| BackgroundMycosis fungoides(MF)is a type of non-Hodgkin’s lymphoma that primarily occurs in cutaneous T lymphocytes,accounting for approximately 60%of cutaneous T cell lymphoma.It is the most common type of skin lymphoma in clinical practice.The etiology and pathogenesis of the disease are still unclear.Early-stage MF presents as localized or generalized patches and plaques,which are easily misdiagnosed as benign inflammatory diseases(BID)such as eczema,pityriasis rosea and pityriasis lichenoides chronica.Advanced-stage MF presents with aggressive progression,possibly involving lymph nodes,blood,and viscera,with poor prognosis and high mortality.Therefore,early identification and diagnosis of MF is of great significance for treatment and improving the prognosis of patients as soon as possible,but there is still a lack of specific disease markers that can be used for early diagnosis of MF.Objective1.To summarize the clinical characteristics and survival rate of mycosis fungoides in a single center of Chinese population;2.To identify the protein expression profile of mycosis fungoides through proteomic research,and screen and validate biomarkers that can be used for the early diagnosis of mycosis fungoides.Method1.MF patients who met the inclusion criteria between October 2012 and January 2018 at our clinic and had a follow-up period of more than one year were retrospectively included.Age at diagnosis,gender,duration between symptom onset and diagnosis,follow-up time(recorded from first diagnosis to last follow-up),TNMB grade at diagnosis,treatment modality,and follow-up data were collected,and diseasespecific survival(DSS)was calculated.2.Skin tissue and peripheral blood samples of mycosis fungoides(including early and advanced stages)from the experimental group and skin samples of inflammatory skin diseases(eczema,psoriasis,etc.)and healthy adults from the control group were collected for data-independent acquisition(DIA)proteomic experiment.By identifying differentially expressed proteins among different groups and bioinformatics analysis,biomarkers that can be used for early diagnosis and differential diagnosis of MF were initially screened.3.A new cohort of mycosis fungoides,inflammatory skin disease and healthy adults was recruited,skin tissues were collected and formalin-fixed and parrffinembedded(FFPE)slides were processed for validation of the selected biomarkers by immunofluorescence staining.Result1.Retrospective study of clinical characteristics:A total of 93 patients with MF were enrolled in the retrospective study,and the mean age at diagnosis was 38.9±1.73 years(range,6-77 years),including 45 males(48.4%)and 48 females(51.6%).The disease-specific survival(DSS)rate was 98.6%in early-stage MF and 88.9%in advanced-stage MF,showing a significant difference(P=0.042).2.DIA mass spectrometry identified 3827,4058,3693,4380 proteins in healthy controls,inflammatory skin disease patients,early-stage MF patients and advanced-stage MF patients,respectively,and 754 differentially expressed proteins in early-stage MF patients compared to controls(inflammatory skin disease patients and healthy adults).There are mainly 8 up-regulated proteins(DYNC1I2,PGM5,PLEC,CLIC2,S100A4,LAMP2,CD14 and COL18A1)and 4 down-regulated proteins(RPL4,RALA,CRABP2 and RPL14)in early-stage MF group compared with healthy controls and BID group.The 12 identified proteins can be used as a potential diagnostic biomarker for early-stage MF.3.Proteins with the largest expression differences between the early-stage MF and BID groups were selected for validation in a new cohort of tissue samples.The immunofluorescence results showed that the up-regulated proteins DYNC1I2,CD14,and COL18A1,and the down-regulated protein CRABP2 in MF can effectively distinguish early-stage MF from the control group.ConclusionThis study retrospectively summarized the disease characteristics of MF in a single center,providing evidence for understanding the clinical characteristics of mycosis fungoides and its variants in Chinese population,and further confirmed that the prognosis of early-stage MF is superior to that of advanced-stage MF.It is necessary to search for disease markers that can be used for the diagnosis of early MF.In this study,for the first time,we characterized and analyzed protein profiles of skin tissue from MF,inflammatory skin disease and healthy adults by DIA proteomics,screened differentially expressed proteins and initially validated biomarkers(CD 14,COL18A1,DYNC1I2 and CRABP2)capable of objective and effective diagnosis of early-stage MF by immunological methods,which may provide an important basis for guiding clinical diagnosis and treatment. |
PDF Full Text Request