Fabienne Liebens

HER2-positive circulating tumor cells in breast cancer.

Purpose Circulating Tumor Cells (CTCs) detection and phenotyping are currently evaluated in Breast Cancer (BC). Tumor cell dissemination has been suggested to occur early in BC progression. To interrogate dissemination in BC, we studied CTCs and HER2 expression on CTCs across the spectrum of BC staging. Methods Spiking experiments with 6 BC cell lines were performed and blood samples from healthy women and women with BC were analyzed for HER2-positive CTCs using the CellSearch®. Results Based on BC cell lines experiments, HER2-positive CTCs were defined as CTCs with HER2 immunofluoresence intensity that was at least 2.5 times higher than the background. No HER2-positive CTC was detected in 42 women without BC (95% confidence interval (CI) 0–8.4%) whereas 4.1% (95%CI 1.4–11.4%) of 73 patients with ductal/lobular carcinoma in situ (DCIS/LCIS) had 1 HER2-positive CTC/22.5 mL, 7.9%, (95%CI 4.1–14.9%) of 101 women with non metastatic (M0) BC had ≥1 HER2-positive CTC/22.5 mL (median 1 cell, range 1–3 cells) and 35.9% (95%CI 22.7–51.9%) of 39 patients with metastatic BC had ≥1 HER2-positive CTC/7.5 mL (median 1.5 cells, range 1–42 cells). In CTC-positive women with DCIS/LCIS or M0 BC, HER2-positive CTCs were more commonly detected in HER2-positive (5 of 5 women) than HER2-negative BC (5 of 12 women) (pu200a=u200a0.03). Conclusion HER2-positive CTCs were detected in DCIS/LCIS or M0 BC irrespective of the primary tumor HER2 status. Nevertheless, their presence was more common in women with HER2-positive disease. Monitoring of HER2 expression on CTCs might be useful in trials with anti-HER2 therapies.

Near-Infrared Fluorescence Lymphatic Imaging to Reconsider Occlusion Pressure of Superficial Lymphatic Collectors in Upper Extremities of Healthy Volunteers

Abstract Background: There are very little scientific data on occlusion pressure for superficial lymphatic collectors. Given its importance in determining the transport capacity of lymphatic vessels, it is crucial to know its value. The novel method of near-infrared fluorescence lymphatic imaging (NIRFLI) can be used to visualize lymphatic flow in real time. The goal of this study was to see if this method could be used to measure the lymphatic occlusion pressure. Methods: We observed and recorded lymph flow in the upper limb of healthy volunteers through a transparent cuff using near-infrared fluorescence lymphatic imaging. After obtaining a baseline of the lymph flow without pressure inside the cuff, the cuff was inflated by increments of 10u2009mm Hg starting at 30u2009mm Hg. A NIRFLI guided manual lymphatic drainage technique named “Fill & Flush Drainage Method” was performed during the measurement to promote lymph flow. Lymphatic occlusion pressure was determined by observing when lymph flow stopped under the cuff. Results: We measured the lymphatic occlusion pressure on 30 healthy volunteers (11 men and 19 women). Mean lymphatic occlusion pressure in the upper limb was 86u2009mm Hg (CI ±3.7u2009mm Hg, αu2009=u20090.5%). No significant differences were found between age groups (pu2009=u20090.18), gender (pu2009=u20090.12), or limb side (pu2009=u20090.85). Conclusions: NIRFLI, a transparent sphygmomanometer cuff and the “Fill and Flush” manual lymphatic drainage method were used to measure the lymphatic occlusion pressure in 30 healthy humans. That combination of these techniques allows the visualization of the lymph flow in real time, while ensuring the continuous filling of the lymph collectors during the measurement session, reducing false negative observations. The measured occlusion pressures are much higher than previously described in the medical literature.

Effect of preventive hormonal therapy on breast density: a systematic qualitative review.

Breast density (BD) is recognized as one of the strongest independent risk factors of breast cancer (BC). Unlike most other risk factors, BD can be modified, suggesting that it may be a biomarker for preventive interventions. We conducted a qualitative systematic review to address the effect of preventive hormonal therapy on BD. Among the 26 relevant studies, 10 assessed the effect of tamoxifen on BD (TAM: n = 2u2009877), 9 that of raloxifene (RLX: n = 1u2009544), and 7 that of aromatase inhibitors (AI: n = 416). The studies were characterized by a large heterogeneity in designs and in methods of BD measurement. BD could be reduced by TAM (10 studies/10). However, the effect of RLX and AI on BD remains unclear due to conflicting results between studies. Consequently, it is crucial to develop practical, accurate, and reproducible methods of measurement in order to be able to compare the effect of preventive hormonal agents on BD and to determine whether change in BD can be used as a predictor of response to therapy.

Abstract P2-12-15: Perception of women after undergoing microbiopsy following a positive breast cander screening

Background: Some authors have challenged the benefit of breast cancer mammographic screening due to high prevalence of overdiagnosis and overtreatment. Purpose: To assess the anxiety and stress of women undergoing a microbiopsy following a false positive breast cancer screening and to evaluate whether this experience refrains them from continuing breast cancer screening. Methods: 296 patients underwent breast microbiopsy for positive breast cancer screening B1, B2 and B3 lesions in our Breast Center between 2011 and 2013. Patients undergoing microbiopsy for B4 and B5 lesions were excluded from the study. The patients were thereafter interviewed by our two psychologists by phone within a time range of 6 to 18 months following the biopsy. Results: 201 women answered the questionnaire (68% response). 5.5% had a breast lesion at risk and 75% had a benign lesion. Half of the women rated their anxiety as high (7-8 on a scalebetween 0 – 10) and 15% as very high (9-10/10) before the biopsy. While waiting for the result, 42% rated their anxiety as high (7-8/10) and nearly a third as very high (9-10/10). But 80% accepted to continue breast cancer screening and 90% would accept to undergo again a microbiopsy if necessary. Conclusion: microbiopsy provides high stress in most patients undergoing it. Nevertheless, 90% of those who had a false positive results accept it and are willing to undergo a biopsy again if necessary. Citation Format: Birgit Carly, Mireille Aimont, Nicolas Beauloye, Fabienne Liebens. Perception of women after undergoing microbiopsy following a positive breast cander screening [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-12-15.

Abstract 839: HER2 expression occurs early in breast cancer dissemination

Background: Systemic spread of disseminated tumor cells from the primary site may occur early in breast tumorigenesis. The role of HER2 in breast cancer (BC) dissemination is not well understood. We evaluated the expression of HER2 on peripheral blood Circulating Tumor Cells (CTCs) from women with various stages of breast cancer (BC). Methods: 81 women with preinvasive breast lesions (Atypical Ductal Hyperplasia, Lobular Carcinoma In Situ and Ductal Carcinoma In Situ), 101 women with early invasive BC and 35 women with metastatic BC were included. CTCs were defined as Cytokeratin 8,18,19+/CD45-/DAPI+ cells detected using the CellSearch® technology. Any staining for HER2 on CTCs using the CellSearch® HER2 profiling kit was considered positive. HER2 detection was calibrated through peripheral blood spiking experiments with MCF7 and SKBR3 cells. Results: > 1 CTCs/22.5mL of blood were detected in 6 of 81 (7%) women with preinvasive breast lesions (range 0-3 CTCs) and 15 of 101 (15%) women with early invasive BC (range 0-8 CTCs), whereas > 1 CTCs/7.5mL of blood were detected in 17 of 35 (49%) women with metastatic BC (range 0-1160 CTCs) (p Conclusion: This is the first report showing that HER2-positive CTCs can be detected in women with preinvasive breast lesions, further supporting the hypothesis that dissemination occurs early in breast tumorigenesis. HER2 expression on CTCs is more commonly observed in earlier stages of breast cancer, suggesting that HER2 plays an important role in early dissemination. The EORTC 90091-10093 trial will test the hypothesis that trastuzumab can eliminate HER2-positive CTCs in patients with HER2-negative early breast cancer. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 839.

HER2+ circulating tumor cells (CTCs): a more sensitive and specific marker for CTCs positivity than total CTCs count in early breast cancer.

CTRC-AACR San Antonio Breast Cancer Symposium: 2008 AbstractsnnAbstract #5023 nnBackground: CTCs have been detected using the CellSearch System in early and metastatic breast cancer (BC). However, no studies have been reported for the detection of HER2+ CTCs using the CellSearch System. In this study we aimed to analyze CTCs for HER2 expression in different stages of BC progression from ductal/lobular carcinoma in situ (DCIS/LCIS) to early and metastatic BC. Patients and Methods: 20ml of peripheral blood per woman was analyzed from healthy women, DCIS/LCIS or early BC patients, whereas 7.5ml of blood was analyzed from metastatic BC patients. The presence of CTCs as well as HER2 expression was assessed with the CellSearchSystem (Veridex, USA). After immunomagnetic enrichment with an anti-Epcam-antibody, cells were labeled with anti-cytokeratin (8,18,19), anti-HER2 and anti-CD45 antibodies. CTCs were defined as Cytokeratin-positive/CD45-negative cells, whereas HER2+ CTCs were defined as Cytokeratin-positive/CD45-negative/HER2-positive cells. Results: The presence of CTCs and HER2+ CTCs in healthy women, DCIS/LCIS, early and metastatic BC is depicted in table. ![][1] Since 1 and 2 CTCs were detected in 7 (14.8%) and 4 (8.5%) of 47 healthy women, respectively, >1CTC or >2CTCs were selected as cutoffs for further analysis. Using the above cutoffs, 7 (18.9%) and 5 (13.5%) of 37 patients with early BC were considered CTC-positive, respectively. Since no HER2+ CTCs were detected in any of the 47 healthy women, a cutoff of ≥1 HER2+ CTCs was chosen. With this cutoff, more women with early BC [11 (29.7%) of 37 women] were considered CTC-positive. Conclusion: HER2+ CTCs seems to be a more sensitive and specific marker for CTCs positivity than total CTCs counts in early breast cancer. Clinical relevance of the above cutoff for CTCs positivity using HER2+ CTCs detection will be performed in ongoing clinical trials of HER2 positive early breast cancer.nnCitation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5023.nn [1]: /embed/graphic-1.gif