bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P3-15-02: A prospective, open label, non-comparative trial to determine the incidence of chemotherapy-induced nausea and vomiting associated with the docetaxel-cyclophosphamide regimen in early breast cancer patients. Results from the GEICAM 2009-02 study
Authors: Carañana V , Ramos M , Damau E , García-Saenz JA , González X , Murillo L , Calvo L , Morales S , González-Farré A , Fernández-Morales LA , Carrasco E , Casas MI , Angulo MdM , Cámara MC , García AI , Llombart A , Jara C
Published: August 2014

Background: Docetaxel-Cyclophosphamide (TC) has become a common chemotherapy regimen for moderate-high risk Early Breast Cancer (EBC) patients. The incidence of chemotherapy induced nausea and vomiting (CINV) with TC together with an adequate standard anti-emetic therapy with 5-HT3 antagonist and corticosteroids are unknown. This study investigates the incidence of emesis control (complete response), defined as no vomiting (any grade of NCI CTCAE version 4.0) and no use of rescue treatment within 120 hours after the first cycle of TC. Secondary objective evaluates the efficacy of Aprepitant in non-responding patients.

Methods: EBC patients with no prior moderate-high risk CINV were included. Patients received Docetaxel 75 mg/m2 plus Cyclophosphamide 600 mg/m2 IV every three weeks. Antiemetic treatment consisted of oral Dexamethasone (D) 8 mg (at night) on day 0; oral D 8 mg x 3 (in the morning, 1 hour before chemotherapy and at night) plus 5-HT3 antagonists on day 1; and oral D 8 mg x 2 (in the morning and at night) on days 2 and 3. Non-responding patients (vomiting or need of rescue therapy in the first cycle) were offered participation in the efficacy phase (Aprepitant 125mg day 1 and 80 mg days 2 and 3 added to the standard antiemetic therapy in cycle 2). In addition to the standard NCI-CTCAE adverse event collection, a patient´s diary (from day 1 to 6) and the FLIE (Functional Living Index-Emesis questionnaire) were used. Assuming a 25% (+/- 6%) of patients resistant to standard antiemetic therapy, 212 patients were estimated.

Results: From May-11 to March-13, 212 EBC patients were included. Median age was 57 years (range 34-82), 29.3% were premenopausal. Twenty-seven patients were excluded from the main analysis because of major protocol violations (25) or consent withdrawal (2). Twenty-four patients (13%; IC95%: 8.2 – 17.8) did not respond to standard antiemetics and entered the efficacy phase with Aprepitant. From these 24 patients, 14 (56%; IC95%: 36.5 – 75.5) achieved a complete response on cycle 2. No adverse events related to Aprepitant were observed.

Conclusions: Proper use of standard antiemetic therapy for early breast cancer patients treated with TC provides a high control rate (87%). Among no responding patients, about half of them were rescued with Aprepitant. Identification of the non-responding patients could lead to a better antiemetic control with Aprepitant from the first cycle.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-15-02.
Abstract P3-15-06: Results of the MARS study on the management of antiangiogenics’ renovascular safety in breast cancer
Authors: Gligorov J , Janus N , Daniel C , Beuzeboc P , Ray-Coquard I , Rey J-B , Jouannaud C , Spano J-P , Thery J-C , Morere J-F , Goldwasser F , Mir O , Scotté F , Oudard S , Azizi M , Dorent R , Deray G , Launay-Vacher V
Published: August 2014

Background:

Anti-VEGF drugs (AVD) are widely used in cancer patients (pts). Hypertension (HTN) and proteinuria (Pu) are class-side-effects of AVD, related to the inhibition of the VEGF pathway. The MARS study has been conducted to assess the renovascular tolerance of these drugs in the clinical setting.

Methods:

This multicentric, prospective, observational study evaluated the renovascular safety of AVD in pts naive from any AVD, conducted in 7 centres in France, from 2009 to 2012, with a follow-up (f/u) of 1 year. Data collected included: gender, age, serum creatinine (SCr), diabetes, HTN, hematuria (Hu) and dipstick Pu, at baseline and at each visit.

Results:

1124 cancer pts were included; 402 breast cancer (BC) pts received bevacizumab (1st line: 14.4%; median durations of treatment: months)). Median age at inclusion was 55 years (19-65). Visceral, bone and cerebral metastasis frequencies were 74.7, 5.1 and 2.5%, respectively. HTN prevalences: 12.4%. Baseline renal assessment retrieved: Pu 23.9%, Hu 16.2%, mean aMDRD 96.4 ml/min/1.73m2 and 14 pts with aMDRD<60. The incidence of de novo Pu and HTN during f/u was 61.7 and 16.8% (Table). 69.7% of pts with Pu at inclusion improved or remained stable. Among pts with de novo Pu, 75.7% afterwards improved/normalized. No grade 4 Pu has been reported. Renal function remained stable with a mean aMDRD of 96.2 at the end of f/u. 7.4% had grade 2-3 SCr increase (no grade 3-4). No thrombotic micro-angiopathy (TMA) was reported.

Conclusion:

These results on the renovascular safety of bevacizumab in BC pts showed that 1) TMA is rare, 2) Grade 3 Pu developed in 4.6% of pts, with no grade 4, 3) less than 17% developed HTN, and 4) aMDRD was stable. Furthermore, in case of a renovascular effect, investigators followed the recommendations from the French Society of Nephrology (Halimi JM. Nephrol Ther 2008) and no treatment withdrawal for unmanageable renovascular toxicity occurred.

Renovascular effects in MARS breast cancer patients treated with bevacizumabRenovascular effectsPrevalence at inclusionIncidence during follow-up (de novo**)Pu* (All grades)23.9%61.7%Pu* (Grade 1)22.2%47.1%Pu* (Grade 2)1.7%10.0%Pu* (Grade 3)0.0%4.6%Pu* (Grade 4)0.0%0.0%Hypertension12.4%16.8%Hu* (All Hu)16.2%42.1%Hu* (Traces/+)11.7%33.2%Hu* (++)2.6%6.3%Hu* (+++)1.9%2.6%SCr increase* (All grades)-80.0%SCr increase* (Grade 1)-68.6%SCr increase* (Grade 2)-7.1%SCr increase* (Grade 3)-0.3%*NCI-CTC v4; **de novo = patients with no event at inclusion and who presented an event during follow-up.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-15-06.
Abstract P3-15-09: Impact of granulocyte colony-stimulating factors on febrile neutropenia risk during early-stage breast cancer treatment
Authors: Ben Abdallah SS , Letarte N , Messier C , Charpentier D , Yelle L , Younan R , Bestawros A
Published: August 2014

Febrile neutropenia (FN) is a significant cause of both mortality and morbidity in patients undergoing adjuvant or neoadjuvant chemotherapy for early-stage breast cancer (ESBC). The risk of FN varies with respect to the type of chemotherapy used and patient-specific characteristics. Granulocyte colony-stimulating factors (G-CSF) are recommended, but not mandatory, as primary prophylaxis for patients at high-risk of FN. This study aimed to evaluate the impact of filgrastim and pegfilgastrim on the incidence of FN during adjuvant and neoadjuvant chemotherapy for ESBC. Risk factors for FN were also assessed.

We conducted a retrospective cohort study that included 297 consecutive patients undergoing adjuvant or neoadjuvant chemotherapy for ESBC between May 2010 and May 2012. Patients on clinical trials were excluded. 173 patients (58%) received primary G-CSF prophylaxis whereas 124 (42%) did not. In patients receiving G-CSF prophylaxis, filgrastim was more commonly prescribed (59.9%), due to reimbursement reasons in Quebec. In total, 38 patients (12.8%) manifested at least one episode of FN. The incidence of FN was significantly reduced in patients receiving primary prophylaxis with G-CSF (p = 0.012).

The most commonly prescribed chemotherapy protocols were FEC-D (5-Fluorouracil, epirubicin, cyclophosphamide followed by docetaxel), AC-T (Adriamycin, cyclophosphamide followed by paclitaxel), and TC (docetaxel and cyclophosphamide). The use of G-CSF as primary prophylaxis was associated with a decrease in FN incidence in patients receiving FEC-D and TC.

Table 1 Incidence of febrile neutropenia in early-stage breast cancer chemotherapy G-CSF primary prophylaxis N = 173Absence of G-CSF prophylaxis N = 124  N (%)N (%) Febrile neutropenia (all chemotherapy)15 (9)23 (19)p = 0.012*TC4/51 (8)5/11 (45)p = 0.01*FEC-D6/59 (10)8/27 (30)p = 0.023*AC-T3/31 (10)8/63 (13)p = 0.668* denotes statistical significance at p < 0.05

One death occurred in a patient receiving FEC-D who had not received prophylaxis. There was no significant difference in the dosing of filgrastim between patients with FN (mean dose received = 5.289 mcg/kg) and those without FN (mean = 5.167mcg/kg) (p = 0.742). Furthermore, previously described risk factors for FN such as age, comorbidities and decreased baseline albumin were not associated with an increased of FN in our study.

Although this was a retrospective study with inherent limitations, our cohort included, to our knowledge, the largest number of patients in a study of G-CSF use in ESBC. We found a statistically significant reduction of FN in patients receiving primary G-CSF prophylaxis. Primary prophylaxis should be routinely offered to patients undergoing chemotherapy for ESBC and in particular, to those receiving docetaxel-containing regimens.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-15-09.
Abstract P4-01-02: The role of FLT PET early assessment of response to endocrine therapy for early stage breast cancer
Authors: Linden HM , Kurland BF , Link JM , Novakova A , Chai X , Gadi VK , Specht JM , Hills D , Gralow JR , Schubert EK , Korde L , Peterson LM , Doot R , Eary J , Shields A , Krohn KA , Mankoff DA
Published: August 2014

Background: In estrogen receptor positive (ER+) tumors, a low proliferative index (Ki-67) two weeks into endocrine therapy predicts response. FLT PET non-invasively measures tumor proliferation in vivo. The pre-operative window is an opportunity to assess impact of systemic therapies. We tested associations between FLT PET qualitative and quantitative measures and Ki-67 following two weeks of aromatase inhibitor (AI) therapy.

Methods: Women with clinical stage I-II ER+ HER2– breast cancer underwent “run-in” of AI monotherapy prior to definitive surgery. Premenopausal women were given GNRH agonist treatment 2 W prior to AI therapy. FLT PET was performed before AI therapy, and 1-7 days before surgery. Ki-67 was measured in baseline core biopsy and surgical specimens.

Results: Fourteen patients (8 postmenopausal, 6 premenopausal) have been enrolled. All have undergone baseline FLT PET imaging; 11 have completed imaging and surgery, including one premenopausal patient with no residual invasive carcinoma following 26 days of AI therapy. The majority harbored ductal carcinomas (n = 9, 5 with lobular histology) with the majority histologic grade ≥ 2 (n = 11). The median number of days exposed to AI was 19 (range, 9-42). Baseline SUVmax ranged from 1.2 to 3.9 (median 2.2), and post run-in SUV (6-64 days later) ranged from 1.2 to 2.8 (median 1.8). Baseline Ki-67 ranged from 6-26.2, median 11.6; surgical Ki-67 post AI therapy ranged from 0- 20.3 median 3.7, with seven below 5%. SUV and flux declined in most patients, as did Ki-67.

Quantitative FLT flux correlated with tumor response assessed by proliferative index (Ki-67) before the “run-in” period, with a stronger correlation at surgery, Pearson correlation coefficients = 0.41 and 0.82, respectively. FLT SUV and qualitative changes were not strongly associated with Ki-67.

Conclusions: Both pre and postmenopausal women with early stage breast cancer showed imaging and tissue response to endocrine therapy. Quantitative, but not qualitative FLT is a promising tool to assess tumor proliferation and response to therapy. Accrual is ongoing and updated results will be reported.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-02.
Abstract P4-01-06: High resolution breast PET for monitoring response of breast cancer to neoadjuvant chemotherapy
Authors: Nelson JC , Lehde NL , Green CJ , Villanueva-Meyer J
Published: August 2014

Purpose:

To evaluate the sensitivity and accuracy of high resolution breast PET for monitoring the response of breast cancer to neoadjuvant chemotherapy.

Patients and Methods:

High resolution breast PET imaging was performed in 29 women before and following the completion of neoadjuvant chemotherapy in this retrospective patient review study. Tumors were divided into three receptor subtype categories: triple negative, human epidermal growth factor receptor 2 (HER2) positive, and estrogen receptor (ER) positive/HER2 negative. Breast PET was used to characterize lesion morphology, size and maximum FDG uptake. Lesion FDG uptake to background glandular FDG uptake (LTB) was used to allow comparison between imaging studies. Tumor response was assessed dichotomously (i.e. presence or abscence of residual tumor in the surgical specimen) and results were compared to breast PET findings. For discrepant findings, retrospective re-review of the breast PET images was completed with full LTB ratio and Grey/Red evaluation.

Results:

The average age of the patients was 52.6 years (range 24 - 72 years). Pathology found residual tumor in 66% (19/29). Breast PET images correctly identified residual disease in 17/19 patients and correctly predicted a complete pathological response in 9/10 patients for an overall accuracy of 90% (26/29). One tumor was missed due to poor positioning and a second (<1mm in size) was not identified. One breast PET showed residual uptake which was found to be sclerosis on pathology. The cancer subtype with the highest FDG uptake before chemotherapy was the triple negatives with a LTB ratio of 17.4, next highest was the HER2+ at 16.7 while the HER2 negative group showed the lowest metabolic activity at 11.1. One patient with two tumors in the same breast showed total response of one tumor to neoadjuvant chemotherapy while the second tumor grew in size.

Conclusion:

High resolution breast PET was highly accurate in identifying tumor response to neoadjuvant chemotherapy.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-06.
Abstract P4-01-15: A high-throughput three-dimensional cell migration assay (BiO assay) for toxicity screening for breast cancer applications
Authors: Tseng H , Sabino LG , Haisler WL , Gage JA , Neeley SK , Souza GR
Published: August 2014

Common in vitro assays for drug toxicity screening are not accurate predictors of human in vivo toxicity as they are performed on two-dimensional (2D) surfaces that do not mimic native cellular environments. Three-dimensional (3D) cell culture systems, like magnetic levitation, offer representative culture environments, with spatial control to design assays for drug toxicity applications. In this study, a label-free 3D in vitro assay for high-throughput toxicity screening (BiO Assay) of breast cancer cells was developed.

Confluent flasks of cancerous and non-cancerous mammary gland epithelial cells (MDA-231 and MCF-10A) were incubated overnight with a magnetic nanoparticle assembly, to which they bind. The next day, these cells were detached from the flask, and with the external application of a magnetic field, levitated to the air-liquid interface, where cells aggregated and interact to form larger 3D structures. These 3D structures were levitated for 24 hours to induce extracellular matrix formation. Afterwards, the structures were mechanically disrupted and patterned into rings using ring-shaped magnets. The magnetic field was removed, drugs, like doxorubicin, were added at varying concentrations, and the rings were allowed to close. This assay was validated against a 2D viability assay. A mobile device was programmed to capture the rings at specified timepoints, and image analysis was performed to track ring closure as a function of drug concentration and time.

MDA-231s and MCF10As were successfully formed into 3D cultures using magnetic levitation, and patterned into rings. We found significantly different results in drug sensitivity between cells grown in 2D and 3D. In conclusion, the BiO Assay is a simple assay that measures drug toxicity in a culture system similar to the native cellular environment.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-15.
Abstract P4-01-22: Measurement of tumour lesion glycolysis by FDG-PET in triple negative breast cancer
Authors: Antony Tharayil I , Harada N , Patel N , Varghese V , Kenny L
Published: August 2014

Background:

Triple negative breast cancer (TNBC) has been associated with a high glycolytic rate. As a surrogate measure, the SUV of FDG uptake is found to be higher both in the TNBC and her2 positive phenotypes. Tumour lesion glycolysis (TLG) is a product of the mean SUV value and tumour volume. We wished to evaluate the role of combined volume and metabolic measurements of glycolysis in patients with TNBC.

Methods: 300 FDG-PET scans were performed in patients with breast cancer, of whom 25 were found to have TNBC over a 4 year period, in 19 patients there were suitable lesions identified for analysis. Patients were scanned on the same Siemens PET/CT scanner in the local hospital, static images were acquired 60 minutes post-injection. SUV was corrected for injected activity and body weight, TLGMAX50 was also calculated considering SUV50 isocontour instead of SUV mean. ∑TLG was the sum of TLG for an individual patient.

Results: 117 lesions in total from 19 different patients were analyzed (mean = 6.15 lesions per patient). SUVMEAN, TLG mean, SUVMAX and TLGMAX were calculated for each lesion. SUVMEAN per patient varied from 2.69 to 15.48 while SUV50 varied from 5.45 to 48.23. ∑TLGMEAN and ∑TLGMAX ranged from 13.06 to 364.072 and 18.29 to 575.627, respectively. There was significant variation between patients and within different tumor lesions within patients. SUVPEAK for all lesions of patients ranged from 4.8 to 36.4.

Conclusions: TLG varies widely in patients with triple negative breast cancer, further studies are required to determine if this will be a useful prognostic/predictive measure in the future.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-22.
Abstract P4-01-24: Positron emission mammography (PEM) does increase specificity in diagnosis of breast cancer
Authors: Müller FHH , Hentschel M , Müller AG , Farahati J
Published: August 2014

Purpose:

To determine the diagnostic performance of PEM, a HR-PET with intrinsic resolution of 1.6 mm in detection of breast cancer (B-Ca).

Methods and Material:

PEM was performed in 122 lesions with suspicious breast lesions 90 min after i.v. application of 3.5 MBq F-18-FDG per KG body weight. A ROI marked the target lesion, measured the maximum PEM uptake value (PUVmax) and was correlated with a corresponding non-target ROI in the contra lateral healthy breast determining the target/non-target ratio. Two independent readers analysed images of all 122 lesions and compared the results to histopathology (11 PEM-Biopsies/8 Breast-Cancers) later. The group analyses for all malignant, benign, corresponding non-target lesions and the mean target/non-target ratio were calculated by paired Student t-Test.

Results:

22 out of 122 lesions were malignant. Mean of PUVmax was measured 4.3±2,7 in malignant lesions and 1.1±0.4 for the contra-lateral healthy breast (p = 0.001). The mean target/non-target ratio in patients with B-Ca of 3.7±1.5 was also significantly higher compared to benign lesions 1.1±0.4 (p = 0.001).

Considering a PUVmax >1.9, PEM was true-positive in all 22 cancers resulting in a sensitivity of 100%, specificity of 97%, PPV of 88% and NPV of 100%.

Detecting of B-Ca by PEM was at it´s best observed with a PUVmax cut-off level of >1.9 combined with a target/non-target ratio of >1.4.

Conclusion:

PEM can detect breast cancer with high accuracy using PUVmax as well as target/non-target ratio with high specificity. The results shown have to be confirmed in larger patient cohort. The higher specificity causes a lower rate of biopsies, surgery, chemotherapy and costs, therefore a evidence based medicine study will start in Germany.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-24.
Abstract P4-03-03: Determination of lymph node status using the primary tumor’s gene expression signature
Authors: Gyorffy B , Sztupinszki Z , Weltz B , Chen S-C , Quay S
Published: August 2014

Introduction:Lymph node status is one of the most important clinical parameters of breast cancer. Axillary lymph node dissection and sentinel lymph node biopsy have considerable morbidity associated with them and a method to accurately predict lymph node positivity would be clinically useful. To date, no gene expression signature has been established that is capable ofestimatin lymph node positivity. Our aim was to develop a new predictor using a large set of patient samples and compare its performance to known clinical variables.

Methods: An integrated database was constructed using publicly available Affymetrix microarrays with known lymph node status. The patients were divided into the four molecular subtypes of basal, luminal A, luminal B and HER2 positive using the St. Gallen criteria and the gene chip based gene expression data of estrogen receptor, HER2 receptor and MKI67. ROC analysis was performed for each gene within each subtype. Then, the top 10 genes of each list were combined, and using the expression data of these 40 genes across all patients, Manhattan or Euclidean distanceswere computed to calculate the distance between each sample. All the patients were ranked, and lymph node status was defined by using the proportional lymph node positivity of “x” nearest ranked patients. Multiple regression was performed to compare the classification using the gene expression data to known clinical variables. Statistical significance was set at p<0.01.

Results: The database includes 2756 patients, of whom 983 are lymph node positive. To optimize the classification, different number of genes (all/top100/top40) and different number of closest patients (x = 1/2/5/10/25/25/100/250/500) and two different distance metrics (Euclidean and Manhattan distance) were assessed. The best performance was achieved using the top40 genes with Manhattan distance to the 100 nearest patients. This setting reached a sensitivity of 0.70, specificity of 0.73 and accuracy of 0.72 across all patients. When compared to ESR1 status, HER2 status, MKI67 expression, grade, size, and agein the multiple regression, only the gene-expression based classification (1.56E-31) and size (1.7E-24) were significant.

Discussion:We have established a pipeline capable of determining lymph node positivity using the gene expression data of 40 genes.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-03-03.
Abstract P4-03-09: Time points to diagnosis and treatment of invasive breast cancer in southwestern Ontario
Authors: Vandenberg TA , Ballantyne B , Diaz Rodriguez LA , Whiston FJ , Stitt LW
Published: August 2014

Background Delays in breast cancer diagnosis and treatment are associated with increased tumour size at presentation, higher incidence of lymph node metastasis, higher relapse rates and lower 5-year survival rates in some studies. Diagnosis and treatment of breast cancer is complex, involving many health care practitioners at multiple locations. New advances in diagnostic and prognostic indicators make the process ever more complex. We reviewed the timing of crucial events in breast cancer diagnosis and treatment in Southwestern Ontario. Methods: Two time periods (2001, 2011) were assessed. Patients with new early invasive breast cancer with specific TNM criteria (T1c or greater plus any N, or any T plus N1-3; M0) were included. Data collection and analysis explored significant time points from first suspicion of malignancy, to diagnosis and definitive treatment, as well as patient demographic information. Results: 300 and 451 eligible patients were identified in 2001 and 2011 cohorts. Distribution of T and N status by cohort was compared by Chi-square test and time from first suspicion to diagnosis by Wilcoxon testing. The proportion of patients in which the time from first suspicion (clinical by lay person or health professional or by imaging) to pathological diagnosis exceeded two months increased from 20.4% in 2001 to 42.1% in 2011. There were no differences in time from first suspicion to diagnosis when analyzed by age (p = 0.54) or location (p = 0.50). Pathological T2-4 status at diagnosis increased from 48.4% in 2001 to 56.8% in 2011, and N2-3 status increased from 7.3% to 12.6%. Patients who had mammograms increased from 52.7% to 59.4%. A positive or suspicious mammogram was the first sign in 36% and 39.1% of cases. There was a trend towards more pathological diagnoses and definitive surgeries at tertiary centers compared to community hospitals. Conclusions: There is a longer time interval from first suspicion of malignancy to pathological diagnosis in 2011 compared to 2001 for both urban and rural populations. The number of non-low risk, non-metastatic cancers at diagnosis increased by 50% over the time interval studied (300 v 451), but number of T2-T4 non-metastatic cancers increased by 77% (145 v 256) and N2,3 cancers by 136% (22 v 52). This is despite more patients receiving mammograms (52.7% v 59.4%). Delays in breast cancer treatment are multifactorial, including both system-, and patient-related factors. This is an under-researched area and more investigation is needed to understand the reasons for the diagnostic delays and more serious cancers at presentation in order to improve outcomes. Reasons for higher proportions of advanced cancers will be discussed.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-03-09.
Abstract P4-04-03: A microgenomic approach using LCM procured breast cancer cells to identify clinically relevant gene signatures that underlie favorable response of ILC to aromatase inhibitors
Authors: Sanders MA , Andres SA , Wittliff JL
Published: August 2014

Based on CGH analyses, invasive lobular carcinoma (ILC) is closely related to low grade invasive ductal carcinoma (IDC), and is genetically unrelated to intermediate and high grade invasive carcinomas (Pathol Res Pract 2005; 201:713). Results from the BIG 1-98 trial, showed that patients with ILC and luminal B (intermediate/high grade) IDC treated with Letrozole, an aromatase inhibitor (AI), had a significant improvement in overall survival, whereas patients with luminal A (low grade) IDC did not (Cancer Res 2012; 72(24 Suppl):Abstract nr S1-1). Despite genetic evidence suggesting a close relationship between ILC and low grade IDC, and disparity between ILC and intermediate and high grade IDC, a clinically relevant gene set underlying a tumor's biologic responsiveness to AIs likely exists. We are deciphering a gene subset that would differ in expression between low grade IDC and ILC, and would be shared by ILC and luminal B IDC. Identification of such a gene set should lead to a more robust marker for identifying AI responsive tumors rather than relying solely on histomorphology and/or immunohistochemistry for the pathologic diagnosis of ILC, since the distinction between ILC and IDC is not always straight forward in pathology practice.

Using 247 de-identified human breast carcinoma biopsies collected under standardized, stringent conditions, total RNA was extracted from carcinoma cells procured by laser capture microdissection to perform microarray analyses of approximately 22,000 genes to identify expression signatures associated with breast cancer characteristics. Of the 247 samples, 169 samples were used for the analysis and included 16 ILCs, 13 low grade IDCs, 55 intermediate grade IDCs and 85 high grade IDCs.

Preliminary analyses identified 1267 genes that were differentially expressed (p<0.01) between cell samples from patients with IDC compared to those with ILC. Gene expression levels in breast cancer cells from patients with low grade IDC compared to those with ILC yielded 200 genes that were statistically differentially expressed (p<0.01). When comparing high grade IDC to ILC, 149 of these genes were not differentially expressed indicating that this gene set may serve as a panel of candidate genes for identifying cancers responsive to AIs. Pathway analysis software (Ingenuity®) identified that these genes were associated with key molecular and cellular functions, e.g., cell-to-cell signaling, cell morphology, cell death and survival, cellular assembly and organization and carbohydrate metabolism. qPCR analyses are used to validate and refine the gene subset distinguishing ILC from low grade IDC. Our approach is revealing subtle molecular features that discriminate these lesions based on clinical behavior. Supported in part by a grant from the Phi Beta Psi Charity Trust.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-04-03.
Abstract P4-04-15: UC San Diego molecular tumor board: Experience in breast cancer
Authors: Parker BA , Schwab RB , Helsten TL , Sarah BG , Ojeda-Fournier H , Datnow B , Schwaederle MC , Coutinho AC , Kurzrock R
Published: August 2014

Background: To realize the promise of personalized medicine, it is important to build the infrastructure to match patients to the most promising therapies using evolving genomic information to inform clinical trial and treatment recommendations.

Methods: A molecular tumor board was established in December 2012 consisting of representatives of medical oncology, surgery, radiation, pathology, radiology, clinical genetics, molecular genetics and diagnostics, basic and translational science, and bioinformatics and pathway analysis. Patients with metastatic breast cancer signed consent for collection of data in a registry format related to clinical history, genomic analysis, treatment recommendations, and response. Tumor specimens from patients with metastatic breast cancer were subjected to a next generation sequencing diagnostic test (Foundation One™) in a CLIA-certified lab (Foundation Medicine) and analyzed for genomic alterations in a 182 gene panel or a 236 gene panel. Clinical cases and genomic alterations were discussed at Molecular Tumor Board where clinical trials and treatment recommendations were considered, with the final decision made by the treating oncologist.

Results: Genomic analyses of metastatic breast cancer specimens from 17 consented patients had at least one genomic alteration and no tumors had the same genomic alterations. 12/17 had ≥ 2 and 4/17 had 6 or 7 genomic alterations. The most common genomic alterations were in PIK3CA observed in tumors from 6/17 patients. 1/17 tumor specimens had only a single genomic alteration for which there are no current targeted drugs in clinical studies or available commercially (GATA3). Radiographic and/or tumor marker response have been observed to date in 3/4 heavily pretreated patients following treatment with a targeted therapy or a targeted therapy informed by the genomic data in combination with other drugs.

Conclusions: The strategy of matching targeted clinical trial and commercially available therapies to genomic alterations of tumor specimens from individual patients with metastatic breast cancer may offer promise to patients with advanced refractory disease. Biopsies of metastatic disease, genomic analysis in CLIA labs, establishment of molecular tumor boards, definition of “matched” versus “non-matched” therapies, capturing data in a registry to create a learning environment, and most importantly, availability of targeted therapies in clinical trials or commercially, are required for this individual patient strategy to inform current and future treatment options for much larger patient populations.

Funding from UC San Diego Moores Cancer Center MyAnswerToCancer philanthropic fund.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-04-15.
Abstract P4-05-02: Phospho-reactome measurements reveal heterogenic kinase signatures
Authors: Coppé J-P , Chen Z , Mori M , Boudreau A , van 't Veer L
Published: August 2014

Background: Treating cancer increasingly relies on targeting kinases, because their oncogenic activity drive tumorigenesis. Discovering which active mechanisms of disease progression can be efficiently targeted, and knowing whether kinase networks circumvent therapeutic interventions, are challenges researchers and clinicians face. Surprisingly however, measuring the phosphorylating activity of kinases, and potentially monitoring the functionality of the entire human phospho-reactome at once, remains largely unexplored. We developed a semi-high throughput assay to monitor the phospho-catalytic activity of kinase enzymes, using their biological targets as phospho-sensors. We successfully used this assay to identify oncogenic phospho-signatures prevalent in breast cancer, and can be used to establish drug-sensitivity profiles in models of kinase-targeted therapies.

Methods: We first defined how to computationally build a library of peptide sensors established from confirmed kinase substrates’ phosphorylation sites. Precisely, we used computational methods to create a unique phospho-repertoire cataloguing 3,408 peptide sequences established from validated human proteins’ phosphorylation sites, curated from 38 public databases. Second, we experimentally used these biologically relevant probes in multiplex assays to quantify the catalytic state of kinases. Specifically, a kinome-representative 242-peptide set was developed into an ATP-consumption screen to identify the activity signatures of EGFR, MAPK, AKT, ABL and SRC family kinases, and explore 642 kinase/substrate nodes. Next, we described analysis methods to derive phospho-signatures from semi-high throughput ATP-consumption measurements. We validated the assay using isogenic culture model of basal-like breast cancer (HMT-3522 S1 and T4-2), and cell lines harboring EGFR/HER2-oncogenic alterations such as MDA-MB-231, MCF7 or T47D.

Results: The differential phosphorylation activity of 25 recombinant, active kinase enzymes was successfully captured. In cancer cell extracts, hyper-activated EGFR, ERK, MEK, AKT, and SRC kinases originally identified by immuno-detection were reliably and specifically detectable using the peptide-based kinase-activity assay. The phospho-sensing assay revealed the heterogeneity of active kinase signaling circuits among different breast cancer cells.

Conclusion: This unique strategy and resources allow to comprehensively measure the catalytic activity of multitude kinases at once, representing a new molecular dimension to characterize biological samples. We will use such new phospho-reactome profiling system to determine the efficacy of new combinatorial therapies, and define how chemotherapeutic interventions lead to the reprogramming of phospho-circuits.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-02.
Abstract P4-05-08: Oncotype Dx assay in BRCA positive ER positive breast cancer patients
Authors: Halpern N , Sonnenblick A , Kaduori L , Uziely B , Goldberg Y , Katz D , Allweis T , Divinsky L , Merlet I , Maly B , Sagi M , Hamburger T , Carmon E , Peretz T
Published: August 2014

Background:

Oncotype DX is a 21-gene RT-PCR assay which quantifies the likelihood of breast cancer recurrence and the potential benefit of chemotherapy in patients with early stage, ER positive, Tamoxifen treated breast cancer. Breast cancer in BRCA carriers is considered more aggressive. The aim of our study was to evaluate whether Oncotype Dx recurrence score distribution is different in breast cancer patients with inherited BRCA mutation.

Methods:

The Oncotype DX assay has been used at Hadassah Medical Center since 2004 on specimens from over 450 patients. We analyzed and compared clinicopathological characteristics and Oncotype Dx recurrence scores of BRCA carriers versus non- BRCA or unknown status of BRCA patients.

Results:

Ten patients had validated inherited BRCA mutation, five of them are BRCA1 carriers and five BRCA2 carriers. There were no significant differences in the clinicopathological characteristics between the two groups. Oncotype Dx recurrence score distribution between low, intermediate and high risk groups was not significantly different. The mean recurrence score was 18.48 for the non- BRCA or unknown status of BRCA patients and 22.8 for the BRCA carriers patients. This difference was not statistically significant.

Conclusion:

Estrogen receptor positive breast cancer tumors from BRCA carriers does not display a significantly different Oncotype Dx recurrence score result distribution.

These preliminary data suggest Oncotype Dx assay might be used to help tailor treatment in this subset of patients, although further follow up is needed.

All patients evaluated for oncotype_DX  All evaluated (except BRCA+) n = 456BRCA positiveP valueAgeMean57.4557.60.7 Median5857  Range56 (25-81)34 (42-76) T stageT167.7%70%0.9 T230.7%30%  T31.6%0% Tumor sizeMean1.71.360.2 Median1.51.26  Range7.8 (0.2-8)1.3 (0.7-2) GradeGrade 1-278.8%60%0.15 Grade 321.2%40% RSLow50%40%0.57 Intermediate39.8%40%  High10.2%20% RSMean18.4822.80.16 Median17.522.5  Range64 (0-64)27 (12-39) 

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-08.
Abstract P4-05-11: A proteasomal signature of breast cancer cells
Authors: Gaczynska M , Mueller B , Rodriguez KA , Osmulski PA
Published: August 2014

The multisubunit, multifunctional protease called proteasome as a part of the essential proteasome-ubiquitin pathway constitutes an attractive target for anti-cancer drugs. A proteasome inhibitor bortezomib is successful in blood cancers treatment, however trials with breast cancers yield disappointingly inconsistent results, likely stemming from the cancers’ diversity. Since bortezomib preferentially induces an apoptosis in cancerous cells, it is likely that differences in the proteasome pathway between the control and cancerous cells, and also between distinct cancers, contribute to this effect. Here we compared components of proteasome assemblies and their peptidase activities in seven breast cancer and two control cell lines in an attempt to find a common proteasomal signature of breast cancer. Defining a common proteolytic denominator for breast cancer cells should facilitate a rational drug design based on proteasomal inhibition. For this purpose, we analyzed cytosolic, nuclear and microsomal extracts from human cultured breast cells: control (MCF10A, Hs 578 Bst) and derived from distinct types of breast cancer: luminal A MCF7 and T47D, triple negative basal A MDA-MB-468, and triple negative basal B MDA-MB-231, Hs 578 T and BT-549. We found a high content of total proteasome in a microsomal fraction with a relatively prominent contribution from immuno proteasomes in all cancer cell lines. These proteasomes are believed to be localized on the outside of endoplasmic reticulum membrane and presumably take part in the endoplasmic reticulum - associated degradation system. Markedly, the pattern of peptidase activities of proteasome and content of subunits of its exchangeable sub-assemblies were specific for each fraction and discriminated between cancerous and control cells. Statistical analysis identified two clusters formed by the cell lines in the microsomal and nuclear fractions and a distinct clustering of the cytosols. Furthermore, three groups of variables, which tend to always cluster together were identified: the 19S regulatory particle subunits, immunoproteasome subunits and peptidolytic activities. Variables that the best quantitatively discriminate between the cell lines at a given level of total proteasome were immunoproteasome subunits and 19S cap subunit Rpt5. Additionally, the housekeeping subunit beta5 in the cytosol, the PA28 activator gamma subunit in the microsomal fraction and chymotrypsin like activity in the nuclear fraction qualitatively were the best markers aiding classification of the cell lines. We speculate that the distinct properties of proteasomes in the microsomal and nuclear fractions may contribute to specific response of the cancer cell lines to the treatment with proteasomal inhibitors. The study shows that a comprehensive analysis of the proteasome system in the established breast cancer cell lines may help to guide diagnostics and therapy design.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-11.
Abstract P4-06-13: Discovering serum biomarkers for predicting accumulation of nanotherapeutics to different tumor and organ microenvironments
Authors: Yokoi K , Tanei T , Godin B , van de Ven AL , Alexander J , Saito Y , Ferrari M
Published: August 2014

Surrogate biomarkers in the circulation for the enhanced permeation and retention (EPR) effect in different tumor and organ microenvironment shall aid in selecting patients who are likely to accumulate higher amount of nanotherapeutics to tumors and thus show better response to the therapeutics. In this study, 4T1 and 3LL murine cancer cell lines, known to have similar sensitivity to pegylated liposomal doxorubicin (PLD) in vitro, were shown to accumulate significantly different quantities of PLD in vivo. PLD accumulation correlated with tumor-specific differences in therapeutic efficacy and vascular permeability, which was modulated by the extent of coverage of tumor-associated endothelial cells by basement membrane. Matrix metalloproteinase (MMP)-9 and its endogenous inhibitor, tissue inhibitor of metalloproteinase (TIMP)-1, played a pivotal role in these phenomena. Differences in vascular permeability and PLD accumulation were also organ-specific and significantly correlated with the relative ratio of MMP-9 and TIMP-1 in the systemic circulation of the tumor-bearing mice. Our findings support the further development of MMP-9 and TIMP-1 as paired serum biomarkers for predicting accumulation of PLD to tumor and the application of these biomarkers could be expanded to other types of nanotherapeutics utilizing the EPR effect for the personalization of cancer therapy.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-06-13.
Abstract P4-07-14: The effects of calorie restriction on microRNA expression profiles and DMBA-induced mammary tumorigenesis in a model of luminal A breast cancer
Authors: Devlin KL , Sanford T , Mambo E , Hursting SD
Published: August 2014

MicroRNA (miR) are small, post-transcriptional regulators that play an integral role in maintenance of cellular functions and whose dyregulation has been shown to promote many types of cancer, including breast cancer. While it has been shown that microRNA are dyregulated through the stages of breast cancer development, it was our goal to define the microRNA expression profiles of the individual stages in a model of luminal A breast cancer in order to identify which are specifically associated with progression. In addition, we aim to establish the effects of calorie restriction, which is known to inhibit breast cancer progression, on relevant microRNA. To address these questions, 100 female Sprague Dawley rats were administered either dimethylbenz(a)anthracene or vehicle control at 50 days of age, then randomized to receive either control (AIN-76A) diet ad-libitum (n = 40) or a 30% CR diet regiment (n = 60). Resultant mammary tumors were allowed to develop for 12 weeks. Calorie restriction significantly increased survival to study endpoint relative to control diet (75% vs 35%, respectively) (p = 0.0047). Furthermore, of the animals that developed tumors, CR significantly decreased median tumor area by 56% compared to control diet (109.4 mm2 vs 250.9 mm2, respectively) (p = 0.0286). To analyze the impact of microRNA modulation on tumor progression, mammary tumors from control animals were microdissected in a stage-dependent fashion, with areas of normal epithelium, intraductal proliferation, ductal carcinoma in-situ, and invasive ductal carcinoma collected and analyzed separately. We have identified 10 microRNAs that are associated with cancer progression in this model (miR-10a, -10b, -21, -92b, -124, -125b, -126, -145, -200a, -658). We are currently analyzing which of these microRNAs are CR-responsive and how their modulation affects tumorigenic indices. The results obtained will provide insights into the mechanisms of breast cancer progression and how calorie restriction inhibits progression through microRNA modulation.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-07-14.
Abstract P4-07-19: MicroRNA miR-24 enhances tumor angiogenesis and metastasis in breast cancer
Authors: Xu Z , Wang X , Fang L
Published: August 2014

MicroRNAs (miRNA) are non-coding single-stranded RNA molecules, which are produced from endogenous transcripts of short stem-loop structures. Through binding to the 3’-untranslated region (3’UTR) of different target messenger RNAs (mRNAs), microRNAs can repress the translation of the target mRNAs. Recent studies have shown that miR-24 can inhibit apoptosis of cancer cells and cardiomyocytes, and can accelerate cell proliferation. Up-regulation of miR-24 has been observed in oral carcinoma and is one of the most abundant miRNAs in cervical cancer. However, the role of miR-24 in breast cancer cell angiogenesis and metastasis is not clear.

We analyzed microRNA miR-24 levels in patients with breast carcinoma and found that miR-24 was higher in breast carcinoma samples than in breast benign tissues. We generated constructs expressing miR-24 and studied its functions using both in vitro and in vivo techniques. We found that the ectopic expression of miR-24 promoted breast cancer angiogenesis and metastasis.

Cell migration assays indicated that expression of miR-24 promoted migration of MT-1 cells and 4T1 cells. We examined proliferation of MT-1 cells and 4T1 cells transfected with miR-24 or the control vector. The experiments showed that the miR-24 transfected cells had high rates of proliferation than the vector-transfected cells. In vivo experiments indicated that the expression of miR-24 enhanced tumor growth, metastasis to lung tissues, and decreased overall mouse survival. Tumor sections were also probed with anti-CD34 antibody to examine angiogenesis. We found that miR-24 expression increased tumor-associated vascularization. We further performed tumor metastatic assays. DNA was isolated from lung tissues and subjected to PCR to indicate metastasis of the cancer cells. Typical metastatic lesions in the lungs are shown.

In the miR-24 expressing cells and tumors, expression of the phosphatases PTPN9 and PTPRF were repressed. We confirmed that miR-24 could directly target both PTPN9 and PTPRF. Consistent with this, we found that the levels of PTPN9 and PTPRF were lower in the patients with metastatic breast carcinoma. Ectopic expression of PTPN9 and PTPRF decreased cell migration, and tumor metastasis.

Our study demonstrates that miR-24 is highly expressed in human breast carcinoma. This finding suggests a role of miR-24 in breast cancer development/progression. Our results suggest that miR-24 plays a key role in breast cancer angiogenesis and metastasis. miR-24 could potentially be a target for cancer intervention.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-07-19.