Abstract P3-01-21: Circulating tumor cells of breast cancer origin identified by fluorescence in situ hybridization and may be an early predictor of therapy failure in early breast cancer

Abstract

Abstract Background: Most modern methods of detecting circulating tumor cells (CTCs) involve identifying cells with epithelial markers. This approach presents challenges, as not all epithelial cells found in circulation originate from the tumor and not all CTCs express epithelial markers. We propose using a size-exclusion filtration system to enrich for CTCs in peripheral blood followed by fluorescence in situ hybridization (FISH) of the filtered cells to identify cells of tumor origin in the early-stage breast cancer patients. We further hypothesize that the presence of CTCs may be indicators of therapy failure in early-stage breast cancer patients. Methods: Patients diagnosed with breast cancer (n = 9) were consented for CTC evaluation. Primary tumor DNA was analyzed by the Affymetrix OncoscanTM genome-wide microarray platform and investigated for somatic copy-number alterations (SCNAs). For each patient, two FISH probes were then identified for two regions of gain or a region of gain and a region of loss from the microarray results. Blood samples from patients were obtained before surgery, radiation therapy, endocrine therapy, and at 6-month or 1-year follow-up visits. Blood samples were filtered using ScreenCell® Cyto V2 devices, and FISH was performed. Cells were categorized as normal (diploid for all FISH probes), suspicious (single SCNA detected by FISH), or CTC (two SCNAs detected by FISH). Patients were identified as having CTCs present in their circulation when ≥2 CTCs were observed or when one CTC and >15 suspicious cells were observed. Results: The microarray data revealed that luminal A tumors ranged from 2-43 SCNAs; luminal B tumors ranged from 15-20 SCNAs; and ER+, PR+, HER2+ tumors ranged from 46-98 SCNAs. Although a correlation appears to exist between tumor genetic complexity and molecular subtype, the degree of complexity was highly varied within each subtype. We found that neither complexity of tumor profile, molecular subtype, nor stage could predict the presence of CTCs in patients. Molecular SubtypeSCNAsCTCsSuspicious Cells OnlyLuminal A2-433/52/5Luminal B15-200/21/2ER+, PR+, HER2+46-982/20/2 In pre-surgical blood samples, we detected CTCs in 63% of patients with stage 1 disease and in 60% of patients with luminal A tumors, 0% of patients with luminal B tumors, and 100% of patients with triple-positive tumors. StageCTCsSuspicious Cells OnlyIA/B5/82/8IIA0/11/1 Although limited in number, ongoing investigation revealed that one of our patients in early follow-up with a luminal A, stage IB tumor was identified to have persistent CTCs at 1-year after starting hormonal adjuvant therapy, suggesting residual tumor burden not detected by standard clinical modalities; this finding also suggests that this patient may be at highest risk for relapse and should be considered for additional therapies. Conclusion: Size-exclusion filtration followed by FISH analysis can accurately identify CTCs in early-stage breast cancer patients. Tumor complexity, molecular subtype, and stage did not predict the presence of CTCs in circulation. Our method for CTC detection may be able to serve as a diagnostic tool for treatment failure. Citation Format: Liu C, Althof PA, Maroni D, Stevens JM, Grabow CE, VanDyke AZ, Price JD, Sanmann JN, Thayer SP. Circulating tumor cells of breast cancer origin identified by fluorescence in situ hybridization and may be an early predictor of therapy failure in early breast cancer [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 P3-01-21.