Abstract P5-17-02: Dissecting the biology of inflammatory breast cancer (BC) through cell free DNA and a circulating tumor cells (CTC)-derived signature
Authors: Gerratana L , Zhang Q , Wang C , Shah A , Davis AA , Ye Z , Zhang Y , Abu-Khalaf M , Flaum L , Strickland K , Rossi G , Behdad A , Gradishar W , Platanias L , Yang H , Cristofanilli M 
Published: May 2019

Abstract
Background: The biological characteristics conferring Inflammatory BC's (IBC) distinctive and aggressive clinical features are currently not fully clarified. The aim of this study is to dissect IBC's biology through the integration of DNA and CTC-based circulating biomarkers.
Methods: This study retrospectively analyzed 251 Advanced BC (ABC) patients (pts) treated and longitudinally characterized for CTCs and circulating tumor DNA (ctDNA) at Thomas Jefferson University (Philadephia, PA) and Northwestern University (Chicago, IL). CTCs were enumerated through CellSearch (Menarini Silicon Biosystems), and characterized for HER2 expression using the CellSearch CXC Kit, while ctDNA was analyzed using the Guardant360 NGS assay (Guardant Health) and its percentage (%ctDNA) was classified based on the previously reported cut-off of 5.7% (Gerratana et al 2018). A subset of 117 pts was further characterized for circulating cell-free DNA (ccfDNA) through Qubit® dsDNA HS quantitation Assay (Thermo Fisher Scientific) and quantitative real-time PCR assay for ALU DNA repeats on chromosome 1.Associations between clinical characteristics, CTCs-derived biomarkers and IBC were explored through Fisher's exact test; survival was tested though Cox regression and log-rank test.
Results: Of the total 251 pts, 115 were diagnosed with IBC. Among the 117 patients characterized for ccfDNA, 70 had IBC. Median ccfDNA was 1.59 for IBC (IQR 1.02-3.19) and 2.37 for non-IBC (nIBC) (IQR 1.13-3.52), P=0.27. Consistent results were observed for %ctDNA levels (median value: 2 vs 1.6). The impact on OS of ccfDNA after log transformation was significant for the total population (HR 1.73 95%CI: 1.11-2.69) but not in IBC pts (HR 1.40 95%CI: 0.84-2.34). On the other hand, ctDNA high pts had a significantly worse OS (nIBC: HR 5.34 95%CI: 1.70-18.81 P=0.004; IBC: HR 4.05 95%CI: 1.91-8.58 P&lt; 0.001). In the ctDNA high subgroup no differences in total number of CTCs were observed between IBC and nIBC, while significantly lower CTCs were observed in ctDNA low IBC pts (P=0.0097). The ctDNA low IBC subgroup had a higher incidence of HER2 positive BC (P=0.003) and a significantly lower incidence of CTCs clusters (P=0.006), HER2 positive CTCs (P=0.041). Notably, no associations were observed with stage at baseline, number of metastatic sites, liver, lung and visceral involvement. On the other hand, the ctDNA_high IBC subgroup was characterized by a lower incidence in liver, bone and visceral involvement (P=0.017, P=0.014 and P=0.03 respectively) and a marginally high incidence in soft tissue involvement (0.084). Moreover, IBC diagnosis conferred a significantly worse prognosis only in the ctDNA low subgroup (OS at 12 months nIBC: 100% vs IBC: 70%; P=0.049), while no differences were observed in the ctDNA_high subgroup (OS at 12 months nIBC: 29% vs IBC: 26%; P=0.767).
Conclusion: ctDNA is able to stratify BC according to aggressiveness independently from the sites and type of metastases, both in the IBC and nIBC subgroups. IBC has a distinctive CTCs/ctDNA-based signature, in particular ctDNAlow pts have a lower incidence of HER2 positive CTCs and CTC clusters. This signature is probably due to predominant lymphatic metastatic spread and aggressive phenotype.
Citation Format: Gerratana L, Zhang Q, Wang C, Shah A, Davis AA, Ye Z, Zhang Y, Abu-Khalaf M, Flaum L, Strickland K, Rossi G, Behdad A, Gradishar W, Platanias L, Yang H, Cristofanilli M. Dissecting the biology of inflammatory breast cancer (BC) through cell free DNA and a circulating tumor cells (CTC)-derived signature [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 P5-17-02.