dc.description.abstract | Fibroblasts are abundant mesenchymal cells present in all tissues in a quiescent state,
which contribute to wound healing when activated. Cytokine transforming growth
factor-β1 (TGF-β1) stimulates fibroblast-myofibroblast differentiation, which induces
extracellular matrix secretion, tissue contraction and promotes cancer cell migration.
Hence, chronic activity of stromal myofibroblasts correlates with a poor prognosis for
cancer and organ fibrosis patients. Therefore, modulating myofibroblast activity may
reduce the severity of these diseases. Previous research suggests blockade of
transmembrane integrin receptors expressed by fibroblasts prevents TGF-β1-
induced differentiation, indicating integrins are attractive therapeutic targets.
However, fibroblasts derived from different organs exhibit heterogeneity, although
their integrin expression and integrin-regulated differentiation has not been directly
compared. The aim of my research was 1) to understand and compare how integrins
regulate TGF-β1-induced activation of fibroblasts derived from normal skin, lung and
breast tissue; 2) to examine the global gene expression of TGF-β1-treated lung
fibroblasts; 3) to identify novel therapeutic targets that modulate TGF-β1-induced
activation of lung fibroblasts using a drug library.
qPCR showed skin, lung and breast fibroblasts differentially expressed TGF-β1-
induced activation markers, including ACTA2, FN1, TIMP3, CTGF and SERPINE1, in
addition to integrin genes for α1, α4, α11 and β3. Small-molecule inhibitors of αv
integrins only reduced the invasion of TGF-β1-exposed skin fibroblasts, but not lung
or breast fibroblasts. siRNA against α11, β3 and β5 decreased TGF-β1-induced
collagen contraction and activation marker expression in skin and lung fibroblasts,
while α1 siRNA prevented collagen contraction by breast fibroblasts only. RNA
sequencing of TGF-β1-treated lung fibroblasts revealed pro-inflammatory and profibrotic
pathways were significantly enriched, while screening TGF-β1-treated lung
fibroblasts with a FDA-approved drug library identified 46 hits that significantly
reduced α-smooth muscle actin and fibronectin expression.
Overall, genes are differentially expressed in TGF-β1-treated skin, lung and breast
fibroblasts, while different integrins in each fibroblast appear to regulate invasion,
TGF-β1-induced collagen contraction and gene expression. RNA sequencing revealed
TGF-β1 promotes the expression of a pro-tumour signature in lung fibroblasts and
several novel therapeutic targets that modulate the activation of lung fibroblasts
have been identified. Understanding these integrin-dependent and independent
mechanisms will facilitate the generation of myofibroblast-targeted treatments for
cancer and organ fibrosis. | en_US |