Background and objectives: The primary objective was to evaluate the benefit of training with virtual reality simulation. The secondary objective was to describe the short-term skill acquisition obtained by simulation training and to determine the factors affecting its magnitude. Materials and Methods: We prospectively performed a three-stage evaluation: face, constructive, and predictive to evaluate the training with a laparoscopic simulator with haptic feedback. The participants (n = 63) were divided according to their level of experience into three groups: 16% residents; 46% specialists and 38% were consultants. Results: Face evaluation demonstrates the acceptance of the design and realism of the tasks; it showed a median score of eight (IQR 3) on a Likert scale and 54% of participants (n = 34) gave the tissue feedback a moderate rating. Constructive evaluation demonstrates the improvement of the participants in the training session and the ability of the designed task to distinguish the experienced from the inexperienced surgeon based on the performance score, at task I (transfer of pegs) and II (laparoscopic salpingectomy). There was an improvement in both tasks with a significant increase in score and reduction in time. The study showed that those with a high score at the pre-test recorded a high score post-test, showing a significant pair-wise comparison (Z) and correlation (p) showing a significant statistical significance (p < 0.001). The predictive evaluation demonstrates the beneficiary effect of training four weeks afterward on the practice of surgeons addressed with five questions. It showed an improvement regarding implementation into daily routine, performance of procedure, suturing, shortening of the operative time, and complication management. Conclusions: Virtual reality simulation established high ratings for both realism and training capacity, including clinical relevance, critical relevance, and maintaining training enthusiasm.

EPIDEMIOLOGY:Vulvar cancer can be classified into two groups according to predisposing factors: the first type correlates with a HPV infection and occurs mostly in younger patients. The second group is not HPV associated and occurs often in elderly women without neoplastic epithelial disorders.HISTOLOGY:Squamous cell carcinoma (SCC) is the most common malignant tumor of the vulva (95%).CLINICAL FEATURES:Pruritus is the most common and long-lasting reported symptom of vulvar cancer, followed by vulvar bleeding, discharge, dysuria, and pain.THERAPY:The gold standard for even a small invasive carcinoma of the vulva was historically radical vulvectomy with removal of the tumor with a wide margin followed by an en bloc resection of the inguinal and often the pelvic lymph nodes. Currently, a more individualized and less radical treatment is suggested: a radical wide local excision is possible in the case of localized lesions (T1). A sentinel lymph node (SLN) biopsy may be performed to reduce wound complications and lymphedema.PROGNOSIS:The survival of patients with vulvar cancer is good when convenient therapy is arranged quickly after initial diagnosis. Inguinal and/or femoral node involvement is the most significant prognostic factor for survival.

Zielsetzung: Untersucht wurde die Kombination von Imatinib und Vinorelbine auf metastasierte Mammakarzinome und heilende Hautwunden (in vivo) in einer Phase I/II Dosiseskalationsstudie. Genexpression und Expression von KIT, PDGF-R-α/-β im Tumorgewebe wurden mit dem klinischen Ansprechen korreliert. Neoangiogenese und Immunantworten in Hautwundenassays wurden gemessen. Materialien und Methoden: 22 Tumorproben wurden auf GIST-typische Mutationen untersucht, die prädiktiv für das Ansprechen auf Imatinib sind. Zusätzlich sollte der Rezeptorstatus (PDGF-R-α-/-β, c-Kit) bestimmt und mit dem Therapieansprechen korreliert werden. Zudem wurde in Hautwunden Assays mit und ohne Imatinib 7 Tage altes Granulationsgewebe in Form von Hautstanzen (Oberarm) entnommen. Bestimmt wurden Gefäßdichte, Endothelzellkinetik und Gefäßreife. Ergebnisse: GIST-typische Mutationen zeigten sich im untersuchten Studienkollektiv nicht, sodass keine prädiktiven Aussagen für das Imatinibansprechen zu machen sind. Aufgrund kleiner Fallzahl ergibt sich für c-Kit, PDGF-R-α-/–β im Tumorgewebe keine Korrelation zwischen Rezeptorexpression oder ggf. nachgewiesener Rezeptormutationsart und klinischem Ansprechen. Die Immunreaktion von CD 4-, CD 8- oder CD 68-Zellen in 7 Tage altem Granulationsgewebe wird durch Imatinib nicht beeinflusst. Die Triple-Immunfluoreszenzfärbungen zeigen, dass Imatinib keinen negativen Einfluss auf die Gefäßquantität (Kapillaranzahl) und Gefäßintegrität hat. Allerdings ist die mittlere Anzahl Ki-67-positiver Gefäßendothelzellen nach Imatinibeinnahme verringert. Zusammenfassung: Prädiktive Mutationen konnten nicht nachgewiesen werden. Auch korrelierten Rezeptorexpression und Therapieansprechen nicht miteinander. Somit wurde kein prädiktiver Marker gefunden. Imatinib hat in dieser Studie keinen negativen Einfluss auf die Immunantwort der CD 4,- 8- und 68-Zellen im Granulationsgewebe der Hautwunden. Die Angiogenese ist in ihrer Qualität und Quantität nicht negativ durch Imatinib beeinträchtigt. Lediglich eine leichte Verminderung proliferativer Endothelzellen der Gefäße ist zu verzeichnen.

BACKGROUND Fibroblast growth factor-2 (FGF-2) supports tumor progression in breast cancer. FGF-2 signaling is modulated by heparan sulfate proteoglycans, such as syndecan-1 (CD138). The exact role of CD138 in ductal carcinoma in situ of the breast (DCIS) is still uncertain. Differential expression depending on grading could suggest a role for syndecan-1 during growth and tumor progression. MATERIALS AND METHODS Samples of 127 cases of breast DCIS associated with follow-up data were included. CD138 staining intensity, number of positive cells, intracellular and tissue localization were examined. RESULTS Median follow-up was 45.4 months and median recurrence-free survival (RFS) 86 months. Age, menopausal status and previous hormone replacement therapy had no significant influence on RFS. Smoking significantly influenced RFS (p=0.008). Endocrine therapy or radiotherapy did not improve RFS. Grading was not correlated with CD138 staining intensity, but was significantly associated with the percentage of CD138-positive cells (low-vs. high-grade, p=0.043). Estrogen receptor (ER) expression did not influence staining intensity of CD138 (p=0.247), but negatively correlated with the proportion of CD138-positive cells (p=0.032). Progesterone receptor (PR) expression significantly influenced the intensity of staining (p=0.010) and the percentage of CD138-positive cells (p=0.004); both were increased in PR-negative cases. CD138 staining intensity and percentage of positive cells did not correlate with RFS. Nuclear grade and syndecan-1 staining localization were significantly associated (p=0.001). ER-positive, and PR-positive DCIS more often exhibited membrane-bound syndecan-1 than ER- or PR-negative cases (p=0.001). Nuclear grade and tissue localization of CD138 correlated significantly (p=0.005). PR influenced CD138 tissue distribution, while ER did not. Syndecan-1 localization did not statistically impact RFS. CONCLUSION In DCIS of different nuclear grades, tissue localization of syndecan-1 is significantly divergent, suggesting a specific effect on biology and progression of DCIS.

Background: Imatinib is a tyrosine kinase inhibitor of BCR-ABL, ABL, PDGFR-α and -β, KIT, and DDR. In solid tumors, it inhibits proliferation and invasiveness and facilitates higher intratumoral cytotoxic drug concentrations. Vinorelbine has good tolerability and efficacy in metastatic breast cancer (MBC). This study evaluates the safety and efficacy of imatinib and vinorelbine in combination. Methods: In a prospective, open-label, phase I/II trial, 400 mg imatinib p.o. daily (corrected from 600 mg) was combined with an escalating dose of vinorelbine i.v. weekly in four dose levels of 10, 15, 20, and 25 mg/m2 (each n ≥ 5) to treat patients with MBC (expressing PDGFR-α and/or -β, and/or KIT). The last patient of each level was treated for >28 days, before enrolment for the next dose level started. Study endpoints were feasibility and tolerability, incidence of hematological and nonhematological toxicity, and clinical efficacy (data cutoff: November 18, 2011). A total of 33 patients have been enrolled, and all dose levels have been fully recruited. One patient is still on study medication. A translational subprotocol is ongoing. Results: All 33 included patients are evaluable for safety (32 within the ITT population). Eleven patients were excluded early from the study (progressive disease, toxicity, and withdrawal of consent). Twenty-two patients participated in the study for >28 days (‘ITT >28'). Within the ITT population, the response rate [complete response (CR) and partial response (PR)] was 9.4% (n = 3), the clinical benefit rate (CBR; CR+PR+stable disease) 50% (n = 16), and the median time to progression (TTP) 155 days. A total of 21.3% of the patients were on study medication for >6 months, and 15.2% for >12 months (mean 140 days, range 15-643). Within ‘ITT >28', the response rate was 13.6%, CBR 72.7%, and median TTP 176 days. The response was independent of the receptor status (PDGFR-α, -β, and KIT). Toxicities were as follows (safety population): 21.6% severe leukopenia, 9.1% severe neutropenia (with 1 febrile neutropenia), 1 case of bowel perforation, 36% diarrhea (3% severe), 84.8% nausea (severe 15.2%), 48.5% vomiting (severe 9.1%), 27.3% infections (severe 6.1%), 12.1% peripheral neuropathy (severe 9.1%), and 36.4% dyspnea (3% severe). Four patients on trial died (nondrug-related). Conclusion: The combination of imatinib and vinorelbine in MBC appeared to be feasible and tolerable. A CBR of 50% (ITT) in pretreated patients suggests that this combination may be active. Although toxicities were frequent, they appeared to be manageable.

Objective: To investigate trends in the performance of hysterectomy at a single certified endoscopic teaching center. Methods: Data were collected retrospectively from 953 patients who underwent hysterectomy between 2002 and 2010 for benign indications at UKSH, Germany. Preoperative risk scores were assigned to patients. Results: The most frequent indications for hysterectomy were uterine myoma, adenomyosis, prolapse, endometrial hyperplasia, menstrual disorders, and endometriosis. The shortest operating time was recorded for vaginal hysterectomy (VH) and the longest for laparoscopically assisted (LAVH). The average uterine weight was highest for abdominal hysterectomy (AH) and lowest for VH. The major postoperative complication rate was 11.8% for laparoscopic supracervical hysterectomy (LSH) and 23.5% for AH. The highest intraoperative complication rate occurred with AH (46.4%) and the lowest with total laparoscopic hysterectomy (TLH; 3.6%). The minor postoperative complication rate was 5.9%. The mean preoperative score was 1.09 +/- 1.51 for AH, 0.75 +/- 0.96 for VH, 1.04 +/- 1.30 for LSH, 1.0 +/- 1.40 for LAVH, and 1.38 +/- 152 for TLH. Conclusion: Laparoscopic hysterectomies have become more common and were associated with decreased complication rates, despite the higher preoperative risk score of these patients. (C) 2014 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

PURPOSE:Nilotinib is a selective tyrosine kinase inhibitor of c-Kit, Abl and platelet-derived growth factor receptor-α/β. To evaluate nilotinib's potential use as a treatment of human ovarian cancer, we tested nilotinib's preclinical activity in ovarian cancer cell lines with different tyrosine kinase expression patterns.METHODS:The effects of nilotinib on ovarian cancer cell growth were studied alone and in combination with carboplatin and paclitaxel. Proapoptotic and antimigratory effects were examined using TUNEL and migration assays.RESULTS:Nilotinib alone and in combination with carboplatin and paclitaxel significantly inhibited cell growth in PDGFR-α-positive ovarian cancer cell lines. The combination of nilotinib with carboplatin and paclitaxel showed synergistic effects on cell proliferation. Nilotinib treatment led to the inhibition of cell migration alone and in combination with carboplatin and paclitaxel. Apoptosis induction occurred in response to nilotinib that increased in combination with carboplatin.CONCLUSIONS:Nilotinib may be a feasible targeted therapy option for the treatment of ovarian cancer.

BACKGROUNDWe have found that the platelet-derived growth factor receptor (PDGFR)/Abl signaling pathway is up-regulated as a determinant of the acquisition of resistance to estrogen deprivation in vitro. We aimed to determine its clinical relevance in aromatase inhibitor (AI)-resistant breast cancer.PATIENTS AND METHODSWe identified a cohort of 45 patients with estrogen receptor-positive breast cancer who had been treated with an AI, subsequently relapsed and had biopsy material available from both the presentation and post-AI recurrent lesion. PDGFRα, PDGFRβ and Abl expression was assessed in formalin-fixed paraffin-embedded sections.RESULTSTumor protein expression of PDGFRα (1.39-fold, P=0.0065), PDGFRβ (4.32-fold, P=0.006) and Abl (1.8-fold, P=0.001) was increased at the point of relapse. Tumor and stromal expression of PDGFRα as well as PDGFRβ was significantly correlated in pre-treatment and relapse samples. High post-treatment tumor and stromal PDGFRβ levels were associated with a short time to treatment failure (TTF). Expression of PDGFRα in relapsing tumor specimens was correlated with Abl expression and Ki67 levels. Furthermore, changes in Abl correlated significantly with changes in ER expression.CONCLUSIONSThese clinical data support a role for enhanced PDGF/Abl signaling in AI-resistant disease and provide a rationale for targeting the pathway in endocrine-resistant breast cancer.

Local treatment of breast cancer with tumor-free surgical margins is the standard procedure in the treatment of T1 and small T2 breast cancers. Surgery is followed by radiation therapy, and adjuvant systemic therapy is offered depending on primary tumor characteristics, such as tumor size, grade of differentiation, number of involved axillary lymph nodes, the status of estrogen (ER) and progesterone (PR) receptors, and the expression of the human epidermal growth factor 2 (HER2) receptor. Although this approach implies a higher risk of ipsilateral breast tumor recurrence, the total risk of recurrence is low (1% per year), with rates of overall survival similar to that after radical procedures. The most peripheral part of epithelial tumors, the tumor margin, is the part which is most likely to remain in loco after surgical resection. Thus, understanding the biology of the invasion front is important as these tumor cells have been reported to lose epithelial properties, such as cohesiveness and keratin expression, and to acquire features of mesenchymal cells. The parallel appearance of tumor cells in different states of cell dedifferentiation implicates a dynamic equilibrium that is determined by the induction of epithelial-mesenchymal transition (EMT). EMT has been suggested to be of prime importance for tissue and vessel invasion. Furthermore, features of EMT are associated with the activity of tumor stem cells (TSC). TSC exist in breast cancer and their appearance varies depending on the used marker profile. Consequently, intratumoral heterogeneity is reflected by the grade of EMT activation. A specific function at the invasion front is hypothesized but has not yet been proven. Nevertheless, the molecular differentiation between the tumor center and the invasion front enhances the importance of tumor-free surgical margins.

INTRODUCTION:Strategies to improve the efficacy of endocrine agents in breast cancer (BC) therapy and to delay the onset of resistance include concomitant targeting of the estrogen receptor alpha (ER) and the mammalian target of rapamycin complex 1 (mTORC1), which regulate cell-cycle progression and are supported by recent clinical results.METHODS:BC cell lines expressing aromatase (AROM) and modeling endocrine-sensitive (MCF7-AROM1) and human epidermal growth factor receptor 2 (HER2)-dependent de novo resistant disease (BT474-AROM3) and long-term estrogen-deprived (LTED) MCF7 cells that had acquired resistance associated with HER2 overexpression were treated in vitro and as subcutaneous xenografts with everolimus (RAD001-mTORC1 inhibitor), in combination with tamoxifen or letrozole. End points included proliferation, cell-cycle arrest, cell signaling, and effects on ER-mediated transactivation.RESULTS:Everolimus caused a concentration-dependent decrease in proliferation in all cell lines, which was associated with reductions in S6 phosphorylation. Everolimus plus letrozole or tamoxifen enhanced the antiproliferative effect and G1-accumulation compared with monotherapy, as well as increased phosphorylation (Ser10) and nuclear accumulation of p27 and pronounced dephosphorylation of Rb. Sensitivity was greatest to everolimus in the LTED cells but was reduced by added estrogen. Increased pAKT occurred in all circumstances with everolimus and, in the BT474 and LTED cells, was associated with increased pHER3. Decreased ER transactivation suggested that the effectiveness of everolimus might be partly related to interrupting cross-talk between growth-factor signaling and ER. In MCF7-AROM1 xenografts, letrozole plus everolimus showed a trend toward enhanced tumor regression, versus the single agents. In BT474-AROM3 xenografts, everolimus alone was equally effective at reducing tumor volume as were the combination therapies.CONCLUSIONS:The results provide mechanistic support for recent positive clinical data on the combination of everolimus and endocrine therapy, as well as data on potential routes of escape via enhanced HER2/3 signaling. This merits investigation for further improvements in treatment efficacy.

INTRODUCTION:The majority of breast tumors at primary diagnosis are estrogen receptor positive (ER+). Estrogen (E) mediates its effects by binding to the ER. Therapies targeting the estrogenic stimulation of tumor growth reduce mortality from ER+ breast cancer. However, resistance remains a major clinical problem. METHODS:To identify molecular mechanisms associated with resistance to E-deprivation, we assessed the temporal changes in global gene expression during adaptation to long-term culture of MCF7 human breast cancer cells in the absence of estradiol (E2), long term estrogen deprived (LTED), that leads to recovery of proliferative status and models resistance to an aromatase inhibitor (AI). The expression levels of proteins were determined by western blotting. Proliferation assays were carried out using the dual platelet derived growth factor receptor (PDGFR)/Abelson tyrosine kinase (Abl) inhibitor nilotinib. Luciferase reporter assays were used to determine effects on ER-mediated transactivation. Changes in recruitment of cofactors to the gene regulated by estrogen in breast cancer 1 (GREB1) promoter were determined by chromatin immunoprecipitation (ChIP). Gene expression data were derived from 81 postmenopausal women with ER+ BC pre-treatment and at two-weeks post-treatment with single agent anastrozole in a neoadjuvant trial. RESULTS:The PDGF/Abl canonical pathway was significantly elevated as early as one week post E-deprivation (P = 1.94 E-04) and this became the top adaptive pathway at the point of proliferative recovery (P = 1.15 E-07). Both PDGFRβ and Abl protein levels were elevated in the LTED cells compared to wild type (wt)-MCF7 cells. The PDGF/Abl tyrosine kinase inhibitor nilotinib, suppressed proliferation in LTED cells in the presence or absence of E. Nilotinib also suppressed ER-mediated transcription by destabilizing the ER and reducing recruitment of amplified in breast cancer-1 (AIB1) and the CREB binding protein (CBP) to the promoter of the E-responsive gene GREB1. High PDGFRβ in primary ER+ breast cancer of 81 patients prior to neoadjuvant treatment with an AI was associated with poorer antiproliferative response. Additionally PDGFRβ expression increased after two weeks of AI therapy (1.25 fold, P = 0.003). CONCLUSIONS:These preclinical and clinical data indicate that the PDGF/Abl signaling pathway merits clinical evaluation as a therapeutic target with endocrine therapy in ER+ breast cancer.

Objectives: Endometriosis is a common gynaecological disease with clinical symptoms such as chronic pain, infertility and intra-abdominal adhesions. Different theories on the pathogenesis of endometriosis and especially its aggressive subtype with infiltrative growth have been discussed. The objective of this study is to evaluate differences in proliferation and invasive properties of invasive colorectal endometriosis, superficial peritoneal endometriosis and endometrial carcinoma (G1 and G2).Study design: Paraffin embedded tissues of peritoneal endometriosis, endometriosis of the intestine and endometrial carcinoma from 97 patients were stained immunohistochemically to assess differences in expression patterns of matrix metalloproteinases (MMP-2, MMP-9) as markers of invasion and the marker of proliferation PCNA. MMP expression was evaluated using the Immuno Reactive Score (IRS) (combining positive cell ratio and staining intensity) and PCNA expression was assessed as the percentage of positively stained cells in representative areas.Results: MMP-2, MMP-9 and PCNA showed differential expression patterns in the different tissues examined. MMP-2 and PCNA expression was stronger in invasive colorectal endometriosis than in superficial peritoneal endometriosis (p = 0.0394). MMP-9, however, was more frequently expressed in peritoneal endometriosis (59.1%) than in colorectal endometriosis (44.4%). This result did not reach statistical significance. When colorectal endometriosis was compared to low grade endometrial carcinoma, proliferation detected by PCNA was significantly higher in endometriosis (p = 0.0008). MMP-2 and MMP-9 showed higher expression in endometrial carcinoma than in endometriosis.Conclusions: There are obvious differences in expression patterns of MMP-2. MMP-9 and PCNA in different stages of endometriosis and in endometrial cancer. These markers can be helpful to evaluate aggressiveness and invasiveness of endometriosis in different localizations. The results obtained could be of relevance for a better understanding of the pathogenesis of endometriosis and the development of an individual therapy concept. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

e13503 Background: Imatinib is a tyrosine kinase inhibitor of bcr-abl, PDGF-R, SCF, c-Kit and abl. In solid tumors it inhibits proliferation and facilitates higher intratumoral cytotoxic drug concentrations. Vinorelbine has good tolerability and efficacy in MBC. This study evaluates the combination of imatinib and vinorelbine. Methods: In aprospective open-label, phase I/II trial 400 mg imatinib p.o. daily (amended from 600 mg) was combined with an escalating dose of vinorelbine i.v. weekly in four dose levels with 10, 15, 20, 25 mg/m² (each n ≥ 5) for pats. with MBC (which express PDGF-R-α and/or -β and/or c-kit). The last pat. of a level was treated > 28 days, before enrolment for the next dose level started. Study endpoints were feasibility and tolerability, incidence of hematological and non-hematological toxicity and clinical efficacy (data cut: 18/11/2011). A translational subprotocol is ongoing. 33 pats. have been enrolled; all dose levels have been fully recruited. 1 patient is still on study medication. Results: 32 pats. are evaluable (ITT population). 11 pats. went off study early (progressive disease, toxicity and withdrawal of consent). 22 pats. were on study >28 days (“ITT>28”). Within the ITT population the response rate (complete (CR) and partial response (PR)) was 9.4% (n=3), the clinical benefit rate CBR (CR+PR+stable disease) 50% (n=16), median time to progression (TTP) 155 days. 21.3% were on study medication >6 months, 15.2% > 12 months (mean 133 days, 15-617 days). Within “ITT>28,” the response rate was 13.6%, CBR 72.7% and median TTP 176 days. Toxicities (ITT population): 21.6% severe leukopenia, 9.1% severe neutropenia (with 1 febrile neutropenia), 1 case of bowel perforation, 36% diarrhea (3% severe), 84.8% nausea (severe 15.2%), 48.5% vomiting (severe 9.1%), 27.3% infections (severe 6.1%), 12.1% periph. neuropathy (severe 9.1%) and 36.4% dyspnea (3% severe). 1 patient on study medication died (non drug related). Conclusions: The combination of imatinib and vinorelbine in MBC appeared to be feasible and tolerable. A CBR of 50% (ITT) in pre-treated pats. proves the efficacy of this combination. Although toxicities were frequent, they appeared to be manageable.

Abstract Background Imatinib mesylate is a tyrosine kinase inhibitor which originally had been developed to block pathognomonic bcr-abl oncoprotein in chronic myeloic leukemia. It is also an inhibitor of the receptor tyrosine kinases of platelet-derived growth factor (PDGF) and stem cell factor (SCF), c-Kit, and ab1. Therefore it also inhibits PDGF- and SCF-mediated cellular events, as tumor cell proliferation in solid tumors. Additionally, inhibition of PDGF- b-receptor relieves tumor hypertension in solid tumors and thereby helps to deliver higher concentrations of cytotoxic drugs into the tumor cell. Vinorelbine is a semisynthetic vincaalcaloid with antitumor activity. As a single agent and as a combination partner of other cytotoxic drugs, it has proven good tolerability and high effectiveness in treatment of patients with metastatic breast cancer (MBC). This study was designed to evaluate the feasability of the combination of imatinib mesylate and the cytotoxic drug of vinorelbine in different concentrations in patients with advanced and MBC. Study design: This is a prospective open-label, single arm phase I/II study combining 400 mg Imatinib Mesylate p.o. daily (amended from 600 mg) with an escalating dose of vinorelbine i.v. weekly for patients with locally advanced or metastatic breast cancer. Patients must have tumors expressing PDGF-receptor-α and/or -β and/or c-kit and have received pretreatment with an anthracyclin containing regimen. Main study endpoints are feasibility and tolerability of this novel combination. In addition to the total toxicity and the incidences of hematological and non-hematological toxicity of grade 3 and 4, the clinical activity together with the clinical response rate and the time to disease progression will be evaluated.The quality of life is examined during the whole course of treatment. Recruitment started for level one with 10 mg/m2 vinorelbine and proceeds to the next higher levels 15, 20 and 25 mg/m2. Each dose level will be filled with at least five patients. Patients of one dose level will be followed for a minimum of 28 days during therapy, before enrolment of patients for the next dose level can start. For translational research multiple skin biopsies will be taken (skin wounding assays). When feasible tumor-biopsies are taken before and during therapy. Expression of tyrosine kinase receptors (c-kit and PDGF-receptor) will be correlated with treatment response. Present accrual and target accrual: 33 patients have been enrolled into the study. Dose levels I-III have been fully recruited. In dose level IV two patients with visceral metastasis are ongoing on study medication. Safety and clinical data will be available after the last patient discontinues study treatment. Information on the study protocol, the translational subprotocol and details on c-kit and PDGF-receptor expression analysis will be presented at the meeting. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr OT3-01-02.

Objective: Many patients with ovarian cancer disease relapse within 6 months after adjuvant chemotherapy, with a limited prognosis. Epigenetic modifications have been shown to play an important role in tumor development and formation. Therefore, global analysis of DNA methylation patterns might reveal specific CpG sites that correlate with progression-free interval (PFI) after therapy. Methods: Twenty samples of advanced ovarian cancer with a predominantly serous papillary histological subtype were subjected to DNA methylation profiling. Illumina HumanMethylation27 BeadChip technology was used for simultaneous analysis of 27,578 CpG sites in >14,000 genes. Results: Differential DNA methylation of various cytosines correlated with PFI. However, this becomes only significant by classification according to PFI with a cutoff of >28 months. Longer survival was associated with hypomethylation at specific CpG sites (e.g. GREB1, TGIF and TOB1) and hypermethylation in other genes (e.g. TMCO5, PTPRN and GUCY2C). Gene ontology analysis revealed that differentially methylated genes were significantly overrepresented in the categories telomere organization, mesoderm development and immune regulation. Conclusion: Epigenetic modifications at specific CpG sites correlate with PFI in ovarian cancer. Therefore, such analysis might be of prognostic value.

Abstract Aim: To determine the relevance of PDGF/Abl signaling pathway as a therapeutic target in endocrine resistance in vitro and in vivo. Rationale: Targeting estrogenic stimulation reduces mortality from ER-positive (ER+) breast cancer (BC) but resistance remains a major clinical problem. To identify the molecular mechanisms associated with resistance to estrogen-deprivation, we previously assessed the temporal changes in gene expression during adaptation to long-term culture of MCF7 human breast cancer cells in the absence of estradiol (E2) (LTED), modeling resistance to an aromatase inhibitor (AI). Platelet-derived growth factor (PDGF)/Abl signaling was the top adaptive pathway at the point of resistance (p=1.15 E-07), but there is limited evidence for it playing a role clinically. Methods: Gene expression data from postmenopausal women with ER+ BC at pre-treatment and at 2-week on-treatment with neoadjuvant anastrozole were interrogated. Immunohistochemical staining using antibodies against PDGFRα, β and Abl was assessed in paired clinical specimen of 45 patients who had received an AI and presented progressive disease. Proliferation assays, immunoblots and ChIP assays were carried out using either the PDGFR/Abl inhibitor nilotinib or siRNA knockdowns in LTED and wild-type MCF7 cells. Results: In vitro: PDGFRβ and Abl expression and phosphorylation were elevated in LTED cells. Cell proliferation was decreased in LTED cells by siRNA knock-down of either PDGFRβ (50% p<0.01), Abl (40% p<0.01) or the combination (70% p<0.01). Inhibition of Abl activity using nilotinib decreased ER protein levels and suppressed ER-mediated transcription by reducing recruitment of AIB1 and CBP to the promoter of E-responsive genes TFF1 and GREB1. Clinical: Data from 81 patients treated with an AI in the neoadjuvant setting, showed increases in PDGFRβ gene expression after two weeks (1.25 fold, p=0.003). Low PDGFRβ at baseline was associated with a better response. Of note, paired pre-AI-treatment and relapse clinical specimens from a cohort of patients treated in the primary or advanced setting revealed increases in tumor protein expression of PDGFRα (1.39 fold, p=0.0065), PDGFRβ (4.32 fold, p=0.006) and Abl (1.8 fold, p=0.001) at the point of relapse. Tumor (T) and stromal (S) expression of PDGFRα as well as PDGFRβ was significantly correlated in pre-treatment and relapse samples (Table 1). High post-treatment tumor and stromal PDGFRβ levels were associated with a short time to treatment failure (TTF) (T: Rs −0.284, p=0.066; S: Rs −0.307, p=0.046). Expression of PDGFRα in relapsing tumor specimens was correlated with Abl expression (Rs 0.342, p=0.027) and Ki67 levels (Rs 0.39, p=0.01). Furthermore, changes in Abl correlated significantly with changes in ER expression (Rs 0.307, p=0.048). Discussion: These in vitro and clinical data support a biological interaction between PDGF/Abl and ER-signaling and suggest that the PDGF signaling pathway warrants clinical evaluation as a therapeutic target in endocrine resistant breast cancer. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-01.

La presente invention concerne des pyrimidylaminobenzamides de formule (I) et d'autres inhibiteurs des recepteurs PDGF-R adequats, seuls ou associes avec au moins un inhibiteur de l'aromatase, utilises pour traiter le cancer du sein hormono-resistant; l'invention concerne egalement un procede de traitement des animaux a sang chaud, notamment les etres humains, souffrant du cancer du sein hormono-resistant en administrant audit animal le necessitant une dose efficace d'un pyrimidylaminobenzamide de formule (I), seul ou associe a un inhibiteur de l'aromatase.

Aromatase inhibitors (AI) have improved the treatment of oestrogen receptor positive (ER+) breast cancer. Despite the efficacy of these agents over 40% of patients relapse with endocrine resistant disease. Here we describe an in vitro model of acquired resistance to long-term oestrogen deprivation (LTED). The LTED cells retain expression of the ER and appear hypersensitive to oestrogen as a result of altered kinase activity. Furthermore analysis of temporal changes in gene expression during the acquisition of resistance highlight growth factor receptor pathways as key mediators of this adaptive process.