Response Of Primary Breast Cancer Research Articles

Prediction of breast cancer and axillary positive-node response to neoadjuvant chemotherapy based on multi-parametric magnetic resonance imaging radiomics models

PurposeAccurate identification of primary breast cancer and axillary positive-node response to neoadjuvant chemotherapy (NAC) is important for determining appropriate surgery strategies. We aimed to develop combining models based on breast multi-parametric magnetic resonance imaging and clinicopathologic characteristics for predicting therapeutic response of primary tumor and axillary positive-node prior to treatment. Materials and methodsA total of 268 breast cancer patients who completed NAC and underwent surgery were enrolled. Radiomics features and clinicopathologic characteristics were analyzed through the analysis of variance and the least absolute shrinkage and selection operator algorithm. Finally, 24 and 28 optimal features were selected to construct machine learning models based on 6 algorithms for predicting each clinical outcome, respectively. The diagnostic performances of models were evaluated in the testing set by the area under the curve (AUC), sensitivity, specificity, and accuracy. ResultsOf the 268 patients, 94 (35.1 %) achieved breast cancer pathological complete response (bpCR) and of the 240 patients with clinical positive-node, 120 (50.0 %) achieved axillary lymph node pathological complete response (apCR). The multi-layer perception (MLP) algorithm yielded the best diagnostic performances in predicting apCR with an AUC of 0.825 (95 % CI, 0.764–0.886) and an accuracy of 77.1 %. And MLP also outperformed other models in predicting bpCR with an AUC of 0.852 (95 % CI, 0.798–0.906) and an accuracy of 81.3 %. ConclusionsOur study established non-invasive combining models to predict the therapeutic response of primary breast cancer and axillary positive-node prior to NAC, which may help to modify preoperative treatment and determine post-NAC surgery strategy.

Association between physical activity and neoadjuvant chemotherapy completion and pathologic complete response in primary breast cancer: the CANTO study.

Regular physical activity is associated with improved symptom control in patients with breast cancer but its association with chemotherapy completion or response is unclear. Using a prospective design, 1075 breast cancer patients receiving neoadjuvant chemotherapy between March 2012 and February 2017 were studied. Physical activity was assessed using the Global Physical Activity Questionnaire [GPAQ-16], quantified in standardised MET-h/wk. Chemotherapy completion was defined as the proportion of patients completing planned treatment course, requiring dose reduction, or requiring dose delay. Response was evaluated by pathologic complete response (pCR). Associations between physical activity and primary outcomes were assessed using multivariable logistic regression models. There was no differences between any chemotherapy completion outcome on the basis of physical activity classification. The percent of patients not completing planned treatment was 5.7% for ≦0.33 MET-h/wk, compared with 6.8% for 0.34-16.65 MET-h/wk, and 4.6% for ≥16.6 MET-h/wk (p = 0.52). No significant relationships were observed between physical activity dose classification and pCR for the overall cohort or upon stratification by clinical subtype. Future studies are required to further investigate the relationship between pre-treatment levels of physical activity and function on treatment completion and response in breast and other cancer populations. NCT01993498.

Relationship between ER expression by IHC or mRNA with Ki67 response to aromatase inhibition: a POETIC study

BackgroundIn clinical practice, oestrogen receptor (ER) analysis is almost entirely by immunohistochemistry (IHC). ASCO/CAP recommends cut-offs of < 1% (negative) and 1–10% (low) cells positive. There is uncertainty whether patients with ER low tumours benefit from endocrine therapy. We aimed to assess IHC and mRNA cut-points for ER versus biological response of primary breast cancer to 2 weeks’ aromatase inhibitor treatment as measured by change in Ki67.MethodsCases were selected from the aromatase inhibitor treatment group of POETIC. We selected the 15% with the poorest Ki67 response (PR, < 40% Ki67 suppression, n = 230) and a random 30% of the remainder categorised as intermediate (IR, 40–79% Ki67 suppression, n = 150) and good-responders (GR, ≥ 80% Ki67 suppression, n = 230) from HER2 − group. All HER2 + cases available were selected irrespective of their response category (n = 317). ER expression was measured by IHC and qPCR.ResultsER IHC was available from 515 HER2 − and 186 HER2 + tumours and ER qPCR from 367 HER2 − and 171 HER2 + tumours. Ninety-one percentage of patients with ER IHC < 10% were PRs with similar rates in HER2 − and HER2 + cases. At or above ER IHC 10% substantial numbers of patients showed IR or GR. Similar proportions of patients were defined by cut-points of ER IHC < 10% and ER mRNA < 5 units. In addition, loss of PgR expression altered ER anti-proliferation response with 92% of PgR − cases with ER IHC < 40% being PRs.ConclusionsThere was little responsiveness at IHC < 10% and no distinction between < 1% and 1–10% cells positive. Similar separation of PRs from IR/GRs was achieved by IHC and mRNA.

Abstract CT074: Pre-existing ESR1 mutations in early-stage primary breast cancer predict failure of endocrine therapy and poor survival

Abstract Background: More than three-quarters of primary breast cancers are positive for estrogen receptor alpha (ER; encoded by the gene ESR1), the most important factor for directing anti-estrogenic endocrine therapy. Although mutation in ESR1 is known as an acquired mechanism of resistance to endocrine therapy (ET), found in 12-55% of metastatic breast cancers treated previously with ET, the impact of ESR1 mutation on therapy response in primary breast cancer is unclear. Patients and methods: In this study we analyzed 3217 real-world and population-based early-stage primary breast cancers (within the SCAN-B study, ClinicalTrials.gov NCT02306096). Tissues were sampled from initial diagnosis prior to any treatment and analyzed for the presence of ESR1 mutations using RNA sequencing. Mutations were verified by SAGAsafe droplet digital PCR. Results: We identified ESR1 resistance mutations in 30 cases (0.9%), of which 29 were ER-positive (1.1%). In ER-positive disease, presence of ESR1 mutation was significantly associated to poor relapse-free survival (RFS) and overall survival (OS) (p=0.011 and p=0.019, respectively), and moreover predicted poor RFS and OS within the patient group that received ET (p=0.007 and p=0.010, respectively). Conclusions: These results indicate that ESR1 mutations at diagnosis of untreated primary breast cancer are rare, however we confirm for the first time that such early mutations predict eventual resistance to standard hormone therapy in the adjuvant setting. If replicated, tumor ESR1 screening may be considered in ER-positive primary breast cancer and, in mutated cases, ER-degraders such as fulvestrant or other therapeutic options may be considered as more appropriate. Citation Format: Malin Dahlgren, Anthony M. George, Christian Brueffer, Sergii Gladchuk, Yilun Chen, Johan Vallon-Christersson, Cecilia Hegardt, Jari Häkkinen, Lisa Rydén, Martin Malmberg, Christer Larsson, Sofia K. Gruvberger-Saal, Anna Ehinger, Niklas Loman, Åke Borg, Lao H. Saal. Pre-existing ESR1 mutations in early-stage primary breast cancer predict failure of endocrine therapy and poor survival [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr CT074.

Abstract P2-09-02: True effect of aromatase inhibitor (AI) treatment on global gene expression (expr) changes in postmenopausal ER+ breast cancer (BC) patients: A POETIC study (CRUK/07/015)

Abstract BACKGROUND Gene expression (expr) analyses are increasingly used for characterising the pharmacodynamic response of primary BC. This includes assessing ER+ BC's dependence on estrogen (E) by measuring gene expr changes after AI-treatment. However, differences in tissue sampling and other preanalytic procedures between samples taken at diagnosis (D) and surgery (S), may lead to systematic artifactual changes that are falsely ascribed to the intervention. To identify genes whose expr is truly affected by AI, we measured global gene expr changes from paired core-cut biopsies at D and S from patients in the POETIC presurgical window trial. METHODS In POETIC, 4486 postmenopausal women with primary ER+ BC were randomised 2:1 to receive perioperative AI (2 weeks pre + 2 weeks post surgery, termed Tr) or no perioperative treatment (termed Con), allowing gene expr changes to be compared between Tr and Con. RNA was extracted from paired RNA-later stored core-cuts of 56 Con and 157 Tr patients and arrayed on Illumina whole genome expr BeadChips. Raw data was extracted, transformed, normalised and batch-corrected. Probes not detected (p&amp;gt;0.01) in &amp;gt;=25% of samples were discarded. Impact of AI on genes was evaluated based on difference of the expr mean changes (log2(S/D)) of the Tr and Con samples. RESULTS In the Con group, expr of 73 genes significantly changed (FDR&amp;lt;5%); 70 of these changed by a similar magnitude in the Tr group, indicating their change was independent of AI therapy but would have been artifactually discovered as changed by AI in the absence on Con. The 8 genes most up-regulated in Tr were all among the 20 genes most up-regulated in Con: many were early-response or stress-associated genes. Three of the 8 most down-regulated in AI were the most down-regulated in Con: all were haemoglobin-related. Expr of some genes was changed in Con (eg MYC increase) but was unaffected in Tr. Such artifactual gene changes in Con tumors conceal true AI-induced changes that would not be detected in the absence of comparison with Con. 615 genes were down-regulated and 472 up-regulated in Tr but not Con. The majority of down-regulated genes were cell cycle or proliferation-associated or E-regulated, including ESR1, PDZK1, GREB1, HSPB1. Functional mapping showed changes in the regulation of cyclins and cyclin dependent kinases impacting on G1/S and G2/M. Of note, up-regulated genes included CDK6 (target for CDK4/6 inhibitors) and CCND2, involved in G1/S checkpoint regulation; SNAI2, TGFB3, TGFBR2, associated with tumour invasion and metastasis; and other genes involved in aryl hydrocarbon receptor, Glioblastoma Multiforme, HIPPO and p53 signalling. CONCLUSION Expr of certain genes is altered by processes involved in presurgical window studies. In the absence of a Con group, these may be wrongly ascribed to an experimental intervention or wrongly considered as unaffected by the intervention (eg MYC in this study). Down-regulation of E-responsive and proliferation genes was an expected response to AI but increased expr of genes such as SNAI2, CCND2 and CDK6 indicates immediate tumour re-wiring and provides mechanistic support for benefit from combination therapy with a CDK4/6 inhibitor. Citation Format: Gao Q, López-Knowles E, Cheang MCU, Morden J, Martin L-A, Sidhu K, Evans D, Martins V, Dodson A, Skene A, Holcombe C, Mallon E, Abigail E, Bliss J, Robertson J, Smith I, Dowsett M. True effect of aromatase inhibitor (AI) treatment on global gene expression (expr) changes in postmenopausal ER+ breast cancer (BC) patients: A POETIC study (CRUK/07/015) [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-09-02.

Tumor slice culture system to assess drug response of primary breast cancer.

BackgroundThe high incidence of breast cancer has sparked the development of novel targeted and personalized therapies. Personalization of cancer treatment requires reliable prediction of chemotherapy responses in individual patients. Effective selection can prevent unnecessary treatment that would mainly result in the unwanted side effects of the therapy. This selection can be facilitated by characterization of individual tumors using robust and specific functional assays, which requires development of powerful ex vivo culture systems and procedures to analyze the response to treatment.MethodsWe optimized culture methods for primary breast tumor samples that allowed propagation of tissue ex vivo. We combined several tissue culture strategies, including defined tissue slicing technology, growth medium optimization and use of a rotating platform to increase nutrient exchange.ResultsWe could maintain tissue cultures for at least 7 days without losing tissue morphology, viability or cell proliferation. We also developed methods to determine the cytotoxic response of individual tumors to the chemotherapeutic treatment FAC (5-FU, Adriamycin [Doxorubicin] and Cyclophosphamide). Using this tool we designated tumors as sensitive or resistant and distinguished a clinically proven resistant tumor from other tumors.ConclusionThis method defines conditions that allow ex vivo testing of individual tumor responses to anti-cancer drugs and therefore might improve personalization of breast cancer treatment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2119-2) contains supplementary material, which is available to authorized users.

S6 kinase signaling: tamoxifen response and prognostic indication in two breast cancer cohorts.

Detection of signals in the mammalian target of rapamycin (mTOR) and the estrogen receptor (ER) pathways may be a future clinical tool for the prediction of adjuvant treatment response in primary breast cancer. Using immunohistological staining, we investigated the value of the mTOR targets p70-S6 kinase (S6K) 1 and 2 as biomarkers for tamoxifen benefit in two independent clinical trials comparing adjuvant tamoxifen with no tamoxifen or 5 years versus 2 years of tamoxifen treatment. In addition, the prognostic value of the S6Ks was evaluated. We found that S6K1 correlated with proliferation, HER2 status, and cytoplasmic AKT activity, whereas high protein expression levels of S6K2 and phosphorylated (p) S6K were more common in ER-positive, and low-proliferative tumors with pAKT-s473 localized to the nucelus. Nuclear accumulation of S6K1 was indicative of a reduced tamoxifen effect (hazard ratio (HR): 1.07, 95% CI: 0.53-2.81, P=0.84), compared with a significant benefit from tamoxifen treatment in patients without tumor S6K1 nuclear accumulation (HR: 0.42, 95% CI: 0.29-0.62, P<0.00001). Also S6K1 and S6K2 activation, indicated by pS6K-t389 expression, was associated with low benefit from tamoxifen (HR: 0.97, 95% CI: 0.50-1.87, P=0.92). In addition, high protein expression of S6K1, independent of localization, predicted worse prognosis in a multivariate analysis, P=0.00041 (cytoplasm), P=0.016 (nucleus). In conclusion, the mTOR-activated kinases S6K1 and S6K2 interfere with proliferation and response to tamoxifen. Monitoring their activity and intracellular localization may provide biomarkers for breast cancer treatment, allowing the identification of a group of patients less likely to benefit from tamoxifen and thus in need of an alternative or additional targeted treatment.

The impact of recurrence score (RS) performed on pretreatment (Tx) core biopsies (CB) in predicting response to neoadjuvant chemotherapy (NCT) in patients (Pts) with ER+ breast cancer (BC).

63 Background: The Oncotype DX 21-gene assay is beneficial in predicting disease-free survival and local regional relapse in hormone receptor BC. The ability of the assay to predict response of primary BC to NCT is less well studied. We hypothesize a (+) association exists between the RS result on pre-Tx CB and % tumor response (%TR) after NCT on final pathology. Methods: Pre-Tx RS was measured on paraffin-embedded CB from 60 pts with ER+ Her2(-) BC who then received NCT and for whom post-Tx pathology was available. %TR is calculated as 100 x (pre-Tx size – post-Tx size)/pre-Tx size. Pre-Tx tumor size was determined using imaging measurements selected in the following preferential order: MRI, US or mammogram. Post-Tx tumor size was calculated as the product of: maximum dimension of tumor-bed/fibrosis and % change in cellularity (compared with pre-therapy biopsy). Partial response (PR) is defined as ≥50% reduction in TR. Results: Mean pre-Tx tumor sizewas 4.8 (range 1-23) cm. 83% of pts were clinically lymph node (-). Nottingham Score was &gt;6 in 93%. Breast conserving surgery was performed in 17 (28%) pts. Median RS was 20 (range 3-69). Median %TR was 42 (range 0-97)%. There were no pCRs. PR was observed in 43% of pts. No association was found between %TR and pre-NCT tumor size, age, Nottingham score or node status (p&gt;0.05). No association for the RS as a continuous or categorical variable with %TR (p&gt;0.05) was seen. There is a non-significant (p=0.07) trend toward increased response to NCT with increasing RS. Response to NCT improved as values of ER (p=0.02) and HER2 (p=0.007), by RT-PCR as reported with the RS, decreased. Conclusions: In the subset of patients with ER+ BC, where NCT was recommended to affect tumor downstaging, RS did not successfully discriminate pts that would respond to NCT as measured by %TR. While there was a trend toward better response with higher scores, the result was nonsignificant. The use of this genetic assay to discriminate between NCT and hormonal therapy requires further study. [Table: see text]

A randomised controlled phase II trial of pre-operative celecoxib treatment reveals anti-tumour transcriptional response in primary breast cancer

IntroductionCyclooxygenase-2 (COX-2) is frequently over-expressed in primary breast cancer. In transgenic breast cancer models, over-expression of COX-2 leads to tumour formation while COX-2 inhibition exerts anti-tumour effects in breast cancer cell lines. To further determine the effect of COX-2 inhibition in primary breast cancer, we aimed to identify transcriptional changes in breast cancer tissues of patients treated with the selective COX-2 inhibitor celecoxib.MethodsIn a single-centre double-blind phase II study, thirty-seven breast cancer patients were randomised to receive either pre-operative celecoxib (400 mg) twice daily for two to three weeks (n = 22) or a placebo according to the same schedule (n = 15). Gene expression in fresh-frozen pre-surgical biopsies (before treatment) and surgical excision specimens (after treatment) was profiled by using Affymetrix arrays. Differentially expressed genes and altered pathways were bioinformatically identified. Expression of selected genes was validated by quantitative PCR (qPCR). Immunohistochemical protein expression analyses of the proliferation marker Ki-67, the apoptosis marker cleaved caspase-3 and the neo-angiogenesis marker CD34 served to evaluate biological response.ResultsWe identified 972 and 586 significantly up- and down-regulated genes, respectively, in celecoxib-treated specimens. Significant expression changes in six out of eight genes could be validated by qPCR. Pathway analyses revealed over-representation of deregulated genes in the networks of proliferation, cell cycle, extracellular matrix biology, and inflammatory immune response. The Ki-67 mean change relative to baseline was -29.1% (P = 0.019) and -8.2% (P = 0.384) in the treatment and control arm, respectively. Between treatment groups, the change in Ki-67 was statistically significant (P = 0.029). Cleaved caspase-3 and CD34 expression were not significantly different between the celecoxib-treated and placebo-treated groups.ConclusionsShort-term COX-2 inhibition by celecoxib induces transcriptional programs supporting anti-tumour activity in primary breast cancer tissue. The impact on proliferation-associated genes is reflected by a reduction of Ki-67 positive cells. Therefore, COX-2 inhibition should be considered as a treatment strategy for further clinical testing in primary breast cancer.Trial registrationClinicalTrials.gov NCT01695226.

Protein phosphatase 1, regulatory subunit 15B is a survival factor for ERα‐positive breast cancer

Breast cancer is a heterogeneous disease at both the clinical and molecular levels. This heterogeneity may give rise to different therapy responses. Molecular profiling has facilitated identification of signatures for stratifying patients who would potentially benefit from given therapies. Previously, we reported on a subset of genes with the potential for predicting response of primary breast cancer to neoadjuvant chemotherapy. Herein, we report that patients with luminal (estrogen receptor α [ERα]-expressing) breast cancer were enriched for nonresponders. To identify novel factors that contribute to the survival of breast cancer cells, a loss-of-function screen was performed with a subset of genes overexpressed in patients with disease resistant to chemotherapy. This approach led us to identify protein phosphatase 1, regulatory subunit 15B (PPP1R15B) as a factor with a potentially essential role in the survival of ERα-positive breast cancer cells. Functional analyses showed that PPP1R15B depletion results in impaired proliferation due to unsuccessful transition of cells from G1 to S phase of the cell cycle, and apoptosis induction. Moreover, our data revealed a regulatory role for PPP1R15B in activating ERα. Furthermore, a high level of PPP1R15B mRNA expression was associated with poor outcome following tamoxifen-based therapy. Accordingly, knockdown of PPP1R15B expression sensitized tamoxifen-resistant MCF-7 breast cancer cells to tamoxifen while reducing ERα abundance in these cells. Our findings reveal a novel role for PPP1R15B in the survival and therapy response of ERα-positive breast cancer and may open new avenues for tumor subtype-specific therapeutic strategies in the era of personalized medicine.

Abstract P4-01-03: quantitative DCE-MRI to predict the response of primary breast cancer to neoadjuvant therapy

Abstract The goal of this study was to determine if quantitative changes in dynamic contrast enhanced MRI (DCE-MRI) following a single cycle of chemotherapy can be used to separate pathologic responders (i.e., no residual tumor in the breast at surgery) from pathologic non-responders. 28 patients with Stage II/III breast cancer were enrolled in an IRB-approved clinical trial where breast MRI scans were acquired before (t1) and after one cycle of therapy (t2). Imaging was performed on a 3.0T MR scanner (Philips Healthcare, The Netherlands) and employed a 3D spoiled gradient echo sequence with a spatial resolution of 6.6 mm3 and a temporal resolution of 16 seconds collected at 25 time points before and after the intravenous injection of 0.1 mmol/kg of gadopentetate dimeglumine (Magnevist, Wayne, NJ). At surgery, 12 patients were responders while 16 patients were non-responders. Both semi-quantitative and quantitative analyses were used to summarize the DCE-MRI data. The semi-quantitative parameters were the signal enhancement rate (SER [1]) and tumor volume (TV). Three pharmacokinetic models, the Tofts-Kety (TK), the Extended Tofts-Kety (ETK), and the fast exchange regime (FXR), were used to estimate the following quantitative parameters: the volume transfer constant (Ktrans), efflux rate constant (kep), vascular volume (vp), and the extravascular extracellular volume fraction (ve) [2]. Each parameter was summarized in two ways: 1) the change in mean from t1 to t2, and 2) the mean at t2. Receiver operating characteristic (ROC) analysis was then performed to determine the ability of each parameter to predict treatment response. The table displays the areas under the ROC curves (AUC) for each parameter. For the early change in parameters, the AUC for TV and SER were 0.48 and 0.66, respectively. The best AUC of the quantitative parameters was 0.73 from kep estimated by the ETK model. The sensitivities/specificities for TV, SER, and kep for predicting pathologic response were 88%/33%, 64%/79%, and 56%/92%, respectively. For the mean parameter values at t2, the AUCs of TV, SER, and kep were 0.50, 0.56, and 0.79, with sensitivities/specificities for predicting pathologic response of 63%/50%, 93%/21%, and 81%/75%, respectively. Our results can be interpreted in light of the ACRIN 6657/I-SPY trial [3] which found that change in TV and SER at an early time point were the most predictive of response with AUCs of 0.72 and 0.71, respectively. Our preliminary results, especially our AUC of 0.79 for kep at t2, suggest that a more quantitative analysis of higher temporal resolution DCE-MRI data may achieve comparable or even superior results. Our ongoing efforts involve combining multiple parameters in a multivariate analysis with apparent diffusion coefficient data from diffusion weighted MRI. [1] Arasu et al. Acad Radiol 2011;18:716–721. [2] Yankeelov &amp; Gore. CMIR 2009;3:91–107. [3] Hylton et al. Radiology. 2012;263:663–672. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-01-03.

Abstract P3-06-08: Ki-67 mRNA as a predictor for response to neoadjuvant chemotherapy in primary breast cancer

Abstract Background: Pathologic complete response (pCR, ypT0/is) is one of the strongest prognostic factors for primary breast cancer (PBC) in the setting of neoadjuvant chemotherapy (NAC). Factors predictive for response to NAC can help to stratify for an individualized therapy. As chemotherapeutic agents are most active in highly proliferative tumors, it was the aim of the present study to evaluate the potential of Ki-67 as a single proliferation marker, quantitatively measured on the mRNA or protein level, to predict response of PBC to NAC. Material and methods: Pretreatment core cut biopsies from n=72 patients with PBC treated within a randomized phase II trial (1) of anthracylin/taxane based NAC were examined. Immunohistochemistry was performed for the Ki67 antigen on an automated IHC platform (Dako Techmate 500). Ki-67 were assessed either by visual scoring (vIHC) or by quantitative image analysis (qIHC). For quantitative IHC analysis tumor proliferation was analyzed after tumor stroma segmentation using the Aperio Image Analysis toolbox. RNA was extracted from formalin-fixed, paraffin embedded (FFPE) routine biopsies using a bead-based extraction method (STRATIFYER XTRAKT kits). Ki-67, TOP2A and RACGAP1 as well as CALM2 as a house keeping gene were measured via a multiplex quantitative RT-PCR (qPCR). Kaplan-Meier survival estimates, patitioning test and correlation analyses were performed using the SAS JMP® 9.0.0 software. Results: There was only a moderate correlation between Ki-67 mRNA or Ki-67 measured by immunohistochemistry (IHC) and histologic grade (Spearman r = 0.52 p &amp;lt; 0.0001; r = 0.23 p = 0.033, respectively). Conventional visual scoring and qIHC correlated well (r = 0,78 p &amp;lt; 0,0001), while qPCR and qIHC correlated moderately (r = 0,50 p &amp;lt; 0,0001). For Ki-67 determined by visual scoring of standard IHC there was an optimal discrimination at a cut-off of 20% stained nuclei with regard to chemotherapy response. Yet, the difference between pCR-rates for high and low Ki-67 on IHC was non-significant, with 16.3% and 12.5% pCRs in the IHC Ki-67 high and low group, respectively. For Ki-67 IHC analysed by quantitative image analysis there was optimal cut-off at 35% stained nuclei resulting in 37,5% versus 8,7% pCR rate and 54% of pCRs classified correctly. For qPCR determination, high Ki-67 mRNA expression was associated with a high pCR-rate of 36.4% as opposed to 5.8% in tumors with low Ki-67 mRNA levels and 82% classified correctly. Conclusion: High Ki-67 mRNA expression measured by RT-qPCR was predictive for the achievement of pCR to NAC and in this respect was superior to Ki-67 determined by vIHC or qIHC. This is consistent with data on Ki-67 IHC as a predictor of pCR on larger neoadjuvant cohorts (1). RT-qPCR based measurements of Ki-67 mRNA ensures an objective and highly reproducible quantification of proliferative activity from FFPE tissue from routine core cut biopsies. As such it seems to be more robust and meaningful compared to protein-based determination of Ki-67 by IHC, both by visual scoring or quantitative image analysis. 1) Schneeweiss et al, Ann Oncol 2011 2) Fasching et al., BMC Cancer 2011 Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-06-08.

Antitumor activity of zoledronic acid in primary breast cancer cells determined by the ATP tumor chemosensitivity assay.

BackgroundThe NeoAzure study has demonstrated that the use of the bisphosphonate zoledronic acid (Zol) in the neoadjuvant setting increases the rate of complete response in primary breast cancer and therefore indicates direct antitumor activity. The purpose of this study was to compare the antitumor effect of Zol with standard chemotherapy in primary breast cancer cells using ATP-tumor chemosensitivity assay (ATP-TCA).MethodsBreast cancer specimens were obtained from patients with breast cancer who underwent primary breast cancer surgery at the Department of Obstetrics and Gynecology, Tübingen, Germany, between 2006 through 2009. Antitumor effects of Zol, TAC (Docetaxel, Adriamycin, Cyclophosphamide) and FEC (5-Fluorouracil, Epirubicin, Cyclophosphamide) were tested in 116 fresh human primary breast cancer specimens using ATP-TCA. ATP-TCA results were analyzed with different cut-off levels for the half maximal inhibitory concentration (IC50), for IC90 and for the sensitivity index (IndexSUM). Each single agent or combination was tested at six doubling dilutions from 6.25, 12.5, 25, 50, 100, and 200% of test drug concentrations (TDC) derived from the plasma peak concentrations determined by pharmacokinetic data. The assay was carried out in duplicate wells with positive and negative controls.ResultsThe median IndexSUM value was lower for Zol than for the combined regimen FEC (36.8%) and TAC (12.9%), respectively, indicating increased antitumor activity of Zol in primary breast cancer cells. The difference regarding Zol and FEC was significant (p < 0.05). The median IC50 value for Zol (8.03% TDC) was significantly lower than the IC50 values for FEC (33.5% TDC) and TAC (19.3% TDC) treatment (p < 0.05). However, the median IC90 value for Zol (152.5% TDC) was significantly higher than the IC90 value obtained with TAC (49.5% TDC; p < 0.05), but similar to the IC90 value for FEC (180.9% TDC). In addition a significant positive correlation was observed for the IndexSum of Zol and the ER status (p < 0.01).ConclusionZoledronic acid has a strong antitumor effect on primary breast cancer cells in vitro which is equal or superior to commonly used chemotherapeutic regimens for treating breast cancer.

P2-08-05: Use of Dynamic Contrast-Enhanced MR Imaging To Predict Pathological Response in Primary Breast Cancer.

Abstract Purpose: To investigate the effect of bevacizumab infusion on vascular parameters assessed by dynamic contrast-enhanced magnetic resonance (DCE-MR) imaging and to test their association with pathological response in primary breast cancer. Materials and Methods: 73 patients (median age, 47 ys; age range, 29–70 ys) with biopsy-proven, previously untreated, primary breast cancer were recruited from October 2009 to November 2010 in this phase II, multicenter and non-randomized clinical trial. Patients (pts) received single infusion of bevacizumab (15 mg/kg) (C1) 3 weeks prior to the beginning of neoadjuvant chemotherapy consisting in 4 cycles of docetaxel (60 mg/mq), doxorubicin (50 mg/mq) and bevacizumab (15 mg/kg) every 21 days (C2-C5) following by surgery. All pts underwent DCE-MR imaging before and 14–21 days after C1. Quantitative and semiquantitative kinetic parameters were calculated at baseline and after C1, including the volume transfer constant (K(trans)), which primarily reflects the wash-in of the contrast agent, the backflow rate contrant (K(ep)), extracellular volume fraction (V(e)) and the initial area under the gadolinium concentration-time curve over 60 seconds (IAUGC(60)). Changes in the DCE-MRI kinetic parameters K(trans), K(ep), V(e) and IAUGC(60) were calculated and Wilcoxon test was used to assess significant effects induced by bevacizumab on kinetic parameters. Pathological response on surgical specimens after C5 was assessed according to Miller and Payne classification. Pts with tumor reduction &amp;gt;30% were considered as responders (G3-G4-G5) whether tumor reduction &amp;lt;30% were considered as no responders (G1-G2). DCE-MR imaging parameters and clinical-pathological characteristics were correlated with pathological response using Mann-Whitney test in univariate and logistic regression in multivariate analyses. Receiver operating curves (ROC) was used to define the best cut-off of the parameter found associated with pathological response. Results: DCE-MRI was performed before (n=72) and after (n=71) C1. K(trans), K(ep), V(e) and IAUCG(60) values were significantly different at the baseline and after C1 (p&amp;lt;0.01). Median changes were, respectively, −51, −101, −52.5 and −4.8. Fifty-two (74%) pts achieved response (G3-G4-G5) after C5 whether 18 (24%) were considered as no responder (G1-G2); for 3 (4%) patients Miller/Payne tumor evaluation was not available. At univariate analysis, negative estrogen receptor (ER) status and higher post-C1 K(ep) (p=.057) showed a trend toward an association with response. At multivariate analysis, only ER status remains a significant predictor of response (p= .04). Area under ROC curve for K(ep) was 0.65 (IC95% 0.05−0.8, p=.057) Conclusion: Bevacizumab affects tumor vasculature, perfusion and permeability as showed by the significantly reduction in all kinetic vasculature parameters obtained in DCE-MR imaging after C1. However, in our population these changes were not associated with pathological response. On the other hand, backflow rate constant, K(ep), a perfusion-related parameter derived from DCE-MRI yielded after C1 major than 80, may be associated with higher pathological response with a specificity of 88% and sensitivity of 90%. Future studies are warranted to confirm these findings. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-08-05.

Antitumor activity of zoledronic acid in primary breast cancer cells determined by the ATP tumor chemosensitivity assay

The NeoAzure study has demonstrated that the use of the bisphosphonate zole-dronate in the neoadjuvant setting increases the rate of complete response in primary breast cancer and therefore indicates direct antitumor activity (Coleman et al. 2010). The purpose of this study was to compare the antitumor effect of zolendronic acid with standard chemotherapy in primary breast cancer cells using ATP-tumor chemosensitivity assay (ATP-TCA).

Antitumor activity of zoledronic acid in primary breast cancer cells determined by the ATP tumor chemosensitivity assay.

e11112 Background: The NeoAzure study has demonstrated that the use of the bisphosphonate zole-dronate in the neoadjuvant setting increases the rate of complete response in primary breast cancer and therefore indicates direct antitumor activity (Coleman et al. 2010). The purpose of this study was to compare the antitumor effect of zolendronic acid with standard chemotherapy in primary breast cancer cells using ATP-tumor chemosensitivity assay (ATP-TCA). Methods: Breast cancer tissues were obtained from patients with breast cancer who underwent primary breast cancer surgery at the Dept of OB/Gyn, Tubingen, Germany, between 2006 through 2009. Antitumor effects of zoledronic acid (Zol), TAC (docetaxel, doxorubucin hydrochloride, cyclophosphamide) and FEC (5-fluorouracil, epirubicin, cyclophosphamide) were tested in 116 fresh human primary breast cancer specimens using ATP-TCA. ATP-TCA results were analyzed with different cut-off levels for the half maximal inhibitory concentration (IC50) and for IC90 and for...

Abstract P2-09-26: Circadian Clock Genes in Primary Breast Cancer: Strong Predictors of Pathologic Response on Neoadjuvant Chemotherapy

Abstract Background: Circadian clocks involve 15 specific genes, which control cell cycle, apoptosis, DNA repair and metabolism. Circadian disruption is associated with both an increased risk of breast cancer, and poor survival in patients (pts) with metastatic cancer (IARC#98; Innominato et al. Cancer Res 2009). Objective: to provide first evidence of clinical relevance of clock genes expression for the therapeutic sensitivity of primary breast cancer. Method: Affymetrix Hgu133plus2 microarrays data were derived from 189 primary breast cancers. All the pts had non metastatic disease and were registered in multicenter Phase II trial of neoadjuvant chemotherapy with epirubicin-cyclophosphamide (4 courses) then docetaxel ± herceptin (4 courses). In this study, negative hormonal receptor status and limited tumor size predicted for pathological complete response rate (pCR) (Pierga JY, et al Breast Cancer Res Treat. 2010).The expression data of 15 core clock genes, within the whole tumor transcriptome, were related both to tumor mitotic index (MI) and estrogen receptors (ESR1), and to chemotherapy-induced pCR. Statistical validation involved T-test statistics, with corrections for multiple testing. Results: In the tumor biopsy obtained before treatment onset, both Per2 and Cry2 expressions correlated positively with ESR1 (r≥0.43; P&amp;lt;0.001) and inversely with (MI (r≤−0.28; P&amp;lt;0.007). In addition, a positive correlation was found for Dec 1 with ESR1 (r=0.65, P&amp;lt;0.001). The mean expressions of Per2, Cry2 and Dec 1 before chemotherapy were lower in the 32 pts who subsequently achieved pathologic complete response (pCR) as compared to the 157 who did not (P&amp;lt;0.001). No other clock gene expression was significantly related to pCR. The odds ratio confirmed the decreased chance of achieving pCR with increased expression of clock genes Per2, Cry2 or Dec1. Clock gene expression and pCR Conclusion: Low expressions of three core genes in the negative loop of the molecular circadian clock strongly predicted for the induction of a pathologic complete response of primary breast cancer with neoadjuvant chemotherapy. While circadian clock disruption seems to constitute a poor prognostic factor in cancer patients, our data suggest that the downregulation of core circadian genes in tumors significantly enhances susceptibility to chemotherapy. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-26.

Early Reduction in Standardized Uptake Value After One Cycle of Neoadjuvant Chemotherapy Measured by Sequential FDG PET/CT is an Independent Predictor of Pathological Response of Primary Breast Cancer

The Breast JournalVolume 16, Issue 6 p. 660-662 Early Reduction in Standardized Uptake Value After One Cycle of Neoadjuvant Chemotherapy Measured by Sequential FDG PET/CT is an Independent Predictor of Pathological Response of Primary Breast Cancer Shigeto Ueda MD, Shigeto Ueda MD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorHitoshi Tsuda MD, PhD, Hitoshi Tsuda MD, PhD Department of Basic Pathology, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorToshiaki Saeki MD, PhD, Toshiaki Saeki MD, PhD Department of Breast Oncology, Tokorozawa Ichou Hospital, Tokorozawa, Saitama, Japan Department of Breast Oncology, Saitama Medical University, International Medical Center, Saitama, JapanSearch for more papers by this authorAkihiko Osaki MD, PhD, Akihiko Osaki MD, PhD Department of Breast Oncology, Tokorozawa Ichou Hospital, Tokorozawa, Saitama, Japan Department of Breast Oncology, Saitama Medical University, International Medical Center, Saitama, JapanSearch for more papers by this authorTakashi Shigekawa MD, Takashi Shigekawa MD Department of Breast Oncology, Tokorozawa Ichou Hospital, Tokorozawa, Saitama, Japan Department of Breast Oncology, Saitama Medical University, International Medical Center, Saitama, JapanSearch for more papers by this authorJiro Ishida MD, PhD, Jiro Ishida MD, PhD Tokorozawa PET Diagnostic Imaging Clinic, Tokorozawa, Saitama, JapanSearch for more papers by this authorKatsumi Tamura MD, Katsumi Tamura MD Tokorozawa PET Diagnostic Imaging Clinic, Tokorozawa, Saitama, JapanSearch for more papers by this authorYoshiyuki Abe MD, PhD, Yoshiyuki Abe MD, PhD Tokorozawa PET Diagnostic Imaging Clinic, Tokorozawa, Saitama, JapanSearch for more papers by this authorJiro Omata MD, Jiro Omata MD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorTomoyuki Moriya MD, PhD, Tomoyuki Moriya MD, PhD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorKazuhiko Fukatsu MD, PhD, Kazuhiko Fukatsu MD, PhD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorJunji Yamamoto MD, PhD, Junji Yamamoto MD, PhD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this author Shigeto Ueda MD, Shigeto Ueda MD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorHitoshi Tsuda MD, PhD, Hitoshi Tsuda MD, PhD Department of Basic Pathology, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorToshiaki Saeki MD, PhD, Toshiaki Saeki MD, PhD Department of Breast Oncology, Tokorozawa Ichou Hospital, Tokorozawa, Saitama, Japan Department of Breast Oncology, Saitama Medical University, International Medical Center, Saitama, JapanSearch for more papers by this authorAkihiko Osaki MD, PhD, Akihiko Osaki MD, PhD Department of Breast Oncology, Tokorozawa Ichou Hospital, Tokorozawa, Saitama, Japan Department of Breast Oncology, Saitama Medical University, International Medical Center, Saitama, JapanSearch for more papers by this authorTakashi Shigekawa MD, Takashi Shigekawa MD Department of Breast Oncology, Tokorozawa Ichou Hospital, Tokorozawa, Saitama, Japan Department of Breast Oncology, Saitama Medical University, International Medical Center, Saitama, JapanSearch for more papers by this authorJiro Ishida MD, PhD, Jiro Ishida MD, PhD Tokorozawa PET Diagnostic Imaging Clinic, Tokorozawa, Saitama, JapanSearch for more papers by this authorKatsumi Tamura MD, Katsumi Tamura MD Tokorozawa PET Diagnostic Imaging Clinic, Tokorozawa, Saitama, JapanSearch for more papers by this authorYoshiyuki Abe MD, PhD, Yoshiyuki Abe MD, PhD Tokorozawa PET Diagnostic Imaging Clinic, Tokorozawa, Saitama, JapanSearch for more papers by this authorJiro Omata MD, Jiro Omata MD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorTomoyuki Moriya MD, PhD, Tomoyuki Moriya MD, PhD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorKazuhiko Fukatsu MD, PhD, Kazuhiko Fukatsu MD, PhD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this authorJunji Yamamoto MD, PhD, Junji Yamamoto MD, PhD Department of Surgery, National Defense Medical College, Tokorozawa, Saitama, JapanSearch for more papers by this author First published: 12 November 2010 https://doi.org/10.1111/j.1524-4741.2010.01011.xCitations: 7 Address corresopondence and reprint requests to: Hitoshi Tsuda, MD, PhD, Department of Basic Pathology, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan, or e-mail: [email protected]. Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume16, Issue6November/December 2010Pages 660-662 RelatedInformation

A simple system for grading the response of breast cancer to neoadjuvant chemotherapy

BackgroundThe response of primary breast cancer to chemotherapy is usually expressed either as a pathological complete remission (pCR) or as ‘no pCR’. A more quantitative measure is called for. Patients and methodsThe ‘neoadjuvant response index’ (NRI) was calculated by adding a breast response score (a number from a five-point scale) to an axillary response score (a number from a three-point scale) and dividing this by the score that would have been obtained in case of a pCR in both breast and axilla. Consequently, the NRI is a number between 0 (representing no response) and 1 (a pCR of both breast and axilla). ResultsThe NRI was calculated in 267 patients who had received neoadjuvant chemotherapy. The average NRI was 0.48 (median 0.40). Forty-one patients (15%) had an NRI of 0; 55 patients (21%) had an NRI of 1 (pCR). ‘Highly endocrine responsive’ tumors responded substantially less than ‘incompletely endocrine responsive’ ones. In triple negatives, an NRI of >0.70 was associated with a better recurrence-free survival than a lower NRI. ConclusionsThe NRI proposed here may be useful to better reflect the efficacy of neoadjuvant systemic regimens than the binary pCR–‘no pCR’ system.

Good clinical response of breast cancers to neoadjuvant chemoendocrine therapy is associated with improved overall survival

BackgroundWe present extended follow-up from a prospective randomised trial evaluating the role of neoadjuvant chemoendocrine therapy in the treatment of operable breast cancer. Patients and methods309 women were randomised to primary surgery followed by eight cycles of adjuvant mitoxantrone, methotrexate with tamoxifen (2MT) or 2MT with mitomycin-C (3MT) versus the same regimen for four cycles before followed by four cycles after surgery. For this analysis the median follow-up of patients was 112 months. ResultsAfter 10 years follow-up there is still no statistically significant difference in disease-free survival (DFS) (71% versus 71%) or overall survival (OS) (63% versus 70%) when comparing adjuvant versus neoadjuvant treatment, respectively. Of 144 evaluable patients in the neoadjuvant arm, 74 achieved a good clinical response and 70 patients achieved a poor clinical response. Good responders had a superior DFS (80% versus 64%, P=0.01) and OS (77% versus 63%, P=0.03) compared to poor responders. ConclusionsAt 10 years, neoadjuvant and adjuvant treatment continue to have equivalent OS and DFS. Good clinical response to neoadjuvant chemotherapy is associated with superior DFS and OS. This supports the use of clinical response of primary breast cancer to neoadjuvant therapy as a surrogate marker of survival benefit.