Extracellular vesicles from mast cells induce mesenchymal transition in airway epithelial cells

• Background: In the airways, mast cells are present in close vicinity to epithelial cells, and they can interact with each other via multiple factors, including extracellular vesicles (EVs). Mast cell-derived EVs have a large repertoire of cargos, including proteins and RNA, as well as surface DNA. In this study, we hypothesized that these EVs can induce epithelial to mesenchymal transition (EMT) in airway epithelial cells. Methods: In this in-vitro study we systematically determined the effects of mast cell-derived EVs on epithelial A549 cells. We determined the changes that are induced by EVs on A549 cells at both the RNA and protein levels. Moreover, we also analyzed the rapid changes in phosphorylation events in EV-recipient A549 cells using a phosphorylated protein microarray. Some of the phosphorylation-associated events associated with EMT were validated using immunoblotting. Results: Morphological and transcript analysis of epithelial A549 cells indicated that an EMT-like phenotype was induced by the EVs. Transcript analysis indicated the upregulation of genes involved in EMT, including TWIST1, MMP9, TGFB1, and BMP-7. This was accompanied by downregulation of proteins such as E-cadherin and upregulation of Slug-Snail and matrix metalloproteinases. Additionally, our phosphorylated-protein microarray analysis revealed proteins associated with the EMT cascade that were upregulated after EV treatment. We also found that transforming growth factor beta-1, a well-known EMT inducer, is associated with EVs and mediates the EMT cascade induced in the A549 cells. Conclusion: Mast cell-derived EVs mediate the induction of EMT in epithelial cells, and our evidence suggests that this is triggered through the induction of protein phosphorylation cascades.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Klinisk medicin (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Clinical Medicine (hsv//eng)

MEDICIN OCH HÄLSOVETENSKAP  -- Klinisk medicin -- Lungmedicin och allergi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Clinical Medicine -- Respiratory Medicine and Allergy (hsv//eng)

Nyckelord

Epithelial cells

Epithelial-mesenchymal transition

Exosomes

Extracellular vesicles

Mast cells

Phosphorylated proteins

Protein microarray

gelatinase B

osteogenic protein 1

transcription factor Slug

transcription factor Snail

transforming growth factor beta1

Twist related protein 1

uvomorulin

A-549 cell line

airway epithelium cell

Article

BMP 7 gene

controlled study

down regulation

epithelial mesenchymal transition

exosome

human

human cell

immunoblotting

in vitro study

intracellular signaling

mast cell

MMP9 gene

phenotype

protein phosphorylation

TGFB1 gene

TWIST1 gene

upregulation

Mast cells

Epithelial cells

Extracellular vesicles

Exosomes

Phosphorylated proteins

Protein microarray

Epithelial-mesenchymal

transition

cancer-cells

intercellular-adhesion

heparan-sulfate

lung

expression

emt

angiogenesis

macrophages

recruitment

activation

Respiratory System

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)