Validated biomarkers predictive of response/resistance to anthracyclines in breast cancer are currently lacking. The neoadjuvant Trial of Principle (TOP) study, in which patients with estrogen receptor (ER)-negative tumors were treated with anthracycline (epirubicin) monotherapy, was specifically designed to evaluate the predictive value of topoisomerase II-alpha (TOP2A) and develop a gene expression signature to identify those patients who do not benefit from anthracyclines. Here we describe in details the contents and quality controls for the gene expression and clinical data associated with the study published by Desmedt and colleagues in the Journal of Clinical Oncology in 2011 (Desmedt et al., 2011). We also provide R code to easily access the data and perform the quality controls and basic analyses relevant to this dataset.

Abstract Objective: To evaluate the combined administration of Docetaxel (D) and Epirubicin (E) versus FEC 100 on 5-year disease-free survival (DFS) among non metastatic lymph node-positive breast cancer patients (pts).Patients and Methods: Main inclusion criteria were: localized unilateral breast cancer, age < 65 years, at least one positive node, no metastasis, normal cardiac, hepatic, haematological and renal functions. Pts were randomized to receive on day 1 every 3 weeks either Arm A: 6 cycles of FEC100 (F and C, each at 500 mg/m², E 100 mg/m²); or Arm B, 6 cycles of ED (E and D, each at 75 mg/m²). G-CSF was mandatory for all subsequent cycles after either febrile neutropenia or treatment delay for neutropenia. Radiotherapy was given after conservative surgery. Hormone therapy was prescribed to pts with positive hormone receptors (HRs). Pts overexpressing HER2 were secondly randomized to sequential one year of trastuzumab or observation. Sample size calculation was based on an expected 10% absolute difference in 3–year DFS for HER2+ pts. Assuming a 20% prevalence of HER2 positivity, a total of 3,000 pts were to be randomised, allowing to detect an absolute difference of 6% in 5-year DFS in favour of one of the two chemotherapy regimens.Results: Between February 2001 and August 2004, 3,010 pts were randomized in France and Belgium. Pts characteristics were well balanced between the 2 arms: median age 50 years, post-menopausal status 48%, breast-conserving surgery 70%, tumor size >2cm 49%, SBR grade III 40%, both HRs negative 20%, both HRs positive 62%, 1-3 involved nodes 67%, HER2+ 19%. Treatment was completed for 96% of pts in both arms. Febrile neutropenia was reported for 2.0% and 6.4% of cycles respectively in Arms A and B. Grade 3-4 NCI-CTC neutropenia were reported for 34% and 9% of pts on day 21(Arms A and B). Other grade 3-4 toxicities were: leucopenia (35 vs 47%), thrombopenia (1.7 vs 0.3%), nausea/vomiting (14 vs 8%) and mucositis (3.2 vs 3.3%). No toxic death was reported.As of April 2009, the median follow-up was 59.3 months. Overall, 576 pts experienced at least one event: 103 loco-regional relapses, 398 metastasis, 46 contralateral breast cancer, and 29 deaths as first event. A total number of 35 second cancers and 288 deaths are registered. The 5-year DFS rates were 79.7% (95%CI: 77.4-81.7) and 81.7% (95%CI: 79.6-83.7) in arms A and B respectively (HR=0.89, 95%CI: 0.76-1.05, p=0.18). Multivariate Cox regression analysis, adjusted for age, tumor size, SBR grade, HRs and stratified for lymph node status, revealed a significant interaction term between treatment and HER2 (p=0.01).Five-year overall survival rates (OS) were 90.3% (95%CI: 88.5-91.8) with FEC and 90.1% (95%CI: 88.3-91.6) with ED (HR=1.07, 95%CI: 0.85-1.35, p=0.54).Conclusion: No advantage in DFS or OS was observed by combination of D to E when compared to standard FEC100. The best use of D in the adjuvant setting is still uncertain. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 602.

BACKGROUND Classical cyclophosphamide, methotrexate, 5-fluorouracil (CMF) including oral cyclophosphamide is still considered an important adjuvant chemotherapy regimen in patients with early breast cancer (BC). Concern has been raised regarding the feasibility of this regimen, especially in postmenopausal patients. PATIENTS AND METHODS 254 pre- and post-menopausal node-positive BC patients aged < or = 70 years received six cycles of CMF in the context of a Belgian multicentric phase III trial of adjuvant chemotherapy. CMF dose and schedule were as follows: cyclophosphamide 100 mg/m2 p.o. on days 1 to 14, methotrexate 40 mg/m2 intravenously (i.v.) on days 1 and 8, 5-fluorouracil 600 mg/ml i.v. on days 1 and 8; cycles q. 28 days. The relative dose intensity (RDI) was calculated as the ratio between the delivered DI and the planned DI. We also analysed the RDI in two subgroups of patients with age > or = 50 years or < 50 years. RESULTS Overall, the percentage of patients ending the six cycles of the planned CMF regimen was 90%. The mean RDI achieved in the population of 254 patients was 90% (range 8% to 129%). The subgroup analysis of patients aged > or = 50 years and < 50 years showed that 81% and 76% of patients, respectively, received > or = 80% of the planned chemotherapy dose intensity (P = 0.33). No statistically significant difference was found between the percentage of patients who received a RDI < 80% and the participating institutions (P = 0.50). CONCLUSIONS The classical CMF regimen was a feasible regimen in the context of a multicentric trial, in which academic institutions as well as community hospitals participated. No substantial differences in RDI and cumulative doses were found in relation to a patient's age and the participating institution.

The authors updated their report on a randomized trial initiated in 1982 comparing, in early breast cancer, high-dose IM Medroxyprogesterone acetate (HD-MPA) adjuvant hormonotherapy during 6 months with no hormonotherapy; node-positive patients also received 6 courses of IV CMF (day 1, day 8; q.4 weeks). 246 node-negative (NN) and 270 node-positive (NP) patients had been followed for a median duration of 13 years. Previous results were confirmed in this analysis on mature data. In NN patients, relapse-free survival (RFS) was improved in the adjuvant hormonotherapy arm, regardless of age while overall survival (OAS) was also increased in younger (less then 50 years) patients. In the whole group of NP patients, no difference was seen regarding RFS or OAS. However, an age-dependant opposite effect was observed: younger patients (< 50) experienced a worse and significant outcome of relapse-free and overall survivals when receiving adjuvant HD-MPA while older patients (> = 50) enjoyed a significant improvement of their relapse-free survival. For both NN and NP patients, differences in overall survivals observed in older women with a shorter follow-up, were no longer detected. © 2001 Cancer Research Campaign http://www.bjcancer.com

PURPOSE: To compare a full-dose epirubicin-cyclophosphamide (HEC) regimen with classical cyclophosphamide, methotrexate, and fluorouracil (CMF) therapy and with a moderate-dose epirubicin-cyclophosphamide regimen (EC) in the adjuvant therapy of node-positive breast cancer. PATIENTS AND METHODS: Node-positive breast cancer patients who were aged 70 years or younger were randomly allocated to one of the following treatments: CMF for six cycles (oral cyclophosphamide); EC for eight cycles (epirubicin 60 mg/m 2 , cyclophosphamide 500 mg/m 2 ; day 1 every 3 weeks); and HEC for eight cycles (epirubicin 100 mg/m 2 , cyclophosphamide 830 mg/m 2 ; day 1 every 3 weeks). RESULTS: Two hundred fifty-five, 267, and 255 eligible patients were treated with CMF, EC, and HEC, respectively. Patient characteristics were well balanced among the three arms. One and three cases of congestive heart failure were reported in the EC and HEC arms, respectively. Three cases of acute myeloid leukemia were reported in the HEC arm. After 4 years of median follow-up, no statistically significant differences were observed between HEC and CMF (event-free survival [EFS]: hazards ratio [HR] = 0.96, 95% confidence interval [CI], 0.70 to 1.31, P = .80; distant-EFS: HR = 0.97, 95% CI, 0.70 to 1.34, P = .87; overall survival [OS]: HR = 0.97, 95% CI, 0.65 to 1.44, P = .87). HEC is more effective than EC (EFS: HR = 0.73, 95% CI, 0.54 to 0.99, P = .04; distant-EFS: HR = 0.75, 95% CI, 0.55 to 1.02, P = .06; OS HR = 0.69, 95% CI, 0.47 to 1.00, P = .05). CONCLUSION: This three-arm study does not show an advantage in favor of an adequately dosed epirubicin-based regimen over classical CMF in the adjuvant therapy of node-positive pre- and postmenopausal women with breast cancer. Moreover, this study confirms that there is a dose-response curve for epirubicin in breast cancer adjuvant therapy.

Background: The predictive role of HER-2 in node-positive breast cancer patients receiving CMF or an anthracycline-based adjuvant therapy remains unclear. In addition, topo-isomerase II alpha (topo II alpha), as the cellular target of anthracyclines, might have value as a predictive marker.Patients and methods: Four hundred eighty-one archival primary tumor samples were collected among 777 patients entered into a multicenter phase III trial comparing classical CMF with epirubicin-cyclophosphamide (HEC) as adjuvant therapy of node-positive breast cancer. HER-2 was evaluated by immunohistochemistry (IHC) using different antibodies (Abs). Topo II alpha was evaluated by IHC using the Ab KiS 1. In each subgroup of patients identified by HER-2 and topo II alpha, adjusted hazard ratios for event-free survival (EFS) and the corresponding 95% confidence intervals have been calculated for the different study comparisons. An interaction test has been performed to investigate the role of HER-2 and topo II alpha as predictive markers.Results: When HER-2 was evaluated by CB-11 and 4D5 mAbs, the EFS adjusted hazard ratios (HR) for the main study comparison HEC vs. CMF were: HER-2 positive: 0.33 (95% confidence interval (95% CI): 0.09-1.27, P = 0.08), HER-2 negative: 1.16 (95% CI: 0.71-1.90, P = 0.56); the P-value for the interaction test was 0.10. When HER-2 was evaluated by TAB-250 + pAb1 Abs, the adjusted HR for the same comparison were: HER-2 positive: 1.06 (95% CI: 0.45-2.52, P = 0.90), HER-2 negative: 0.99 (95% CI: 0.58-1.68, P = 0.97); the P-value for the interaction test was 0.84. With regard to topo II alpha, the adjusted HR for the EFS comparison HEC vs. CMF were: topo II alpha positive: 0.66 (95% CI: 0.32-1.36, P = 0.25), topo II alpha negative: 1.26 (95% CI: 0.63-2.50, P = 0.51); the P-value for the interaction test was 0.13.Conclusions: This study suggests that in node-positive breast cancer patients randomly treated with CMF or an epirubicin-based regimen, the predictive value of HER-2 may vary according to the Abs used in the immunohistochemistry assay. In addition, the study supports the concept that topo II alpha might be involved in the determination of tumor responsiveness to an anthracycline-based adjuvant therapy.

366 patients fully resected from a Dukes B2 or C colorectal cancer were randomised to receive 6 courses of systemic chemotherapy comprising either 5-fluorouracil (5 FU) alone (arm A : 450 mg/m(2)/day - 5/21 days) or combined folinic acid (FOL) and 5 FU (arm B : respectively 200 mg/m(2) racemic form or 100 mg/m(2)-l-form and 370 mg/m(2)/day - 5/21 days). 173 patients had also been initially randomised to receive one course of intraportal chemotherapy just after surgery or no portal treatment. Oral levamisole (150 mg/day; 3 days every other week) was given to all patients for one year. A significantly higher incidence of leuco-granulocytopenia was observed in the arm A (5 FU alone) inducing more frequent dose delays and adaptations as well as levamisole's withdrawal. Then dose-intensities and dose-intensity products were lower in this arm but the does intensity expressed in mg/m(2)/week remained higher (631 +/- 107 vs 557 +/- 99; p < 0.001). The median follow-up in the study was 4.5 years. Relapse free (RFS) and overall survivals (OAS) were prolonged in the 5 FU alone group peculiarly in those patients who had not been randomised for portal treatment. Curves diverged progressively with longer follow-up (at 8 years; RFS in arm A : 67-71 % vs 59-53 % in arm B; OAS in arm A : 72-74 % vs 56-46 % in arm B). Patients suffering from a colon or a Dukes C cancer benefited the most from the treatment with 5 FU alone. The results are discussed in the light of other recent adjuvant trials. Well dosed 5 FU over a short period of time without folinic acid may be a valuable and inexpensive adjuvant treatment for colorectal cancer. Levamisole may no longer be recommended in this setting.

Certain familial breast and/or ovarian cancers, specially those diagnosed early, are dominantly heritable and have been linked to mutations in BRCA1 and BRCA2 genes. We have tested 30 women selected from 25 different families with specific criteria. Blood samples were always taken with the informed consent and preliminary interview of the patient by a physicologist specialized in presymptomatic testing. Mutation detection were performed by protein truncation test (PTT), gradient gel electrophoresis (DGGE) and subsequent sequencing. The results showed four frameshift mutations among which three induced truncation of the BRCA1 protein and one of the BRCA2 protein. One of the BRCA1 mutations and the only BRCA2 mutation are prevelant among caucasians. Interestingly, one BRCA1 mutation is shared both by Dutch and French families and another one has not yet been reported. Furthermore, a new unclassified varient was identified. Conclusion: by using specific selection criteria, we have been able to detect BRCA mutations in four out of the 25 families tested. One of the mutations seems to be found only in Belgium. Genetic counselling is being offered to their relatives.

PURPOSE To determine whether recombinant human interleukin-3 (rhIL-3) reduces bone marrow depression and improves chemotherapeutic schedule adherence in ovarian cancer patients receiving first-line combination chemotherapy. PATIENTS AND METHODS In a randomized multicenter study, 185 patients received carboplatin (dose based on projected area under the concentration-time curve [AUC]=4) and cyclophosphamide (750 mg/m2) day 1, every 3 weeks for six cycles. Patients were randomized to receive rhIL-3 (5 microg/kg) or placebo once daily subcutaneously on days 3 to 12. RESULTS Adherence to chemotherapeutic regimen, mean chemotherapy cycle length, tumor response rate, and median survival at 24 months did not differ between groups. The number of side effects-primarily allergic reactions, flu-like symptoms and fever-were higher in the rhIL-3 group, which resulted in 21 discontinuations compared with one in the placebo group. Compared with placebo, the rhIL-3 group had higher platelet counts day 1 of cycles 2 to 6. The number of patients with World Health Organization (WHO) grade IV thrombocytopenia or number of platelet transfusions did not differ. Leukocyte counts differed only in cycles 1 and 2 between groups. The leukocyte nadir occurred earlier in the rhIL-3 (day 12) than in the placebo group (day 15, P=.006). Leukocytes and neutrophils were only higher in the rhIL-3 group day 1 of cycle 2. In cycles 4 and 5, more patients with WHO grade IV neutropenia received rhIL-3 (P < .005). Eosinophil counts were higher day 1 of cycles 2 to 6 in the rhIL-3 group (P < .0001). CONCLUSION rhIL-3 had stimulatory hematopoietic effects. This did not result either in reduction of platelet transfusions or in improvement of chemotherapeutic schedule adherence. There were more side effects in the rhIL-3 group than in the placebo group. rhIL-3 at 5 microg/kg/d is, therefore, not of clinical benefit in this chemotherapeutic regimen.