Published online Oct 21, 2019. doi: 10.3748/wjg.v25.i39.5918
Peer-review started: August 1, 2019
First decision: August 27, 2019
Revised: September 6, 2019
Accepted: September 27, 2019
Article in press: September 27, 2019
Published online: October 21, 2019
Intestinal inflammation and epithelial injury are the leading actors of inflammatory bowel disease (IBD), causing an excessive pro-inflammatory cytokines expression. Tristetraprolin (TTP), an mRNA binding protein, plays a role in regulating the inflammatory factors, recognizing specific sequences on the 3’ untranslated region of cytokine mRNAs. TTP activity depends on its phosphorylation state: the unphosphorylated TTP degrades pro-inflammatory cytokine mRNAs; on the contrary, the phosphorylated TTP fails to destabilize mRNAs furthering their expression. The phospho-TTP forms a complex with the chaperone protein 14-3-3. This binding could be one of the factors that promote intestinal inflammation as a cause of disease progression.
To assess if TTP phosphorylation has a role in paediatric IBD.
The study was carried out on a cohort of paediatric IBD patients. For each patient enrolled, a specimen of inflamed and non-inflamed colonic mucosa was collected. Furthermore, the experiments were conducted on macrophages differentiated from blood samples of the same patients. Macrophages from healthy donors’ blood were used as controls. Co-immunoprecipitation assay and immunoblotting analyses were performed to observe the formation of the phospho-TTP/14-3-3 complex. In the same samples TNF-α expression was also evaluated as major factor of the pro-inflammatory activity.
In this work we studied indirectly the phosphorylation of TTP through the binding with the chaperone protein 14-3-3. In inflamed and non-inflamed colon mucosa of IBD paediatric patients immunoblot assay demonstrated a higher expression of the TTP in inflamed samples respect to the non-inflamed; the co-immunoprecipitated 14-3-3 protein showed the same trend of expression. In the TNF-α gene expression analysis higher levels of the cytokine in inflamed tissues compared to controls were evident. The same experiments were conducted on macrophages from IBD paediatric patients and healthy controls. The immunoblot results demonstrated a high expression of both TTP and co-immunoprecipitated 14-4-3 protein in IBD-derived macrophages in comparison to healthy donors. TNF-α protein levels from macrophages lysates showed the same trend of expression in favour of IBD paediatric patients compared to healthy controls.
In this work, for the first time, we describe a relation between phospho-TTP/14-3-3 complex and IBD. Indeed, a higher expression of TTP/14-3-3 was recorded in IBD samples in comparison to controls.
Core tip: We report a preliminary study describing the role of tristetraprolin (TTP)/14-3-3 protein complex in a small cohort of paediatric patients affected by inflammatory bowel disease (IBD). The experiments were conducted on IBD inflamed and non-inflamed colonic tissues and in IBD-derived macrophages in comparison to healthy donors. This study demonstrates for the first time the relation between the phosphorylation of TTP and IBD in paediatric patients. Indeed, a higher expression of the TTP/14-3-3 protein complex was recorded both in inflamed compared to non-inflamed colon specimens and in IBD-derived in comparison to healthy donors’ macrophages.