Clinical Significance Of Heat Shock Protein 90α And Circulating Tumor DNA In The Evaluation Of Therapeutic Effect Of Sorafenib In Hepatocellular Carcinoma

Objective To investigate the diagnostic and therapeutic monitoring efficacy of plasma heat shock protein 90a(Hsp90a)in patients with hepatocellular carcinoma(HCC)receiving targeted therapy with sorafenib tosylate tablets,and to explore the clinical significance of circulating tumor DNA(ctDNA)in predicting the therapeutic effect of sorafenib on HCC patients.Method Twenty-two patients who were treated with Sorafenib tosylate at the Tumor Diagnosis and Treatment Center of Naval General Hospital from March 2017 to December 2018 and who had been diagnosed with HCC according to relevant histopathological criteria were enrolled during the hospitalization follow-up period.Retained plasma samples.The patients were evaluated for efficacy by using the Response Evaluation Criteria in Solid Tumors Version 1.1(RECIST 1.1),and the Hsp90 a and ctDNA in plasma samples were monitored dynamically.Enzyme-linked immunosorbent assay(ELISA)was used to detect plasma HSP90α in 22 HCC patients and 40 healthy subjects.The value of HSP90α in the clinical diagnosis and therapeutic effect of HCC was investigated.According to the length of progression free survival(PFS),12 patients with HCC who met the sequencing quality control criteria were divided into excellent effect group(7 cases)and inferiority group(5 cases),ctDNA and leucocyte genomic DNA were isolated from peripheral blood of 12 HCC patients at two time points beforeand after treatment.The mutation profiles of 620 selected genes were obtained by targeted depth sequencing using NGS method.The frequency of each gene mutation was combined with the actual clinical data of the patients for biosignal analysis,and the key genes affecting the difference in the efficacy of sorafenib were dynamically observed and the types of genes associated with the poor prognosis of sorafenib were initially explored.Result The plasma Hsp90α level in the sorafenib group was significantly higher than that in the healthy control group(P<0.001).The level of Hsp90α was correlated with BCLC stage,TNM stage and extrahepatic metastasis.In TNM stage,the level of Hsp90α in stage II and stage IV patients had significant difference(P<0.0165).In the diagnosis of HCC,the AUC of Hsp90α was 0.969,the diagnostic efficacy was better than AFP(AUC=0.926),the AUC of Hsp90α combined with AFP was 0.994,which was higher than that of single diagnostic index.There was no statistical difference in plasma Hsp90α levels between AFP-negative and AFP-positive patients(P>0.05).After treatment with sorafenib,the level of plasma Hsp90α increased in patients with disease progression(P < 0.05),but there was no significant change in the level of plasma Hsp90α in patients with stable disease(P > 0.05).We compared the blood samples before and after treatment and found that the mutation frequency of 19 genes(SDHA,OTUD7 A,TMPRSS2,NAV3,AKT1,EIF2S2,DICER1,MAP3K1,RAD9 A,TCF3,IRS1,PIK3C2 G,CTCF,TSHZ2,ITPKB,KDM5 C,STAG2,AMER1,CDC27)can separate the “superior effect group” and the “inferiority group”.The dynamic changes of the 19 gene mutations can predict the therapeutic effect.The difference of BRD4 and FANCG gene mutation abundances can effectively stratify the patients in the “superior effect group” and “inferior group”.Most patients with poorefficacy had higher BRD4 and FANCG gene mutation abundances(p=0.014).In addition,ARID1 B,BRD4,FANCG,KMT2 B,and TP53 mutations in the pre-treatment blood were associated with shorter PFS,BRD4 and TP53 were significantly associated with PFS(BRD4 mutation vs.wild type: 8 weeks vs.32 weeks,p = 0.001;TP53mutation vs.wild type: 8 weeks vs.32 weeks,p = 0.009).Conclusion We found that in patients with HCC treated with sorafenib,plasma Hsp90α may be used as an indicator of disease diagnosis,and its level may be positively correlated with tumor burden.Hsp90α combined with serum AFP for adjuvant diagnosis will help to improve the sensitivity and specificity of diagnosis.In addition,dynamic changes in plasma Hsp90α can monitor the efficacy of sorafenib targeted therapy,which may be more important for patients with negative AFP.The mutation of ctDNA can predict the efficacy and prognosis of sorafenib in the treatment of advanced HCC.Dynamic monitoring of the frequency of ctDNA mutations in peripheral blood may be an effective method for monitoring the efficacy of sorafenib targeted therapy.The clinical significance of BRD4,FANCG,TP53 and other 19 gene mutations needs to be further studied by expanding the sample size.