Investigate The Role Of STAT3Pathway In NP

Background and Objectives Nasal polyposis (NP) is a nasal mucosaillness. It is a commonly encountered in clinical otorhinolaryngology. On thebasis of the European position paper on rhinosinusits and nasal polyposis(EP3OS) document， NP is considered as a subgroup of chronicrhinosinusits(CRS). Because of the high recurrence rate and the treatmentare unsatisfactory, NP has become a global health issues which bringseriously degrade the quality of human life and considerable economicburden.In recent decades, the study associate with the etiology of NP has beenthe hotspot of research in the field of rhinology, and the etiology of NP hasbeen studied widely. Although it is remain poorly understood, it is sure thatthe formation of NP is a multi-step process that a variety of factors involvedin the development, and the allergic, chronic inflammatory are known to beinvolved in the origin of NP. The Th cell, especially the Th17cell, plays acrucial role in the pathogenesis of NP in Chinese patient, and the atopy mayplay a key role in the etiology of NP in Chinese patient via exerting theinfluence on Th17cell response. Among the STATs family, STAT3is a very pivotal member he tyrosineresidues of STAT3have become phosphorylated by JAK upon receptoractivation by extracellular signals, and then form a dimer which transferredinto the nucleus and bind specific DNA fragment to play the role oftranscription regulation. In recent years, studies have found that STAT3signaling pathway is involved in the process of Th17cell proliferation andactivation via IL-6stimulation, and the STAT3-mediated Th17response hasbeen found to play a dominanted role in the pathogenesis of immune andinflammatory disease such as Uveitis, Crohn’s disease and asthma. So inChina, whether the abnormal activation of STAT3signaling pathway ispresent in NP that enhanced TH17response existed? Whether atopy makesimpact on Th17response in NP via exerting influence on STAT3signalingpathway?To explore these problems, we partitioned NP patients in the light ofthe atopy or non-atopy, moreover, the abnormal activation of STAT3signaling pathway was designed as the entry point. In order to evaluated therole of STAT3pathway in NP, we studied the characteristics of inflammatoryimmune response in NP patients with atopy or not, and we explored whetherthe STAT3pathway is associate with the role of atopy in NP. This study willopen a new avenue to study the pathogenesis of NP, improve the theory thatChinese NP patients have Th17-bias response; this research can provide newtargets for the research of NP and provide a new breakthrough for the treatment of NP.Part one Expression of STAT3signaling pathway related factorsin NP patients with and without atopyObject: we aimed to assess whether the abnormal activation of STAT3pathway is exist in patients with NP, we evaluated the expression of STAT3signaling pathway related factors and the expression of its downstream Th17cell related factors in NP patients with and without atopy.Method: Forty patients (20atopic,20non-atopic) and10Inferiorturbinate tissue as the control group were included. Among the atopic NPpatients, nonatopic NP patients and controls, the levels of STAT3pathwaycomponents, including IL-6, soluble IL-6receptor (sIL-6R), phosphoSTAT3(pSTAT3), and suppressor of cytokine signaling3(SOCS3), were assessed.The Th17milieu was examined by measuring the levels of retinoidacid-related orphan receptor C (RORc), CD4+RORc+cell and IL-17. Finally,we investigated the correlation between STAT3signaling pathway relatedfactors and Th17cell related factors.Results: The results demonstrated that as well as the levels of STAT3pathway components, the levels of RORc, CD4+RORc+cell and IL-17weresignificantly higher in both NP groups than in the control. Furthermore,significantly higher levels of pSTAT3, RORc, CD4+RORc+cell and IL-17, and significantly lower levels of SOCS3were found in the atopic group thanin the non-atopic group. In addition, IL-6and sIL-6R levels were notsignificantly different between the NP groups. pSTAT3exhibitedsignificantly positive correlations with RORc, CD4+RORc+cell and IL-17.Conclusion: We found that the abnormal activation of STAT3pathwayis existing in NP. And atopic NP has more severe abnormal activation ofSTAT3pathway and Th17response. These consequent indicated that theSTAT3pathway must be playing a key role in the develepment of NP inChinese patients, and atopy may contribute to NP by affecting the STAT3pathway. Furthermore, the different levels of SOCS3between two NP groupmay explains the significant difference in the level of pSTAT3between theNP groups, as well as the lack of a difference in the expression levels of IL-6and sIL-6R.Part two The study of STAT3inhibitor on inhibition of STAT3signal transduction pathway in NPObject: Treated with AG490(STAT3inhibitor) in NP single cellsuspension in different concentrations, and then observe the Th17responsein NP after inhibition of STAT3pathway. To confirmed the role of STAT3pathway in NP in Chinese patients in vitro.Method: We evaluated the frequency of Th17in NP after inhibition ofSTAT3pathway48h among twenty NP patients (10atopic NP,10Nonatopic NP) that be divided into low concentration group (50μM AG490), highconcentration group (100μM AG490) and control group(PBS). Then weevaluated the levels of RORc and IL-17among the three groups.Results: After be treated with AG490for48h, the levels of Th17celland its related factors were significantly decreased, furthermore, the levelsof those factors were significantly decreased in high concentration group(100μM AG490) when compared to low concentration group (50μMAG490).Conclusion: These results indicated that the AG490blocked theSTAT3pathway in vitro result in the significantly decreased level of Th17response. This part of the study further proves that the STAT3pathway playcrucial role in NP in Chinese patients via making impact on Th17response.Part three House dust mite (HDM) induced activation of STAT3pathway in the Chinese patients with nasal polyposisObject: This study in order to analysis the levels of STAT3pathwaykey factors after stimulation by HDM allergen in NP patients, to assess thepossible correlation between STAT3pathway and atopy, to confirm thatatopy may play role in the pathogenesis of NP via the impact on STAT3pathway.Methods: We obtained single cell suspension from atopic NP patients,non-atopic NP patients, and controls. Then we used phytohemagglutinin (PHA) or house dust mite extracts (HDM) plus PHA to stimulate it. Theresulting level of pSTAT3was detected by Western Blot. The level ofSOCS3was measured by Western Blot and real-time PCR.Results: The levels of pSTAT3and SOCS3in both stimulation groupswere significantly higher in NP patients versus controls. In addition, there ishigher level of pSTAT3and lower level of SOCS3in atopic group versusnonatopic ones. Moreover, in atopic NP, HDM+PHA Intervention resulted ina significant increase of pSTAT3and significant decrease of SOCS3versusthose in PHA stimulated ones, whereas the levels of pSTAT3and SOCS3innonatopic NP and controls have no significant different.Conclusion: The results indicated that HDM could induce moreabnormal activation of STAT3pathway in atopic NP, it confirmed that atopymay play role in NP by aggravate abnormal activation of STAT3pathway. Inaddition, the decreased level of STAT3inhibitor in stimulation by HDMillustrated that atopy may induce more serious abnormal activation of STAT3pathway by induce the decline of STAT3inhibitor, and those explained whythe levels of IL-6, sIL-6R and the levels of downstream related factors weregrowth and decline inconsistent.