bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of phosphoserine phosphatase-like in breast disease in African American women.
Authors: Field LA , Love B , Kane J , Deyarmin B , Hooke JA , Ellsworth RE , Shriver CD
Published: December 2010

Abstract #2095

Background: Although breast cancer incidence is higher among Caucasian women (CW), mortality is higher in African American women (AAW). AAW also tend to be diagnosed at a younger age and with larger and more aggressive tumors. Previously, we have found that gene expression patterns in AAW and CW differ in both breast tissues from women without breast disease as well as those from women with invasive breast cancer. Characterization of these differentially expressed genes is critical to understanding breast pathogenesis in AAW.  Methods: Total RNA was isolated from breast tissue from 29 AAW and 26 CW without evidence of breast disease using the RNeasy Lipid Tissue Midi kit (Qiagen) and from laser microdissected primary breast tumors from 23 AAW and 24 CW using the RNAqueous Micro kit (Ambion). Based on earlier microarray results, genes were selected for validation by quantitative real-time PCR using TaqMan Gene Expression Assays (Applied Biosystems). All TaqMan assays were performed on duplicate samples using the iCycler Real-Time Detection System (Bio-Rad). Relative expression levels were determined using the Comparative Ct Method. A Wilcoxon test was used to determine significance.  Results: Among those genes that were confirmed to be differentially expressed in breast tissues from AAW and CW was phosphoserine phosphatase-like, or PSPHL. PSPHL showed significantly higher expression in both disease-free samples as well as primary tumors from AAW when compared to the same sample type from CW. In tissues from disease-free women, the relative level of PSPHL expression was significantly higher in AAW (median = 17.15) compared to CW (median = 0; P<0.000001). Similarly, primary tumors from AAW also had higher levels of PSPHL expression (median = 42.22) compared to those from CW (median = 0; P<0.0005).  Discussion: The current study has confirmed that expression of PSPHL is significantly higher in both disease-free and invasive breast tissues from AAW than those from CW. Recently, PSPHL has also been shown to have higher expression in prostate tumors from African American men than those from Caucasian men. The function of PSPHL is unknown; however, its increased expression has been associated with aberrant cellular proliferation and cell cycle progression in Fanconi anemia. Higher expression of PSPHL in both normal and diseased breast tissues may, therefore, increase cellular proliferation, contributing to the initial development and/or aggressiveness of breast tumors in AAW.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2095.
Tolerability of lapatinib given concurrently with paclitaxel and trastuzumab as part of adjuvant therapy in patients with resected HER2+ breast cancer: initial safety data from the Mayo Clinic cancer research consortium trial RC0639.
Authors: Johnson BS , Dueck AC , Dakhil SR , Stella PJ , Nikcevich DA , Franco SX , Wender DB , Schaefer PL , Colon-Otero G , Diekmann BB , Perez EA
Published: December 2010

Abstract #2109

Background: Despite the impressive results of the recently released trastuzumab adjuvant therapy trials, 15% of patients with HER2 overexpressing or amplified breast cancer developed tumor relapse at 4 years. Lapatinib is a small molecule reversible TKI that inhibits both ErbB1 and ErbB2. The current study was developed to assess the cardiac safety and feasibility of adding lapatinib to paclitaxel and trastuzumab following as part of adjuvant therapy.  Methods: A single-arm phase II study of doxorubicin (A, 60 mg/m2 day 1) and cyclophosphamide (C, 600 mg/m2 day 1) [q2w or q3w for 4 cycles]; followed by paclitaxel (P, 80 mg/m2 days 1, 8, 15), trastuzumab (T, 4 mg/kg loading dose then 2 mg/kg days 1, 8, 15), and lapatinib (L, 1000 mg days 1-21) [12 weeks]; followed by T (6 mg/kg day 1) and L (1000 mg days 1-21) [40 weeks] was conducted. The primary endpoint was the incidence of congestive heart failure. The current unplanned safety analysis was undertaken due to the observance of a high rate of G3/4 diarrhea.  Results: From April 2007 to June 2008, 98 pts were enrolled and initiated study treatment. Median age was 51 (range 32-72). Among 83 pts with adverse event (AE) data available, 50 (60%) pts have experienced a G3/4 non-hematologic AE. During post-AC treatment, among 53 pts with AE data available, 31 (58.5%) patients have experienced a G3/4 non-hematologic AE with 24 (45%) patients reporting G3/4 diarrhea. Median cycle of onset of G3/4 diarrhea was cycle 5 (first cycle of PTL) with 16 (64%) cases first reported during cycle 5 and 5 (20%) cases first reported during cycle 6. Among 57, 46, 38, and 32 pts receiving treatment with PTL during cycles 5-8, 65%, 57%, 61%, and 72% of patients received the full L dose, respectively. 31 patients have ended active treatment with 10 due to patient refusal and 8 due to adverse events.  Conclusions: Preliminary data suggest that L given concurrently with P and T at a dose of 1000 mg per day induces an unacceptable rate of moderate to severe diarrhea. Careful monitoring of diarrhea as well as L dose reduction and initiation of loperamide at first occurrence of diarrhea are recommended. The dose of L when given concurrently with P and T has been amended to 750 mg per day in the current study and safety data for the 1000 mg and 750 mg per day cohorts will be presented. Implications for the ongoing ALTTO study will also be presented.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2109.
Guideline-conform systemic treatment improves survival of breast cancer patients.
Authors: Kurzeder C , Wöckel A , König J , Blettner M , Koretz K , Döhner H , Kreienberg R
Published: December 2010

Abstract #2115

Purpose: The influence of guidelines for systemic therapy of early-stage breast cancer on survival of patients is still not clear. We investigated to which extend adherence to the St Gallen expert consensus recommendations can improve outcome of the disease.  Methods: Clinical characteristics and survival data of 2454 patients with early breast cancer which received primary therapy between 1992 and 2005 at a cancer referral center were collected by patient chart review, queries to local death registries and physicians involved in follow-up care. Patients were assigned one of three risk categories and an algorithm based on the St Gallen Expert Consensus 2005 on primary therapy of early-stage breast cancer was developed to estimate risk reduction by compliance to consensus recommendations using different Cox proportional hazard models.  Results: According to the St Gallen consensus recommendations, indication for endocrine therapy was classified as mandatory in 78.3% (n=1922) of patients and as optional in 3.7% (n=92). A significant risk reduction by guideline-compliant endocrine therapy in patients with mandatory indication (HR = 0.62, CI 0.48 - 0.81) and also in patients with optional indication for endocrine therapy (HR = 0.20, CI 0.04 - 0.90) was estimated after adjustment for prognostic factors, co-existing morbidity and adjuvant treatment measures. Based on the consensus, indication for chemotherapy was classified as mandatory in 42.5% (n=1044) of the patients and as optional in 53.7% (n=1318). A significant risk reduction was observed for both subgroups of patients after adjustment for prognostic factors, co-existing morbidity and adjuvant treatment measures (HR = 0.80, CI 0.59 - 1.09 and HR = 0.53, CI 0.33 - 0.86). Model based estimates of overall survival for the subgroup of patients for whom endocrine therapy is mandatory and chemotherapy is optional is shown in figure 1, even more pronounced effects were found in patients for whom both endocrine therapy and chemotherapy are mandatory.        Conclusion: Systemic adjuvant therapy in accordance with the St Gallen consensus recommendations 2005 is clearly associated with prolonged survival. Chemotherapy and endocrine therapy are beneficial when applied in accordance with the indications of St. Gallen consensus recommendations.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2115.
Adjuvant therapy decisions in breast cancer: comparison of a multi disciplinary team's decisions with the recommendations of web-based computer programme “Adjuvant Online”.
Authors: Nowak VA , Ravichandran D , Austin A , Ah-See M
Published: December 2010

Abstract #2117

Introduction: Adjuvant therapy decisions in breast cancer impact on survival, recurrence and quality of life. We compared the hormonal and chemotherapy recommendations of a multi-disciplinary team (MDT) with the “best treatment” recommended by an evidence-based online computer programme “Adjuvant Online” (AO).  Materials and Methods: We prospectively monitored a breast cancer MDT for a period of one year. Among 218 breast cancer patients discussed, patients who had DCIS, neoadjuvant therapy, primary hormonal therapy, multifocal disease and micrometastases only in the axillary lymph nodes and recurrent disease were excluded leaving 122 suitable for input into AO. The MDT recommendation and actual treatment received were recorded. Ten-year cancer-related death and relapse rates were calculated using AO and estimates of proportional risk reduction with MDT recommended therapy and AO “best treatment” were made.  Results: Median age was 61 years (range 28-86). MDT recommended endocrine therapy (ET) to all patients with ER/PR positive cancers (n=103). Among 30 women aged 55 years or less with ER/PR positive cancers, 25 had tamoxifen, 4 had an aromatase inhibitor (AI) and one switched from tamoxifen to an AI. AO suggested AI or tamoxifen-AI switch as the best treatment for post-menopausal women. Among 73 women aged over 55 years with ER/PR positive cancers, 46% (n=34) received tamoxifen as per local cancer-network guidelines, which recommended tamoxifen for low-risk (T1 N0 M0) post-menopausal breast cancer. Others received an AI (n=38) except one patient who declined ET. There were 54 patients with >10% risk of cancer-related death in 10 years. Among these 36 were offered and 29 received chemotherapy. Chemotherapy was not offered to 17 patients due to age and/or comorbidity. Only 1 suitable patient with >10% risk was not offered chemotherapy by the MDT. Among those who had chemotherapy, only 31% had 3rd generation chemotherapy as recommended by AO, the main reason being national guidelines in the United Kingdom limiting the use of this regimen.  Discussion: There are differences between the adjuvant therapy decisions by the MDT and the “best treatment” recommended by Adjuvant Online. This study shows that in the majority of these cases the differences are due to either local or national guidelines that are in force in the United Kingdom. Cancer MDTs in the UK National Health Service are expected to adhere to these guidelines. However, calculation of the risk of death and recurrence rates using AO may be useful to facilitate decision making of the MDT by giving quantitative prognostic estimates and this could be a useful adjunct to help patients make informed decisions.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2117.
Combination of oncolytic adenoviral therapy with chemotherapy for enhanced breast cancer cell killing.
Authors: Stoff-Khalili MA , Nedeljkovic-Kurepa A , Jung JS , Glover BV , Wappenschmidt B , Rhiem K , Bosse K , Mallmann P , Curiel DT , Schmutzler RK , Mathis MJ
Published: December 2010

Abstract #2129

Oncolytic adenoviruses are emerging agents for treatment of cancer by tumor-restricted virus replication, cell lysis and virus spread. A promising oncolytic adenovirus agent, known as Ad5-24-RGD, harbors a 24-bp deletion in the E1A gene that abrogates the binding of E1A to the retinoblastoma tumor suppressor (Rb) and presents enhanced infectivity of primary cancer cells due to insertion of an Arg-Gly-Asp (RGD) motif into the fiber knob. Thus, Ad5-24-RGD has improved cancer cell infection efficiency due to expanded tropism toward alpha-v integrins. It also replicates selectively in cancer cells with Rb/p16 mutations. As with conventional therapy regimes, oncolytic virotherapy, by itself, has limited success in complete tumor eradication in both preclinical animal models and clinical studies. Combination of anticancer agents with different modes of action remains a mainstay in cancer treatment. We undertook one approach towards this end by combining oncolytic adenoviral therapy with chemotherapy. In this study, we investigated a combination treatment of breast cancer cells with Ad5-4-RGD and Docetaxel, a microtubule-stabilizing taxane that is being used in the clinic for the treatment of breast and prostate cancers and small cell carcinoma of the lung. Our results indicate a synergistic effect between Docetaxel and Ad5-24-RGD in breast cancer cell killing at a lower dose than either agent alone. These results suggest that viral replication was not inhibited by this chemotherapy treatment and that chemotherapy could reduce the amount of viral particles needed to help eradicate the tumor. Administration of lower viral loads would simultaneously improve safety and decrease immunogenicity of the vector. Likewise lower doses of chemotherapy agents would decrease toxicity and side effects. The inclusion of oncolytic adenoviruses into multimodal cancer treatment together with chemotherapy has a potential to become powerful therapeutic regimen.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2129.
Effects of the combined blockade of EGFR and ErbB-2 on signal transduction and regulation of cell cycle regulatory proteins in breast cancer cells.
Authors: D'Alessio A , De Luca A , Maiello MR , Palumbo C , Rachiglio A , Gallo M , Normanno N
Published: December 2010

Abstract #2130

Co-expression of EGFR and ErbB-2 occurs in a subset of primary breast carcinoma, and it is associated with worse prognosis. We previously demonstrated that treatment of breast cancer cells that express both receptors with a combination of the EGFR tyrosine kinase inhibitor gefitinib (G) and the anti-ErbB-2 monoclonal antibody herceptin (H) results in a synergistic anti-tumor effect. Such combination failed to show significant activity in a phase I/II clinical trial. However, patients enrolled in this trial were not selected for expression of EGFR. We explored the molecular mechanisms involved in the synergism of G and H in breast cancer cells. Treatment of both SK-BR-3 and BT-474 cells with a combination of G+H produced a more significant reduction of AKT and p42/p44-MAPK (MAPK) activation as compared with treatment with a single agent. The combination also produced a more prolonged blockade of both signalling pathways. In agreement with these findings, treatment with G+H induced a more pronounced accumulation of cells in the G0/G1 phase of the cell cycle as compared with treatment with a single drug. A sub-G0/G1 peak, suggestive of apoptosis, was evident in cells treated with the combination but not with G or H. The combination of G+H produced a more significant increase in the levels of p27kip1 and of hypophosphorylated forms of pRb2, and a more significant decrease in the levels of Cyclin D1 and survivin as compared with a single agent. Analysis of p27kip1 protein stability, in presence of the protein synthesis inhibitor cycloheximide, revealed that both G and the combination of G+H significantly increased the stability of p27kip1, with the combination showing higher effects. Furthermore, breast cancer cells treated with the combination expressed higher levels of p27kip1 mRNA as compared with cells treated with a single drug. The combination also induced a significant increase in the nuclear levels of the FKHRL-1 transcription factor that was not observed in cells treated with G or H alone. Treatment of breast cancer cells with the dual EGFR/ErbB-2 inhibitor lapatinib produced an increase in the levels of p27kip1 and of hypophosphorylated pRb2, and a decrease in the levels of Cyclin D1 and survivin comparable to that observed following treatment with the combination of G+H. In addition, an increase in p27kip1 mRNA and in the nuclear levels of FKHRL-1 was induced in breast cancer cells by treatment with lapatinib. These findings suggest that the synergistic anti-tumor effects deriving from EGFR and ErbB-2 blockade are mediated by significant alterations in the expression of cell cycle regulatory proteins, including transcriptional and post-transcriptional regulation of p27kip1 expression.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2130.
A multi-center phase II study of three doses of TAS-108 in postmenopausal women with advanced breast carcinoma following first or second line endocrine therapy.
Authors: Buzdar A , Tan-Chiu E , Schwartzberg L , Perez A , Ellis M , Garin A , Ingle J , Carlson R
Published: December 2010

Abstract #2131

Introduction: TAS-108 is an oral steroidal anti-estrogen agent that selectively inhibits ERα and is mainly metabolized by CYP3A4. The purpose of this study is to investigate the efficacy and safety of TAS-108 administrated orally in three dose levels in patients with locally advanced, locally recurrent inoperable or metastatic breast carcinoma (BC) in four countries (USA, Russia, Mexico and Chile).  Methods: Postmenopausal women with confirmed ER and/or PgR positive BC who had previously responded to one or two standard endocrine therapies, with or without one prior chemotherapy were randomly assigned to three doses of TAS-108, 40 mg, 80 and 120 mg daily. Using a modified Panageas' optimal two-stage trinomial design, the enrollment of 60 evaluable patients (first-stage: 19) was required to each individual dose group. Tumor response was assessed every 8 weeks according to RECIST criteria. Adverse events (AEs) were graded by CTC-AE v3.0.  Results: A total of 146 patients with mean age of 63 years old were enrolled with 61 patients in the 40 mg group and 66 in the 80 mg group. The 120 mg group was terminated at the end of stage 1 with 19 patients enrolled due to lack of efficacy. The mean duration of study treatment was 172 days for the 40 mg group and 160 days for the 80 mg group. Partial response (PR) was documented in 6 (10%) patients in the 40 mg group and 4 (6.7%) patients in the 80 mg group. The rate of disease stabilization (CR+PR+SD) reported by the investigators was 43% (95%CI, 31%,56%) in the 40 mg group and 45% (95%CI, 32%,58%) in the 80 mg group. Adjudicated clinical benefit (CR+PR+SD more than 24 wks) was observed in 22% of patients in the 40 mg group and 20% of patients in the 80 mg group. Clinical benefit was achieved in 25% of patients with 1 prior hormonal therapy and 15% of patients with more than one line of prior hormonal therapy. Median time to progression (TTP): 15 weeks for the 40 mg group and 15.9 weeks for the 80 mg group. Median duration of clinical benefit was 32 weeks in the 40 mg group and 64 weeks in the 80 mg group. In the 40/80/120 mg groups, the commonly reported treatment-related AEs included nausea (15%/11%/16%), fatigue (10%/11%/16%), headache (10%/6%/16%), hot flushes (10%/5%/32%), diarrhea (2%/6%/5%), constipation (3%/3%/5%), and arthralgia (2%/5%/11%). No endometrial cancer and treatment-related deaths occurred during the study.  Conclusions: TAS-108 has demonstrated anti-tumor activity in this population and was generally well tolerated. The 40 mg dose was chosen as the recommended dose for future clinical evaluation in patients with advanced breast cancer.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2131.
Effects of novel retinoic acid metabolism blocking agent (VN/14-1) on N-methyl-N-nitrosourea (MNU)-induced mammary carcinoma and uterus in the rat model.
Authors: Goss PE , Njar VC , Qi S , Hu H , Gediya LK , Puranik P , Khandelwal A
Published: December 2010

Abstract #2133

Background: All-trans-retinoic acid (ATRA) and other retinoids play key roles in prevention and therapy of many proliferative diseases including cancers. VN/14-1 [4-(±)-(1H-Imidazol-1-yl)-(E)-retinoic acid], which is a new generation novel retinoic acid metabolism blocking agent (RAMBA), works by inhibiting the breakdown of ATRA. The inhibitory effects of VN/14-1 on the growth of human breast cancer cells and human breast tumors in the nude mouse model have been previously demonstrated. The purpose of this study was to evaluate the effects of VN/14-1 on the N-methyl-N-nitrosourea (MNU)-induced rat mammary carcinoma model, as well as on the uterus in immature ovariectomized (OVX) rats.  Methods: (1) VN/14-1 (5, 10, and 20 mg/kg/d) was given by oral gavage to grouped female Sprague Dawley (SD) rats bearing MNU-induced mammary carcinoma for 8 weeks, after which tumor weight and volume, as well as histology were measured. (2) VN/14-1 (10 and 20 mg/kg/d) and b-estradiol (10 mg/kg/d) were given alone or in combination, by oral gavage (VN/14-1) and subcutaneous injection (b-estradiol), to immature OVX SD rats for 3 days, after which uterine weight and histology were measured.  Results: (1) At the end of the treatment period, the administration of 5, 10 and 20 mg/kg/d VN/14-1 caused significant reductions of 19.1, 34.4 and 44.3%, respectively, in mean tumor weight compared with the control animals (all p < 0.05). The cumulative tumor growth was also significantly slower in groups receiving 5, 10 and 20 mg/kg/d compared to the control group in a dose-dependent manner. (2) Immature OVX rats given VN/14-1 at doses of 10 and 20 mg/kg, had reduction in uterine wet weight of up to 56% compared to OVX controls (P < 0.001). OVX rats given VN/14-1 of 10 and 20 mg/kg in combination with β-estradiol had reduction in uterine wet weight of up to 58% compared to the OVX rats given β-estradiol alone (P < 0.001). The adverse toxic effects such as fatigue and anorexia were occurred in the groups at high dose of 20 mg/kg.  Conclusions: RAMBA VN/14-1 was able to inhibit the growth of tumors in the MNU-induced ER positive rat mammary tumor model and antagonized the stimulatory effect of β-estradiol on the uterus. The studies suggest VN/14-1 might be an effective novel therapy for ER positive breast cancer.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2133.
Safety of the anti-IGF-1R antibody CP-751,871 in combination with exemestane in patients with advanced breast cancer.
Authors: Ryan PD , Neven P , Dirix LY , Barrios CH , Miller WH , Fenton D , Abraham MF , Paccagnella L , Gualberto A , Goss PE
Published: December 2010

Abstract #2136

Background: Aromatase inhibitors (AIs) are established first-line treatment for postmenopausal estrogen receptor (ER)+ metastatic breast cancer. However, not all patients benefit from AIs and those whose tumors initially respond eventually relapse. One hypothesized mechanism for tumor insensitivity to hormonal agents seems to be cross-talk between the ER pathway and other growth factor signaling pathways, in particular the insulin-like growth factor receptor type 1 (IGF-1R). In xenograft breast cancer models, CP-751,871 administration increased tumor growth inhibition induced by tamoxifen. Thus our trial addresses the effect of combining AI with an IGF-1R antagonist.  Methods: A phase II, multicenter, randomized, two-arm, comparative, two-stage trial to determine PFS of CP-751,871, a fully human IgG2 monoclonal antibody targeting IGF-1R, in combination with exemestane versus exemestane alone as first-line treatment in patients with hormone receptor positive, advanced breast cancer. Secondary endpoints include clinical benefit (CR, PR, or SD ≥6 months), safety/tolerability and PK. Patients included in the study are post menopausal, ≥18 years with locally advanced/metastatic breast cancer (stage IIIB or IV), ECOG PS 0–2, and adequate hematological, biochemical, and cardiac functions. CP-751,871 is given by intravenous infusion at a dose of 20 mg/kg every 21 days while 25 mg exemestane was given p.o. daily.  Results: To date 37 patients have been dosed with CP-751,871 plus exemestane. Median age is 60.5 years (range 34–84). Patients received a median of 9.5 treatment cycles (range 1–22). One GR 4 CP-751,871-related AE (hoarse voice) was reported, which resolved after 3 days without intervention. GR 3 CP-751,871-related toxicities included 8.1% hyperglycemia (n=3), 8.1% GGT elevation (n=3), 5.4% allergic reaction (n=2), 5.4% hearing loss (n=2), 5.4% weight loss (n=2), and 2.7% anorexia (n=1). GR 2 CP-751,871-related AEs >10% were headaches, muscle cramps, and nail changes. Both hyperglycemia and hypersensitivity reactions were manageable (with oral hypoglycemic drug/insulin and antihistamine), while GGT elevation seems to be reversible.  Conclusions: CP-751,871 in combination with exemestane is well tolerated. The most frequent GR 3 side effects are either well managed with medications or appear to be reversible. Therefore, CP-751,871, due to its safety profile, is a good targeted agent to combine with standard hormonal therapy. The stage I portion of the study to determine efficacy and toxicity is ongoing.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2136.
The taccalonolides are a novel class of microtubule stabilizers that circumvent clinically relevant taxane resistance mechanisms.
Authors: Risinger AL , Jackson EM , Polin LA , Helms GL , Joe PA , Luduena RF , Mooberry SL
Published: December 2010

Abstract #2142

The taccalonolides are a novel class of microtubule stabilizing agents isolated from the tropical plant Tacca chantrieri. Their unique structure and inability to directly bind to or stabilize tubulin in vitro strongly suggest that the taccalonolides cause microtubule stabilization through a distinct mechanism of action. The ability of four taccalonolides, A, E, B and N, to circumvent multidrug resistance mechanisms associated with taxane resistance was studied. The four taccalonolides were able to overcome multidrug resistance mediated by expression of the P-glycoprotein ATP-binding cassette drug transporter in vitro. Furthermore, the taccalonolides A and E were highly active against tumors in a P-glycoprotein expressing Mam17/ADR murine model of breast cancer that is resistant to both doxorubicin and paclitaxel. In addition to P-glycoprotein, the expression of the βIII isotype of tubulin has also been linked with taxane treatment failure. HeLa cells expressing high levels of βIII-tubulin were resistant to paclitaxel, docetaxel, epothilone B and vinblastine. Interestingly, these βIII-tubulin expressing cells were more sensitive to each of the four taccalonolides than the parental cell line. These data demonstrate that the taccalonolides have advantages over the taxanes in clinically relevant drug resistant cell lines and tumor models. Therefore, the taccalonolides represent a novel family of microtubule stabilizing agents with clinical potential.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2142.
MG28, a novel fatty acid synthase inhibitor, overcomes resistance to anti-HER therapies in breast cancer.
Authors: Puig T , Urruticoechea A , Oliveras G , Aguilar H , Turrado C , Cufí S , Ortega-Gutiérrez S , Benhamú B , López-Rodríguez M , Colomer R
Published: December 2010

Abstract #2143

Introduction: Resistance to anti-HER2 therapy is a common feature in Her2 positive breast cancer. The green tea polyphenol epigallocatechin 3-gallate (EGCG) is an effective antitumoral therapy through the blockade of FASN. We have developed polyphenolic compounds that inhibit FASN with greater efficacy than EGCG. Here we report the anticancer activity of MG28, one of the new synthesized FASN inhibitors, alone or in combination with trastuzumab, lapatinib, erlotinib and cetuximab in a HER-amplified breast cancer models.  Experimental design: We analysed the cytotoxic effects of MG28 alone and in combination with trastuzumab, lapatinib, erlotinib, or cetuximab in HER2+ and EGFR+ human breast cancer cells. Combinations of MG28 with targeted agents were tested for synergism, additivity and antagonism using the isobologram analysis. We further analysed changes in FASN protein levels, in apoptosis [as assessed by cleavage of poly(ADP-ribose) polymerase (PARP)], and in activation of HER2, AKT and MAPK.  Results: Combination index analysis showed synergistic cytotoxic effects of MG28 combined with trastuzumab, erlotinib or lapatinib (p < 0.001) and antagonic effects with cetuximab (p < 0.05) in HER2 overexpressing cells. MG28 plus lapatinib was significantly correlated with increased apoptosis and inhibition of phosphorilated HER2 and MAPK. Response to co-exposure of MG28 plus erlotinib was significantly correlated with increased apoptosis and its ability to inhibit HER2, MAPK and AKT phosphorilation. HER2+ cells showed partial resistance to trastuzumab and combined treatment of trastuzumab with MG28 reversed significantly this resistance and thus correlated with increasing cleavage of PARP and its ability to inhibit phosphorylation of HER2 and AKT.  Conclusion: FASN inhibition seems to be an effective approach to overcome resistance to anti- HER therapies. Our results provide a new therapeutic strategy for further in vivo – combined treatments using drugs that target HER2, EGFR and FASN.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2143.
A phase I-II trial of Ispinesib, a kinesin spindle protein inhibitor, dosed every two weeks in patients with locally advanced or metastatic breast cancer previously untreated with chemotherapy for metastatic disease or recurrence.
Authors: Gomez H , Castaneda C , Philco M , Pimentel P , Falcon S , Escandon R , Saikali K , Conlan M , Seroogy J , Wolff A
Published: December 2010

Abstract #2148

Kinesin Spindle Protein (KSP) is a mitotic kinesin essential for cell cycle progression. Ispinesib, a selective KSP inhibitor, blocks mitotic spindle assembly with cell cycle arrest in mitosis and subsequent cell death. When dosed on a q21d schedule, the maximum tolerated dose (MTD) was 18mg/m2 and neutropenia was the dose-limiting toxicity (DLT), with nadir at 7-10 d and recovery by d15. Activity was observed in a Phase II trial of ispinesib dosed at 18 mg/m2 q21d in patients (pts) with locally-advanced (LA) or metastatic breast cancer (MBC) after anthracycline and taxane failure (response rate 4/45 [9%]). This trial evaluates safety and efficacy of ispinesib in LA or MBC, given on d1 and d15 q28d, thus increasing dose density.  Methods: This is a multicenter Phase I-II trial. In Phase I, DLT and MTD of ispinesib given d1 and d15 q28d will be determined. Eligibility criteria: LA or MBC; no prior chemotherapy (CT) except neoadjuvant or adjuvant and ≥ 1 year elapsed since CT; no CNS or leptomeningeal metastases; ECOG 0-1. This is a standard 3+3 dose escalation trial design, starting at 10 mg/m2 and escalating based on tolerability in Cycle 1. Pharmacokinetic data are collected on d1 and d15 of Cycle 1. Phase II of this trial will evaluate efficacy (response rate by RECIST) of ispinesib at the MTD.  Results: Phase I of the trial is ongoing. To date, 13 pts have been treated at 3 dose levels: 10 (n=2; 2 cycles, n=1; 6 cycles), 12 (n=1; 1 cycle, n=2; 3 cycles) and 14 mg/m2 (n=7; Cycle 1 ongoing). Among the first 6 pts, 4 were Stage IV, 3 had prior neoadjuvant and 2 adjuvant CT; 2 were chemo-naïve. Four had prior anthracycline and 3 prior taxane. Biomarker status was ER+, PR+, HER2- (n=3), ER+, PR-, HER2- (n=1), ER-, PR-, HER2+ (n=3), ER-, PR-, HER2- (n=4), and unknown (n=2). Mean age was 55 yr; for the first 2 dose levels (n=6 pts), the most frequent toxicity was neutropenia (n=4). Other events included mild GI toxicity. No neuropathy or alopecia has been reported. The only grade 3 or 4 toxicity was neutropenia (n=2). At the 14 mg/m2 dose level, one DLT has been reported: a grade 3 transaminase (AST) elevation in a patient with liver metastases which resolved with a dose delay of 4 days. As a result, this cohort was expanded to include 6 evaluable pts. One additional patient at the 14mg/m2 dose had a dose delay of 5 days due to non-DLT neutropenia.  Conclusions: Ispinesib appears to be well tolerated on a q2w dosing schedule at doses tested to date. A dose-density equal to that administered in the prior Phase II trial (0.86 mg/m2/day) was tolerated with the new q2w schedule. Dose escalation is ongoing.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2148.
N-ethoxybenzylimidazoles: a novel acid-sensitive linker for controlled release of therapeutics from drug delivery systems.
Authors: Luong A , Issarapanichkit T , Yang JC
Published: December 2010

Abstract #2156

Background:  Drug delivery systems (DDS) are useful for cancer therapies because they can target tumors and cancer cells more specifically than cancer therapeutics alone. An important component of a DDS is the triggering mechanism for drug release. This presentation will highlight a new class of N-linked imidazoles as potential acid-sensitive cleavable linkers for use in cancer DDSs. These acid-sensitive linkers are designed to exploit the lower extracellular pH of some tumors and the endosomes and lysosomes within cells to trigger the controlled release of therapeutic agents from drug delivery vessels. Cleavage of N-ethoxybenzylimidazoles (NEBIs) exhibit a 10-fold increase in the rate of hydrolysis in mild aqueous acidic solutions (at pH = 5.5) compared to solutions at normal, physiological pH. The rate of hydrolysis can be tuned to range from minutes to months through the addition of electron donating or withdrawing groups.  The NEBI linker can be used to conjugate a cytotoxic agent to a carrier. Carriers for this DDS include molecules such as tumor specific antibodies, polymers/nanoparticles, peptides, and ligands, that can target cancer cells or accumulate around tumors. Cytotoxic agents include FDA approved small molecules and chemotherapeutics.  Methods and Materials:  The NEBI was developed into a bifunctional crosslinker containing a carboxylic acid and an azide for conjugation of cancer therapeutics to carriers. For simplicity, we used Human Serum Albumin (HSA) as our model carrier and doxorubicin as our model cytotoxic agent. The NEBI was conjugated to doxorubicin through a simple amide coupling. The NEBI was then conjugated to an alkyne containing HSA via “Click Chemistry.”  Results:  We were able to load approximately 1-2 doxorubicin molecules per HSA. Imaging studies have shown that our HSA conjugates to doxorubicin via a NEBI linker (HSA-NEBI-dox) localize in lysosomes. Cytotoxicity studies show HSA-NEBI-dox is cytotoxic while HSA conjugates to doxorubicin via a stable linkage is not.  Discussion:  We were able to develop methods for conjugating doxorubicin to HSA. The techniques used here can be applied to conjugate other carriers and other cancer therapeutics. We will continue to develop the NEBI linker and demonstrate how the linker can be used to conjugate trastuzumab to doxorubicin. We chose use trastuzumab, a monoclonal antibody, as a carrier because it is known to bind to the HER2 receptor. HER2 is over expressed in 25%-30% of breast cancers. Trastuzumab not only targets cancer cells that are over expressing the HER2 receptor, but binding of trastuzumab to the HER2 receptor can trigger receptor mediated endocytosis, which can lead to controlled release of doxorubicin within the cell.   

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2156.
Novel thermal tumor ablation for breast cancer in mice using magnetic nanoparticles.
Authors: Motomura K , Ishitobi M , Komoike Y , Koyama H , Inaji H , Inoue M , Nagae H , Nagano I
Published: December 2010

Abstract #2158

Background: Thermal ablation therapy using lasers, ultrasound and microwaves is being investigated as a less invasive alternative to surgery for breast cancer. This study examined the efficacy of magnetic nanoparticles, which produce heat when exposed to an alternating magnetic field, for the destruction of breast cancer in mice.  Material and Methods: Tumors were induced on both sides of the back of BALB/C mice by subcutaneous injection of MDA-MB-231 breast cancer cells. Magnetic nanoparticles were injected in and around one of the tumors in a mouse, while saline were injected in another tumor (control). Mice were exposed to an alternating magnetic field for 40 min. The temperature of the tumor injected with nanoparticles reached 45°C and was maintained at that temperature by adjusting the magnetic field intensity. Histopathological response was evaluated one to two months after thermal ablation therapy.  Results: In 8 (67%) of 12 mice, complete tumor regression was observed in tumors treated with nanoparticle injection, while there was no regression in control tumors in any of the mice. There was no irreversible damage seen in normal tissue surrounding the nanoparticle injected tumors.  Discussion: Since this procedure can destroy cancer cells specifically without injuring normal tissue, it may be applicable to the treatment of human breast cancer. Moreover, it may be possible to develop this procedure into a systemic therapy for tumor-specific targeting.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2158.
Micrometastases and isolated tumor cells: relevant and robust or rubbish? (MIRROR): preliminary results of the MIRROR study from the Dutch breast cancer trialists' group (BOOG).
Authors: de Boer M , van Deurzen CH , van Dijck JA , Borm GF , van Diest PJ , Adang EM , Nortier HW , Rutgers EJ , Seynaeve C , Menke-Pluymers MB , Bult P , Tjan-Heijnen VC
Published: December 2010

Abstract #23

Background  The sentinel lymph node procedure (SNP) has largely replaced axillary lymph node dissection (ALND) in patients with early breast cancer (BC). Intensified examination of the sentinel node (SN) results in the detection of isolated tumor cells (ITCs) and micrometastases, whereas its relevance is still debated. This retrospective cohort study is the first study on this topic which includes only patients who underwent a SNP, with central review of all SN slides, and with separate analyses for the impact of administration of adjuvant systemic therapy.  Methods  Patients operated for BC in all Dutch hospitals in the years 1998-2005, having favourable tumor characteristics (> 35 year; tumor size 1-3 cm and differentiation grade I-II OR tumor size ≤ 1 cm irrespective of grade), having undergone a SNP with pN0(i-), pN0(i+), or pN1mi as final N-stage were selected. Patients were classified in cohort I in case of pN0(i-) and no adjuvant systemic therapy (AST), in cohort II in case of pN0(i+)/pN1mi and no AST, and in cohort III in case of pN0(i+)/pN1mi with AST. SNs were centrally reviewed and restaged according to 6th TNM classification. The 5-year disease-free survival (DFS) was analysed for the 3 different cohorts.  Results  So far, data are available for 1,744 of the 3,240 selected patients (cohort I n=935; cohort II n=340, cohort III n=469) with a median follow-up of 5.5 years. At diagnosis median age was 58, 56 and 56 years (p < 0.01) and median tumor size was 1.2, 1.4 and 1.5 cm (p < 0.0001). Differentiation grade and hormone receptor status were equally distributed among the three cohorts. Adjuvant systemic therapy in cohort III consisted of chemotherapy (10%), hormonal therapy (63%), or both (27%). The 5-yr DFS is 84% in cohort I, 73% in cohort II, and 86% in cohort III (p = 0.003 cohort I vs II; p < 0.0001 cohort II vs III). Correction for baseline characteristics did not lead to a relevant change in the results.  Conclusion  BC patients with ITCs or micrometastases as final N-stage after SNP have a significantly worse 5-yr DFS as compared to patients having no ITCs or micrometastases, independent of primary tumor characteristics. Adjuvant systemic therapy significantly improves the 5-yr DFS in the group of patients with ITCs or micrometastases in the lymph node(s). At the meeting, updated analyses will be available for the entire MIRROR study population.  Support: The Netherlands organization for health research and development (ZonMw) and all Dutch CCCs.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 23.
Determination of HER2 amplification by in situ hybridization: when should chromosome 17 also be determined?.
Authors: Bartlett JM , Campbell FM , Mallon EA
Published: December 2010

Abstract #3002

Introduction: Aberrations of chromosome 17 (aneusomy) are common in breast cancer and therefore have a critical impact on the assessment and reporting of HER2 gene amplification in a significant sub-set of cases. There is an ongoing debate as to the proportion of cases for which assessment of chromosome 17 copy number may be important. According to current guidelines1,2, amplification of HER2 is considered to be a HER2/chromosome 17 ratio ≥2.0 and a ratio <2.0 is regarded as non-amplified. For HER2 gene copy number assays, it has been assumed that copy numbers of >6.0 reflect amplification and a result of <4.0 HER2 gene copies per nucleus is always associated with lack of amplification; cases with 4-6 copies per cell are thought to require validation by testing of a parallel section for chromosome 171,2. To our knowledge, this assumption has not been verified experimentally.  Methods: HER2 and chromosome 17 were measured by dual color FISH in 1711 breast cancer samples referred to the authors laboratories between 2000-2008. Using HER2 copy number and chromosome 17 data the impact of chromosome 17 testing upon accuracy of diagnosis of gene amplification was assessed.  Results: At a HER2 copy number of 2 to <3, 16 of 488 cases (3.3%) had HER2 amplification; and at a copy number of 3 to <4, 32 of 195 cases (16.4%) were amplified. The proportion of cases with HER2 amplification increased considerably at HER2 copy numbers of 4 to <7: 50.0% at 4 to <5; 67.5% at 5 to <6, and 77.3% at 6 to <7. Virtually all cases were amplified at HER2 copy numbers of ≥7.  Conclusion: Werecommend that all cases with observed HER2 copy numbers of 2 to 7 should also be analyzed for chromosome 17 in order to accurately determine HER2 gene amplification. This would require analysis of chromosome 17 in 48.3% of all breast cancer cases based upon the study population. Current guidelines1,2 recommend chromosome 17 measurement only in cases with a HER2 copy number of 4 to <6, which represents only 6.6% of all breast cancer cases in the current study. Although single color ISH is becoming more widely used with the availability of CISH, the importance of chromosome 17 measurement cannot be ignored. It is essential that HER2 testing is of high quality, so that optimal patient management can be provided.  1. Ellis IO, Bartlett J, Dowsett M et al: Updated recommendations for HER2 testing in the UK. J.Clin.Path. 57(3), 233-237 (2004).  2. Wolff AC, Hammond ME, Schwartz JN et al: American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer. Journal of Clinical Oncology 25(1), 118-145 (2007).

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3002.
Contribution of Mib1 to the SBR grade for prediction of metastatic recurrence in breast cancer.
Authors: MacGrogan G , deMascarel I , Debled M , Tunon de Lara C , Brouste V , Bonnefoi H , Mauriac L
Published: December 2010

Abstract #3003

Background: The Scarff Bloom and Richardson (SBR) histologic grade, which assesses tumor differentiation, nuclear pleomorphism and tumor proliferation through a mitotic score, is known to be an independent prognostic factor in breast cancer. Ki-67 is a nuclear protein that is expressed by cells that are engaged in the cell cycle and may be a more accurate proliferation marker than the mitotic score used in SBR grade. The aim of our study was to look at the prognostic value of ki-67 (detected with Mib1 antibody) in breast cancer and if it could improve the prognostic value of SBR grade (SBR G).  Materials and Methods: We analyzed ki-67 expression by immunohistochemistry using the Mib1 antibody on tissue microarrays comprising four 0.6mm diameter tissue cores of 1033 consecutive invasive ductal carcinomas (290 SBR G1, 470 SBR G2, 273 SBR G3) operated on between 1989 and 1993 (median follow-up: 167 months, 280 metastatic events). Using the median and 75th percentile values of Mib1 expression in the series (10 % and 20%, respectively) as cut offs, we defined a three tiered Mib1 score and used it to replace the mitotic score in the SBR G (Mib1 G). The prognostic values of Mib1 score alone, SBR G, Mib1 G and combined SBR/Mib1 G using the higher of the 2 grades to assign tumors to a risk category were assed by univariate analysis using the Log-rank test with the Kaplan Meier method.  Results: 53/290 (18%) and 53/470 (11%) of SBR G1 and 2 tumors were reclassified in combined SBR/Mib1 G2 and G3, respectively. In the combined SBR/Mib1 G three groups of tumors could be individualized according to their metastatic risk.    Patients with SBR G1/Mib1 G1 tumors had the lowest metastatic risk. Patients with SBR G1/Mib1 G2 tumors, SBR G2/Mib1 G1 tumors and SBR G2/Mib1 G2 tumors had an intermediate risk. Finally, patients with SBR G2/Mib1 G3 tumors, SBR G3/Mib1 G2 and SBR G3/Mib1 G3 tumors had the highest metastatic risk. Mib1 score, Mib1 G, SBR G, and combined SBR/Mib1 G were all strongly associated with Metastasis-free Interval (p=3.10-10, p=8.10-11, p=2.10-12 and p=4.10-13, respectively).  Discussion: In the combined grade patient assignment to risk category (low, intermediate or high) is made on the higher of the two grades. Metastatic risk was more accurately estimated when SBR and Mib1 grades were combined compared to SBR G or Mib1 G separately. Mib1 may complement the prognostic information provided by histologic grade.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3003.
Clinical significance of the entity 'flat epithelial atypia' on core needle biopsies of breast.
Authors: Dabbs DJ , Bhargava R , Tseng G , Chivukula M
Published: December 2010

Abstract #3006

Background: Flat epithelial atypia (FEA) is a newly emerging entity of uncertain clinical significance. Defined by the World Health Organization (WHO) in 2003, FEA differs from columnar cell hyperplasia by the presence of nuclear atypia and from atypical ductal hyperplasia (ADH) by the absence of complex architecture. As the term FEA is relatively new, the clinical relevance and the outcome data are sparse. The aim of this study is to evaluate the clinical and pathologic significance of this entity in order to construct the proper management of these lesions.  Design: All core-needle biopsies (CNB) diagnosed as ADH, which includes pure FEA at our institution, from January 2006-April 2008 were retrieved from our pathology files. Hematoxylin and eosin (H&E) slides of five levels on each case were reviewed. Statistical analysis was performed with calculation of p-value.  Results: Total number of CNB performed from 2006-2008 was 8054. 99% (8051/8054) were stereotactic guided and 1% (3/8054) were ultrasound/MRI guided biopsies. All the CNB were performed for microcalcifications. The mean age of the patients is 54 years (range 29-83). Incidence of ADH was 4% (338/8054). We have reviewed 203 cases so far and the study is ongoing. 32 cases were discarded due to either presence of DCIS or IC in the CNB. Upon review, 9% (18/203) cases were classified as pure FEA and 91% (185/203) cases as FEA+ADH. The upstaging to a significant lesion in pure FEA group is 14% in comparison to 12% in FEA+ADH group (p=0.9471), is not statistically significant. Lobular neoplasia is seen in association with pure FEA group in 33% (6/18) and 11% (21/185) in FEA+ADH group. In 6% (11/185) of cases in FEA+ADH group, we observed FEA evolved into ADH at the same site at an average of 3-4 levels.  Conclusions: 1.The incidence of FEA in our CNB targeted for calcifications is 9% 2. The upstaging in the follow-up resections in FEA in comparison to ADH+FEA shows no statistical difference (14% versus 12%). 3. Since FEA and ADH commonly occur together on the same slide, it is prudent to examine deeper tissue levels when pure FEA is encountered on CNB.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3006.