Effect of the abrogation of TGF-β1 by antisense oligonucleotides on the expression of TGF-β-isoforms and their receptors I and II in isolated fibroblasts from keloid scars

Abstract

Abstract. Disequilibrium of dermal wound repair can result in continued accumulation of ECM and excessive scar formation. In susceptible genetically predisposed individuals, keloid formation can be observed. Keloid disease represents a benign dermal fibroproliferative tumor that is unique to humans. TGF-ß is known to play a key role in the pathogenesis of this disease which is still not fully understood. The isoforms TGF-ß1 and TGF-ß2 have profibrotic properties, whereas TGF-ß3 may have antifibrotic functions. TGF-ß exerts its influence by binding to type I and type II TGF-ß receptors, thereby forming a complex and activating specific downstream effector molecules. The aim of this study was to investigate the effect of TGF-ß1 targeting by antisense oligonucleotides on the RNA synthesis and protein expression of TGF-ß isoforms and their receptors in keloid-derived fibroblasts. In tissue samples with normal fibroblasts (NFs) serving as control samples, expression of TGF-ß1 and -ß2 was decreased when compared to keloid fibroblasts (KFs), while expression of TGF-ß3 and of TGF-ßRII was significantly higher in NFs. In the ELISA assay, abrogation of TGF-ß1 led to a significant decrease in TGF-ß1 and -ß2 (p<0.05). Expression of TGF-ß2 mRNA was reduced. Expression of TGF-ß3 mRNA revealed contrary patterns in KFs from different patients while expression of TGF-ßRI was found to be equal during the measurement period. TGF-ßRII mRNA expression was increased after 48 and 72 h respectively. There is growing evidence for a regulatory mechanism between TGF-ß1 and its receptors. Our findings support this theory by suggesting interrelations between the different TGF-ß isoforms and their receptors. Abnormal response of KFs to TGF-ß might reflect a modification in the regulatory pathway that occurs at the receptor level or during intracellular transduction. Improving the understanding of TGF-ß in keloid disease could lead to the development of clinically useful therapeutic modalities for treatment of keloid disease or even allow identification of preventive strategies.

Tacrolimus abrogates TGF‐β1‐induced type I collagen production in normal human fibroblasts through suppressing p38MAPK signalling pathway: implications on treatment of chronic atopic dermatitis lesions

“…TGF‐β exerts its function by binding to TGF‐βRI and TGF‐βRII. Moreover, TGF‐β1 regulates TGF‐βRI and TGF‐βRII expressions in an autocrine positive‐feedback fashion . In agreement with previous studies demonstrating that TGF‐β1 stimulation enhanced the expression of TGF‐β receptors in certain types of cells including human lung fibroblasts, our results also showed an increased expression of TGF‐β receptor I and II in cultured fibroblasts after TGF‐β1 treatment.…”

“…Moreover, TGF-b1 regulates TGF-bRI and TGF-bRII expressions in an autocrine positive-feedback fashion. [45][46][47] In agreement with previous studies demonstrating that TGF-b1 stimulation enhanced the expression of TGF-b receptors in certain types of cells including human lung fibroblasts, 48,49 our results also showed an increased expression of TGF-b receptor I and II in cultured fibroblasts after TGF-b1 treatment. Addition of tacrolimus markedly inhibited the TGF-b1 enhanced TGF-bRI and TGF-bRII expressions in cultured fibroblasts.…”

TGF-beta1 Gene Polymorphism in Association with Diabetic Retinopathy Susceptibility: A Systematic Review and Meta-Analysis

“…Previous research has revealed that TGFβ1 and TGFβ2 were profibrotic, but TGFβ3 might be antifibrotic [16]. Because there is little information on TGFβ2 and TGFβ3 with DR, a pertinent analysis of isoforms other than TGFβ1 in this meta-analysis has not been done.…”

Review papers Some molecular aspects of the fibroproliferative process in keloids

“…Indukowaną TGF-β1 proliferację fibroblastów keloidowych oraz syntezę kolagenu hamują również retinoidy, genisteina i takrolimus [19,80]. Innymi kierunkami modulowania ekspresji TGF-β1 w procesie gojenia ran są: śródskórna iniekcja ludzkiego rekombinowanego TGF-β3 (avotermin) -izoformy TGF-β o działaniu antyfibrotycznymoraz terapie genowe wykorzystujące antysensowe oligonukleotydy TGF-β1 i przeciwciała dla TGF-β1 [81][82][83].…”