Les progres spectaculaires de la genetique et de la biologie moleculaires du cancer ont rendu possible le developpement d'une nouvelle classe de produits: les therapies ciblees du cancer. A la difference des chimiotherapies anticancereuses classiques, la conception des traitements cibles repose sur la comprehension des mecanismes moleculaires de la cancerogenese et de la machinerie proteique de la cellule cancereuse. Parmi ces nouveaux traitements, on distingue les inhibiteurs des voies de transmission des signaux de proliferation cellulaire et de neoangiogenese, les inducteurs de l'apoptose, les correcteurs de la regulation epigenetique, les stimulateurs de la reponse immunitaire et les vehicules cibles de drogues toxiques. La convergence des avancees en genomique et en biologie moleculaire du cancer, en developpement technologique et dans les traitements cibles ouvre la voie de la medecine personnalisee en cancerologie.

Abstract Objective: To evaluate the long-term impact on disease-free survival (DFS) and on overall survival (OS) of the sequential administration of docetaxel (D) following FEC100 among patients (pts) with node positive, operable breast cancer.Patients and Methods: Pts with localized, resectable, non pre-treated, unilateral breast cancer were randomly assigned to receive either Arm A: 6 cycles of FEC100 (5FU/epirubicin/cyclophosphamide 500/100/500 mg/m² day 1, every 3 weeks), or Arm B: 3 cycles of FEC100 followed by 3 cycles of docetaxel (D) 100 mg/m² (day 1, every 3 weeks). First chemotherapy cycle was to be started no more than 42 days after surgery. Radiotherapy was mandatory after conservative surgery and hormone therapy was given for 5 years if tumors were positive for at least one hormone receptor. Main inclusion criteria were: age < 65 years, at least one positive node, no metastasis and normal cardiac, hematologic and renal functions. The main end-point of this prospective, non blinded, randomized, multicentre phase III trial was the 5-year DFS. These results have been already published (Roché et. al. J Clin Oncol. 2006 24(36):5664-71). Since this first analysis, survival data have been updated.Results: Between June 1997 and March 2000, 1999 pts were recruited. Main pts characteristics were well balanced between the 2 arms: median age 50 years, conservative surgery 57%, grade III 39%, both HR negative 21%, both HR positive 60%, 1-3 involved nodes 62%. Treatment was completed for 95% and 93.4% of pts in arms A and B, respectively.As of 15 April 2009, with a median long term follow-up of 92.8 months, 639 pts have experienced at least one event: 124 loco regional relapses, 421 metastasis, 68 contra lateral breast cancer, and 26 deaths as first event. A total number of 71 second cancers and 383 deaths have been registered.8-year DFS rates were 65.8% with FEC and 70.2% with FEC-D. Cox regression analysis adjusted for age and number of positive nodes showed a 15% reduction in the relative risk of relapse with FEC-D (HR = 0.846 [95%CI 0.724 – 0.988], p=0.03).8-year OS rates were 78% with FEC and 83.2% with FEC-D. Cox regression analysis adjusted for age and number of positive nodes showed a 25% reduction in the relative risk of death with FEC-D (HR = 0.754 [95%CI 0.616 – 0.922], p=0.006).Conclusion: Initial report of 5-year benefit in DFS and OS with sequential administration of FEC followed by D are fully confirmed at 8 years. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 603.

Background: To determine the incidence of reversible amenorrhea in women with breast cancer undergoing adjuvant anthracycline- based chemotherapy with or without docetaxel.Methods: We studied the incidence and duration of amenorrhea induced by two chemotherapy regimens: ( i) 6 cycles of 5- fluorouracil 500 mg/ m(2), epirubicin 100 mg/ m2 and cyclophosphamide 500 mg/ m2 on day 1 every 3 weeks ( 6FEC) and ( ii) 3 cycles of FEC 100 followed by 3 cycles of docetaxel 100 mg/ m2 on day 1 every 3 weeks ( 3FEC/ 3D). Reversible amenorrhea was defined as recovery of regular menses and, where available ( 101 patients), premenopausal hormone values ( luteinizing hormone ( LH), follicle-stimulating hormone ( FSH) and estradiol) in the year following the end of chemotherapy.Results: One hundred and fifty- four premenopausal patients were included: 84 treated with 6FEC and 70 with 3FEC/ 3D. The median age was 43.5 years ( range: 28 - 58) in the 6FEC arm and 44 years ( range: 29 53) in the 3FEC/ 3D arm. Seventy- eight percent of patients were treated in the context of the PACS 01 trial. The incidence of chemotherapy- induced amenorrhea at the end of chemotherapy was similar in the two groups: 93 % in the 6FEC arm and 92.8 % in the 3FEC/ 3D arm. However, in the year following the end of chemotherapy, more patients recovered menses in the 3FEC/ 3D arm than in the 6FEC arm: 35.5 % versus 23.7 % ( p = 0.019). Among the 101 patients for whom hormone values were available, 43 % in the 3FEC/ 3D arm and 29 % in the 6FEC arm showed premenopausal levels one year after the end of chemotherapy ( p < 0.01). In the 3FEC/ 3D group, there was a statistically significant advantage in disease-free survival ( DFS) for patients who were still amenorrheic after one year, compared to patients who had recovered regular menses ( p = 0.0017).Conclusion: Our study suggests that 3FEC/ 3D treatment induces more reversible amenorrhea than 6FEC. The clinical relevance of these findings needs to be investigated further.

10517 Background: The objective of this study was to determine the incidence of reversible amenorrhea in women with early-stage breast cancer undergoing adjuvant anthracyclin-based chemotherapy with or without docetaxel. METHODS We studied the amenorrhea duration induced by 2 chemotherapy regimens. The first arm (6FEC) consisted in the administration of 6 cycles of FEC100 (5-Fluorouracil 500 mg/m2, Epirubicin 100 mg/m2, Cyclophosphamide 500 mg/m2, day 1, every 3 weeks) and the second arm (3FEC/3D) in the administration of 3 cycles of FEC100 followed by 3 cycles of Docetaxel (100 mg/m2, day 1, every 3 weeks). Some of these patients (N = 121, 78%) were treated in the PACS01 trial that was designed to evaluate the impact of the sequential addition of docetaxel to FEC100 on disease-free survival for patients with node positive, operable breast cancer. Other patients (N = 33.22%) were patients treated with the same regimens after the inclusion period of PACS01. Reversible amenorrhea was defined as a recovery of regular menses and -when available (101 patients)- premenopausal hormonal values (minimum 2 tests at 3 months interval) in the year following the end of chemotherapy infusion. RESULTS 154 pre-menopausal patients were included: 84 were treated in the 6FEC and 70 in the 3FEC/3D. Median age was 43.5 (28-58) and 44 (29-53), respectively. The incidence of chemotherapy-induced amenorrhea at the end of chemotherapy was not different between the two groups, 6FEC: 93% and 3FEC/3D: 92.5%. In the year following the end of chemotherapy, more patients recovered regular menses in the 3FEC/3D arm compared to the 6FEC arm: 35.5% (25/70) versus 23.7% (20/84) (p = 0.019). For the 101 for which hormonal values were available, more patients recovered premenopausal hormonal values in the 3FEC/3D arm: 42% (18/42) versus 29% (17/59) (p < 0.01). CONCLUSIONS Our study suggests that it could be interesting to record the menopausal status at different time points after the end of chemotherapy. In addition, the survival advantage in the PACS01 trial in favor of the 3FEC/3D arm was observed only for women over the age of 50 and not for women younger than 50. In this context, the clinical relevance of reversible amenorrhea warrants further investigation. No significant financial relationships to disclose.

Purpose The PACS 01 trial compared six cycles of fluorouracil, epirubicin, and cyclophosphamide (FEC) with a sequential regimen of three cycles of FEC followed by three cycles of docetaxel (FEC-D) as adjuvant treatment for women with node-positive early breast cancer.Patients and Methods Between June 1997 and March 2000, 1,999 patients with operable node-positive breast cancer were randomly assigned to either FEC every 21 days for six cycles, or three cycles of FEC followed by three cycles of docetaxel, both given every 21 days. Hormone-receptor-positive patients received tamoxifen for 5 years after chemotherapy. The primary end point was 5-year disease-free survival (DFS).Results Median follow-up was 60 months. Five-year DFS rates were 73.2% with FEC and 78.4% with FEC-D (unadjusted P = .011; adjusted P = .012). Multivariate analysis adjusted for prognostic factors showed an 18% reduction in the relative risk of relapse with FEC-D. Five-year overall survival rates were 86.7% with FEC and 90.7% with FEC-D, demonstrating a 27% reduction in the relative risk of death (unadjusted P = .014; adjusted P = .017). The incidence of grade 3 to 4 neutropenia, the need for hematopoietic growth factor, and incidence of nausea/vomiting were higher with FEC. Docetaxel was associated with more febrile neutropenia in the fourth cycle, stomatitis, edema, and nail disorders. Though rare overall, there were fewer cardiac events after FEC-D (P = .03), attributable mainly to the lower anthracycline cumulative dose.Conclusion Sequential adjuvant chemotherapy with FEC followed by docetaxel significantly improves disease-free and overall survival in node-positive breast cancer patients and has a favorable safety profile.

BACKGROUND : It is not known whether increasing the dose of filgrastim after mobilizing chemotherapy improves collection of peripheral blood progenitor cells (PBPC) and leads to faster hematopoietic engraftment after autologous transplantation. STUDY DESIGN AND METHODS : A randomized, open‐label, multicenter trial was carried out in patients with breast cancer, multiple myeloma, and lymphoma, in which patients were randomized to receive 5 or 10 μg per kg per day of filgrastim after standard chemotherapy to mobilize PBPCs. After high‐dose chemotherapy, the components from the first two leukapheresis procedures were returned, and all patients received 5 μg per kg day of filgrastim after transplantation. RESULTS : A total of 131 patients were randomized, of whom 128 were mobilized (Group A, 5 μg/kg, n = 66; Group B, 10 μg/kg, n = 62) and 112 were transplanted. Only six patients were not transplanted because of insufficient CD34+ cell numbers. The median number of CD34+ cells collected in the first two leukapheresis procedures tended to be higher in Group B than in Group A (12.0 vs. 7.2 × 10 6 /kg, NS), but after transplantation there was no significant difference in median times to platelet (9 days in both groups) or neutrophil (8 days in both groups) engraftment or the number of platelet transfusions (three in both groups). A subsequent subgroup analysis separating patients transplanted after first‐ or second‐line chemotherapy also showed no measurable impact of filgrastim dose on the median CD34+ cell yield or on platelet engraftment in either subgroup. CONCLUSION : PBPC mobilization with chemotherapy and 5 μg per kg of filgrastim is very efficient, and 10 μg per kg of filgrastim does not provide additional clinical benefit.

Utilization of chemotherapy for treatment of tumors is mainly limited by its hematological toxicity. Because of the low level expression of drug resistance genes, transduction of hematopoietic progenitors with multidrug resistance 1 (MDR1) or multidrug resistance-associated protein 1 (MRP1) genes should provide protection from chemotherapy toxicity. Successful transfer of drug resistance genes into hematopoietic cells might allow the administration of higher doses of chemotherapy and, therefore, increase regression of chemosensitive tumors. In addition, this approach can be used to select in vivo transduced cells by their enrichment after administration of cytotoxic drugs. Our group has studied the potential value of MRP1 to protect hematopoietic cells. The interest in the use of MRP1 as an alternative to MDR1 gene transfer for bone marrow protection lies in its different modulation. Indeed, classical P-gp reversal agents, tested in clinic to decrease MDR1 tumor resistance, have little or no effect on MRP1 function. This would allow, in the same patient, the use of reversal agents to decrease P-gp tumor resistance without reversing bone marrow protection of the transduced hematopoietic cells provided by MRP1. We constructed two different MRP1-containing vectors with either the Harvey retroviral long terminal repeat (LTR) or phosphoglycerate kinase (PGK) as promoters and generated ecotropic producer cells. MRP1 transduced fibroblasts were more resistant to doxorubicin, vincristine, and etoposide and their chemoprotection was increased after selection with chemotherapeutic agents in the presence of glutathione, a co-factor for MRP1 function. Lethally irradiated mice were engrafted with bone marrow (BM) cells transduced with MRP1 vectors (PGK promoter). We demonstrated that high expression of MRP1 in murine hematopoietic cells reduces doxorubicin-induced leukopenia and mortality. In addition, in vivo selection of MRP1-transduced BM cells was achieved following doxorubicin administration and allowed a better chemoprotection after the second chemotherapy cycle.

There is not yet a consensus on the reliability of the methods that should be used for the detection of rare disseminated tumor cells from non-hematological malignancies.In this review, we will discuss the advantage and drawbacks of the classical approach of immunocytochemistry and the molecular detection by reverse transcriptase polymerase chain reaction (RT-PCR).The interpretation of the biological significance of circulating tumor cells and the pitfalls of the detection techniques are the main causes of discrepancy between the conclusions of different tumor-cell detection (TCD) studies.

A total of 101 cancer patients with 121 febrile neutropenia episodes were randomised to receive empirical treatment with i.v. meropenem (1g/8 h) or ceftazidime (2 g/8 h). After 3 days, 89% of patients were on unmodified therapy in the meropenem group, compared with 83% in the ceftazidime group. Of the evaluable episodes (n = 106), the success rate with unmodified empirical therapy until the end of the treatment course was slightly higher with meropenem than with ceftazidime (48% vs 38%, P=0.39). Furthermore, initial success with further infections was observed in 22% of episodes treated with meropenem and in 13% of episodes treated with ceftazidime. Glycopeptides were used as first modification in 28% and 39% of meropenem and ceftazidime recipients, respectively. Both treatments were well tolerated and there were no reports of drug-related nausea/vomiting or seizures. No significant differences in response rate or in tolerability were observed when analysing only the first febrile episodes. In conclusion, meropenem seems to be as efficacious and well tolerated as ceftazidime and may be associated with a lesser requirement for the addition of glycopeptides.

The primary objective of this study was to evaluate the safety of infusion of CD34 + cells, selected using a clinical scale magnetically activated cell sorting device, assessed by time to hematological engraftment and incidence of adverse events. Secondary objectives included evaluation of device performance in terms of purity and recovery of the CD34 + cell product. Breast cancer patients suitable for transplantation received cyclophosphamide and filgrastim for mobilisation, followed by three leukaphereses. The products of the first two leukaphereses underwent CD34 + cell selection. The product of the third leukapheresis was cryopreserved unmanipulated. Following high-dose cyclophosphamide, thiotepa and carboplatin, selected CD34 + cells were infused. In 54 patients who received selected cells only, the median time to platelet recovery and neutrophil recovery was 11 days (range 5–51) and 9 days (range 5–51), respectively. There were no adverse events associated with infusion of selected cells. A total of 126 leukapheresis samples was available before and after selection for central CD34 + analysis. The median purity was 96.1% (27.4–99.4) and the median recovery was 52.3% (15.2–146.3). These data show that cells selected using magnetically activated cell selection provide safe and rapid engraftment after high-dose therapy. Bone Marrow Transplantation (2000) 25, 243–249.

We measured the concentration of CD34+ cells in peripheral blood (PB) ½ h prior to and ½, 1, 3, 6, and 12 h following hematopoietic stem cell (HSC) infusion in 34 breast cancer patients treated with high-dose chemotherapy (HDC). The decrease in these concentrations over time enabled us to determine the clearance kinetics of CD34+ cells from PB. The absolute number of CD34+ cells in PB generally peaked at ½ h after infusion, then rapidly declined from 1 to 3 h post infusion and continued to fall until 12 h post transplant, but more slowly. In univariate analysis, CD34+cells/kg infused, CFU-GM/kg infused, the CD34+ count at ½ h, and the 12-h clearance of CD34+ cells from PB were predictors of hematologic recovery, as were each of the two phases of clearance when the slope was divided into rapid and slow phases (from ½ to 3 and from 3 to 12 h post transplant, respectively). We then stratified our population by the number of CD34+ cells/kg infused. In group 1, patients received ⩽7.5 × 106 CD34+ cells/kg; in group 2, >7.5 × 106 CD34+ cells/kg. After adjusting for CD34+ cells injected, age, and purged or unpurged graft in multivariate analysis, the 12 h clearance remained a predictor of hematologic recovery in group 1. In addition, the second phase of clearance (from 3 to 12 h after infusion) was an even better predictor than the 12 h clearance. In group 2, however, no statistically significant correlation was observed, even with the number of HSC injected. Results suggest that rapidity of clearance of CD34+cells from PB is an independent indicator of hematologic recovery in patients receiving lower doses of CD34+ cells. When the cell dose injected is over a threshold, PB clearance correlations with hematologic recovery are masked.

Some heavily pretreated cancer patients fail to mobilize enough peripheral blood stem cells (PBSC) after stimulation with chemotherapy and hematopoietic growth factors. For these patients the best way to obtain an adequate PBSC collection is unknown. Here we report 6 heavily pretreated cancer patients who failed to mobilize sufficient PBSC after stimulation with chemotherapy and G-CSF 5 microg/kg/day. In these cases, we used G-CSF 10 microg/kg/day alone for six days at least 3 weeks after the last chemotherapy. After three consecutive leukaphereses starting on day 5, five patients had adequate PBSC collections. With 6 days of G-CSF 10 microg/kg/day alone, 2.8 x 10(6) (+/- 1) CD34+ cells/kg were collected. This was significantly higher than the number of CD34+ cells/kg collected after chemotherapy and G-CSF 5 microg/kg 0.3 x 10(6) (+/- 0.1) [P = 0.05]. Four patients received high-dose chemotherapy with PBSC support. Hematologic recovery observed in these patients was as expected. In conclusion, G-CSF 10 microg/kg alone can mobilize progenitor cells into peripheral blood when previous mobilization with chemotherapy and G-CSF 5 microg/kg fails.

In order to investigate the protein synthesis in megakaryocyte polyploidization, phorbol myristate acetate (PMA, 5 x 10(-9) M), a differentiation marker known to induce megakaryocyte polyploidization, was added to human megakaryocytic cell lines (DAMI, HEL and K562) and the expression of platelet/megakaryocytic integrins, the numbers of nucleolar organizer regions (AgNORs) and the total protein content were estimated. Following exposure of PMA, the expression of the platelet membrane glycoprotein GPIIIa and thrombospondin and transferrin receptors was augmented in the three cell lines. The number of AgNORs shifted from 16.4 +/- 4.3, 24.4 +/- 2.5 and 13.6 +/- 3.1 for unstimulated cells to 20.0 +/- 5.3, 38.7 +/- 7.9 and 16.8 +/- 2.3 for PMA-treated DAMI, HEL and K562 cells, respectively. Furthermore, after treatment with PMA, the numbers of AgNORs clusters or nucleoles increased significantly to 179%, 238% and 154% of controls in DAMI, HEL and K562 cell lines, respectively. Finally, addition of PMA culture for four days, significantly increased the protein contents to 153%, 171% and 254% of controls for DAMI, HEL and K562 cell lines, respectively (p < 0.05 by t-test). In conclusion, the increase in the total protein content and in the number of AgNORs by PMA, suggests that PMA-induced-megakaryocyte polyploidization occurs by enhanced protein production.