Chemokines as predictive biomarkers for immune checkpoint inhibitor (ICI) efficacy in triple negative breast cancer (TNBC)

Background: TNBC, although an aggressive breast cancer (BC) subtype, is highly immunogenic and the only BC subtype where the ICI pembrolizumab is approved. However, predictive biomarkers for pembrolizumab benefit are limited. The chemokines CXCL9 and CXCL10 attract CD8⁺ T cells into the tumor microenvironment (TME) and are associated with chemotherapy benefit, but little is known about their role in prediciting pembrolizumab benefit in TNBC. We investigated the association of CXCL9, CXCL10 and their cognate receptor CXCR3 with TME and ICI efficacy. Methods: 3,038 TNBC samples were analyzed via NGS (592-gene panel, NextSeq; WES/WTS, NovaSeq; Caris Life Sciences, Phoenix, AZ). Tumor mutational burden (TMB) totaled somatic mutations per tumor (high. 10 mt/MB). Immune cell fractions were estimated using WTS deconvolution (Quantiseq). CXCL9/CXCL10/CXCR3-high (H) and -low (L) tumors were classified by RNA expression above or below the 50th percentile. Real-world overall survival (OS) was derived from insurance claims and calculated from start of pembrolizumab to last contact using Kaplan-Meier. Statistical significance was assessed using chi-square and Mann-Whitney U with multiple comparison adjustments (q,.05). Results: TNBC expressed higher levels of CXCL9 and CXCL10 (median (TPM): 5.3 and 14.7) compared to N = 1,082 HER2+ (4.8 and 10.3, p, 0.05) and N = 4,918 HR+HER2- (2.7 and 7, q,.05) BC. CXCR3 expression was higher in TNBC compared to HR+HER2- (1.9 vs 1.7, q,.05), but no difference when compared to HER2+ (1.9 vs 2, q = 0.97) BC. CXCL9/CXCL10/CXCR3-H TNBC had higher median OS post pembrolizumab [CXCL9-H vs -L: 26.5 vs 15.7 months (mo), HR: 0.65 (95% CI 0.5–0.84); CXCL10-H vs -L: 26.0 vs 20.6 mo, HR 0.74 (0.57–0.95); CXCR3-H vs -L: 32.6 vs 18.3 mo, HR 0.68 (0.52–0.88), all p, 0.05]. CXCL9-H, CXCL10-H and CXCR3-H had higher PD-L1 positivity (22C3), TMB high, higher T cell inflamed score, TP53 mutations, elevated B and CD8⁺T cells infiltration, but not neutrophils, and higher expression of immune checkpoint genes (Table). Conclusions: High CXCL9/CXCL10/CXCR3 expression is associated with longer survival in patients with TNBC post pembrolizumab, and characterized by an immune-enriched TME. Further investigation is needed to evaluate this chemokine axis in TNBC and its potential as a therapeutic target to enhance ICI efficacy. Research Sponsor: NIH (NCATS, NCI); K08CA279766-01A1.