Introduction: The detection and monitoring of ctDNA in metastatic breast cancer showed ability to predict treatment resistance and outcome. But the mechanisms has been a challenge to clinicians. Immune escape and immune tolerance has also been reported to cause BCa progress. Herein, we report a novel finding of the association between plasma IL-2 and the ctDNA in advanced BCa patients who received the systemic therapies, and it is potential utilization in clinic.

Methods: This study enrolled 43 patients with stage III/IV BCa at the Northwestern Memorial Hospital (2016-2017) that had longitudinally detection of ctDNA and circulating tumor cells (CTCs) before (baseline, BL) or 3 months after (first evaluation, FE) systemic therapies respectively. Duplicate whole blood samples (7.5ml/each) were collected in EDTA tubes from these patients. Plasma ctDNA was analyzed using the Guardant360 NGS-based assay (Guardant Health) and CTC enrichment and enumeration were performed in FDA approved semi-automated fluorescence CELLTRACKS ANALYZERII® System (Menarini Silicon Biosystems) by using CELLSEARCH® CXC Kit (Menarini). ELISA (Fisher) for IL-2 was performed by using patients' plasma. Database of IL-2, ctDNA and CTCs was generated and linked with clinical database. Kruskal-Wallis test was used for statistics. We previously reported cut-off of 5.7 was used to dichotomize the prognostic value of ctDNA percentage (%ctDNA) in 2018 ASCO. Matched pairs variations between IL2 levels at BL and at FE were tested through Wilcoxon signed-ranks test. Associations between %ctDNA and IL2 levels were explored through Kruskal-Wallis test. The prognostic impact of IL2 was tested through Cox regression.

Results: CTCs ≥ 5 were found in 23 patients at BL and 21 patients in FE respectively. There were 12 patients that had increase CTCs, and 31 patients with similar or less CTCs FE after systemic therapies. Decreased in CTCs was associated with increased IL-2 (P=0.004).The FE analysis showed that IL-2 dropped significantly in patients with CTC stably ≥5 (from 95.84pg to 79.46pg) after therapies (P<0.001). Furthermore, baselineIL-2 levels were significantly higher in patients with % ctDNA levels ≥5.7 (97.15pg) compared to patients with %ctDNA levels <5.7 (68.64pg) (P=0.0027). No other associations were highlighted in respect to age or number of ctDNA alterations. There was no significant variations between BL and FE levels of IL2 were observed according to BCa subtypes nor in respect to baseline %ctDNA ≥5.7 or CTCs ≥5. Compared with low level of BL IL-2 (<78.3pg) group, high level of BL IL-2 (≥78.3pg) had a significant negative impact on overall survival (OS) (P=0.037) in univariate analysis.

Conclusions: Our findings indicated that aggressive BCa with high level ctDNA mutation are associated with high level of IL-2 and immune response in patients with advanced disease. In addition we confirm a reverse correlation between change of IL-2 and change of CTCs potentially indication of immune escape. In summary, the study shows a dynamic relation between IL-2 level and tumor burden (ctDNA) and immune escape (CTCs) suggesting another potential biomarker to monitor interaction between tumor and immune environment.

Citation Format: Zhang Q, Gerratana L, Zhang Y, Flaum L, Shah A, Davis A, Behdad A, Gradishar W, Platanias L, Cristofanilli M. Association between interleukin 2 (IL-2) and circulating tumor DNA (ctDNA) is a novel biomarker for patients with metastatic breast cancer (BCa) after systemic therapies [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-01-14.