bluuebunny/crossref_metadata_2025 huggingface dataset:
Resident Macrophages Influence Stem Cell Activity in the Mammary Gland.
Authors: Gyorki D. , Lindeman G. , Visvader J.
Published: December 2010

Background: Increasing evidence suggests that macrophages play a role in the normal development of certain organs, including the breast1. Intriguingly, macrophages are often found in the stroma of breast tumours, where they may promote tumour growth and metastasis. The mechanism underpinning the requirement for macrophages during normal development may involve their interactions with mammary stem cells (MaSCs). Recently, a population enriched for MaSCs has been prospectively identified in mice2. We have used 2 approaches to determine whether macrophages are required for normal MaSC function.Methods and Results: The role of macrophages in mammary development was studied in mice with an inactivating recessive mutation in the CSF-1 gene (CSFop/op). MaSCs from CSFop/op mice exhibit significantly reduced clonogenicity in vitro with a reduction in colony size and number in CSFop/op mice. Transplantation assays confirmed a reduced regenerative capacity of the MaSCs derived from CSFop/op mice when transplanted into wild-type recipients. Furthermore, in reciprocal experiments, we found that a macrophage-deficient micro-environment does not support stem cell growth with an inability for wild-type stem cells to repopulate the fat pad of CSFop/op mice.To support these findings, an alternative strategy was used to eliminate the macrophages from the stem cell microenvironment by co-transplanting stem cells with liposomes containing clodronate or saline. Clodronate-containing liposomes are toxic to macrophages after ingestion. There was a greater than 80% impairment in stem cell repopulating frequency as well as a marked decrease in outgrowth potential when cotransplanted with clodronate-containing liposomes.Discussion: These studies indicate a major role for macrophages in supporting stem cell growth during mammary development. This may relate to a role in the stem cell niche. Understanding the mechanisms by which mammary stem cells interact with macrophages during mammary development will shed light on why macrophage infiltration in tumours portends a poor prognosis in breast cancer.1. Gouon-Evans V, Lin EY, Pollard JW. Requirement of macrophages and eosinophils and their cytokines/chemokines for mammary gland development. Breast Cancer Res. 2002; 4:155-64.2. Shackleton M, Vaillant F, Simpson KJ, et al. Generation of a functional mammary gland from a single stem cell. Nature. 2006; 439:84-8.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 49.
Utilization of Serine-Derivatized Gadonanotubes as Magnetic Nanoprobes for Intracellular Labeling of MCF-7 Human Breast Cancer Cell.
Authors: Chan B. , Hassan A. , Hartman K. , Hartman K. , Ananta J. , Ananta J. , Mackeyev Y. , Hu L. , Pautler R. , Wilson L. , Wilson L. , Lee A.
Published: December 2010

The use of contrast agent enhances the image and gives a more accurate diagnosis by magnetic resonance imaging (MRI). However, safety of the patients with renal dysfunction raised the concern of increase in use. Comparing to the current clinical contrast agent gadopentetate dimeglumine (Gd-DTPA or Magnevist®), gadolinium-filled ultra-short carbon nanotubes (GNs) showed superior relaxivity. At different magnetic fields, GN increased relaxivity by 40-, 4.6- and 1.6-fold at magnetic fields of 1.5, 7 and 9.4T, respectively. More importantly, GN showed the ability in internalization that Gd-DTPA lacks. GN entered MCF-7 human breast cancer cells and allowed imaging of the cells by MRI. Detectable amount of gadolinium was found only in GN-treated MCF-7 cells by ICP-OES analysis. MRI of the kidney and liver showed GN was more dispersed suggesting that GN may be able to enter the interstitial space while Gd-DTPA was remained in the vasculature. Taken together, we found GN may be developed as a better contrast agent for MRI.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5005.
Non-Invasive Measurement of Pathological Heterogeneity of Cancer Tissues Using Water State Information from Diffuse Optical Spectroscopic Imaging.
Authors: Chung S. , Chung S. , Cerussi A. , Hsiang D. , Tromberg B. , Tromberg B. , Tromberg B.
Published: December 2010

Background: In our previous work, we have shown that biochemical tissue water state correlates with histopathological scores of breast cancer (R=-0.96). Based on the result, we extended our research to test whether the tissue water measurement can differentiate heterogeneous pathological structures of cancer tissues of different grades. Diffuse Optical Spectroscopic Imaging (DOSI) measures water molecular vibrational states associated with macro-molecular complexes in tissues such as proteins in a non-invasive way without compression of the breast. For DOSI measurements, a large region over breast tumors is scanned using a hand-held probe and spectral information on each point is spatially mapped to generate an image.Subjects and Methods: DOSI was employed to measure 19 primary cancers (18 IDC and 1 ILC). The spectral features of tissue water absorption from 935 nm to 998nm were compared to those of a pure water spectrum measured at body temperature in order to acquire the Bound Water Index (BWI: the residual between tissue and pure water spectra). In the spectroscopic images, the average and standard deviation of the pixels less than 20% of the range of the BWI in the image were calculated based on our previous finding that carcinoma tissues have lower BWI (more free water). The same parameters were calculated on normal tissues from the same patients in order to compare the tissue heterogeneity. The standard deviation was divided by the average value of each tissue type, and this value is communicated as a Heterogeneity Index (HI).Results: In our in vivo clinical studies, the cancer tissues showed 51% higher heterogeneity than the normal tissues (cancer HI: 0.046±0.018, normal HI: 0.024±0.018, p<0.001). Furthermore, the low (N=4), medium (N=9) and high grade (N=6) tumors, defined by the Nottingham Bloom-Richardson histopathological scores (3-5: low, 6-7: med and 8-9: high), demonstrated increasing heterogeneity in the water disposition with the tumor grade. The medium and high grade tumors had 10.5 and 28% more heterogeneity than the low grade tumors in average (low grade HI: 0.041±0.015, med: 0.045±0.02 and high: 0.052±0.017).Discussion: The tissue water measurement using DOSI showed its ability to measure increased pathological heterogeneity in more invasive cancer tissues. This result supports our earlier finding that the BWI can report on histopathological grades of cancer tissues. These pathological properties were acquired in a completely non-invasive way with non-ionizing low power light. The results of this study suggest that the water state measurement using DOSI may be used as a non-invasive optical biopsy with high specificity.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5008.
Molecular Breast Imaging as a Cost Effective Diagnostic Tool for Breast Cancer.
Authors: Larsen A. , Hughes J. , Frazier T.
Published: December 2010

Introduction: Molecular breast imaging (MBI), also referred to as breast specific gamma imaging (BSGI), utilizes nuclear medicine imaging with technetium 99-sestamibi to demonstrate metabolic activity of breast tissue. A focused gamma camera is used to assist in detecting lesions concerning for malignancy based on abnormal radiotracer uptake. MBI is performed with the patient in a sitting position and requires less breast compression than mammography. MBI is safe for patients with implanted metallic devices and renal disease. Facility charge for MBI is approximately $600 vs. $2500 for magnetic resonance imaging (MRI).Methods: IRB approval was obtained prior to study initiation. Forty four (44) patients underwent MBI with technetium 99-sestamibi after mammography and or ultrasound detected abnormalities for which biopsy was recommended.Results: Forty four (44) women ranging in age from 28 to 82 (median age 52) were found to have abnormal mammogram and or ultrasound findings requiring image guided biopsy. Prior to biopsy, MBI was performed. Fifteen MBI's were interpreted as demonstrating areas concerning for malignancy. Nine of the 44 patients (20.5%) demonstrated malignancy on biopsy pathology. Eight of these nine patients had positive MBI findings concerning for malignancy (88.9%). The patient whose MBI did not demonstrate findings consistent with malignancy had two sites of DCIS. The positive predictive value of these results was 53.3% and the negative predictive value was 96.55%.Conclusions: MBI shows promise in evaluating patients with abnormal mammogram or ultrasound findings. MBI is available at a much lower cost than MRI ($600 vs. $2500), which would have resulted in savings of $83,600 in this study alone. MBI can be used in patients for whom MRI is not an option such as claustrophobic patients, patients with renal disease limiting use of gadolinium contrast and patients who for other reasons such as body habitus and implanted metallic devices are not candidates for MRI. Further studies evaluating MBI are warranted to determine what role it may play in providing additional diagnostic data about patients with abnormal mammograms or ultrasounds. The role of MBI in settings where breast MRI is not available, is not technically feasible or is not an option due to cost also warrants further evaluation.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5014.
Correlation between Parameters of Optical Diffusion Imaging and Prognostic Factors of Breast Cancer.
Authors: Moon J. , Kim H. , Shin H.
Published: December 2010

Background: Total hemoglobin (HBT) and oxygen saturation (SO2) of optical diffusion imaging might be useful to predict extensive intraductal component (EIC) and lymphovascular invasion (LVI) in patients with breast cancer.To determine if the parameters of optical diffusion imaging correlate with the prognostic factors of breast cancer.Method and materials: A total of 95 patients who underwent surgery due to breast cancer were included in our study from March to October 2008. We obtained HBT, SO2 and synthesis diagnosis index (SDI) from optical diffusion imaging. Prognostic factors including estrogen receptor, progesterone receptor, HER-2/neu, HER-1, CK5/6, p53 status, LVI, EIC, lymph node metastasis and histologic grade were reviewed from pathologic report. We correlated HBT, SO2 and SDI on optical diffusion imaging with prognostic factors using Pearson correlation coefficient test.Result: Negative EIC showed correlation with low level of HBT and positive EIC showed correlation with high level of HBT (p=0.022). Negative LVI showed correlation with high level of SO2 and positive LVI showed correlation with low level of SO2 (p=0.038). Other prognostic factor shows no significant correlation with HBT and SO2. SDI shows no significant correlation with any prognostic factors.Conclusion: HBT and SO2 on optical diffusion imaging correlated with EIC and LVI, which enable prediction of EIC and LVI in patients with breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5018.
Correlation between Histopathological Diagnosis and Real-Time Tissue Elastography Scores.
Authors: Nishiyama Y. , Sakaguchi U. , Matsushita K. , Kawagoe A.
Published: December 2010

Background“Hardness” is important characteristics of carcinoma. Recent invention of Real-time Tissue Elastography (EG) has made scaling the hardness possible. At 31st SABCS, we reported that B-mode ultrasound(US)+mammography(MMG)+EG demonstrated high sensitivity even for the malignancy of the mass-forming lesions of 1.1 cm or smaller in diameter. This results suggested that possibility of a mass-forming lesion being malignant is ignorable low when it is judged as “not malignant” by this combined method. In this study, we verified correlation between EG score and histopathological diagnosis.Method416 patients with mass-forming and non-mass-forming breast lesions (118 malignant, 298 benign) whose tissues were diagnosed by histopathologically from October 2007 to March 2009 were studied. The hardness was scored on a scale 1 to 5 on Tsukuba elasticity scoring system.ResultsIn mass-forming breast lesions, the mean overall score of malignant diseases was 4.1 (invasive carcinoma 4.3, noninvasive carcinoma 3.5), and the mean overall score of benign diseases was 2.4, and thus scores of malignant diseases were significantly high (< 0.0001). On the other hand, in non-mass-forming breast lesions, the mean overall score of malignant diseases was 2.3 (invasive carcinoma 2.5, noninvasive carcinoma 2.4), and the mean overall score of benign diseases was 2.0, and thus no differences were seen between them. Furthermore, for examination by mass-forming lesions combined with non-mass-forming lesions the mean score of noninvasive carcinomas was 2.5.ConclusionIt was found that in mass-forming lesions, high scores suggest invasive carcinoma and low scores suggest benign diseases. However, as in non-mass-forming lesions, both invasive and noninvasive carcinomas almost show no high EG scores, it appeared necessary to look into invasive examinations such as biopsy for the diagnosis of non-mass-forming lesions.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5026.
MDCT Is Very Useful for Evaluating the Spread of DCIS.
Authors: Takayanagi H. , Kimijima I. , Yoshida K.
Published: December 2010

Introduction: Ductal carcinoma in situ (DCIS) may occasionally spread widely in the duct-lobular segment, thereby complicating the complete resection of the lesion in breast-conserving surgery. In order to perform a safe operation for DCIS, the lesion area must be precisely evaluated in the preoperative imaging studies. We performed multidetector-row computed tomography (MDCT) for patients with DCIS and investigated the potential of MDCT for detecting DCIS lesions.Patients and methods: A total of 74 patients with DCIS underwent MDCT. The size of the DCIS lesion in each patient was measured in the volume-rendering view, and these sizes were compared with the lesion sizes obtained by mapping from the pathological tests. The differences between the lengths obtained from pathological tests and those obtained from MDCT (L-path-L-CT) were calculated, and the relationships between the differences and some factors were investigated.Results: Among the 74 DCIS patients who underwent MDCT, DCIS was detected in 61 patients (82.4%). Among these 61 cases, 45.8% were cases of comedo-type DCIS, but the detection rates for comedo and noncomedo cases were not different. The L-path-L-CT values for histological grade I DCIS (1.55 cm) and grade II & III DCIS (0.23 cm) were statistically different. Higher-grade DCIS was more detectable than low-grade DCIS (p = 0.013). Among the other factors, the MDCT assessments and the pathological assessments for comedo-type DCIS and Her-2-positive cases were more consistent than the corresponding results for noncomedo DCIS and Her-2-negative cases. However, these differences were not significant.Conclusion: MDCT shows high efficiency in detecting DCIS. More aggressive types of DCIS, such as higher grade, comedo type, and Her-2-positive DCIS were more precisely detected by MDCT. In comparison with magnetic resonance imaging (MRI), the patient's position during MDCT testing is more similar to that during surgery. Therefore, MDCT is a highly useful presurgical imaging technique for the assessment of DCIS.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5029.
A Randomized Controlled Multicenter Trial of a Nutritional Formula in Women with Mastalgia Associated with Fibrocystic Breast Changes.
Authors: Mansel R. , Voss A. , Seth A. , Rushlow K.
Published: December 2010

Objective: Nodularity is a problem for clinical and radiological evaluation of the breast. A randomized, controlled, double-blinded trial was performed in women with cyclic breast pain (mastalgia) associated with fibrocystic breast changes to determine the effect of a liquid nutritional formula on breast pain and nodularity.Study Design: Following a one month baseline assessment, women were randomly assigned to receive 4 ounces per day of an experimental (EXP) or control liquid nutritional formula (CONT) for three months. The EXP contained 1 gram of gamma linolenic acid (GLA) (as borage oil), 750 mcg of iodine (as potassium iodide), and 70 mcg of selenium (as sodium selenate). The CONT was an isonitrogenous, isocaloric formula prepared with high-oleic safflower oil in place of borage oil, and without additional GLA, iodine, or selenium. Women were asked to record breast pain, supplemental medications, and menstrual signs daily using an interactive voice-response system (IVRS). Physical breast exams were performed by a healthcare professional at baseline and at the end of the study for a determination of nodularity.Results: A total of 503 women were screened for eligibility from 21 centers distributed across the United States. From this group, 188 women were randomized to receive the EXP or CONT intervention. The groups were similar at baseline. Peak breast pain scores were reduced from baseline similarly in the EXP (-32.2%) and CONT (-33.1%) groups (p=0.64). Among women who continued to take non-prescription pain medication for management of breast pain, the amount of medication was significantly less in the EXP group (p=0.0143). In addition, there was a greater reduction in breast nodularity in women taking the EXP compared to the CONT (p=0.03 vs. p=0.64). Both groups reported similar average weight at baseline, but women in the EXP lost an average of 2.6 lbs. of body weight, while those in the CONT gained an average of 3.6 lbs by the end of the study. Both formulas were well tolerated and willingness to consume the nutritional intervention was 82.7% in the CONT and 90.4% in the EXP as calculated by the number of voluntary withdrawals from the study and discontinuation of treatment for any reason. Women were considered compliant if the consumed at least 70% of the study product (daily reported by IVRS).Conclusion: These results indicate that the daily liquid nutritional formula containing GLA, iodine, and selenium may provide a safe nutritional alternative for reducing both nodularity and nonprescription medication use associated with cyclic breast pain in women with fibrocystic breast changes. Reduction in nodularity may improve physician and mammographic evaluation of the breast.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5034.
A Multicenter Randomized Phase II Study of Sequential Epirubicin/Cyclophosphamide Followed by Docetaxel with or without Celecoxib or Trastuzumab According to HER2 Status, as Primary Chemotherapy for Localized Invasive Breast Cancer Patient.
Authors: Pierga J. , Delaloge S. , Giacchetti S. , Brain E. , Savignoni A. , Sigal-Zafrani B. , Mathieu M. , Bertheau P. , Guinebretière J. , De Cremoux P. , Spyratos F. , Marty M.
Published: December 2010

Purpose: To assess anti tumor activity of sequential epirubicin/cyclophosphamide followed by docetaxel with the randomized addition of celecoxib In Her2-ve patients or trastuzumab in Her2+ve patients versus no additional treatment, in terms of pathological complete response (pCR), defined as absence of residual invasive breast carcinoma and of nodal involvement.Patients and methods: 340 patients with stage II and III breast adenocarcinoma, with tumors ineligible for breast conservative surgery received 8 sequential 3 weekly cycles with epirubicin (75mg/m2)-cyclophosphamide (750mg/m2) for 4 cycles followed by docetaxel (100 mg/m2) for 4 cycles. According to HER2 status (IHC + FISH) they were randomized to receive together with docetaxel, in HER2 negative group (220 patients) celecoxib 800 mg/day during cycles 5-8 or no additional treatment and in HER2 positive group (120 patients) trastuzumab (8mg/kg then 6mg/kg) infused together with docetaxel or no additional treatment. All patients except eight with HER2+ tumor received adjuvant trastuzumab for a cumulative duration of 12 months.Results: In the HER2 negative group, pCR (grade 1 and 2 of Chevallier's classification) was observed in 11.5% and 13% of patients treated without and with neoadjuvant Celecoxib respectively. In the HER2+ group, pCR rate reached 26% in those having received neoadjuvant trastuzumab versus 19% in the others. There was no cardiac toxicity and no toxic death. Triple negative breast cancers experience the highest pCR rate of 30%.Conclusion: It is the first report on the effect of the addition of celecoxib to chemotherapy in neoadjuvant setting in breast cancer, showing that celecoxib does not improve the pCR rate. Addition of trastuzumab does, yet not to the extent reported with a protracted co-administration before surgery. Expression of hormonal receptors appears to be the major prognosticator for pCR. Molecular studies of gene expression profiling should allow improving such prediction.Supported by PHRC AOM/2OO2/02117, Pfizer inc., Roche, sanofi-aventis.ISRCTN10059974

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5054.
High Resolution Oligonucleotide Array-CGH and Hot Spot Mutations To Select Patients for Targeted Therapies.
Authors: De La Cruz J. , Scott V. , Richon C. , Job B. , Dessen P. , Mathieu M. , Delaloge S. , Soria J. , Campone M. , Dalenc F. , Bachelot T. , Lazar V. , Andre F.
Published: December 2010

Background. A high number of genomic alterations (gene amplification, mutations) have been reported in breast cancers, and could potentially be targeted in early clinical trials. In the present molecular screening program, we aim at performing CGH array and hot spot mutations for 200 patients with metastatic breast cancer, in order to drive these patients in specific early clinical trials.Patients and Methods. Patients were selected to present a metastatic breast cancer under treatment in four French Centers. DNA was extracted by Qiagen method from either primary breast cancer or metastatic lesions. The hybridizations were done on 4*44K Agilent Array CGH. The analysis was done with Analytics v3.4.40 software. The file reported amplified zones (log2ratio>0.84 for biopsy, 1.58 for FNA) and focused on predefined genes that encode a candidate target. Hot spot mutations cover PI3K, AKT, PTEN genes. The primary endpoint was to determine how many patients were driven to a specific clinical trial because of molecular analyses. The secondary endpoints included PFS and response rates.Results. As june 12th, 109 patients were included in the molecular screening program. CGH array was feasible and could be interpreted within one month in 85 (78%) patients. Array-CGH identified high level amplification in 57 patients (67%). As expected, the two most frequent amplicons included ERBB2 (17q11-12, 24%) and FGFR1 (8p11, 16%).The accuracy to detect ERBB2 amplification was 90%. A significant number of patients presented rare gene amplifications including ERBB4 amplification (n=1), CCND1/FGF4 amplification (n=6), FGFR2 (n=2), EGFR (n=1), VEGFB (n=4), BCR (n=1)Interestingly, only 4 patients presented TOP2A amplification. Hot spot mutations for PI3KCA, AKT, PTEN is being performedConclusion. This preliminary analysis shows that array CGH allows to identify patients who are candidate for targeted agents in phase I/II trials. Accrual will continue until 200 patients. Selected patients will be included in clinical trials in second semester of 2009 after a target validation by IHC.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5067.
Brk/PTK6 & Bcl-x Proteins Contribute to Reduced Sensitivity to Chemotherapy.
Authors: Burmi R. , Pennington C. , Edwards D. , Harvey A.
Published: December 2010

Background: Brk/PTK6 is an intracellular protein tyrosine kinase which is frequently overexpressed in the majority of breast cancers and has been shown to increase the anchorage-independent survival of breast cancer cell lines by altering Bcl-xL:Bcl-xS ratios. A Brk binding partner has also been found to modulate the alternative splicing of the Bcl-x gene, and altered Bcl-xL/xS levels are implicated in cell sensitivity to chemotherapeutic agents. We hypothesised that Brk may decrease cell response to chemotherapy through regulation of Bcl-x.Materials & Methods: To determine the clinical relevance and coexpression of Brk and Bcl-xL/S, quantitative PCR was performed on a clinical breast cancer biopsy material (n= 46). Gene knockdown studies using siRNA for either Brk or Bcl-xL were performed in T47D breast cancer cell line. Cells were subsequently cultured onto a 96 well plate in quadruplicate and subject to dose range of the chemotherapeutic drug, doxorubicin or paclitaxel for 72 hours before being tested for viability using the MTT assay. Protein levels for Brk, Bcl-xL/xS in response to siRNA treatments were determined using western blotting.Results: Quantitative PCR analysis of clinical breast cancer series reveal that Bcl-xL and Bcl-xS expression were associated with high grade tumours (p=0.028 and p=0.009 respectively). Moreover, both Bcl-xL and Bcl-xS were found to be significantly associated with Brk expression within these clinical samples (p<0.0001 and p=0.0026 respectively). T47D breast cancer cells with suppressed Brk levels showed a significant increase in sensitivity to doxorubicin and paclitaxel compared to control cultures (p=0.035 for doxorubicin and p<0.05 for paclitaxel at 0.625μM and 0.25nM respectively). Reduced Brk levels in T47D cells resulted in reduced levels of Bcl-xL protein. T47D cells with suppressed Bcl-xL levels also showed a significant increase in the sensitivity to doxorubicin compared to control cultures (p=0.039 at 0.31μM).Conclusion: These data suggests that Brk expression decreases the sensitivity of currently used chemotherapies through a mechanism that may involve regulation of Bcl-xL/xS protein levels. Thus targeting Brk may represent an important strategy for improving patient responses to chemotherapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5068.
A TCRMimic Monoclonal Antibody Generated from a Directly Discovered Epitope of p68 RNA Helicase Dramatically Debulks Large Established Tumors through Recognition of a Specific MHC/Peptide Complex.
Authors: Verma B. , Weidanz J. , Weidanz J.
Published: December 2010

Debulking large tumors using monoclonal antibodies (mAb) that specifically recognize unique T cell epitopes represents a novel approach to cancer therapy.Using a mAb designated as a TCRm we recently demonstrated the in vivo suppression of tumor growth. The TCRm had specificity for a peptide/MHC class I molecule expressed on the surface of the tumor cells. In this study we use an innovative direct epitope discovery strategy that led to the identification of peptide YLLPAIVHI in the context of HLA-A*0201 (YLL/A2) in human breast carcinoma cell lines. The peptide is derived from the DEAD Box RNA helicase protein p68, a protein overexpressed and highly phosphorylated in malignant cells. A TCRm antibody (RL6A) was generated and characterized for binding specificity, sensitivity and affinity for the YLL/A2 target. The TCRm specifically stained cells that co-expressed HLA-A2 and p68 but not cells lacking expression of p68 protein or HLA-A2 alone. It binds to cognate peptide/HLA-A2 with an affinity constant (KD) of 5.69 x 10-10 M and recognized specifically the YLL/A2 epitope in human breast cancer tissue but not in normal control breast tissue.Mice implanted in the mammary fat pad with human breast tumor cells grew tumors of considerable size, yet when treated with RL6A TCRm there was a marked reduction in tumor size. Examination of frozen tumor tissue sections revealed significantly greater numbers of apoptotic cells in mice treated with RL6A TCRm than with control antibody. Investigation in vitro showed that TCRm engagement of specific YLL/A2 epitope led to the induction of apoptosis providing the first experimental evidence that MHC signaling could directly mediate tumor cell death. Together, these data indicate that TCRm targeting of specific peptide/MHC complexes unique to tumor cells is effective not only in slowing the growth of tumors but also in dramatically reducing the size of established tumors and suggests a possible novel approach to the treatment of cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5072.
Combination of a Novel Growth Hormone Releasing Hormone Antagonist JMR 132 and Tamoxifen Enhances the Inhibitory Effects on Growth of ZR 75 and T47D Estrogen Dependent Human Breast Cancer Cells In Vitro and In Vivo.
Authors: Buchholz S. , Koester F. , Schally A. , Krishan A. , Papadia A. , Ortmann O. , Seitz S.
Published: December 2010

Background: Tamoxifen is the oldest selective estrogen receptor modulator (SERM) developed to antagonize the estrogen receptor function on human breast cancer cells. JMR 132 is a novel GHRH Antagonist proven to be potent inhibitor of tumor growth in estrogen receptor negative human breast cancer cell lines. A combination of both substances could induce a more potent growth inhibition, as compared to the single drugs.Methods: To determine inhibitory effects of the GHRH antagonist JMR 132 and Tamoxifen alone and in combination, estrogen receptor positive cell lines ZR 75 and T 47D were incubated with substances at concentrations of 2.5µM, 5µM and 10µM. The cell viability after a period of 72 h was measured by MTT assay. To assess if the treatment arrested cell growth at a specific phase of the cell cycle or simply caused cell death, we performed a flow cytometric DNA cell cycle analysis on ZR 75 cells exposed to JMR 132, Tamoxifen and their combination. In an in vivo model, the cell line ZR 75 was xenografted subcutaneously into nude mice. Treatment groups were control, JMR 132 at a dose of 10µM, Tamoxifen at a dose of 100µgr per mouse q3 per week and the combination of both substances. All given for a period of 4 weeks.Results: In vitro 1o µM JMR 132 and Tamoxifen significantly decreased the proliferation of the ZR 75 and T47D cell lines, as compared to controls (p< 0.05). The combination of 1o µM inhibited the growth of ZR 75 was significantly by 14%, as compared to JMR 132 alone. The combination also significantly suppressed the growth of T47D by 14% and 22% respectively, as compared to Tamoxifen and JMR 132. In in vivo treatment with the combination of JMR 132 and Tamoxifen induced a significant inhibition of tumor growth of ZR 75 xenografts after 14 days compared to the single agents groups and inhibition remained significant until the end of the study on day 28. The flow cytometric DNA cell cycle analysis revealed a cell cycle arrest in the S Phase which led to apoptosis of the cells.Conclusion: Treatment of ZR 75 and T47D estrogen receptor positive human breast cancer lines with GHRH antagonist JMR 132 and its combination with Tamoxifen induced potent inhibitory effects in vitro and in vivo and led to subsequent cell death. Further studies are being conducted to elucidate the mechanisms of action of these effects.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5073.
An Evaluation of the Safety and Efficacy of Lapatinib Plus Trastuzumab Plus Paclitaxel in First-Line HER2+ Metastatic Breast Cancer.
Authors: Esteva F. , Franco S. , Hagan M. , Somer R. , Dombroski C. , Florance A. , Turner S. , Stein S. , Perez A.
Published: December 2010

Background: Trastuzumab given concomitantly with paclitaxel is an established first-line metastatic breast cancer (MBC) treatment; however, approximately 30% to 40% of patients show no response as defined by RECIST to this combination. Lapatinib is an oral dual tyrosine kinase inhibitor of EGFR and HER2 and approved, in combination with capecitabine, after trastuzumab-based therapy for HER2+ MBC. The differing mechanisms of action may allow synergy between lapatinib and trastuzumab when given concurrently with paclitaxel for MBC.Methods: This is an open-label phase I safety study. The primary objective of the study was to assess the safety and tolerability of lapatinib when administered with concurrent paclitaxel and trastuzumab. Patients were enrolled into 1 of 3 cohorts. Cohort 1 received lapatinib (1000 mg/d), paclitaxel (80 mg/m2 weekly for 3 weeks of a 4-week cycle), and the standard weekly dose of trastuzumab. Cohort 2 received lapatinib (1000 mg/d), paclitaxel (70 mg/m2), and trastuzumab. Cohort 3 received lapatinib (750 mg/d), paclitaxel (80 mg/m2), and trastuzumab. Patients: were 3+ by IHC or FISH amplified for HER2; had not received prior treatment in the metastatic setting; and had an ECOG performance status of 0 or 1. Safety was assessed weekly and tumor response per RECIST every 8 weeks. Treatment continued until disease progression or toxicity. All patients received prophylactic loperamide per protocol amendment.Results: Open-label safety enrollment is complete and preliminary safety data are available for all 63 patients. The most frequently reported AEs for all cohorts were diarrhea (81%), fatigue (71%), rash (68%), nausea (59%), alopecia (56%), and vomiting (37%). Dehydration was the most frequently reported serious AE (SAE) (n=6; 10%) and most were in cohort 1 (5 patients). Diarrhea of any grade was reported for 97% of patients in cohort 1, 93% of patients in cohort 2, and 50% of patients in cohort 3. No grade 4 or 5 diarrhea events were reported. The proportion of patients with a maximum toxicity of grade 3 diarrhea was 62% in cohort 1, 43% in cohort 2, and 20% in cohort 3. Two patients (both in cohort 1) reported diarrhea as a serious AE and a total of 4 patients (6%) had lapatinib withdrawn due to diarrhea (none in cohort 3).Conclusions: Preliminary safety results show a high incidence of grade 3 diarrhea when lapatinib was given at a dose of 1000 mg/d concurrently with paclitaxel and trastuzumab. This incidence was significantly reduced with a lapatinib dose of 750 mg/d in combination with paclitaxel and trastuzumab when given with loperamide. Final safety and efficacy data, including overall response and progression-free survival, from this open-label safety study will be presented.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5085.
Pharmacokinetic (PK) Interactions between Lapatinib (L) and Vinorelbine (VNR) in a Phase I Study in Locally Advanced or Metastatic Breast Cancer (LAMBC) Patients(Pts) Overexpressing HER2. GEP-01 – A Study of the FNCLCC-Group of Early Phase Trials.
Authors: Lokiec F. , Brain E. , Isambert N. , Dalenc F. , Tresca P. , Bonneterre J. , Rezaï K. , Roché H. , Penel N. , Jimenez M. , Diéras V. , Fumoleau P.
Published: December 2010

Background: Lapatinib is effective in overexpressing HER2 LAMBC pts. Following anthracyclines, taxanes and capecitabine, VNR is an active agent in metastatic setting. Both drugs use the same main metabolic pathway: Cytochrome P450 3A4. We investigated the combination of L + VNR, seeking the recommended dose for further phase II studies, and looking for the PK interactions between the 2 drugs.Methods: Women with HER2+ LAMBC, in progression after ≤ 2 lines of trastuzumab-based treatment were treated with a 7 day (D-7 to D0) loading dose of L before starting VNR on a D1 and D8 q3w iv regimen. L was given po continuously. G-CSF as primary prophylaxis of febrile neutropenia (FN) was not permitted. Dose levels (DL) (L mg/VNR mg/m²) ranged from 750/20 to 1250/30 with 3 patients per DL. PK samples were collected on 7 points on D1 of cycle 1 for L and VNR dosages. PK time points analysis was performed on plasma for L and whole blood for VNR. For both drugs the analytical method was Ultra Performance Liquid Chromatography (UPLC) coupled with tandem mass spectrometry (MS-MS) as detection. VNR PK parameters were estimated by nonlinear mixed effect modeling (NONMEM) and L PK parameters were estimated by non compartmental software (MicroPharm).Results: Seventeen pts and 68 PK time points were available for VNR analysis. VNR was ascribed to a tri-compartment model. When considering all VNR data together, VNR clearance (CL) was 37.8±16.7 L/h, while it showed a gradual decrease according to specific dose of L: 47.5±20.4 L/h for L 750 mg (3 patients) vs 35.6±4.3 L/h for pooled data for L 1000 and 1250 (14 patients). This trend was not significant likely due to low number of patients treated at lower dose of L, while no significant difference was observed between L 1000 and L 1250. The mean elimination half life time (t1/2) of VNR was 0.12 h. Six patients and 42 PK time points were analyzed for L. L CL were respectively 83.8 ± 41.3 and 57.1 ± 5.5 L/h for patients who received 750 mg (3 patients) and 1000 mg (3 patients) of L with 20 mg of VNR. The mean elimination half life time (t1/2) of L was 2.89 h.Conclusions: A potential PK interference occurred between VNR and L. When the dose of L increased, both VNR and L CL decreased, likely due to CYP450-3A4 interactions. This trend should be confirmed by the PK analysis of L and VNR in more patients, including an intermediate DL (1250/22.5) still ongoing.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5091.
A 28-Day Intrathecal Toxicity Study with Trastuzumab in Nonhuman Primates.
Authors: Braen A. , Perron J. , Tellier P. , Catala A. , Kolaitis G. , Geng W.
Published: December 2010

Introduction: Trastuzumab is indicated for the treatment of patients with breast cancer whose tumors overexpress the HER2 protein. Brain metastases have been observed in breast cancer patients, being the highest in women with HER2-overexpressing tumors. The blood-brain barrier, however, may prevent trastuzumab from reaching appropriate concentrations in the cerebrospinal fluid (CSF). Case studies have been reported in which trastuzumab was intrathecally administered for the treatment of metastatic brain cancer in humans. However, no extensive preclinical safety evaluation has been performed for intrathecal application after repeat-dosing in a trastuzumab cross-reactive species using doses equivalent to or higher than those used in patients. To evaluate the potential of local effects, a 28-day toxicology study with weekly intrathecal administration of trastuzumab was performed in cynomolgus monkeys, a trastuzumab cross-reactive species. Preservative-free trastuzumab was used in this preclinical safety study, as caution has been raised for the intrathecal use of benzyl alcohol-containing solutions. Methods: Five male and five female monkeys were evaluated per dose group, of which two monkeys/sex/group were evaluated after a 4-week recovery period. Animals were assigned to 3 groups which received 4 weekly 30-minute intrathecal infusions at 0, 3, or 15 mg per dose at an infusion rate of 1.5 mL/hour. Endpoints evaluated included clinical signs, body weight, appetite, neurological examinations, clinical pathology, gross pathology, and histopathology of the brain (with inclusion of meninges), spinal cord, and infusion site. In addition, CSF and serum samples were collected for toxicokinetic evaluations. Results and Discussion: No test article-related effects on clinical signs, body weight, appetite, neurological examinations, clinical pathology, gross pathology or histopathology were noted. Of note is the absence of demyelination as was discussed as an uncertainty of intrathecally administered proteins. The doses used of 3 and 15 mg per dose resulted in 0.051 and 0.24 mg/g brain doses, respectively. Doses administered in clinical case reports ranged from 5 to 100 mg per dose. For a 1500 g human brain weight, these intrathecal doses would be equivalent to 0.0033 and 0.067 mg/g brain weight. At the high dose of 15 mg, maximum CSF concentrations (Cmax) were as high as 403 µg/mL. The applied doses and CSF concentrations achieved in the current study greatly exceeded those reported in patients (6.4 µg/mL at an intrathecal dose of 20 mg). Conclusion: The results of this 4-week toxicology study in monkeys with the results of the clinical case studies support future studies to evaluate intrathecal application of trastuzumab in patients with brain metastasis in HER2-positive breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5101.
Outcome of HER2-Positive (HER2+) Metastatic Breast Cancer Patients (MBC) Treated with Trastuzumab (T): An Institutional Based Review.
Authors: Guilhaume M. , Dieras V. , Fall M. , Pierga J. , Beuzeboc P. , Cottu P. , Simondi C. , Courbard M. , Mignot L. , Livartowski A.
Published: December 2010

Background: HER2+ status is associated with poor prognosis, high incidence of visceral and brain metastasis. However the addition of trastuzumab to chemotherapy (CT) significantly improves survival in early and advanced breast cancer. The purpose of this retrospective study was to explore the pattern of outcome in a cohort of MBC patients treated with T-based chemotherapy in a single institution. T was approved in Europe in 2000 and in 2001 all pts had access to T according HER2+ status. Methods: Women with de novo or recurrent breast cancer treated with trastuzumab at Institut Curie between 2001 and 2006 with HER2+ status (IHC 3+ or FISH +) were identified from the Institut Curie database. Disease was classified in two groups: patients who received T upfront and those who received T after one or several CT regimens. Overall survival (OS) was defined as the time from the date of the first metastasis to the date of death or last follow-up and was estimated using the Kaplan-Meier product method. Results: The final analysis included 244 patients. Median age was 53.4 yrs (29-80). Median time from primary and first metastasis was 22 mths (0-238). Visceral metastasis were present in 153 pts (63%) and 125 pts (51%) presented multiple sites. One hundred pts (42%) developed brain metastasis during the course of disease. One hundred and sixty five pts (68%) received T as first line, 79 pts (32%) after a median of one line of CT (median 1, range 1-5). One hundred and twenty four pts (52%) received more than 3 regimens. The median overall survival was 53 mths (4-113), similar in both groups. However there is a major bias: pts with very aggressive disease not treated upfront with T not have not been offered delayed T and don't appear in the analyzed population. Patients who developed brain metastasis had a median survival of 41 mths (11-90). Complete characteristics of pts will be presented. Conclusions: The introduction of T has altered the natural history of HER2+disease. Even outside a clinical trial, our results show that the addition of T to CT improves the prognosis of MBC patients with HER2+ disease. Prolongation of T after progression with other CT appears beneficial, even in pts with a high disease burden. The high incidence of brain metastases remains an issue in such a population.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5107.
Observational GHEA Study: Adjuvant-Trastuzumab-Treatment of HER2-Positive Breast Carcinoma.
Authors: Campiglio M. , Tagliabue E. , Balsari A. , Bufalino R. , Ferri E. , Menard S.
Published: December 2010

Background: GHEA (Group HErceptin in Adiuvant treatment), an Italian multicentric observational study, was designed with the aim to investigate on adjuvant trastuzumab-therapy of HER2-positive breast carcinomas in conventional clinical setting.Material and Methods: Eligible patients should have been treated with trastuzumab in adjuvant setting and were collected among 35 oncologic Italian centers.Results: The patient registered so far in the GHEA database (500 pts) had a median age of 52 years and 64% were in post-menopause at the time of trastuzumab therapy. Primary breast tumor were mainly of ductal type (93%) of high grade (59% were GIII) and small in size (58% T1, 35% T2, 4% T3 and 3% T4). 12% of the samples included in GHEA were classified as T1micro, T1a and T1b, three categories generally not included in clinical trials. ER and PgR positive tumors were 58% and 51%, respectively and more than 53% presented in situ component. Positive lymph-nodes were present in 53% of cases. 15% of the patients received neo-adjuvant therapy prior surgery and adjuvant treatment. In the neo-adjuvant therapy, 95% of patients received anthracycline–based regimens and 72% were treated with taxanes. In adjuvant setting 89% of patients received anthracycline–based regimens and only 43% were treated with taxanes. 88% of the patients completed 1-year-trastuzumab treatment. Median exposure time of the series was 26 months. To date, 6% of the patients presented distant metastasis (i.e. bones, liver, lung, SNC) and half of them progressed during trastuzumab treatment.Discussion: The purpose of the present study is to gather information from a large number of patients related to the effect of adjuvant trastuzumab treatment in conventional clinical setting, the treatment of tumor categories not included in clinical trails and the therapies used at the relapse of these patients. All together, these data are warranted to design further clinical trials to better define guide-line for trastuzumab adjuvant therapy and for the treatment of these patients at relapse.Supported by AIRC and Roche.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5112.
Pertuzumab and Trastuzumab: Re-Responses to 2 Biological Agents in Patients with HER2-Positive Breast Cancer Which Had Previously Progressed during Therapy with Each Agent Given Separately: A New Biological and Clinical Observation.
Authors: Baselga J. , Cortes J. , Fumoleau P. , Petrella T. , Gelmon K. , Verma S. , Pivot X. , Ross G. , Szado T. , Gianni L.
Published: December 2010

Background: Pertuzumab, a fully humanised IgG1 monoclonal antibody, is a human epidermal growth factor receptor 2 (HER2)-dimerisation inhibitor directed to the dimerisation epitope of HER2 (trastuzumab binds to the juxta-membrane epitope). Pertuzumab has demonstrated promising activity when given with trastuzumab to patients with HER2-positive metastatic breast cancer (mBC) which had progressed during therapy with trastuzumab in a 2-step Phase II study.1Methods: The protocol was amended to include a 3rd cohort of patients to determine the activity of pertuzumab when given without trastuzumab.2 Patients recruited into this 3rd cohort were allowed to have trastuzumab re-introduced in combination with pertuzumab if there was inadequate response to pertuzumab alone or response followed by relapse.Results: Twenty-nine patients were recruited into this 3rd cohort. Patients had reached their 3rd line of treatment for mBC. To date, 15 patients have had trastuzumab re-introduced after disease progression on trastuzumab therapy and pertuzumab monotherapy. Among these 15 patients, at the time of this analysis there have been 3 patients with confirmed responses. There are also 4 patients who had not yet undergone 8 cycles of assessments to reach the overall best response end point, of which at least 2 were experiencing stablisation of disease. Updated data on activity and toxicity will be presented.Conclusions: We believe this is the first time that anti-tumour activity has been reported in patients when 2 biological agents have been used together after the disease has progressed during therapy with each agent alone. There are several mechanisms which might explain this phenomenon. Trastuzumab prevents proteolytic cleavage of the extracellular domain of HER2, keeping the receptor in situ. The addition of a second antibody to a separate epitope increases the potential for antibody-dependent cell-mediated cytotoxicity and prevents dimerisation between HER2 and other HER family members, such as HER1 and HER3. The combined antibodies might increase the efficiency of inhibition of signal transduction. There are wide-ranging and potentially significant biological and clinical implications.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5114.