Application Of Urine Proteomics In The Diagnosis And Efficacy Evaluation Of Early-stage Mycosis Fungoides

BackgroundMycosis fungoides(MF)is the most common cutaneous T-cell lymphoma(CTCL).The etiology and pathogenesis of MF remain unclear.MF is usually misdiagnosed as atopic dermatitis or psoriasis in early stage due to its nonspecific erythema or plaques,thus leading to delayed treatment.There is no effective treatment for advanced MF,with a median survival time of less than 1.5 years.Therefore,new diagnostic biomarkers for MF are essential for early detection and evaluation of efficacy.Objectives1.To evaluate the efficacy and safety of interferon α-2a combined with phototherapy for the treatment of early-stage MF;2.Proteomics technology was used to detect the urine of early-stage MF patients before and after treatment,identify the protein expression profiles of MF,search and verify biomarkers conducive to the early diagnosis and efficacy evaluation of MF.Methods1.Twenty patients with early-stage MF were enrolled and then administered with subcutaneous injection of interferon alpha-2a combined with phototherapy.The efficacy was evaluated according to changes of BSA,mSWAT,as well as histopathology of skin lesions before and after treatment,and urine samples were collected from patients before and after treatment.2.Proteomics was used to detect the urine of MF cases before treatment and healthy controls.The differentially expressed proteins and related signaling pathways between different groups were identified for bioinformatics analysis,and the biomarkers that could be used for early diagnosis of MF were searched.3.ELISA was used to detect the urine of MF cases before and after treatment,and the urine of cases with atopic dermatitis,psoriasis,vitiligo and basal cell carcinoma before treatment and the urine of healthy controls were compared to verify the urine biomarkers.Results1.A total of 13 cases with MF completed the whole study.After 6 months of treatment of interferon α-2a combining with phototherapy,11 cases achieved CR(84.62%),2 cases achieved PR(15.38%),and the ORR reached 100%.The histopathology of skin lesions showed a significant decrease in the number of atypical lymphocytes in both epidermis and dermis.No serious adverse reactions occurred.2.Proteomic analysis of the urine of MF cases before treatment and healthy controls showed 41 differentially expressed proteins,of which 11 proteins were up-regulated and 30 proteins were down-regulated.The analysis of differentially expressed proteins showed that these proteins were significantly enriched in phospholipase D,PI3K-AKt and MAPK signaling pathways.3.ELISA showed that before treatment,the levels of Serpin B5,EGF,and RhoA in the urine of MF patients were significantly lower than those in the healthy controls.After 1 and 3 months of treatment,the levels of Serpin B5 and EGF in the urine of MF cases were still significantly lower than those in the healthy controls.However,after 6 months of treatment,there was no significant difference between the levels of Serpin B5,EGF,and RhoA in the urine of MF patients and the healthy controls.With the remission of skin lesions,the urine Serpin B5,EGF and RhoA levels also gradually approached the levels of healthy controls.However,there was no significant difference in urine Serpin B5,EGF,and RhoA levels between cases with atopic dermatitis,psoriasis,vitiligo,and basal cell carcinoma before treatment and the healthy controls.And the AUC values of Serpin B5,EGF,and RhoA were 0.817,0.900,and 0.933,respectively.Conclusions1.Combination of interferon α-2a and phototherapy is safe and effective against earlystage MF.2.In this study,proteomics was used for the first time to identify the urine protein profiles of MF and healthy controls,analyze the function of proteins related to the onset and progression of MF and the involved signaling pathways,and screen out the differentially expressed proteins.The urine of MF,inflammatory skin diseases,neoplasm skin diseases and healthy controls were detected by immunological methods,and the early biomarkers(Serpin B5,EGF and RhoA)that could be helpful for diagnosis and evaluation efficacy of MF were preliminatively verified,which provided an important basis for guiding clinical diagnosis and treatment.3.This study demonstrated the application value and potential of urine proteomics in MF research,and provided ideas for the follow-up investigation into the pathogenesis of MF and the development of auxiliary diagnosis and disease monitoring means.