Paper One:Discordance Of PD-L1 Expression Between Primary And Recurrent Triple-negative Breast Carcinoma Has A Prognostic Impact Paper Two:Thioredoxin Reductase: A Novel,Independent Prognostic Prognostic Marker In Patients With Hepatocellular Carcinoma

Background:PD-1/PD-L1 inhibitors against triple negative breast cancer achieved good therapeutic effect in clinical trials.Expression of the programmed death igand 1 is used to select patients and analyze responses to PD-1/PD-L1–directed immunotherapy.Although the application of PD-1/ PD-L1 inhibitor is mainly for patients with metastasis or recurrence current ly,the specimen of primary tumor is easy to obtain,so predictive tissue biomarker research is usually conducted by analyzing the primary tumor.To date,several studies reported a discordance of PD-L1 status between primary and metastatic melanoma and clear cell renal cell carcinoma.Breast cancer is characterized by heterogeneity.So heterogeneity of PD-L1 maybe exist in TNBC.Purpose: Aim of this study is to assess the discordance rate in PD-L1 expression between primary TNBC and matched recurrences,and to evaluate the prognostic impact of the change.This maybe make more TNBC patients benefit from PD-1 pathway inhibition.Methods: This study included 65 cases of primary tumor specimens of TNBC and 111 corresponding local recurrences.The PD-L1 expression was examined with immunohistochemical taining.Discordance of PD-L1 expression in primary and recurrent tumors was analyzed.The prognostic impact of the discordance was evaluated.Results: 40%(26/65)primary TNBC specimens were PD-L1 positive in tumor cells and 36.9%(24/65)in tumor infiltrating lymphocytes(TILs).24.6%(16/65)patient specimens were discordant in PD-L1 expression between primary tumors and recurrences.7.1%(2/28)multiple recurrences from the same primary tumor were discordant in tumor cell PD-L1 positivity.PD-L1 expression was associated with negative prognostic features such as histologic grade,lymph node status(p<0.05).Recurrent patients who gained PD-L1 expression were associated with significantly shorter Post-recurrence survival(PRS)when compared with those who maintained PD-L1 negativity by survival analysis.Conclusion: we observed discordance of PD-L1 expression between primary TNBC specimens and recurrent lesions.The poorer outcome observed for patients with PD-L1 gain may be due to a biologic switch to a more aggressive phenotype.This finding should guide oncologist to select PD-1/PD-L1 inhibitors for advanced TNBC.The selection criteria for enrolling breast cancer patients for immune checkpoint inhibitors would not only depend on the primary tumor,but also on the metastatic samples.Future studies are needed in order to investigate whether a tight adherence to therapeutic strategies based on the biomarker profile of the recurrent disease might improve patient's prognosis.Background: Several tumor markers for HCC,such as alpha fetoprotein(AFP)retinol-binding protein 4,desgamma-carboxy prothrombin time(DCP)and AFP-leptin 3(AFP-L3),have been identified as potential candidates to evaluate the prognosis of patients with HCC.However,they are not always sufficient for prediction of recurrence and prognosis.In this regard,the search for innovative biomarkers capable of identifying high-risk patients and of modulating cancer treatment options is still actively ongoing.Secretion of Trx R under conditions of oxidative stress and inflammation has been observed from many normal and neoplastic cells.The level of Trx R in tumor cells is often 10-fold or even greater than in normal tissues,and tumor proliferation seems to be crucially dependent on an active thioredoxin system.In sharp contrast,the possible presence of Trx R in serum has remained much less elucidated.Objective: The aims of this study were to determine if Trx R was detected in the serum,and to establish the long-term prognostic value of early measurement of serum Trx R levels in Chinese patients with HCC after curative resection.Methods: All consecutive patients underwent curative hepatic resection for HCC at department of gastrointestinal surgery of the First Affiliated Hospital of Chongqing Medical University from January 2007 to December 2010 were enrolled into the study.None of the patients had received any previous anti-cancer treatment.In addition,300 age and sex matched controls(100 patients with liver cirrhosis [LC],100 patients with chronic liver disease [CLD],and 100 healthy individuals)were also included.The preoperative serum Trx R levels were simultaneously measured in the patients using standard methods at 1 day before the hepatectomy.Venous blood samples were taken in the morning's fasting state.After at least 30 min,but within 2 h,the tubes were centrifuged at 20? for 15 min at 1,200 g,and the sera were stored frozen in plastic vials at-80? until the time of consecutive analyses.The controls samples were collected and stored in the same way as the HCC samples.Serum levels of Trx R was measured in duplicate using a solid-phase sandwich ELISA that uses two highly specific antibodies to human Trx R protein.Other biomarkers were also tested by standard laboratory method.Results: The serum Trx R levels were significantly(P<0.0001)higher in HCC patients as compared to controls [16.1(IQR,9.0-23.2 U/ml)vs.5.1(IQR,3.0-7.1 U/ml)].Serum Trx R levels were also significantly higher in HCC when compare with LC and CLD.There was no significantly difference among the three groups of control cases.There was a significant correlation between Trx R and Child-Pugh class,tumor stage or tumor size,Hs-CRP,venous invasion(r =0.501,P<0.0001;r = 0.364,P<0.0001;r=0.378,P<0.0001;r=0.230,P<0.0001;r=244,P<0.0001respectively).Patients with recurrence,serum Trx R levels were higher compared with those in patients with a recurrence-free [22.5(IQR,11.4-26.9)U/ml vs.11.8(IQR,7.5-20.1)U/ml;p<0.0001).With an AUC of 0.837(95% CI,0.794–0.881),Trx R showed a significantly greater discriminatory ability as compared with Hs-CRP(AUC,0.649;95% CI,0.586–0.712;P<0.001)and AFP(AUC,0.584;95% CI,0.519–0.659;P<0.001),while was in the range of Child-Pugh class(AUC,0.827;95% CI,0.799–0.878;P=0.658).Interestingly,the combined model(Trx R/AFP/Hs-CRP)improved those factors alone(AUC of the combined model,0.883;95% CI,0.819–0.947).At 3 years,non-survivors had significantly higher Trx R levels than survivors(25.6[IQR,21.0-31.5] U/ml vs.11.3[IQR,7.2-17.9] U/ml;P < 0.0001;).with an AUC of 0.901(95% CI,0.869–0.933),Trx R showed a significantly greater discriminatory ability as compared with Child-Pugh class(AUC,0.802;95% CI,0.717–0.869;P <0.001),Hs-CRP(AUC,0.679;95% CI,0.618–0.740;P<0.001)and AFP(AUC,0.586;95% CI,0.525–0.647;P<0.001).Interestingly,the combined model(Trx R/AFP/Hs-CRP)also improved those factors alone(AUC of the combined model,0.933;95% CI,0.879–0.977).Conclusion: The search of reliable and efficient biomarkers for a prognostic evaluation of HCC is still an open issue.To the best of our knowledge,this is the first study to report the clinically prognosis value of Trx R for HCC in a cohort of Chinese sample.In this study,we found that elevated serum Trx R levels independently predicted worse survival and recurrence in patients with HCC underwent curative hepatic resection.Trx R was utilized as prognostic indicators of HCC which appeared to be more evident when compare with Hs-CRP and AFP.Although the current study suggests that serum Trx R level may be a useful prognostic biomarker,further studies with larger patient populations are needed to validate its prognostic value and determine the optimum cut-off value.Furthermore,the biologic significance of circulating Trx R in HCC patients remains to be clarified.