweezers in a minimally invasive state, methods for manipulating a wide range of cell types have yet to be established. To mimic organ-like structures, the construction of 3D cellular assemblies with variety of cell types is essential. Our recent studies have shown that the presence of nonspecific soluble polymers in aqueous solution is the key to creating stable 3D cellular assemblies efficiently. The present study further expands on the construction of 3D single cell assemblies using two different cell types. We have successfully generated 3D cellular assemblies, using GFP-labeled adipose tissue-derived stem cells and endothelial cells by using optical tweezers. Our findings will support the development of future applications to further characterize cellular interactions in tissue regeneration.

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