Dataset Information

Human keratinocyte-derived extracellular vesicles activate the MAPKinase pathway and promote cell migration and proliferation in vitro.

ABSTRACT:

Background

Wound healing is a complex biological process and complete skin regeneration is still a critical challenge. Extracellular vesicles (EVs) play essential roles in cell communication and cell regeneration, and recent studies have suggested that EVs may contribute to wound healing, though the molecular mechanisms behind this contribution remain unclear. For these reasons, we decided to use EVs isolated from human keratinocytes (HaCaT) in vitro to determine the potential mechanism of action of EV-derived wound healing.

Method

Scratch assays were used to determine cell migration and proliferation. Scratched cells were exposed to EVs in multiple conditions to determine how they affect wound healing. Statistical analysis between groups was carried out to using Student's two-sided t test. A p value of ResultWe found that proteomic analysis of purified EVs shows enrichment of proteins associated with cell communication and signal transduction, such as MAPK pathways, and keratinocyte and fibroblast cultures exposed to EVs had higher levels of proliferation, migration, and ERK1/2 and P38 activation. Moreover, we found that treatment with specific ERK1/2 and P38 signaling inhibitors PD98059 and SB239063 impaired EV-mediated cell migration, which suggests that ERK1/2 and P38 signaling is essential for EV-induced wound healing.

Conclusion

HaCaT cell-derived EVs accelerate the migration and proliferation of human keratinocytes and fibroblasts and may promote wound healing via the activation of MAPKinase pathways. These findings may be key in developing new methods to treat wounds and accelerate wound healing in the future.

SUBMITTER: Glady A

PROVIDER: S-EPMC7852286 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Publications

Human keratinocyte-derived extracellular vesicles activate the MAPKinase pathway and promote cell migration and proliferation in vitro.

Glady Azela A Vandebroek Arno A Yasui Masato M

Inflammation and regeneration 20210202 1

<h4>Background</h4>Wound healing is a complex biological process and complete skin regeneration is still a critical challenge. Extracellular vesicles (EVs) play essential roles in cell communication and cell regeneration, and recent studies have suggested that EVs may contribute to wound healing, though the molecular mechanisms behind this contribution remain unclear. For these reasons, we decided to use EVs isolated from human keratinocytes (HaCaT) in vitro to determine the potential mechanism ...[more]

PMID: 33526070

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Project description:BackgroundGlioblastomas are lethal brain tumors under the current combinatorial therapeutic strategy that includes surgery, chemo- and radio-therapies. Extensive changes in the tumor microenvironment is a key reason for resistance to chemo- or radio-therapy and frequent tumor recurrences. Understanding the tumor-nontumor cell interaction in TME is critical for developing new therapy. Glioblastomas are known to recruit normal cells in their environs to sustain growth and encroachment into other regions. Neural progenitor cells (NPCs) have been noted to migrate towards the site of glioblastomas, however, the detailed mechanisms underlying glioblastoma-mediated NPCs' alteration remain unkown.MethodsWe collected EVs in the culture medium of three classic glioblastoma cell lines, U87 and A172 (male cell lines), and LN229 (female cell line). U87, A172, and LN229 were co-cultured with their corresponding EVs, respectively. Mouse NPCs (mNPCs) were co-cultured with glioblastoma-derived EVs. The proliferation and migration of tumor cells and mNPCs after EVs treatment were examined. Proteomic analysis and western blotting were utilized to identify the underlying mechanisms of glioblastoma-derived EVs-induced alterations in mNPCs.ResultsWe first show that glioblastoma cell lines U87-, A172-, and LN229-derived EVs were essential for glioblastoma cell prolifeartion and migration. We then demonstrated that glioblastoma-derived EVs dramatically promoted NPC proliferation and migration. Mechanistic studies identify that glioblastoma-derived EVs achieve their functions via activating PI3K-Akt-mTOR pathway in mNPCs. Inhibiting PI3K-Akt pathway reversed the elevated prolfieration and migration of glioblastoma-derived EVs-treated mNPCs.ConclusionOur findings demonstrate that EVs play a key role in intercellular communication in tumor microenvironment. Inhibition of the tumorgenic EVs-mediated PI3K-Akt-mTOR pathway activation might be a novel strategy to shed light on glioblastoma therapy. Video Abstract.

Project description:Despite decades of research, the pathogenesis of acute respiratory distress syndrome (ARDS) remains poorly understood, thus impeding the development of effective treatment. Diffuse alveolar damage (DAD) and lung epithelial cell death are prominent features of ARDS. Lung epithelial cells are the first line of defense after inhaled stimuli, such as in the case of hyperoxia. We hypothesized that lung epithelial cells release 'messenger' or signaling molecules to adjacent or distant macrophages, thereby initiating or propagating inflammatory responses after noxious insult. We found that, after hyperoxia, a large amount of extracellular vesicles (EVs) were generated and released into bronchoalveolar lavage fluid (BALF). These hyperoxia-induced EVs were mainly derived from live lung epithelial cells as the result of hyperoxia-associated endoplasmic reticulum (ER) stress. These EVs were remarkably different from epithelial 'apoptotic bodies', as reflected by the significantly smaller size and differentially expressed protein markers. These EVs fall mainly in the size range of the exosomes and smaller microvesicles (MVs) (50-120 nm). The commonly featured protein markers of apoptotic bodies were not found in these EVs. Treating alveolar macrophages with hyperoxia-induced, epithelial cell-derived EVs led to an increased secretion of pro-inflammatory cytokines and macrophage inflammatory protein 2 (MIP-2). Robustly increased macrophage and neutrophil influx was found in the lung tissue of the mice intranasally treated with hyperoxia-induced EVs. It was determined that EV-encapsulated caspase-3 was largely responsible for the alveolar macrophage activation via the ROCK1 pathway. Caspase-3-deficient EVs induced less cytokine/MIP-2 release, reduced cell counts in BALF, less neutrophil infiltration and less inflammation in lung parenchyma, both in vitro and in vivo. Furthermore, the serum circulating EVs were increased and mainly derived from lung epithelial cells after hyperoxia exposure. These circulating EVs also activated systemic macrophages other than the alveolar ones. Collectively, the results show that hyperoxia-induced, lung epithelial cell-derived and caspase-3 enriched EVs activate macrophages and mediate the inflammatory lung responses involved in lung injury.

Dataset Information

Human keratinocyte-derived extracellular vesicles activate the MAPKinase pathway and promote cell migration and proliferation in vitro.

Background

Method

Conclusion

Publications

Human keratinocyte-derived extracellular vesicles activate the MAPKinase pathway and promote cell migration and proliferation in vitro.

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