Maria Bramley

Effects of oestrogens and anti-oestrogens on normal breast tissue from women bearing BRCA1 and BRCA2 mutations

There is considerable interest in whether anti-oestrogens can be used to prevent breast cancer in women bearing mutations in the BRCA1 and BRCA2 genes. The effects of oestradiol (E2), tamoxifen (TAM) and fulvestrant (FUL) on proliferation and steroid receptor expression were assessed in normal breast epithelium taken from women at varying risks of breast cancer and implanted into athymic nude mice, which were treated with E2 in the presence and absence of TAM or FUL. Tissue samples were taken at various time points thereafter for assessment of proliferative activity and expression of oestrogen and progesterone receptors (ERα and PgR) by immunohistochemistry. Oestradiol increased proliferation in the breast epithelium from women carrying mutations in the BRCA1/2 genes, those otherwise at increased risk and those at population risk of breast cancer. This increase was reduced by both TAM and FUL in all risk groups. In the absence of E2, PgR expression was reduced in all risk groups but significantly more so in the BRCA-mutated groups. Subsequent E2 treatment caused a rapid, complete induction of PgR expression in the population-risk group but not in the high-risk or BRCA-mutated groups in which PgR induction was significantly delayed. These data suggest that the mechanisms by which E2 induces breast epithelial PgR expression are impaired in BRCA1/2 mutation carriers, whereas those regulating proliferation remain intact. We conclude that early anti-oestrogen treatment should prevent breast cancer in very high-risk women.

Abstract PD4-02: A study to determine the optimal method of detection and threshold for lymphoedema intervention: a multi-centre prospective study

Introduction Lymphoedema, a complication of nodal surgery in 30-40% of patients, reduces quality of life for sufferers. This prospective, multi-centre study compared multi-frequency bioimpedance spectroscopy (BIS, ImpediMed) with a validated perometer method to determine which test is more sensitive for detecting lymphoedema after axillary clearance and identify the factors predicting lymphoedema development. Material and methods Participants (n = 629) undergoing axillary clearance at 9 UK centres underwent pre-operative and arm volume measurements post-surgery (1, 3, 6, 9 & 12 months, then 6 monthly) by arm perometry, BIS measurements (L-Dex) and recorded self-reported symptoms via questionnaires. Follow-up was a minimum of two years from surgery. Change in arm volume was calculated using relative arm volume change (RAVC) with >10% increase defined as lymphoedema. The predictors of lymphoedema development and optimal method for its detection were assessed using Cox Regression, Log Rank and Kaplan-Meier survival analyses. Results In total, 629 women underwent axillary surgery, with a median age of 56 (range 22 to 90) years; 80% were ER positive and received endocrine therapy, 78% received radiotherapy and 65% received chemotherapy. Lymphoedema was detected by 24 months in 124 (20%) women by perometry. Using the LDex >10 cut-off score, bioimpedance sensitivity was 71% and specificity was 89% (PPV 47%) compared to RAVC changes. Women who had an RAVC >5%- 0.000001). Twenty-six per cent of ER negative patients developed lymphoedema compared to 19% ER positive cancer patients. The type (taxane versus no taxane) and whether chemotherapy was neo-adjuvant or adjuvant did not predict lymphoedema development. Univariate analysis revealed BMI (p=0.003), ER negativity (p= 5%- 5%- Conclusions This is the first report; ER negative cancer is associated with an increased risk of lymphoedema after axillary node clearance. Arm measurements should be taken from baseline in all patients undergoing axillary surgery and increases greater than 3% should lead to further surveillance to prevent lymphoedema development. Perometer measurement is the optimal technique for measuring and predicting the development of lymphoedema. A threshold RAVC of >5%- (Funded by NIHR Programme Grant). Citation Format: Bundred NJ, Ashton S, Riches K, Ashcroft L, Evans A, Todd C, Bramley M, Hodgkiss T, Purushotham A, Keeley V. A study to determine the optimal method of detection and threshold for lymphoedema intervention: A multi-centre prospective study [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr PD4-02.

Abstract P2-06-02: Breast cancer stem-like cell activity correlates with tumour progression to metastasis but not with clinical or tumour characteristics

Introduction: Breast cancers exhibit cellular heterogeneity, containing both stem-like and more differentiated cells. The activity of cancer stem cells (CSC) is likely to be dependent on the microenvironment or niche. Using 158 patient tumour samples, correlations between niche-independent breast CSC activity and clinical and tumour characteristics were tested. Methods: 104 early breast cancer surgical samples and 54 unrelated metastatic samples from pleural or ascitic fluid were harvested. To test CSC activity, isolated cells were grown in both primary (formation) and secondary (self-renewal) mammosphere (MS) culture. Tumour initiating activity was also tested by transplanting breast cancer fragments or cells into the sub-cutaneous flanks of NSG mice (n=84 early and n=10 metastatic). Results: No correlation was found between MS growth, MS formation (%), MS self-renewal (%) or in vivo tumour initiation and breast cancer sub-type, grade, node status or Nottingham prognostic index. 33% of the samples that formed MS in vitro initiated tumours in vivo while only 9% that failed to form MS initiated tumour growth. Metastatic compared to early BC samples grew MS more frequently (53/54 compared to 81/104), and had a higher primary MS formation efficiency (1% vs 0.6%; P Conclusions: In summary, niche-independent breast CSC activity measured in vitro by MS assay and in vivo by xenograft growth is not directly correlated with standard clinical parameters. However, both in vitro and in vivo CSC activity are increased in metastatic samples. These results suggest that breast CSC activity is independent of other prognostic indicators but may predict for poor outcome tumours. Relapse free survival data are maturing and will be presented with analysis of primary tumour ALDH1 expression. Citation Format: Sacha J Howell, Denis Alferez, Katherine Spence, Rachel Eyre, Fran Shaw, Bruno Simoes, Angelica Santiago-Gomez, Maria Bramley, Mohamed Absar, Zahida Saad, Sumohan Chatterjee, Cliona Kirwan, Ashu Gandhi, Anne C Armstrong, Andrew M Wardley, Gillian Farnie, Robert B Clarke. Breast cancer stem-like cell activity correlates with tumour progression to metastasis but not with clinical or tumour characteristics [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 P2-06-02.

Abstract P6-08-07: Optimal method of detection and threshold for early intervention to prevent lymphoedema: A multi-centre prospective study

Introduction Women who undergo axillary surgery are at risk of developing lymphoedema. Early detection is recommended by measuring arm volume from a baseline before surgery to enable early intervention. The optimal measurement method to enable early detection and time to intervention are unclear. This prospective multi-centre study compares multi-frequency bioimpedance spectroscopy (BIS, ImpediMed) with the validated perometer method to determine which test is more sensitive for detecting the optimal threshold to prevent lymphoedema. Methods Participants (N = 960) undergoing axillary clearance at 9 UK centres have pre-operative and regular arm volume measurements post-surgery (1, 3, 6, 9 & 12 months, then 6 monthly), by the validated arm perometry compared with BIS (L-Dex) measurements as well as self-reported symptoms questionnaire. Change in arm volume was calculated using relative arm volume change (RAVC). The predictors of lymphoedema development and optimal method were assessed. Results Currently 612 patients, median age 55 (range 24 to 90) years, have 6 month follow-up data and 327 have 18 month follow-up data. Seventy six percent were ER positive and received endocrine therapy, 84% percent received radiotherapy and 67% received chemotherapy in addition to surgery. Lymphoedema by 18 months was detected in 19% (n=79) of women by perometry (≥10% RAVC) and a change in L-Dex of 10 was observed in 31% of women. A moderate correlation between perometer and BIS at 3 months (r=0.40) and 6 months (r=0.60), with a sensitivity of 73% and specificity of 84% was found. Univariate analysis revealed a threshold for early intervention to prevent lymphoedema was RAVC ≥5%- Conclusions The optimal threshold for early intervention to prevent progression to lymphoedema is ≥5%- (Funded by NIHR Programme Grant). Citation Format: Nigel J Bundred, Charlotte Stockton, Katie Riches, Linda Ashcroft, Abigail Evans, Anthony Skene, Maria Bramley, Tracey Hodgkiss, Arnie Purushotham, Vaughan Keeley, BEA Investigators. Optimal method of detection and threshold for early intervention to prevent lymphoedema: A multi-centre prospective study [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 P6-08-07.

Abstract P3-09-09: Comparison of multi-frequency bioimpedance with perometry for the early detection of lymphoedema after axillary node clearance

Introduction Women who undergo axillary clearance are at risk of developing lymphoedema. Early detection is recommended by arm volume measurements from a baseline before surgery but the optimal test is unclear. This prospective multi-centre study compares multi-frequency bioimpedance spectroscopy (BIS, ImpediMed) with the validated perometer method to determine which test is more sensitive for detecting lymphoedema within 24 months of surgery. Results from 441 women with up to six months follow-up are reported here to determine whether the timing of arm measurement affects results. Methods Participants (N = 441) undergoing Axillary Clearance underwent pre-operative and subsequent regular measurements post-surgery (1, 3, 6, 9 & 12 months, then 6 monthly), of arm volume by perometry and BIS measurements as well as self-reported symptoms of swelling, numbness or heaviness. The primary endpoint of lymphoedema was defined as ≥10% increase in volume compared to the contralateral arm by perometry. Results We report the data from 441 patients with 6 month follow-up data, their median age is 55 years ranging from 27 to 90 years. Eighty percent of patients were ER positive and received endocrine therapy as well as surgery. Eighty percent also underwent radiotherapy to the breast or chest wall, while 70% received chemotherapy in addition to surgery. Mean percentage increase in arm volume by perometry at 6 months was 2.03% with a moderate correlation between perometry and BIS at 3 months (r = 0.38) and 6 months (r = 0.37). In 441 patients with 6 months follow-up, the gold standard perometry detected lymphoedema in 44 (10%) patients by 6 months compared to the contralateral arm, whereas BIS measured using the unit L-Dex was positive (showed an increase of an L-Dex of 10) in 103 (21%) patients. Of the 99 patients with 18 months follow-up, 24% have lymphoedema as detected by perometry. When compared with the baseline measurements for perometry and BIS, the month 1 measurements detected fewer cases of lymphoedema by 6 months, 11 (42%) fewer for perometry and 18 (30%) fewer for BIS. 25% of patients reported symptoms before surgery. While 100% of those with lymphoedema at 6 months post-surgery reported symptoms, 23% with no lymphoedema also reported at least one symptom at 6 months. Conclusions Pre-operative measurements should be used as baseline to enable accurate monitoring of lymphoedema development. Symptoms alone are not an accurate predictor of current or future lymphoedema and arm sleeves should not be prescribed for symptoms without measuring arm volume change compared to the contralateral arm. The modest correlation between perometry and BIS at 6 months suggests arm volume measurements remain necessary before and after ANC for monitoring, although longer term follow-up data is required to determine the most sensitive method of predicting lymphoedema.(Funded by NIHR Programme Grant). Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-09-09.

Orbital Metastases as the Primary Presentation of Lobular Breast Cancer

Invasive lobular carcinoma can be diagnostically challenging and may present with vague symptoms. Orbital metastases of any breast cancer are rare. We present a unique case of Invasive lobular carcinoma presenting primarily with orbital metastases. A 54-year-old female patient presented with an 18-month history of exopthalmos, ptosis, lagopthalmos, dermatochalasis, and widespread subcutaneous nodules (Fig. 1). An orbital and staging CT scan revealed diffuse thickening of her right eyelid with a 1.5-cm retro-orbital mass incorporating the anterolateral ocular muscles, consistent with metastases to the eyelid and extraocular muscles. The mass was incorporating the ocular muscles, but was not invading the eye (Fig. 2). Biopsy of a