Immune Checkpoint and EMT-Related Molecules in Circulating Tumor Cells (CTCs) from Triple Negative Breast Cancer Patients and Their Clinical Impact

Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype. There are few targeted therapies for these patients, leading to an unmet need for new biomarkers. The present study aimed to investigate the expression of PD-L1, CTLA-4, GLU, and VIM in CTCs of TNBC patients. Ninety-five patients were enrolled in this study: sixty-four TNBC and thirty-one luminal. Of these patients, 60 were in the early stage, while 35 had metastatic disease. Protein expression was identified by immunofluorescence staining experiments and VyCAP analysis. All the examined proteins were upregulated in TNBC patients. The expression of the GLU(+)VIM(+)CK(+) phenotype was higher (50%) in metastatic TNBC compared to early TNBC patients (17%) (p = 0.005). Among all the BC patients, a significant correlation was found between PD-L1(+)CD45(-)CK(+) and CTLA-4(+)CD45(-)CK(+) phenotypes (Spearman test, p = 0.024), implying an important role of dual inhibition in BC. Finally, the phenotypes GLU(+)VIM(+)CK(+) and PD-L1(+)CD45(-)CK(+) were associated with shorter OS in TNBC patients (OS: log-rank p = 0.048, HR = 2.9, OS: log-rank p < 0.001, HR = 8.7, respectively). Thus, PD-L1, CTLA-4, GLU, and VIM constitute significant biomarkers in TNBC associated with patients' outcome, providing new therapeutic targets for this difficult breast cancer subtype.

Automated summary

This study investigated the expression of PD-L1, CTLA-4, GLU, and VIM in circulating tumor cells (CTCs) of triple negative breast cancer (TNBC) patients. All the examined proteins were upregulated in TNBC patients. GLU(+)VIM(+)CK(+) phenotype was more prevalent in metastatic TNBC, and significant correlation was found between PD-L1(+)CD45(-)CK(+) and CTLA-4(+)CD45(-)CK(+) phenotypes, suggesting the importance of dual inhibition in breast cancer. Moreover, GLU(+)VIM(+)CK(+) and PD-L1(+)CD45(-)CK(+) phenotypes were associated with shorter overall survival in TNBC patients, indicating their potential as prognostic biomarkers.