Markers Associated With Tumor Recurrence In Patients With Breast Cancer After Neoadjuvant Chemotherapy

Background:Breast cancer patients underwent neoadjuvant chemotherapy(NAC)have different risk of tumor recurrence after NAC,including non-p CR and p CR patients.Exploring markers associated with tumor recurrence after NAC would be helpful for learning the mechanism of metastasis and optimizing individualized treatment.However,revealing the risk factors of tumor recurrence in patients with NAC,especially in the p CR patient population,is still an unresolves challenge.Therefore,we retrospectively analysis clinicopathological data of patients with NAC and explore predictive factors of recurrence after NAC.Furthermore,proteomics and bioinformatics study were used to identify potential prognostic biomarkers in p CR patient,which subsequently support exploration of adjuvant treatment escalation in the high-risk patients.Methods : First,clinicopathological data of 1,913 patients diagnosed with breast cancer between 1995 and 2020 and received NAC were retrospectively collected.Univariate and multivariate Logistic regression analysis were used to determine the factors associated with p CR.Univariate and multivariate Cox regression analysis were used to assess the associations of clinicopathological factors with non-p CR and p CR patients’ outcome.Recurrence-free survival(RFS)were analyzed using the Kaplan–Meier survival analysis and log-rank test.P values tested two-sided ≤ 0.05 were considered significant.Second,we applied proteomic study of tumors to explore differentially expressed proteins(DEPs)between tumors from the p CR patients with tumor recurrence and tumors from paired p CR patients without tumor recurrence.The corresponding differential expressed genes(DEGs)of DEPs were identified using uniprot database.PPI network analysis of DEGs was used to identify the hub genes.The prognostic value of the DEGs was evaluated using two online databases(Kaplan-Meier Plotter and bc-Gen Ex Miner).The prognostic genes as well as being identified as hub genes,were considered as potential prognostic biomarkers of p CR patients.Publicly available data from two NAC databases(GSE32603 and GSE25066)in Gene Expression Omnibus(GEO)was used to verify the prognostic value of the identified biomarkers.Results: Among the 1,913 included patients,with median follow-up of 24.9 months(IQR,11.6-46.0),420 had tumor p CR and 1,437 patients had residual tumor after NAC.Multivariate analysis showed that clinical T stage(p<0.001),clinical N stage(p<0.001),histological grade(p=0.002),ER(p<0.001),HER2(p<0.001)and NAC regimens(p<0.001)were independent factors for achieving p CR after NAC.The p CR patients have better prognosis: The 5-year risk of tumor recurrence of p CR patients was lower than that of non-p CR patients(11% vs 44%,HR=0.21,95%CI:0.144-0.296,p<0.001).ER status(HR=0.537,95%CI: 0.409-0.706,p<0.001),clinical T/N stages(HR=1.369,95%CI: 1.046-1.792,p=0.022 for clinical T stages and HR=2.766,95%CI: 2.030-3.767,p<0.001 for clinical N stages),pathological T/N stages(HR=1.914,95%CI: 1.045-1.996,p=0.026 for pathological T stages and HR=2.091,95%CI: 1.405-2.607,p<0.001 for pathological N stages)were independent prognostic factors for RFS after NAC in non-p CR patient,while clinical T/N stages(HR=2.57,95%CI: 1.01-6.51,p=0.047 for clinical T stages and HR=3.48,95%CI 1.37-8.83,p=0.009 for clinical N stages)were independent prognostic factors for RFS in p CR patient.Finally,with proteomic study and GEO database,we found that aldehyde dehydrogenase(ALDH)3A2 was correlated with tumor recurrence in breast cancer patients with NAC,as well as being a potential predictor for tumor recurrence in the p CR patients(tumor recurrence rate: low expression vs high expression,20.7% vs 4.5% [data from GSE32603 with a median follow up of 3.78 years];10.9% vs0% [data from GSE25066 with a median follow up of 2.74 years]).Conclusions: Clinical T/N stage,tumor grade,ER,HER2 and NAC regimens were independent predictors for achieving p CR after NAC.ER,clinical and pathological T/N stages were strongly correlated with tumor recurrence in non-p CR patients after NAC.Clinical T/N stage and tumor expression of ALDH3A2 were potential markers for predicting tumor recurrence in the p CR patients.