New insights into the regulation of TGF‐β/Smad and MPK signaling pathway gene expressions by nasal allergen and methacholine challenge test in asthma

Abstract

Background Asthma is a heterogeneous chronic inflammatory disease of the bronchi, the course of which is significantly influenced by extrinsic factors (specific and non‐specific). Methods The aim of this study was to evaluate the effect of these factors represented by nasal allergen challenge (specific factors) and methacholine challenge test (non‐specific) on changes in mRNA expression of genes encoding the TGF‐β (TGF‐β1 and TGF‐β3)‒Smad (mitogen‐activated protein kinase 1/3 [MPK1/3], Smad1/3/6/7) signaling pathway in asthmatic patients. Results Seventy‐five subjects were included in the study, of whom 27 were applied an intranasal allergen provocation and 48 a methacholine provocation. There were 9 men and 18 women in the intranasal provocation group, and 17 men and 31 women in the methacholine test group. We found that both examined the types of challenges contributed to changes in the relative expression of genes of the TGF‐β (TGF‐β1 and TGF‐β3)‒Smad (MPK1/3, Smad1/3/6/7) signaling pathway in asthmatic patients. A decrease was noted for MAPK1, MAPK3, Smad3, Smad6, and Smad7 genes and an increase of up to 2.5 times for TGF‐β1 gene. Conclusions Our experiment allows us to conclude that the change in the mRNA expression of the TGF‐β1–MPK1/3 and Smad3/6/7 genes occurs after an intranasal allergen and bronchial methacholine challenge.

Assessment of the effectiveness of the peptide inhibitor homologous to the transforming growth factor β cytokine blocking the TGFβRI/TGFβRII receptor complex—pilot study

“…The complex thus formed leads to the phosphorylation of the kinase domain of TGFβRI, which in turn results in the activation of the Smad pathway. [9][10][11] This is therefore a targeted pathway for research into Analyses of the functional role of selected TGF-β cytokine homologous molecules conducted so far as part of the pilot study have demonstrated that newly designed peptide inhibitors for the region I inhibit gene expression of Smad-dependent and Smad-independent pathways, which indicates that they are potent to block the TGFβRI/ TGFβRII receptor complex in vitro (Figures 7 and 8, Tables 2-5).…”

“…Expression analysis of the studied genes was performed in the Department of Medical Biochemistry of the Medical University of Lodz, Lodzkie Voivodeship, Poland. 10,[22][23][24] Expression was analyzed for the following genes:…”

“…The complementary DNA (cDNA) was subjected to real-time quantitative PCR using gene-specific primers for the studied genes and 18 S rRNA with the use of the TaqMan Sonds® & Master Mixes for qRT-PCR (Biotium, Inc.). 10,[22][23][24][25][26][27] The analysis of expression of selected genes was performed using a Real-Time PCR Optical Thermocycler manufactured by Biometra (Biometra Biomedizinische Analytik GmbH) using commercial TaqMan probes. TaqMan probes are hybridization probes designed to increase the specificity of Real-Time PCR reactions.…”

“…Therefore, the inhibition of TGFβRI (also known as activin receptor-like kinase 5, Alk5) seems to be a good strategy for the treatment of many abovementioned disorders manifesting with fibrosis. [9][10][11] To sum up, TGF-β is a pluripotent cytokine involved in a range of cellular functions including cell growth, proliferation, differentiation and death. The TGF-β-Smad signaling pathway is also muddled in the pathophysiology of many diseases including organ fibrosis, such as asthma or COVID-19 (Coronavirus disease), but also plays a significant role in the development of several cancers.…”

“…Hyperactivity of TGF‐β‐Smad pathway by specific TGFβRI/TGFβRII receptors (two highly conserved single transmembrane receptors with intracellular serine/threonine kinase domains) leads to the development of chronic pathway inflammation and miofibroblast activation. Therefore, the inhibition of TGFβRI (also known as activin receptor‐like kinase 5, Alk5) seems to be a good strategy for the treatment of many abovementioned disorders manifesting with fibrosis 9–11 …”