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2017 ; 12
(9
): e0184499
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Liver myofibroblasts of murine origins express mesothelin: Identification of
novel rat mesothelin splice variants
#MMPMID28898276
Fausther M
; G Lavoie E
; Dranoff JA
PLoS One
2017[]; 12
(9
): e0184499
PMID28898276
show ga
Liver myofibroblasts are specialized effector cells that drive hepatic fibrosis,
a hallmark process of chronic liver diseases, leading to progressive scar
formation and organ failure. Liver myofibroblasts are increasingly recognized as
heterogeneous with regards to their origin, phenotype, and functions. For
instance, liver myofibroblasts express cell markers that are universally
represented such as, Itg?V and Pdgfr?, or restricted to a given subpopulation
such as, Lrat exclusively expressed in hepatic stellate cells, and Gpm6a in
mesothelial cells. To study liver myofibroblasts in vitro, we have previously
generated and characterized a SV40-immortalized polyclonal rat activated portal
fibroblast cell line called RGF-N2 expressing multiple mesothelin mRNA
transcripts. Mesothelin, a cell-surface molecule expressed in normal mesothelial
cells and overexpressed in several cancers such as, mesothelioma and
cholangiocarcinoma, was recently identified as a key regulator of portal
myofibroblast proliferation, and fibrosis progression in the setting of chronic
cholestatic liver disease. Here, we identify novel mesothelin splice variants
expressed in rat activated portal fibroblasts. RGF-N2 portal fibroblast cDNA was
used as template for insertion of hemagglutinin tag consensus sequence into the
complete open reading frame of rat mesothelin variant coding sequences by
extension PCR. Purified amplicons were subsequently cloned into an expression
vector for in vitro translation and transfection in monkey COS7 fibroblasts,
before characterization of fusion proteins by immunoblot and immunofluorescence.
We show that rat activated portal fibroblasts, hepatic stellate cells, and
cholangiocarcinoma cells express wild-type mesothelin and additional splice
variants, while mouse activated hepatic stellate cells appear to only express
wild-type mesothelin. Notably, rat mesothelin splice variants differ from the
wild-type isoform by their protein properties and cellular distribution in
transfected COS7 fibroblasts. We conclude that mesothelin is a marker of
activated murine liver myofibroblasts. Mesothelin gene expression and regulation
may be critical in liver myofibroblasts functions and fibrosis progression.