bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P6-14-03: Statin and aspirin use is not associated with a reduced risk of VTE's in breast cancer patients
Authors: Shai A , Rennert HS , Ballan Haj M , Lavie O , Steiner M , Rennert G
Published: August 2014

Introduction: Statins and aspirin have been shown to reduce the risk of venous thromboembolic events (VTE's) in the general population in randomized trials.

Aim: To assess whether statins and aspirin reduce the incidence of deep vein thrombosis and pulmonary embolism in patients diagnosed with breast cancer.

Methods: The Breast Cancer in Northern Israel Study (BCINIS) is an on-going population-based case-control study of consecutive breast cancer cases and matched controls diagnosed in the northern part of Israel since 2000.

Only cases insured by the Clalit Health Services (64%) were included in this analysis. Data on medication use and VTE were extracted from the computerized database. Patients taking Warfarin or LMWH were excluded.

Statistical analysis was performed using SPSS (v 18). Use of medications was analyzed as a time dependent covariate in a Cox Regression model.

Results: Of 3552 patients 261 (7.3%) had a VTE during a median follow up of 4.7 years.

In a multivariate analysis age (HR 1.03, 95% CI 1.02–1.04), BMI (HR 2.12, 95% CI 1.43–3.14 for BMI >30), chemotherapy (HR 3.87, 95% CI 2.50–5.98) and metastatic disease (HR 2.27, 95% CI 1.10–6.53) were associated with an increased risk for VTE's.

Statins, mainly simvastatin and pravastatin, were used by 55.7% of the patients, and 45.7% used aspirin. Neither statins nor aspirin were associated with a significantly reduced risk for a VTE. After controlling for age, BMI, stage, chemotherapy and tamoxifen use the HR for statins was 0.80, CI 0.56–1.13, p = 0.2, and the HR for aspirin was 1.004, CI 0.74–1.76, p = 0.98.

Conclusion: In contrast to the findings in the general population, statin and aspirin use were not associated with a reduced risk for VTE's in our cohort of breast cancer patients. Our results might be explained by an alternate mechanism of VTE formation in breast cancer and the use of low potency statins (simvastatin and pravastatin) in this cohort.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-14-03.
Abstract P6-14-04: Rosehip extracts prevent triple negative breast cancer cell proliferation by regulation the phosphorylation of p70S6 Kinase.
Authors: Coburn TL , Cagle PD , Shofoluwe AI , Martin PM
Published: August 2014

Triple Negative Breast Cancer (TNBC) is an aggressive form of breast cancer, characterized by its lack of the human epidermal growth factor receptor-2 (HER-2), the estrogen receptor (ER), and the progesterone receptor (PR). Due to the lack of these hormone receptors, TNBC has abnormal therapeutic results; leading to the need for unique treatment methods. TNBC has a partial response to chemotherapy, which is associated with the lack of clinically established targeted therapies. Plant extracts have shown to be useful as chemopreventive agents due to their disruption of pro-survival and proliferative mechanisms on cancer cells. Rosehip extracts have shown potential anti-proliferative activity against cancer cells in vitro. This study investigated the effect of rosehip extracts in TNBC cell lines (HCC1806 and MB157) and basal-like breast cancer cell lines (HCC70 and HCC1500). To determine the inhibitory role of rosehip extracts in cell proliferation, MTT assays were conducted, following treatment with varying concentrations of rosehip extracts (1mg/ml, 250ug/ml, 25ug/ml, and 25ng/ml) for 48 hours. The data suggests that rosehip extracts, can significantly decrease cell proliferation in all the breast cancer cell lines assayed. Preliminary studies suggest that this is accomplished without toxic effects. Western blot analysis was used to investigate which signaling pathway rosehip extracts were affecting. Western Blot analysis suggests that rosehip extracts are inhibiting p70 S6 kinase phosphorylation and its ability to act as a kinase for S6 ribosomal protein. Based on these findings, rosehip extracts prevent cell proliferation by interfering with the p70 S6 kinase signaling.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-14-04.
Abstract PD01-02: Increased expression of phosporylated mTOR in metastatic breast tumors compared to primary tumors in patients who received adjuvant endocrine therapy
Authors: Hoefnagel LDC , Beelen KJ , Opdam M , Vincent A , Linn SC , van Diest PJ
Published: August 2014

BACKGROUND: Activation of the PI3K pathway as an adaptive change in response to estrogen depletion results in acquired endocrine therapy resistance in vitro. Metastatic breast cancer patients with previous exposure to endocrine therapy do derive substantial benefit from the addition of an mTOR inhibitor to endocrine therapy compared to endocrine therapy alone. This suggests that compensatory PI3K pathway activation might play a role in the development of clinical resistance to endocrine therapy. We evaluated the activation of the downstream PI3K pathway protein mTOR in primary breast tumors and matched metastatic lesions.

METHODS: From a previously described series of breast cancer patients from whom both primary tumor tissue as well as metastatic tumor tissue was collected, we selected estrogen receptor α (ERα) positive patients who had received endocrine therapy (N = 34) and who did not receive endocrine therapy (N = 37). The difference in expression of phosphorylated mTOR (p-mTOR) between primary and metastatic tumor was assessed by immunohistochemistry, which was scored by using the proportion of tumor cells with sub-membranous p-mTOR expression. We assessed whether this p-mTOR difference between primary and metastatic tumor was associated with clinico-pathological factors (location of metastasis, lymph node status, T-stage, grade, progesterone receptor status) or varied between patients who did and did not receive endocrine therapy, using Mann-Whitney tests. In addition we performed a linear regression model including the same clinico-pathological factors.

RESULTS: In patients who had received endocrine therapy we observed an increase in p-mTOR expression in metastatic tumor lesions compared to the primary tumor (median difference 45%). This was significantly different from the mTOR changes observed in patients who did not receive endocrine therapy (median difference between metastatic and primary lesion 0 %) (p = 0.003). The difference remained significant in the multivariate regression model (p = 0.001). None of the other factors was significantly associated with a differential p-mTOR change.

CONCLUSION: In patients who received previous adjuvant endocrine therapy, the increase in p-mTOR expression in metastatic tumor lesions compared to the primary tumor is significantly higher than in patients who did not receive endocrine therapy. Compensatory activation of the PI3K pathway might therefore be a clinically relevant resistance mechanism resulting in secondary endocrine therapy resistance.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD01-02.
Abstract PD02-05: Comparison of fluorescence in situ hybridization (FISH) and dual-ISH (D-ISH) in the determination of HER2 status
Authors: Mansfield AS , Sakai Y , Walsh FJ , Wiktor AE , Sukov WR , Dogan A , Jenkins RB
Published: August 2014

Introduction: The appropriate assessment of HER2 amplification status is critical to determining which patients should receive HER2-directed therapy. An alternative to FISH is D-ISH (Ventana Medical Systems Tucson, Arizona) which utilizes chromogenic HER2 and chromosome 17 probes. Both FISH and D-ISH utilize formalin-fixed, paraffin-embedded (FFPE) human breast cancer tissue specimens. D-ISH has two principle advantages over FISH: it can be visualized using light microscopy and after analysis the specimens can be archived.

Methods: We tested 250 samples with FISH and with D-ISH. Fifty specimens were controls: 25 cases with no HER2 or CEP17anomaly and 25 cases with HER2 amplification. There were also 200 test subjects of 50 cases each of chromosome 17 aneusomy, HER2 deletion (HER2/CEP17 ratio <0.8), HER2 duplication (HER2/CEP17 ratio 1.3–1.8), and equivocal results (HER2/CEP17 ratios between 1.8 and 2.2). FISH (Abbott Molecular) and D-ISH analysis were performed using methods described in the appropriate clinical kit package inserts, with the following exception: Four cases with CEP17 and HER2 co-amplification were also included and for FISH analysis these cases were reflexed to the use of chromosome 17 control probe (D17S122).

Results: Four samples failed testing by D-ISH. There was a 63% (155/246) concordance between FISH and D-ISH by anomaly (aneusomy, duplication, deletion, etc.) and an 83% concordance by amplification status (non-amplified, equivocal, and amplified). D-ISH detected 18 (62%) of F-ISH-amplified cases. D-ISH resulted in lower estimates of HER2/CEP17 ratios than FISH, and many cases that were equivocal by FISH were normal by D-ISH. D-ISH did not detect HER2 amplification in any of the four HER2 and CEP17 co-amplified cases.

Conclusions: We observed a lower concordance rate between FISH and D-ISH than the ∼95% rate in the D-ISH package insert. This lower concordance rate is likely a result of how this study was designed: it was biased toward common chromosome 17 anomalies and difficult specimens. Such specimens comprise about 60% of our current clinical HER2 biomarker practice. D-ISH may underestimate the HER2/CEP17 ratio, or FISH may overestimate this ratio. D-ISH also does not reliably detect HER2 amplification when the centromere of chromosome 17 is also co-amplifed. Compared to FISH, in this study cohort D-ISH had a significant false negative rate for the detection of HER2 amplification.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD02-05.
Abstract PD03-02: Evidence for the anti-cancer action of metformin mediated via tumor AMPK, Akt and Ki67, in a preoperative window of opportunity trial
Authors: Hadad SM , Dowling RJO , Chang MC , Done SJ , Purdie CA , Jordan LB , Dewar J , Goodwin PJ , Stambolic V , Thompson AM
Published: August 2014

Background: Metformin reduces the overall risk of cancer incidence by 31%, improves cancer-related mortality and enhances response to neoadjuvant chemotherapy in type 2-diabetics receiving metformin. There is evidence that metformin works through activation of Adenosine Monophosphate Protein Kinase (AMPK), an energy-sensing kinase that maintains cellular energy homeostasis. In a pre-operative window-of-opportunity randomized trial we have demonstrated that metformin significantly reduces proliferation and blunts the insulin response in primary operable breast cancer. This study further analysed cancer tissue from the trial to dissect the molecular mechanisms involved.

Methodology: Non-diabetic women with operable invasive breast cancer were randomised to receive pre-operative metformin or no drug. Forty seven patients had core biopsy at diagnosis then were randomized to metformin (metformin 500mg o.d. for 1 week increased to 1g b.d for a further week continued to surgery) or no drug, and 2 weeks later had core biopsy at surgery. Insulin receptor, phospho-AMPK (pAMPK), phospho-Akt (pAkt) and Ki67 immunohistochemistry was performed on formalin-fixed paraffin-embedded cores and scored blinded to treatment. Paired t-test was used for analysis.

Results: Significant up-regulation of pAMPK (p = 0.04) and down-regulation of pAkt (p = 0.04) in metformin treated patients was demonstrated compared to the control group. No change in insulin receptor expression was identified but, as previously reported, there was a fall in ki67. Changes were independent of Body Mass Index. Seven patients (7/24) receiving metformin withdrew because of gastro-intestinal upset and were excluded from the immunohistochemical analyses.

Conclusion: These findings suggest that metformin works in vivo in breast cancer patients via up-regulation of tumor pAMPK and down-regulation of pAkt and proliferation. Since down-regulation of pAMPK is a feature of breast cancer, this suggests mechanistic evidence for the therapeutic effect of metformin.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD03-02.
Abstract PD05-01: Next generation genomic sequencing (NGS) identifies molecular targets in inflammatory breast cancer (IBC)
Authors: Cristofanilli M , Alpaugh KR , Ross J , Binghman C , Wu H , Stephens P , Lipson D , Palmer G
Published: August 2014

BACKGROUND: Inflammatory breast cancer (IBC) is the most aggressive form of invasive breast cancer and is associated with poor outcome in spite of optimal multidisciplinary management. The disease is associated poor response to neoadjuvant chemotherapy and a peculiar pattern of recurrence (chest wall, nodes, CNS). There currently few distinct molecular alterations for effective therapeutic targeting in the advance setting. The availability of comprehensive cancer genomic testing using Next-Generation Sequencing (NGS) technology may identify a number of genomic alterations not assessed by traditional methods and provide opportunities for more effective treatment selection particularly in aggressive and uncommon diseases such as IBC.

METHODS: Twenty-three patients with refractory metastatic IBC were included in this analysis. All patients underwent confirmatory biopsy and/or tissue collection. Tissue from metastastic lesions of patients with recurrent IBC was submitted as slides or FFPE blocks. NGS was performed by Foundation Medicine, Cambridge, MA. The entire coding sequence of 182 cancer-related genes (3,230 exons) plus 37 introns from 14 genes often rearranged in cancer were sequenced to high depth (>500X) All classes of genomic alterations were detected.

RESULTS: The site distribution and frequency of disease recurrence included: soft tissue (19) lung (4), bone and liver (3), and brain (2). The disease subtype classification based on the expression of ER, PR and HER-2 were as follow: ER−, PR−, Her-2−, Basal-like or TN (65%); ER+ and/or PR+, HER-2− (grade3) or HER-2+, Luminal B (22%); ER−, PR−, HER2+ (13%). Genomic alterations were identified in all patients with various frequencies. Genetic mutations included: P53 (DNA-binding domain, splice site), PI3KCA (H1047R, R441N, E545K, K111E,); RB1; PTEN (D107Y); EGFR (L858R); ERBB2 (V777L, S310F) in a patient with TN disease; ESR1 D538G. Gene amplifications included: MYC (26%); CCND1 (22%); ERBB2 (in a patient with TN disease), MLC-1, CCNE1, CK4, K-RAS. One patient had evidence of discordant genomic abnormalities in two simultaneous sites of recurrence (skin and pleura) suggesting disease heterogeneity. Three patients were effectively treated with genomic alterations-guided therapies.

CONCLUSIONS: Genomic sequencing of advanced IBC is feasible, identifies potential therapeutic targets and can advance our understanding of the molecular alterations of this aggressive disease.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD05-01.
Abstract PD05-03: What is the appropriate sample (s) on which to perform sequencing for mutational analysis to guide the selection of targeted therapy?
Authors: Alpaugh RK , Bingham C , Fittipaldi P , Banzi M , Palmer G , Cristofanilli M
Published: August 2014

BACKGROUND: Success of targeted therapy requires expression of the protein. Tumor tissue source can include diagnostic biopsy, surgical samples from initial or follow-up surgeries, fluids e.g. pleural or ascites and circulating tumor cells (CTC). The goal of using CTCs was 1. To determine whether CTC can be used as a “liquid” tumor biopsy and enable gene sequence information at the single cell level and 2. To determine the heterogeneity represented in the circulation compared to that seen in solid tumor by examining single cells (or a small cluster of cells) for the presence of a specific mutation which was detected in tissue tumor source.

METHODS: We performed sequencing for mutational analysis on tissue(s) from patients with inflammatory breast cancer (IBC). Tumor sources varied from mastectomy tissue, metastatic site(s) e.g. liver or skin from chest wall disease, pleural fluid and CTC isolated into pure single cell populations (or groups of cells) using Silicon Biosystems DEPArray. Ampli1™ WGA kit was used for CTC amplification. Of the 22 patients sequenced, mastectomy primary tumor was examined in 3, metastatic site skin chest wall disease in 15, other metastatic site in 4, pleural fluid in 2 and CTC collected to investigate p53 mutations in 8.

RESULTS: To date 22 patients have had mutational data performed, 14/22 had mutations in p53, 4/22 in RB1, 2/22 in each PI3K and ERBB2, 1/22 in each of BRAC1, BRAC2, ATM, KRAS, Notch 1, MEN1 and ESR1. Numerous amplifications were noted including AKT1, RPTOR, MLC1, MYC, CCND1 and ERBB2. For one patient's chest wall biopsy compared to two single CTCs and a cluster of 10 CTCs the same TP53C229fs*10 mutation was detected revealing the same 2bp deletion. No 2bp deletion was found in single white blood cells. Whereas, another patient which showed a TP53 S215G mutation in her skin biopsy of chest wall disease, only amplifications of AURKA, CCND1, IGF1R, MDM2 and SRC in pleural tumor cells were detected and no mutations in three single CTC, two single pleural tumor cells and in single white blood cells were seen. Primary tumor tissue is being sort for both of these patients. Mutational data reviewed to date suggest that IBC is not one disease but a multiplicity of diseases, revealing a variety of target(s). Aberrations are not consistent across tissue source.

CONCLUSIONS: Successful treatment outcomes using standard of care chemotherapy combined with target therapies will require not one, but a panel, of tissue sources for sequencing to guide the selection of appropriate targeted therapies.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD05-03.
Abstract PD06-04: St. Gallen 2011 clinicopathological subtyping of breast cancer: impact of different proliferation assessment methods
Authors: Focke CM , Gläser D , Finsterbusch K , Decker T
Published: August 2014

Rationale: The St Gallen Consensus 2011 strongly recommends subtyping of breast cancer (BC). However, because there is no data from phase III trials to substanciate adjuvant therapy benefit gene expression arrays were not generally accepted to guide systemic therapy decisions. Instead, the consensus introduced clinicopathological BC subtypes (cpBCST) defined as sets of risk and predictive biomarkers which are established worlwide (Table 1).

To discriminate Luminal A vs. Her2 negative Luminal B BCs the St Gallen Consensus recommends Ki67 for defining proliferation. Although, because no standardized methodology or cut-off definition for Ki67 is available so far ‘some alternative measure of proliferation’ was suggested. We analyzed the influence of different proliferation assessment methods on BC subtype distribution.

Methods: cpBCST assessment including proliferation measurement by different variants of Ki67-Labeling-Index (KI67LI) was performed on 225 BCs. For comparison we used the Mitotic Activity Index (MAI) as the only prospectively evaluated and best reproducible proliferation measurement (Baak et al 2008). The Ki67-LI was based on counting any nuclear staining of distinctly defined target areas: (1) 100 tumor cells within the hot spot (Ki67-100), (2) 1020 tumor cells in 3 HPF in the tumor periphery including the hot spot (Ki67-1020periphery), and (3) 1020 tumor cells in 3 HPF including hot spot, cold spot, and an intermediate area (Ki67-1020spectrum). Staining quality and counting accuracy was tested as excellent by the 2011 circulation of the German Quality Initiative in Pathology. According to St Gallen, a cut-off <14% was used. The MAI was measured in 10 HPFs with a total area of 1.59 mm2 using a cut-off ≥10 as evaluated by Baak et al. 2008.

Results:

Conclusions: The delimination of Luminal A and Luminal B Her2 negative BCs according to the St. Gallen Consensus 2011 is highly influenced by the method of proliferation assessment. Using the recommended cut-off <14% all estimations of Ki67LI came out with Luminal A/Luminal B ratios (10–34% vs. 45–69%) which are much lower than those published (44–53% vs. 9–29%). As a consequence, this could lead to more cytotoxic treatment. Therefore, the biological relevance of KI67LI seems to be not unequivocal. Unlike this, proliferation measurement by MAI resulting in a Luminal A/Luminal B ratio of 52% vs. 29% seems more reliable. The differences may be due to the fact that mitotic figures reflect only the M phase, whereas Ki67 stains cells in nearly all phases of the cell cycle.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD06-04.
Abstract PD06-07: Evaluation of an optimal cut-off point for the Ki-67 index as a prognostic factor in primary breast cancer.
Authors: Tashima R , Nishimur R
Published: August 2014

Background: The proliferation biomarker Ki-67 is considered to be a prognostic factor for breast cancer and has been investigated in several studies. However, there is no standard cut-off point for the Ki-67 index. Therefore, we retrospectively investigated an optimal cut-off point in terms of the prognostic factor in primary breast cancer.

Patients&Methods: Immunohistochemical (IHC) analysis of the proliferation marker Ki-67 index was performed on 4338 patients with primary breast cancer until March 2012 at Kumamoto City Hospital. Out of these patients, there were 2375 consecutive cases with ER and/or PgR positive and HER2 negative tumors since 1997. Items examined were ER, PgR, HER2, tumor size, nodal status, nuclear grade, and p53 overexpression. The Kaplan-Meier test was used to calculate prognosis (cumulative disease-free survival (DFS) and overall survival (OS)) and tested with the log-rank procedure. Cox's proportional hazard model was used to perform a univariate and multivariate analyses of the factors related to DFS. In addition, an analysis was conducted to determine various cut-off values for the Ki-67 index.

Results: The median Ki-67 value was 21%, with a mean of 26.2%. Univariate and multivariate analysis revealed that the Ki-67 index was an independent prognostic factor for DFS and OS. A multivariate analysis revealed the following hazard ratio (HR) and cut-off values for the Ki-67 index; HR was 1.92 in cases with a cut-off value of 20%, 1.91 in 35%, 1.87 in 40%, 1.89 in 45% and 1.79 in 50%, respectively. These findings suggested that the optimal cut-off point was 20%. In the cases with luminal type (ER and/or PgR positive and HER2 negative) breast cancer, a higher Ki-67 value (20% ≤) correlated with larger tumor size, positive nodes, higher grade, and overexpression of p53. Moreover, patients with a higher Ki-67 value showed significantly lower DFS/OS rates. A multivariate analysis also revealed that the Ki-67 index is a significant prognostic factor in luminal type breast cancer.

Conclusion: The optimal cut-off value for the Ki-67 index is 20%. Moreover, this proliferation marker is a significant prognostic factor in evaluating primary breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD06-07.
Abstract PD07-03: INCREASED PATHOLOGIC COMPLETE RESPONSE RATE AFTER A LONG TERM NEOADJUVANT LETROZOLE TREATMENT IN POSTMENOPAUSAL ESTROGEN AND/OR PROGESTERONE RECEPTOR-POSITIVE BREAST CANCER
Authors: Allevi G , Strina C , Andreis D , Zanoni V , Bazzola L , Bonardi S , Foroni C , Milani M , Cappelletti MR , Generali D , Berruti A , Bottini A
Published: August 2014

Background: Neoadjuvant endocrine therapy has been increasingly employed in clinical practice to improve surgical options for postmenopausal women with bulky hormone receptor-positive breast cancer. Recent studies indicate that tumor response and in particular the pathologic complete response (CR) in this setting may predict long-term outcome. The letrozole approval in neoadjuvant setting is based on a study that included 4 months of letrozole treatment only. Prospective studies have not investigated treatment duration beyond 6 months, and retrospective studies suggest that useful responses can occur after this period. The purpose of this study was to determine the clinical and pathologic response of 2.5 mg daily letrozole used with different schedule in breast cancer patients (pts) unfit for chemotherapy.

Patients and Methods: This single Institution retrospective study comprised 102 postmenopausal women with primary breast cancer (clinical stage ≥T2, N0-1), who were divided into 3 subgroups (34 each in single subgroup) according to the different duration of neoadjuvant letrozole treatment: 4 months (subgroup A), 8 months (subgroup B) or 12 months (subgroup C) of treatment. The subgroups were comparable in terms of baseline characteristics.

Results: Pts and tumor characteristics included: median age 77.3 years (range 53.9–95.4); 100% estrogen receptor-positive, 81.4% progesterone receptor-positive; median Ki67 13.5% (range 2–90%). All pts were evaluable for clinical and pathologic response. A total of 77 pts (75.5%) achieved an objective response at individual end: 40 (39.2%) clinical CR and 37 (36.3%) partial responses (PR). The clinical CR were significantly confined to subgroups C (20/34 pts, 58.8%) and B (16/34 pts, 47.1%) than to subgroup A (4/34 pts, 11.8%) (p for trend = 0.00001). Objective response rate was 46.1% (47/102 pts) at month 4, 83.8% (57/68 pts) at month 8, and 94.1% (32/34 pts) at month 12 compared with baseline. As expected, letrozole induced a significant reduction of Ki67 expression after treatment in any single subgroup, without significant differences between them. With very interest, the pathologic CR rate was significantly higher in subgroup C (7/34 pts, 20.6%) than in subgroups A (1/34 pt, 2.9%) and B (1/34 pt, 2.9%) (p for trend <0.02).

Conclusions: One-year neoadjuvant letrozole therapy can lead to a high pathologic CR rate. This observation suggests that pathologic CR may be a reliable endpoint after endocrine therapy, but a long-term drug exposure is needed. The optimal letrozole treatment duration in neoadjuvant setting is a matter of future research.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD07-03.
Abstract PD08-05: Spanning the Continuum to assess, serve and navigate Latinas with breast cancer: A Tale of Six Projects
Authors: Ramirez AG , Holden AE , Gallion K , SanMiguel SA , Munoz E , Penedo FJ , Perez-Stable EJ , Talavera GG , Carrillo JE , Fernandez ME
Published: August 2014

Background Breast cancer is the most commonly diagnosed cancer among Latinas: 14,200 diagnosed in 2009. Groundbreaking work has shown that patient navigation may assist minority patients to negotiate the Cancer Care Continuum. Here we report the efforts of Redes En Acción: The National Latino Cancer Research Network at the Institute for Health Promotion Research in San Antonio, Texas in applying these findings to Latinas.

Methods We conducted six studies to assess and address the needs and self-reported barriers to care of Latina breast cancer patients, leading to the development and controlled trial of a theory-driven, culturally appropriate patient navigation model and program. We currently are evaluating health-related quality of life and barriers to genetic testing in Latina breast cancer survivors.

Results Latinas with breast cancer face significant obstacles across the cancer care continuum, originating from multiple domains, including traditional values, timely access to care and cultural orientation. These have a direct influence on interaction with the system or indirectly via influences on existing barriers including socioeconomic marginalization and affective response to the stress of any of these.

Conclusion Although not considered as such, the Cancer Care Continuum implicitly demands consideration of the cultural underpinnings of behavior that govern interaction with it. Further research is required to understand these underpinnings. Simultaneously, culturally-sensitive services must be incorporated into the medical care system in order to ensure its successful function.

Acknowledgement This research was enabled by grants from the San Antonio Cancer Institute/Cancer Therapy and Research Center, San Antonio, Texas (P30-CA054174), Susan G. Komen for the Cure (POP 2000 704), and the National Cancer Institute via Redes En Acción (U01-CA86117 and U54 CA153511-01).

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD08-05.
Abstract PD09-05: Single nucleotide polymorphism of XRCC1 which participates in DNA repair mechanism predicts clinical outcome in relapsed or metastatic breast cancer patients treated with S1 and oxaliplatin chemotherapy: Results from multicenter prospective study (TORCH_KCSG BR07-03)
Authors: Im S-A , Oh D-Y , Keam B , Lee KS , Ahn J-H , Sohn J , Ahn JS , Kim JH , Lee MH , Lee KE , Kim HJ , Lee K-H , Han SW , Kim S-Y , Kim SB , Im Y-H , Ro J , Park H-S
Published: August 2014

Background: S1 and oxaliplatin (SOX) combination chemotherapy is an effective regimen in anthracycline and taxane pretreated metastatic breast cancer (MBC) patients with manageable toxicities (KCSG BR07-03, SABCS 2011 #Abst P3-16-06). The aim of this study was to investigate the association of the single nucleotide polymorphisms (SNPs) and clinical outcome in MBC treated with SOX chemotherapy.

Patients and Methods: A total of 87 MBC patients previously treated with or resistant to anthracycline and taxane chemotherapy were enrolled in this prospective multicenter trial. The patients received S-1 80mg/m2/day (day 1–14) and oxaliplatin 130 mg/m2 (day 1) every 3 weeks till progression. Among the 87 patients, 77 patients were available for SNP analysis. Germline DNA from peripheral blood (PB) mononuclear cells was extracted. SNPs in 4 genes from pathways that may influence cellular sensitivity to S1 and oxaliplatin (TS, ERCC, XPD, and XRCC) were genotyped from PB sample using PCR-restriction fragment length polymorphism.

Results: Overall response rate (RR) was 38.5% (95% CI: 27.7–49.3) and disease control rate was 67.9% (95% CI:57.5–78.3) to SOX. Median time-to-progression (TTP) and overall survival (OS) were 6.0 mo (95% CI: 5.1–6.9 mo) and 19.4 mo (95% CI: not estimated), respectively. XRCC1 Arg194Trp SNP which participates in DNA repair mechanism showed correlation with the clinical outcome. RR was tend to higher in XRCC1 Arg194Trp CC genotype compared with CT or TT genotype (50.0 % vs 35.1% or 12.5%, P = 0.121). TTP of patients with CC genotype in XRCC1 Arg194Trp was significantly longer than the TTP of patients with CT or TT genotype (median TTP: 6.4 mo in CC, 5.9 mo in CT, 3.0 mo in TT, P = 0.007) as well as overall survival (OS) (median OS: not reached in CC, 13.9 mo in CT, 7.1 mo in TT, P = 0.006). After adjusting for hormone receptor status, performance status, and visceral involvement, prognostic value of XRCC1 Arg194Trp SNP remained significant (Hazard Ratio=1.322 and 4.484, P = 0.016). Other SNPs were not significantly associated with survival or toxicities.

Conclusion: XRCC1 Arg194Trp SNP is associated with clinical outcome of MBC patients treated with SOX chemotherapy. Further studies of the relationship between germline polymorphisms in XRCC1 and functional mechanism researches are warranted.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD09-05.
Abstract PD09-08: The potential role of TLK2 as a therapeutic target in breast cancer.
Authors: Kim J-A , Cao X , Tan Y , Schiff R , Wang X
Published: August 2014

Invasive breast cancer (IBC) is a heterogeneous disease with distinct molecular and histological features as well as diverse biological behaviors. It is thus critical to find specific gene aberrations that can drive breast cancer initiation and progression as potential therapeutic targets. Our lab has established bioinformatics workflows to identify therapeutic oncogene targets. We have nominated Tousled-Like Kinase 2 (TLK2) as a candidate therapeutic target from 44,932 human genes based on integrative copy number data from The Cancer Genome Atlas (TCGA). TLK2 is a nuclear serine/threonine kinase that serves as a cell cycle checkpoint regulating chromosome assembly, and is inactivated by phosphorylation through the ATM dependent manner in the response to the double strand breaks during the S-phase. Interestingly, we observed 12–14% of focal TLK2 gene amplifications and translocations in invasive breast cancers, which are more frequent in luminal B tumors. Chromosomal aberrations may leads the deregulation of TLK2 kinase activity and results in overcoming the cell cycle checkpoint upon DNA damage such as IR or chemotherapeutic drugs treatments in breast cancer, thus destabilizing the cancer genome. Indeed, our preliminary data show that TLK2 knockdown substantially inhibits cell proliferation in breast cancer cells overexpressing TLK2. Further, this response is sustained in their derivative strains resistant to hormone or anti-Her2 therapies, suggesting the potential of TLK2 inhibition to sensitize these therapies as well. Overall, I expect that this study will provide compelling evidence regarding the role of TLK2 in breast cancer tumorigenesis and genomic instability, and establish its potential as a therapeutic target. The possibility of developing specific TLK2 inhibitors can be well demonstrated by the previous success of drugs against other cell cycle checkpoint kinases that are currently under clinical trials. In addition, molecular assays detecting TLK2 derangements may be applied in the clinics to identify patients who might benefit from future TLK2 inhibitor. If TLK2 endows resistance or sensitivity to other therapies, assays detecting TLK2 aberrations may also have important predictive value for personalization of these treatments.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD09-08.
Abstract PD10-06: CXCL13 mRNA predicts Docetaxel benefit in Triple Negative tumors
Authors: Wirtz RM , Leinonen M , Bono P , Isola J , Kellokumpu-Lehtinen P-L , Kataja V , Turpeenniemi-Hujanen T , Jyrkkiö S , Eidt S , Schmidt M , Joensuu H
Published: August 2014

Background: Presence of immune cell infiltrates in tumor may influence outcome of patients with node negative breast cancer (BC) (Schmidt et al 2008). High expression of CXCL13 (B lymphocyte chemoattractant chemokine) was strongly associated with favorable survival in one recent series consisting of BC patients treated with adjuvant chemotherapy (Razis et al. 2012). Here we examined the clinical significance of breast tumor CXCL13 mRNA levels within the context of the FinHer trial, where patients were randomly assigned to receive 3 cycles of either docetaxel or vinorelbine followed by 3 cycles of FEC. Patients with HER2-positive BC had a second randomization for trastuzumab vs. control. Since intratumoral B cells are of particular importance for obtaining response to chemotherapy in triple negative BC (TNBC) and of limited value in luminal BC (Schmidt et al. 2012), and trastuzumab treatment is a confounding factor in HER2+ BC, we focused on TNBC.

Methods: RNA was extracted from FFPE tumor tissue of 917 (90.8%) out of the 1010 patients who participated in the FinHer trial. CXCL13 mRNA expression was measured using RT-qPCR. The molecular subtypes (luminal, HER2-enriched and triple-negative) were determined using IHC and central chromogenic in situ hybridization (CISH) testing. Prognostic significance of factors was assessed using univariate and multivariate analyses. Distant disease-free survival (DDFS) between groups was compared using the log-rank test.

Results: Tumor CXCL13 mRNA expression showed a bimodal distribution and correlated negatively with tumor ESR1 mRNA levels (r = −0,31; p < 0.0001). 146 (15.9%) out of the 917cancers were classified as TNBC. High tumor CXCL13 mRNA levels above the median were strongly associated with favorable 5-yr DDFS in TNBC compared to low levels (85% vs. 60%; p < 0.0001). The prognostic value of tumor CXCL13 was evident in the subset of women with TNBC treated with vinorelbine (5-year DDFS 90% vs. 40%; p < 0.0001), whereas no significant association was found in the docetaxel arm (5-year DDFS 80% vs. 80%; p = 0.76). When only patients with TNBC and low CXCL13 mRNA levels were assessed, patients treated with docetaxel had better DDFS compared to those treated with vinorelbine (5-year DDFS 80% versus 40%; p < 0.0064). There was no DDFS difference between treatment arms in patients exhibiting high CXCL13 expression.

Conclusions: Tumor CXCL13 mRNA expression is a favorable prognostic factor for women with TNBC treated with adjuvant chemotherapy providing further evidence that the host immune response may influence outcome of patients with early BC. The clinical significance of the host B cell immune response may vary substantially pending on the type of chemotherapy administered.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD10-06.
Abstract PD10-09: CYP2D6 and adjuvant tamoxifen: Impact on outcome in pre- but not postmenopausal breast cancer patients
Authors: Margolin S , Lindh J , Thorén L , Xie H , Koukel L , Dahl M-L , Eliasson E
Published: August 2014

Background: The therapeutic effect of tamoxifen in adjuvant treatment of hormone receptor positive breast cancer might be impaired in patients with cytochrome P450 2D6 (CYP2D6) genotypes that predispose to decreased formation of potent anti-estrogenic tamoxifen metabolites. However, previous studies have shown inconsistent findings in this regard. In light of two recent studies failing to show an impact of CYP2D6 genotype on outcome in postmenopausal patients, we hypothesized that deficient CYP2D6 might be of greater importance in premenopausal patients with high levels of circulating estrogen. We therefore aimed to study the effect of CYP2D6 activity in both pre- and postmenopausal patients who were adherent to tamoxifen treatment for at least one year.

Methods: 382 patients, from a population-based cohort of unselected breast cancer patients that were prescribed adjuvant tamoxifen for five years, constituted the base of the study. Information on menopausal status, tumor characteristics, treatment data including compliance and outcome was retrieved from medical records. Comprehensive CYP2D6 genotyping was performed and functionally translated into constitutive, metabolic activity.

Results: In patients adherent to tamoxifen for at least one year (n = 313) there was an association of reduced CYP2D6 activity (≤50% of normal) to both recurrence (p = 0.02) and breast cancer-specific mortality (p = 0.03). In a multivariable analysis including CYP2D6 activity, age at diagnosis, tumor size, lymph node status, grade, adjuvant chemotherapy and concomitant use of CYP2D6 inhibitors, CYP2D6 remained an independent predictor of recurrence (HR = 0.39, 95% CI: 0.18–0.85, p = 0.02) and breast cancer specific survival (HR = 0.33, 95% CI 0.12–0.90, p = 0.03). Gradually decreasing CYP2D6 activity paralleled increasing risk of recurrence and breast cancer related mortality. The effect of CYP2D6 derived mainly from premenopausal patients with an association to both recurrence (p = 0.01) and breast cancer specific survival (p = 0.04). There was no such association in the postmenopausal group.

Conclusion: In a prospectively collected cohort of tamoxifen-treated breast cancer patients, an association between CYP2D6 genotype and outcome was evident in patients that were adherent to tamoxifen treatment for at least a year. Importantly, this effect derived from premenopausal patients only.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD10-09.
Abstract S1-10: Association between the 21-gene recurrence score (RS) and benefit from adjuvant paclitaxel (Pac) in node-positive (N+), ER-positive breast cancer patients (pts): Results from NSABP B-28
Authors: Mamounas EP , Tang G , Paik S , Baehner FL , Liu Q , Jeong J-H , Kim S-R , Butler SM , Jamshidian F , Cherbavaz DB , Sing AP , Shak S , Julian TB , Lembersky BC , Wickerham DL , Costantino JP , Wolmark N
Published: August 2014

Background: The RS result predicts outcome in N- and N+, ER+ pts treated with adjuvant endocrine therapy. RS also predicts benefit from adjuvant chemotherapy (CT) and pts with a high RS result receive most of the benefit. We evaluated the association between RS and paclitaxel (Pac) benefit in N+, ER+ pts from NSABP B-28.

Methods: B-28 compared 4 cycles of doxorubicin/cyclophosphamide (AC) vs. AC followed by Pac × 4 (AC→Pac). Pts ≥50 yrs and those<50 yrs with ER+ and/or PR+ tumors also received 5 yrs of tamoxifen starting with CT. From 8/95 to 5/98 3060 pts were accrued. The present study includes 1065 pts, ER+ by central tissue microarray IHC assay, tamoxifen-treated, and with a RS result from the Oncotype DX 21-gene assay by standard processes. Sub-distribution analyses were used for loco-regional recurrence (LRR) to account for competing risks, including distant recurrence (DR), second primary cancers, and death. Kaplan-Meier estimates and Cox models were used for other clinical outcomes.

Results: Median follow-up time was 11.2 yrs. Of the 1065 pts, 386 (36%) had low RS (<18); 364 (34%) intermediate RS (18–30); and 315 (30%) high RS (≥31). RS was a significant predictor of LRR, DFS event, DR, and death in univariate analyses both in pts treated with AC and in those treated with AC→Pac (Table). Pts with low RS had similar outcomes at 10 yrs when treated with AC→Pac vs. AC (Table). The majority of Pac benefit was observed in pts with intermediate/high RS (Table). Interaction tests between RS and Pac benefit were not statistically significant (LRR: p = 0.75, DFS: p = 0.65, DR: p = 0.93, death: p = 0.30).

Conclusions: RS significantly predicts risk for LRR, DR, DFS event, and death in N+, ER+ pts treated with AC or AC→Pac adjuvant CT. Pts with a low RS value have similar outcomes whether treated with AC or with AC→Pac and most of Pac benefit is evident in pts with intermediate/high RS. Although there was no significant interaction between RS and Pac benefit, these results support previous findings of lack of CT benefit in pts with a low RS result.

Supported by: NCI grants U10-CA-12027, -69651, -37377, -69974, U24-CA-114732, and CA-75362; Susan G. Komen for the Cure® grants; and Bristol-Myers Squibb Pharmaceutical Research Institute

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr S1-10.
Abstract S1-8: Prolactin-humanized mice: an improved animal recipient for therapy response-testing of patient-derived breast cancer xenotransplants
Authors: Rui H , Utama FE , Yanac AF , Xia G , Peck AR , Liu C , Rosenberg AL , Wagner K-U , Yang N
Published: August 2014

Eighty percent of newly diagnosed breast cancer represents estrogen receptor(ER)-positive luminal subtypes. Many patients with luminal breast cancer develop antiestrogen resistant disease. It has historically been particularly difficult to establish ER-positive breast cancer lines from primary breast cancer in the laboratory or in mice. Murine and bovine prolactins, the major lactogens in current laboratory experimental in vivo and in vitro conditions, fail to activate human prolactin receptors because of species incompatibility. In fact, murine prolactin is a potent antagonist for human prolactin receptors. Because ER-positive, luminal breast cancers also express prolactin receptors, we hypothesized that lack of human lactogenic activity under experimental conditions selected against establishment of ER-positive breast cancer in the laboratory. We therefore genetically engineered mice to express physiological levels of human prolactin in place of mouse prolactin and backcrossed the mice for ten generations into the immunodeficient NSG strain. The resulting hPRL.NSG mice have a greatly improved take rate for ER positive, luminal type of breast cancer, suggesting key tumor-promoting roles for prolactin in luminal breast cancer. A panel of novel transplantable human breast cancer lines has been established in hPrl.NSG mice, the majority of which are ER-positive. The transplantable lines maintain key histopathological characteristics and expression of major marker proteins of the primary patient tumors. Intriguingly, initial tumor establishment and growth rates of breast cancer xenografts were consistently greater in the hPrl.NSG mice than in wildtype NSG mice. Furthermore, tumors grown in hPrl.NSG were more responsive to tamoxifen than size-matched tumors grown in wildtype NSG mice. At least two new tumor lines examined so far develop spontaneous distant metastases in hPrl.NSG mice, with evidence of prolactin-dependent progression of ER-positive disease. Collectively, these observations validate the hPrl.NSG mice as an improved recipient for preclinical modeling of human breast cancer in vivo, both for therapeutic targeting of prolactin-pathways and other growth and survival pathways, as well as overcoming anti-estrogen resistance.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr S1-8.
Abstract S4-4: Independent validation of Genomic Grade in the BIG 1-98 study
Authors: Sotiriou C , Ignatiadis M , Desmedt C , Azim Jr HA , Veys I , Larsimont D , Lyng M , Viale G , Leyland-Jones B , Ditzel H , Giobbie-Hurder A , Regan M , Piccart M , Michiels S
Published: August 2014

Background: We have reported that a 97-gene signature, the Genomic Grade Index (GGI) can separate histological grade (HG) 2 breast tumors into low vs high GGI tumors with different outcomes. We have also developed a quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) tool including 6 reporter and 3 control genes that can reliably evaluate Genomic Grade (GG) in paraffin embedded tumors.

Methods: We evaluated the qRT-PCR GG in women treated with either tamoxifen or letrozole monotherapy in the Breast International Group (BIG) 1–98 study with 8.1 year median follow-up. The association between continuous GG and distant recurrence-free interval (DRFI) was evaluated in Cox regression models stratified for the 2 vs 4 arm randomization option, chemotherapy and hormonotherapy use, with and without adjustment for clinicopathological characteristics. Estrogen, progesterone receptor (ER, PR), Ki67 and HG were centrally reviewed. The clinicopathological model included age and log2 tumor size, ER and PR as continuous variables, nodal status (N0 vs 1–3 vs >3), HG (1 vs 2 vs 3) and HER2 (negative vs positive). Similar analyses were performed for log2 Ki67 as continuous variable. Added prognostic value was assessed using the likelihood ratio statistic.

Results: We obtained GG in 883 (62%) of 1428 samples. Non evaluable results were due to pre-analytical issues or technical failure. Among the 883 patients, 521 (59%) had N0 disease, 435 (49%) were treated with tamoxifen and 84 (10%) had distant recurrences. The GG classified 502 patients with HG2 tumors as either GG1 (N = 202, 40%), equivocal (N = 220, 44%) or GG3 (N = 80, 16%). In univariate analysis, increasing GG and Ki67 were significantly associated with lower DRFI (Table). When the 773 N0/1-3 patients were analyzed based on HG, the Kaplan-Meier estimate for 10-year DRFI was 98% (95% confidence intervals 96–100%) for the 123 HG1, 92% (88–95%) for the 456 HG2 and 84% (78–90%) for the 194 HG3 patients. In the N0/1-3 population, 10 year DRFI based on GG was 95% (96–100%) for the 290 GG1, 92% (89–95%) for the 309 GG equivocal and 84% (77–90%) for the 178 GG3 patients. Interestingly, when the 456 HG2 and N0/1-3 patients were analyzed based on GG, the 10-year DRFI was 95% (92–100%) for the 185 HG2/GG1 patients, 92% (88–96%) for the 202 HG2/GG equivocal patients, 86% (76–96%) for the 69 HG2/GG3 patients. In all patients either GG or Ki67 as continuous variable significantly improved the clinicopathological model in predicting distant recurrence (Table).

Conclusion: Either Genomic Grade or centrally reviewed Ki67 provides independent prognostic information for risk of distant recurrence beyond clinicopathological characteristics in patients treated with endocrine therapy.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr S4-4.