bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P4-06-17: Antitumor activity of the novel mithramycin analog EC8042 in triple negative breast cancer
Authors: Pandiella A , Montero JC , Cuenca D , Re-Louhau F , Nuñez LE , Moris F , Ocana A
Published: August 2014

Background: Triple negative breast cancer (TNBC) represents around 15% of all breast tumors. Therapeutic strategies in TNBC are mainly based on classical chemotherapeutic agents, with limited success in the metastatic setting. Here we have explored the action of a novel mithramycin analog named EC-8042 in TNBC.

Methods: we used a panel of cell lines representative of triple negative tumors including BT549, HBL100, HCC-1037, HS578T, MDA-MB213, HCC-1187, HCC-3153 and HCC-70. Proliferation studies with EC-8042 were measured by MTT assays. Propidium iodide (PI) staining followed by FACS was used for cell cycle analyses and AnnexinV-fluorescein for analysis of apoptosis. Proteins involved in cell cycle control and apoptosis were evaluated by western blot. Mitochondrial membrane potential was assessed by measuring the fluorescence of tetramethylrhodamineethylester (TMRE) dye.

Results: The IC50 values obtained were in the nM range, indicating that the drug had a potent effect on these cell lines. Mechanistically, the drug caused accumulation of cells in the S phase of the cell cycle at doses of 100 and 500 nM in MDA-MB231. Biochemically, EC-8042 caused an increase in cyclin E, cyclin A, cyclin D3, cyclin B, and p27, and decreased cyclin D1 and Rb phosphorylation at serine 780. These data are consistent with an action of EC-8042 on the cell cycle. Moreover, the increase in wee1 and pY15-CDK1 are in line with an effect of the drug blocking cell cycle progression through the S phase. Annexin V staining indicated that treatment with 500 nM of EC8042 provoked apoptosis, which was also revealed by PARP processing. The treatment reduced the amounts of MCL1, BCLXL, and XIAP, but did not affect BCL2 levels.

Conclusion: our results indicate that EC-8042 has a strong antitumoral action on TNBC cells, which is caused by increased apoptosis and a decrease in cell cycle progression, and opens the possibility for further development of this drug in TNBC.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-17.
Abstract P4-06-18: Targeting basal-like breast cancer through downstream effectors of oncogene cooperation
Authors: McMurray HR , Walters EA , Heyer DM , Candelaria PV , Wang S , Grose VA , Llop JR , Zhang Y
Published: August 2014

Basal-like breast cancer is a highly aggressive and lethal form of cancer, not amenable to treatment by molecularly targeted agents or conventional chemotherapy effective against other forms of breast cancer. Thus, discovery of novel intervention targets regulating tumorigenesis in basal-like breast cancer cells is critical. The success of molecularly targeted therapies against some forms of cancer has led to a search for additional cancer cell-specific targets that will permit effective tumor inhibition with minimal damage to normal cells. These efforts are largely focused on identification of novel oncogenes and tumor suppressors by genome sequencing or synthetic lethalities found via genome-wide RNAi screens. We employ a novel approach to identify cancer cell vulnerabilities among genes disregulated downstream of cooperating oncogenic mutations, which drive malignant transformation in large part through synergistic modulation of non-mutated downstream effector genes, the ‘cooperation response genes’ (CRGs) [McMurray et al., Nature, 19 June 2008]. After assessing how common was CRG disregulation in different epithelial cancers, via comparative genomic analysis of CRG expression patterns in tumor vs. normal tissue, we found that 34 CRGs (∼35%) were significantly disregulated in breast cancer. Because of the need for novel intervention targets in basal-like breast cancer, we focused our functional analysis there. We have tested a subset of CRGs disregulated in breast tumors for their contribution to malignancy in basal-like breast cancer cells, re-setting CRG levels toward normal cell levels by stable cDNA expression or shRNA-mediated knock-down and testing tumor formation capacity of perturbed cells. Our studies reveal that CRGs play an essential role in controlling tumor formation of basal-like breast cancer cells, with significant reductions in tumor volume, and in some cases reduced tumor incidence, upon CRG modulation. Disruption of cell behavior appears to be limited to cancer cells, as non-transformed mammary cells harboring analogous perturbations retain capacity to form acinar structures in 3D culture. Several FDA approved pharmacological agents, identified to reverse the CRG expression pattern, display inhibitory effects on basal-like breast cancer cell lines, with minimal effects on non-transformed cells. CRGs thus represent cancer cell-specific vulnerabilities in basal-like breast cancer, providing novel and unexpected opportunities for intervention against this difficult-to-target disease.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-18.
Abstract P4-07-02: Growth differentiation factor 15, a novel mediator of progression in HER2-positive breast cancer
Authors: Nahta Rita , Joshi Jayashree P , Griner Samantha E , Adams Amy L , Andre Fabrice
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-07-02.
Abstract P4-07-03: Rapamycin and Dalotuzumab in combination inhibit parental and endocrine resistant breast cancer cells.
Authors: Beckwith H , Fettig-Anderson L , Yueh N , Douglas Y
Published: August 2014

The PI3K/Akt/mTOR pathway is involved in breast cancer resistance to endocrine therapy. Inhibitors of mTOR have been shown to have benefit for patients with ER positive tumors that have developed endocrine therapy resistance. Inhibition of mTOR triggers a negative feedback loop resulting in enhanced phosphorylation of Akt and subsequent breast cancer cell proliferation via the IGF signaling pathway. Therefore, blockade of both IGF-1R and mTOR may be necessary to completely suppress this pathway. Our laboratory studied the effect of dual inhibition of mTOR and IGF-1R with rapamycin and dalotuzumab respectively on parental and endocrine resistant MCF-7 breast cancer cell tumorigenesis.

Immunoblotting demonstrates parental MCF-7L cells treated with rapamycin alone demonstrate increased phosphorylation of Akt. Inhibition of phosphorylation of Akt can be achieved with combined treatment with rapamycin and dalotuzumab at concentrations of 5 nM and 2 µg/mL respectively. In addition to inhibition of pAkt, combined treatment inhibits phosphorylation of S6K1 and the translational repressor protein, 4eBP1. Furthermore, MTT proliferation assay and soft agar assay demonstrate inhibition of parental cell proliferation and colonization respectively with combined rapamycin and dalotuzumab treatment. In order to examine the effect of combination therapy in endocrine resistant cell lines, we established tamoxifen resistant (TamR) and long term estrogen deprived (LTED) cell lines. TamR cells were generated by culturing MCF-7L cells in the presence of tamoxifen for more than 1 year. LTED cells were generated by culturing MCF-7L cells in the absence of estrogen for more than 6 months. Proliferation of TamR and LTED cells treated with rapamycin at a concentration of 5nM and dalotuzumab at a concentration of 2µg/mL in combination was significantly inhibited. We conclude that combination of IGF1R and mTOR inhibition might be necessary to inhibit endocrine resistant breast cancers. Ongoing clinical trials will test this combination of drugs.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-07-03.
Abstract P4-07-05: MEDI3379, an antibody against HER3, is active in HER2-driven human breast tumor models
Authors: Xiao Z , Rothstein R , Carrasco R , Wetzel L , Kinneer K , Chen H , Tice D , Hollingsworth R , Steiner P
Published: August 2014

HER3 (ERBB3) is a member of the EGFR/HER family of receptor tyrosine kinases (RTK), consisting of EGFR, HER2, HER3 and HER4. Unlike the other HER family members, HER3 lacks intrinsic tyrosine kinase activity and therefore needs to form heterodimers with EGFR, HER2 or other kinase-proficient RTKs to be functionally active. Dimerization is induced by overexpression of EGFR or HER2 in a ligand-independent (LI) manner. In this process HER3 acts as a driver in divergent cancer types including HER2-positive breast cancer (BC) via induction of the PI3K-AKT pathway. Alternatively, heregulin (NRG1/HRG), the major HER3 ligand, induces a conformational shift in HER3 which leads to dimer formation with a partner RTK in a ligand-dependent (LD) manner.

We have developed an antagonistic human monoclonal antibody against HER3, termed MEDI3379, and tested it in multiple breast cancer cell lines. We observed effective suppression of constitutive pHER3 and pAKT with MEDI3379, leading to anti-proliferation effects in cell culture models. Preclinical evaluation of MEDI3379 demonstrated antitumor activity in several orthotopic BC xenografts in nude mice which did not express HRG. For example, the BC xenograft model BT474 with amplified HER2 responded to MEDI3379 (65% dTGI). In conclusion, our findings with targeting of HER3 in mouse models support continued development of MEDI3379 for cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-07-05.
Abstract P4-08-02: Reactivation of oncogenic signaling through mTOR inhibitors-induced feedback adaptations.
Authors: Rodrik-Outmezguine V , Chandarlapaty S , Poulikakos P , Scaltriti M , Baselga J , Rosen N
Published: August 2014

mTORC1 inhibitors (or rapalogs) relieve feedback inhibition of receptor tyrosine kinases (RTKs) and cause mTORC2-dependent phosphorylation of AKT S473 and activation of AKT kinase and signaling. The relief of feedback inhibition of mTORC2/AKT signaling has been shown to abolish the antitumor activity of rapalogs and has often been proposed as a potential mechanism of resistance to these compounds.

mTOR kinase inhibitors that inhibit both mTORC1 and mTORC2 complexes have now been developed to circumvent these problems. mTOR kinase inhibitors block mTORC1 and mTORC2 and thus, do not cause the mTORC2 activation of AKT observed with rapamycin. We now show, however, that these novel classes of drugs have a biphasic effect on AKT. Inhibition of mTORC2 leads to AKT S473 dephosphorylation and a rapid but transient inhibition of AKT T308 phosphorylation and AKT signaling. However, inhibition of mTOR kinase, as observed with rapalogs, also relieves feedback inhibition of RTKs leading to subsequent PI3K activation and rephosphorylation of AKT but only at T308, which is sufficient to reactivate AKT activity and signaling. Thus, catalytic inhibition of mTOR kinase leads to a new steady state characterized by profound inhibition of mTORC1 and accumulation of activated AKT phosphorylated on T308 but not S473.

We now confirm that the reactivation of AKT signaling is a mechanism of acquired resistance to rapalogs. We observe an induction of phospho-RTKs in rapamycin-resistant cells leading to both AKT and ERK signaling pathway activation enhancing cell survival. Combined inhibition of mTOR and the induced RTKs can prevent AKT reactivation and fully abolishes AKT signaling. These results reveal the adaptive capabilities of oncogenic signaling networks, highlighting the possible need for combinatorial approaches to block feedback-regulated pathways.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-08-02.
Abstract P4-08-06: Notch-dependent Regulation of Novel Genes Associated with Trastuzumab Resistance
Authors: Osipo C , Baumgartner A , Zlobin A , O'Toole M
Published: August 2014

Background: We have shown previously that Notch-1 is critical for the development and maintenance of trasuzumab or lapatinib resistance in HER2 positive breast cancer cell lines. Here we sought to identify the critical genes that are regulated by Notch to promote anti-HER2 targeted resistance.

Methods: We measured expression of 84 known breast cancer-associated transcripts using a real-time PCR array in both trastuzumab sensitive and resistant BT474 cell lines. A 4 fold increase or decrease in transcripts levels were measured to be statisitically significant. To confirm the change in levels, we designed primers to each gene of interest and repeated the PCR at least three independent times. Furthermore, we specifically asked whether Notch-1 or it's transcriptional mediator, CBF-1, directly regulated any of the identified genes in HER2 positive breast cancer cells using a genetic knocked down approach. An ANOVA for multiple comparisons was used to compute statistical significance.

Results: We identified more than 4 fold increase in APC, ABCG2, ABCB1, Gata-3, Bcl-2, Id2, and HRG-2 transcripts in trastuzumab resistant versus sensitive cells. Conversley, Gli1 was downregulated in resistant versus sensitive cells. Interestingly, siRNA directed against Notch-1 or CBF-1 decreased ABCG2 and Gata-3, but conversely increased ABCB1 and HRG-2 in resistant cells.

Conclusions: Our findings demonstrate APC, ABCG2, ABCB1, Gata-3, Bcl-2, Id2, and HRG-2 transcripts are signficantly increased in trastuzumab resistant breast cancer cells compared to sensitive cells. More importantly, Notch-1 or it's direct transcriptional activator CBF-1 may contribute to traztuzumab resistance by directly regulating critical genes that contribute to multidrug resistance: ABCB1 and ABCG2 and/or genes associated with stem cell survival: Gata-3 and HRG-2.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-08-06.
Abstract P4-08-09: Targeting Thyroid Receptor b in Estrogen Receptor Negative Breast Cancer
Authors: Gu G , Covington K , Rechoum Y , O'Malley B , Mangelsdorf D , Minna J , Webb P , Fuqua S
Published: August 2014

Background: The treatment of estrogen receptor (ER)-negative breast cancer (BC) is a major clinical problem due to the lack of useful therapeutic targets. Nuclear receptors (NRs) are potential targets in these patients because they regulate global transcriptional events and many already have agonists/antagonists available.

Material and Methods: We used microarray analysis of 227 ER-negative tumors to identify NR targets, and performed hierarchical clustering using 41 NRs. Expressed receptors were scored using prediction analysis of microarrays (PAM) across clustered groups. Cell lines were matched to subtypes using previously described data (Neve et al. 2006). Candidate gene expression levels were confirmed by qRT-PCR using TaqMan probes. pGIPZ lentiviral vectors encoding shRNA were used to knockdownselected candidates. MTT and soft agar assays were used to measure chemosensitivity and growth following treatment with Docetaxel (Doc), Doxorubicin (Dox), or Cisplatin (Cis). Statistical analysis was performed using Red-R.

Results: The 41 NRs clustered tumors into 5 groups. For each group we selected genes representing the highest ranked discriminators, and examined their effects in cell lines matching each groups' gene signature. Thyroid hormone receptor b (THRβ) was selected from group V. The expression levels of this receptor were confirmed by qRT-PCR and Western blot analysis.

Knockdown of THRβ in ER-negative HCC2185 cells rendered cells more resistant to all chemotherapeutics by using MTT assay. Similar results were confirmed in ER-negative MDA-MB-453 and HCC202 cells. Knockdown of THRβ enhanced colony forming potential in anchorage-independent soft agar assays in MDA-MB-453 and HCC202 cells. Statistical analysis using clinical data from Sabatier et al. (BCRT 2011) showed that patients with low THRb have a worse clinical outcome. In order to translate these findings into the clinic, we treated cells with a specific THRβ agonists, GC-1 and KB-141. GC-1 inhibited cell growth in growth assays, and synergistic effects were observed when cells were treated with GC-1 and Docetaxel in combination. Re-expression of ERα protein was observed in ER-nagative cells lines after treatment with GC-1 and KB141, suggesting that modulation of THRβ may also extend hormonal therapy to this hormonally insensitive group of patients.

Conclusion: Clinical targeting of NRs in ER-negative BCs is a novel strategy since receptors can be specifically targeted with ligands. Our data suggest that chemotherapy response in ER-negative patients overexpressing THRβ could be enhanced with a THRβ agonist. Similarly, functional re-activation of ERα by activating THRβ might extend hormonal therapies to these patients as well.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-08-09.
Abstract P4-09-03: Comparative proteomic analysis identifies a 15-protein signature related to clinical outcome of triple negative breast cancer
Authors: Liu Ning Qing , De Marchi Tommaso , Stingl Christoph , Timmermans Annemieke M , Look Maxime P , Braakman René BH , Smid Marcel , Foekens Renée , Trapman Anita MAC , Luider Theo M , Martens John WM , Foekens John A , Umar Arzu
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-09-03.
Abstract P4-09-04: Gene expression profile that predict outcome of Tamoxifen-treated estrogen receptor-positive, high-risk, primary breast cancer patients: a DBCG study.
Authors: Lyng MB , Lænkholm A-V , Tan Q , Vach W , Gravgaard KH , Knoop A , Ditzel HJ
Published: August 2014

Background: Tamoxifen significantly improves outcome for estrogen receptor-positive (ER+) breast cancer, but the 15-year recurrence rate remains 30%. The aim of this study was to identify gene profiles that accurately predicted the outcome of ER+ breast cancer patients who received adjuvant Tamoxifen mono-therapy.

Methods: Post-menopausal breast cancer patients diagnosed no later than 2002, being ER+ as defined by >1% IHC staining and having a frozen tumor sample with >50% tumor content were included. Tumor samples from 108 patients treated with adjuvant Tamoxifen were analyzed for the expression of 59 genes using quantitative-PCR. End-point was clinically verified recurrence to distant organs or ipsilateral breast. Gene profiles were identified using a model building procedure based on conditional logistic regression and leave-one-out cross-validation, followed by a non-parametric boostrap (1000x re-sampling). The optimal profiles were further examined in 5 previously-reported datasets containing similar patient populations that were either treated with Tamoxifen or left untreated (N = 623).

Results: Three gene profiles were identified, the strongest being a 2-gene combination of BCL2-CDKN1A exhibiting an accuracy of 75% for prediction of outcome. Independent examination using 4 previously-reported microarray datasets of Tamoxifen-treated patient samples (N = 503) confirmed the potential of BCL2-CDKN1A (average accuracy: 68%, range: 53%–85%). The predictive value was further determined by comparing the ability of the genes to predict recurrence in an additional, previously-published, cohort consisting of Tamoxifen-treated (n = 58, p = 0.015) and untreated patients (n = 62, p = 0.25).

Conclusion: A novel gene expression profile predictive of outcome of Tamoxifen-treated patients was identified. The validation suggests that BCL2-CDKN1A exhibit strong predictive potential.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-09-04.
Abstract P4-11-02: Novel BRCA1 and BRCA2 genomic rearrangements in Southern Chinese breast/ovarian cancer patients
Authors: Kwong A , Ng EKO , Law FBF , Wa A , Wong CLP , Wong CHN , Kurian AW , West DW , Ford JM , Ma ESK
Published: August 2014

Background and Aims: Germline mutations in the two breast cancer susceptibility genes, BRCA1 and BRCA2 account for a significant portion of hereditary breast/ovarian cancer. Most of the BRCA mutations reported in Southern Chinese patients were point mutations, small deletions, and insertions. The spectrum of large genomic rearrangement (LGR) is largely unknown. Here we perform the first study on the LGR of BRCA genes in a Hong Kong Chinese population. We aimed to determine the spectrum of BRCA LGRs in Southern Chinese patients with breast cancer.

Methods: A total of 555 clinically high-risk breast and/or ovarian cancer patients were recruited from the Hong Kong Hereditary Breast Cancer Family Registry, diagnosed from March 2007 to November 2011. Multiplex ligation-dependent probe amplification (MLPA) for detecting BRCA LGRs together with comprehensive BRCA1 and BRCA2 gene sequencing of all coding exons were performed. cDNA sequencing of the LGRs was performed to locate the breakpoint of the deletions.

Results: Overall BRCA1/2 mutation prevalence among this cohort was 12.4% (69/555). Among the 69 mutations identified, 4 novel LGRs (2 in BRCA1 and 2 in BRCA2) were detected only by MLPA but not full gene sequencing. Overall the LGR genes accounted for 5.8% (4/69) of all BRCA mutations in our cohort, 6.9% (2/29) of all BRCA1 mutations and 5% (2/40) of all BRCA2 mutations.

Conclusions: These findings highlight the LGR spectrum of BRCA1 and BRCA2 genes in Southern Chinese breast cancer patients. LGR testing together with BRCA1/2 full gene sequencing is superior to other methods for comprehensive BRCA1/2 analysis in clinical settings.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-11-02.
Abstract P4-11-03: Single nucleotide polymorphism testing for breast cancer risk assessment: patient trust and willingness to pay
Authors: Howe R , Omer Z , Hanoch Y , Miron-Shatz T , Thorsen C , Ozanne EM
Published: August 2014

Background: The field of breast cancer risk assessment is advancing rapidly with recent discoveries about risk conferring single nucleotide polymorphisms (SNPs). While these discoveries can promote personalized medicine, they are often brought to the market with direct to consumer (DTC) testing before data can support their widespread use and before reliable options for dealing with testing outcomes can be offered. In this context, and knowing that patients often misunderstand risk information, it is unclear how patients will respond to these options.

Methods: We surveyed high risk women's interest in SNP testing. Participants were recruited from the Cancer Genetics Network (CGN), a national network of cancer centers that maintains a database of individuals with a family history of cancer. Participants were asked to answer questions regarding their interest in SNP testing including: whether they trust it, how much they would be willing to pay for testing, how they prefer to be tested, and how they would proceed with information identifying them as below or above average risk.

Results: 189 women without a history of breast cancer or SNP testing completed the questionnaire. The average age of the participants was 49, ranging from 30 to 65. All participants had at least one relative with breast or ovarian cancer. 13% had previously tested positive for a BRCA mutation, and 33% had received BRCA testing. Most women (90%) did not know what SNP testing was prior to the survey. Once SNP testing was described, 68% of women were interested in DTC SNP testing; at the same time, only 38% of the participants reported that they trusted DTC SNP testing.

Conclusion: While our results show that women are interested in DTC SNP testing, their willingness to pay is lower than the DTC cost (∼$300). Involvement of genetic counselors and providers in SNP testing discussions may be needed to overcome the current lack of trust of DTC testing among patients. Many women showed interest in lifestyle interventions, suggesting that these interventions should be incorporated as part of standard follow-up recommendations. When identified by SNP testing as “below average” risk, women do not seem to trust the results enough to forego regular mammograms. As DTC testing becomes more common, and as more SNP tests become available, it will be necessary for the medical community to address patients' interest in these tests and to assist in interpreting results.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-11-03.
Abstract P4-11-05: Optimal age to start preventive measures in women with BRCA1/2 mutations or high familial breast cancer risk
Authors: Tilanus-Linthorst MMA , Lingsma H , Evans GD , Kaas R , Manders P , Hooning MJ , van Asperen CJ , Thompson D , Eeles R , Oosterwijk JC , Leach MO , Steyerberg EJ
Published: August 2014

Background: Women from high risk breast cancer families consider preventive measures including screening. Guidelines on screening differ considerably regarding starting age. We investigated whether age at diagnosis in affected family members is predictive for age at diagnosis of a relative.

Methods: We analyzed the age of breast cancer detection of 1304 first and second degree relatives of 314 BRCA1, 164 BRCA2 and 244 high-risk participants of the Dutch MRI-SCreening study. The within and between family variance in age at diagnosis was analyzed with a random effect linear regression model. We compared the starting age of screening based on risk-group (25 years for BRCA1, 30 years for BRCA2 and 35 years for familial risk), on family history, and on the model, which combines both. The findings were validated in 63 families from the UK- MARIBS study.

Results: Mean age at diagnosis in the family varied between families; 95% range of mean family ages was 35–55 in BRCA1−, 41–57 in BRCA2− and 44–60 in high-risk families.14% of the variance in age at diagnosis was explained by family history, 7% by risk group.

Approaches to determining start of screening based on the model and on risk-group were similar in terms of cancers missed and years of screening. The approach based on familial history only, missed more cancers and required more screening years in both the Dutch and UK datasets.

Conclusion: Age at breast cancer diagnosis is in part dependent on family history which may assist planning starting age for preventive measures.

Keywords: BRCA1, BRCA2, breast cancer, prevention, age at onset

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-11-05.
Abstract P4-12-03: Towards a risk prediction model for breast cancer that utilizes breast tissue risk features
Authors: Pankratz VS , Degnim AC , Visscher DW , Frank RD , Vierkant RA , Ghosh K , Aziza N , Vachon CM , Frost M , Radisky DC , Hartmann LC
Published: August 2014

Background: Optimal early detection and prevention strategies for breast cancer are predicated on our ability to accurately identify women at increased risk for disease. Current breast cancer risk prediction models rely on clinical and epidemiologic features. As demonstrated with other cancers, such as those of the cervix and colon, cancer risk assessment is enhanced when the tissue at risk is examined for evidence of premalignant abnormalities. Here, we present a multivariate model for individualized breast cancer risk prediction that incorporates tissue features identified from biopsies from women with benign breast disease (BBD).

Methods: We used our cohort of more than 9000 women diagnosed with BBD at the Mayo Clinic from 1967–1991, and identified a nested case:control series of 377 women who developed cancer (cases) matched with 734 women who did not (controls). We identified a set of variables that either alone or in two-way interactions contributed to a multivariable risk prediction model. We combined this relative risk model with the baseline incidence of breast cancer and the baseline hazard of death from our full BBD cohort to develop a risk prediction tool that computes absolute risk estimates for breast cancer. We calculated 5-year risk predictions for these cases and controls using our model and using the Gail model and compared c-statistics from these risk predictions. We validated our model using an independent set of 378 breast cancer cases and 728 matched controls drawn from our BBD cohort.

Results: Cases and controls in the model development data set differed significantly on a number of features when examined individually. Those included in a multivariable relative risk model for breast cancer among women with BBD are shown below.

After including interactions between lobular involution, and histologic impression, with age at first live birth/number of children, and degree of involution with sclerosing adenosis, we devised a model that predicts the absolute risk of breast cancer. Predicted five-year risks in the model development series, we observed a c-statistic of 0.66, higher than the c-statistic of 0.61 for the Gail model. With risk prediction estimates from our validation series of cases and matched controls, the c-statistic of our model decreased to 0.59 and the c-statistic for the Gail model was 0.55.

Conclusion: We have identified an initial risk prediction model for breast cancer in women with BBD. The identified model includes basic demographic information as well as a number of histologic features of the tissue samples. Its initial performance is promising, providing improved discrimination over the Gail model in our replication data set. Further work is needed to identify features which might provide additional predictive information for breast cancer risk.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-12-03.
Abstract P4-12-04: Association of single-strand breaks (SSBs) in normal breast DNA with estimates of breast cancer risk
Authors: Chatterton RT , Sahadevan M , Heinz RE , Sukumar S , Stearns V , Fackler MJ , Lee O , Sivaraman I , Kenney K , Khan SA
Published: August 2014

Background: Formation of single strand breaks in DNA is a constant process, estimated to occur 10,000 times per day in each cell, either from endogenous biosynthetic errors or from interference by endogenous or exogenous agents. Base excision repair in some cases may be impaired or may be exceeded by the rate of DNA damage, leading to cancer. Nick translation of with labeled nucleotides can be used as a quantitative indicator of SSBs.

Methods: Healthy women were recruited through the Love-Avon Army of Women and breast clinics of Northwestern and John Hopkins Universities. Digital or digitized mammograms were evaluated for percent density using Cumulus software. The medians (ranges) were age 51 (36 to 60); BMI 28.2 (18.7 to 51.3); life-time Gail estimate 12.5 (5.6 to 28.3); % breast density 16.5 (2.4 to 52.5); Masood score 13 (0 to 18). Breast tissue was obtained by random fine needle aspiration (rFNA). Specimens were rinsed into ice-cold phosphate buffered saline and were stored at −80°C. Thawed samples were centrifuged at 2200 g for 60 min. A kit from Norgen Inc., was used to separate DNA, RNA, and protein from the pellet. The lipids were extracted with ethyl acetate-hexane (3:2) from the supernatant fluid, and triglycerides were precipitated from cold 90% methanol, leaving a purified lipid fraction containing the steroids. Steroids were then fractionated by HPLC on a C18 column, and estradiol was analyzed by a radioimmunoassay. Blood was obtained at the same time as the rFNA samples and was frozen prior to analysis. An aliquot containing 200 ng of DNA (260/280 ratio 1.3 to 2.3) was taken for the nick translation assay. Incorporation of free nucleotides at SSBs with dCTP labeled with3H was catalyzed by Polymerase I from E. coli. A quality control preparation prepared from calf thymus DNA and a reagent blank without DNA was included with each set of 6 samples. Incorporation of3H-dCTP was linear with dose of DNA and reached a maximal at 30 min. Separation of free from incorporated3H-dCTP was accomplished by gel chromatography on an 80 × 15 mm column. The concordance of interassay values was 0.96.

Results: Incorporation of nucleotides into DNA ranged from 0.03 to 8.59 pmol/μg, median 0.63 pmol/μg DNA. The association with other factors associated with breast cancer is shown in the Table. A significant correlation, was found with % breast density, life-time risk by the Gail model, but not serum estradiol concentrations.

Conclusions: Assessment of SSBs by the nick translation procedure may be a useful indicator of breast cancer risk. Future studies will relate this method with actual risk as assessed by analysis of pre-diagnosis specimens with subsequent occurrence of breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-12-04.
Abstract P4-12-07: Diabetes Increases the Risk of Breast Cancer
Authors: Hardefeldt PJ , Edirimanne S , Eslick GD
Published: August 2014

Background: The hyperinsulinaemia associated with type 2 diabetes and the known proliferative effects of insulin lead to the hypothesis that diabetes mellitus increases the risk of breast cancer. The aim of this meta-analysis was to collate and analyse all primary observational studies investigating the risk of breast cancer associated with diabetes. In addition, we aimed to complete sub group analyses by both type of diabetes and gender of study participants to further clarify the origin of any such association between the two.

Method: Studies were obtained from a database search of MEDLINE, EMBASE, PubMed, Current Contents Connect and Google Scholar with additional cross checking of reference lists. Databases were searched using subject headings, keyword searches and text word searches wherever possible using the search terms ‘diabetes’ AND ‘breast cancer’ OR ‘breast carcinoma’ OR ‘breast neoplasm’. Inclusion criteria required a confirmed diagnosis of both diabetes and breast cancer, reporting of an odds ratio or data to calculate an unadjusted odds ratio (and 95% confidence interval) and the use of an internal control group as the comparator. Collated data was assessed for heterogeneity and a pooled odds ratio calculated.

Results: 34 studies were included in the meta-analysis with 31 studies investigating breast cancer in women and 5 studies investigating breast cancer in men. Overall, we found a significantly increased risk of breast cancer associated with diabetes in both women (OR 1.25, 95% CI 1.13–1.38) and men (OR 1.40, 95% CI 1.10–1.79). The pooled odds ratio was consistent across both case control studies (OR 1.25, 95% CI 1.06–1.46) and cohort studies (OR 1.26, 95% CI 1.09–1.45). Heterogeneity was significant (I2 = 73.40, p < 0.001) and publication bias was present (p = 0.03). The 31 studies investigating breast cancer in women were sub grouped by type of diabetes with the association strengthened by a diagnosis of type 2 diabetes (OR 1.32, 95% CI 1.09–1.61) but nullified with gestational diabetes (OR 1.06, 95% CI 0.79–1.40). There were insufficient studies to calculate a pooled odds ratio of the risk of breast cancer associated with type 1 diabetes.

Conclusion: Diabetes increases the risk of breast cancer in both women and men. This association is strengthened in women with type 2 diabetes and supports the hypothesis that diabetes is an independent risk factor for breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-12-07.
Abstract P4-13-02: Comparing data quality of client intake forms by interview mode: results of a pilot study on the use of audio computer-assisted self-interview (ACASI) in the Avon Breast Health Outreach Program
Authors: Hallum-Montes R , Senter L , D'Souza R , Hurlbert M , Gates-Ferris K , Anastario M
Published: August 2014

Background: Numerous studies have documented the relative advantages that computer assisted self-interview (CASI) technology holds over traditional interviewing methods in collecting research data on stigmatized and/or risky health behaviors. However, few studies have explored the broader implications of using CASI to collect basic client-level data during routine health promotion and disease prevention screenings.

Aims: The present study aims to address this gap in research by comparing the quality of client-level data collected via audio-computer assisted self-interview (ACASI) with data collected via face-to-face and self-administered paper-based interviews among a sample of clients served through the Avon Breast Health Outreach Program (Avon BHOP).

Methods: A total of 303 clients served through the Avon BHOP were sampled from three U.S. sites between November 2011 and March 2012. Clients were randomly assigned to complete the Avon BHOP client intake form (CIF) via one of three interview modes: face-to-face interview (FFI), self-administered paper-based interview (SAPI), or ACASI. We conducted logistic regression and determined odds ratios with 95% confidence intervals to test our hypothesis that use of ACASI would result in significantly higher rates of CIF completion than either FFI or SAPI. We compared rates of completion of the entire CIF, as well as completion of each of the three CIF sections: Demographics, Breast Health Information, and Correlates of Breast Health.

Results: Clients were significantly more likely to complete the entire CIF via ACASI than either FFI or SAPI interview modes. The greatest differences were observed when comparing rates of completion between ACASI and SAPI, as clients were almost five times more likely to complete the CIF via ACASI as opposed to SAPI (OR = 4.7, p < 0.001). Additionally, when comparing rates of completion of the three CIF sections on Demographics, Breast Health Information (e.g. mammogram history), and Correlates of Breast Health (e.g., smoking behavior, alcohol consumption), we found that clients were between 3.2 and 7.2 times more likely to complete the sections via ACASI as opposed to SAPI.

Discussion: Overall, we found that use of ACASI resulted in significantly higher rates of overall form completion and lower rates of missing data than use of SAPI or FFI, with the greatest identified disparity in form completion between ACASI and SAPI. This study has important implications for breast health specialists or any health practitioners who regularly rely on self-administered questionnaires and/or face-to-face interviews to collect important health information. We recommend that where feasible, ACASI be utilized as an effective means of collecting high quality client-level data.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-02.
Abstract P4-13-12: Identifying women at increased risk for breast cancer using the electronic health record in an integrated health system
Authors: Leader JB , Bengier A , Darer J , Stark A , Vogel VG
Published: August 2014

Women at increased risk for breast cancer (BC) are eligible to take selective estrogen receptor modulators (SERMs) to reduce their risk; Food and Drug Administration (FDA) approval of tamoxifen or raloxifene for BC risk reduction and American Society of Clinical Oncology (ASCO) guidelines for the use of SERMs recommend the two drugs for any woman over the age of 35 years with a 5-year risk of 1.67% or greater, but identifying those women can be both challenging and costly. We used an electronic database (Centricity RIS-IC) from the Geisinger Health System (GHS) Department of Radiology with 77,000 women ages 35–90 years to calculate 5-year and lifetime risks of developing invasive BC using National Cancer Institute's (NCI) Breast Cancer Risk Assessment Macro (BrCa RAM). BrCa RAM calculates risk based on patient age, number of biopsies, did a biopsy ever display atypical hyperplasia (Yes/No), age at menarche, age at first live birth, number of first degree relatives with breast cancer, and patient race. Demographic information (age, race, sex) was obtained from the electronic health record (EpicCare), pathology information (number of biopsies, atypical hyperplasia) was obtained from the pathology application (CoPath), and personal history (number of first degree relatives with breast cancer) were obtained from RIS. Age at menarche and age at first live birth could not be obtained, but makes a small relative contribution to the risk of BC. Sensitivity analysis explored implications of missing data; imputing ages for age at first live birth and age at menarche showed that the absence of this data did not overestimate the five-year and lifetime risks. There were 5,897 patients with calculated 5-year breast cancer risk 2; mean age was 65.8 years, mean 5-yr risk of BC = 3.05% (max 18.2%). The number of patients by 5-year risk score category were: risk 2–2.5% (n = 1728); 2.5%–3% (n = 3188); 3%+ (n = 981). There were 4,196 patients with a GHS primary care physician (PCP); 5,086 patients had seen any Geisinger physician within the past year; 4,113 women had seen their PCP in the past year. Only 239 patients ever received a prescription for tamoxifen or raloxifene, and some received raloxifene for prevention or treatment of osteoporosis and not for BC risk reduction. Only 40 were currently taking tamoxifen or raloxifene. These data from an integrated health system with an electronic health record validate the under-utilization of SERMs for primary BC risk reduction. Strategies are being designed to increase their use in GHS by using the risk score to identify the population and attempt to intervene using a risk modification clinical program.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-12.
Abstract P4-13-14: Lung cancer after treatment of breast cancer:retrospective study from Curie Institut
Authors: Sebbagh S , Cosquer M , Kirova YM , Livartowski A
Published: August 2014

Background: Few studies have evaluated the effects of adjuvant radiotherapy (RT)of breast cancer(BC). The relation between the risk of lung carcinoma and radiotherapy have been controversial.

Methods and materials: We retrospectively studied 127 patients treated at the Institut Curie with non metastatic breast cancer and lung carcinoma between 2000 and 2011(2/3 of BC apperead befor lung). Confirmation Diagnosis bronchial cancer obtained by: biopsy: histological data:architecture, IHC (HR, HER 2, TTF1), EGFR, Kras statut clinical and radiological Correlation. Comparison with breast tumor

Results: BC: median age at diagnosis 54 years, predominatly invasive ductual carcinoma(IDC), lumpectomy 78%, mastectomy 21%. Lung cancer: median age at diagnosis: 63 years, 67 smokers. histology: 52 % Adenocarcinoma, 18.1 % scamous cell carcinoma, 18.1% large cell carcinoma, 13.4 % small cell carcinoma. EGFR mutation in 4.3%. 109 patients underwent RT (3 cases of lung cancer befor BC):Region: internal mammary chain: 46, supracalvicular: 42, axillary: 21. Technique: lateral decubitus position: 44, dorsal decubitus position: 57. Interval between breast and lung cancer: 0–3 years: 24.4%, 3–5 years: 15%, 6–10years: 16.5%, 11–20 years: 28%, >20 years: peak of incidence of lung cancer in the 3 years of diagnosis of breast cancer: 24.4 %. There was no apparent relation between treatment of BC and relative risk of developing lung carcinoma. 2nd peak between 11–20 years: 32 % patients, suggest that RT may increase risk of lung carcinoma (latency period for radiation induced second malignancy).

Conclusion: This study suggest that adjuvant RT is associated with a real but small risk of developing lung carcinoma.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-14.
Abstract P4-14-03: Nipple-sparing mastectomy and intra-operative nipple biopsy: To freeze or not to freeze?
Authors: Guth AA , Blechman K , Samra F , Shapiro R , Axelrod D , Choi M , Karp N , Alperovich M
Published: August 2014

Background: Advances in breast cancer screening and treatment have fostered the use of surgical procedures that increasingly optimize cosmetic outcome, while ensuring oncologic safety remains the primary concern of the oncologic surgeon. The role of nipple-sparing mastectomy (NSM) for risk-reducing surgery and breast cancer is evolving. It can be difficult to demonstrate involvement of the nipple-areolar complex (NAC) preoperatively, and and in this report we examine the utility of intraoperative subareolar frozen section (FS).

Methods: Records of patients undergoing NSM at the NYU Langone Medical Center from 2006–2011 were reviewed retrospectively. Use of subareolar FS was at surgeon's discretion.

Results: 237 NSM were performed (146 prophylacytic, 91 theraputic). FC was not utilized in 58 mastectomies (28 prophylactic), with 2 (+) on paraffin. Among the remaining 180 mastectomies, 11 biopsies were (+)(7.2%); 5 subareolar biopsies were (+) on FS and paraffin histologic slides (PS)(2.8%); 6 were negative on FS and (+) on PS. Among the 3 prophylactic NSM with (+) subareolar biopsies there was 1 (+) FS, 1 (−) FS, and 1 with no FS performed. There were no false (+) FS. Four of the 5 patients with (+)FS underwent simultaneous excision of the NAC. The 5th patient had atypia on FS and DCIS on PS, and returned to the OR during the same hospitalization for removal of NAC. The remaining patients underwent subsequent excision of the NAC either during planned 2nd stage reconstruction or following completion of chemotherapy. 8 NAC were free of disease and 5 were positive for in situ or invasive disease. There has been no local recurrence in these patients to date.

Conclusions: The rate of NAC involvement is low, 5.5% in this series, and FS, while utilized in the majority of these cases, detected only 46% of subareolar disease. While FS can direct intraoperative decision making, the predictive power is low, and these considerations must be addressed in preoperative patient discussions. Furthermore, among those patients with (+) subareolar biopsies, only 39% had residual disease on NAC excision. Thus, optimal oncologic management of the NAC must still be considered an unresolved issue.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-14-03.