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Table S1: Genes significantly differentially expressed in samples with pathological complete response compared to those without in the Combination arm. Table S2: DAVID analysis of 720 differentially expressed genes in tumors that achieved pathological complete reponse compared to those that did not, in the combination arm. Table S3 : Genes significantly differentially expressed in samples with pathological complete response compared to those without in the Chemotherapy arm. Table S4: DAVID analysis of 1243 differentially expressed genes in tumors that achieved pathological complete reponse compared to those that did not, in the chemotherapy arm. Table S5 : List of genes with significant lower expression in week 12 samples treated with combination therapy compared to those treated with chemotherapy Table S6: DAVID analysis of 42 differentially expressed genes in week-12 tumors in the combination arm compared to chemotherapy arm. Table S7 : Differentially expressed genes between week-0 and week-12 in Basal-like tumors treated with combination therapy Table S8 : Differentially expressed genes between week-0 and week-12 Luminal B tumors treated with combination therapy Table S9 : Differentially expressed genes between week-0 and week-12 Luminal A tumors treated with combination therapy Table S10: Differentially expressed genes between week-0 and week-12 Basal-like tumors treated with chemotherapy Table S11 : Differentially expressed genes between week-0 and week-12 Luminal B tumors treated with chemotherapy Table S12 : Differentially expressed genes between week-0 and week-12 Luminal A tumors treated with chemotherapy Table S13: Pathways significantly altered between Chemotherapy and Combination therapy arms from week-0 to week-12 LuminalB samples Table S14: Pathways significantly altered between anthracycline and taxane treatment in the Luminal A samples in chemotherapy arm

Purpose: Chemotherapy-induced alterations to gene expression are due to transcriptional reprogramming of tumor cells or subclonal adaptations to treatment. The effect on whole-transcriptome mRNA expression was investigated in a randomized phase II clinical trial to assess the effect of neoadjuvant chemotherapy with the addition of bevacizumab.Experimental Design: Tumor biopsies and whole-transcriptome mRNA profiles were obtained at three fixed time points with 66 patients in each arm. Altogether, 358 specimens from 132 patients were available, representing the transcriptional state before treatment start, at 12 weeks and after treatment (25 weeks). Pathologic complete response (pCR) in breast and axillary nodes was the primary endpoint.Results: pCR was observed in 15 patients (23%) receiving bevacizumab and chemotherapy and 8 patients (12%) receiving only chemotherapy. In the estrogen receptor–positive patients, 11 of 54 (20%) treated with bevacizumab and chemotherapy achieved pCR, while only 3 of 57 (5%) treated with chemotherapy reached pCR. In patients with estrogen receptor–positive tumors treated with combination therapy, an elevated immune activity was associated with good response. Proliferation was reduced after treatment in both treatment arms and most pronounced in the combination therapy arm, where the reduction in proliferation accelerated during treatment. Transcriptional alterations during therapy were subtype specific, and the effect of adding bevacizumab was most evident for luminal-B tumors.Conclusions: Clinical response and gene expression response differed between patients receiving combination therapy and chemotherapy alone. The results may guide identification of patients likely to benefit from antiangiogenic therapy. Clin Cancer Res; 23(16); 4662–70. ©2017 AACR.

Supplementary figures as referred to in the manuscript

I denne artikkelen evaluerer vi hvordan strengere klimapolitikk påvirker norske lån til utlandet. Ved å benytte syndikerte lånedata fra Norge og klimaindeksen fra Germanwatch for både Norge og mottakerlandene i perioden 2007 til 2017, finner vi to stiliserte resultater. Først avdekker vår analyse på aggregatnivå at en strengere innenlands klimapolitikk i Norge fører til en reduksjon i andelen lån som norske banker utsteder til utenlandske låntakere. For det andre indikerer våre resultater at strengere klimapolitikk i mottakerlandet ikke påvirker størrelsen på utenlandslån fra norske banker. Samlet sett peker våre resultater på mangel på bevis for regulatorisk arbitrasje i det norske markedet for syndikerte lån. In this paper, we study how climate policy stringency affects cross-border lending in Norway. Using syndicated loan data from Norway and Climate Change Performance Index for both Norway and recipient countries from 2007 to 2017, we find two stylized results. First, our aggregate-level analysis reveal that a more stringent domestic climate policy in Norway tends to reduce the proportion of loans that Norwegian banks extend to foreign borrowers. Second, when looking at cross-country evidence, our results indicate that Norwegian banks cross-border lending amount does not seem to respond to changes in international climate policy regulation stringency. Altogether, our results point to no evidence of regulatory arbitrage in the Norwegian syndicated loan market.

Oppgaven tar for seg innvandring sin påvirkning på lønnsnivået i Norge. Ved hjelp av en regresjonsanalyse på et datasett fra 2015-2019, ser vi påvirkningskraften til innvandring på lønn. Analysen konkluderer med at vi ikke finner konklusive svar som tilsier at innvandring påvirker lønnsnivået. The thesis looks at immigrations impact on the norwegian avarage wages. By using a regression analysis on a dataset spanning from 2015-2019, we look to find immigration's effect on norwegian wages. The analysis concludes that there is not any conclusive evidence that says immigration affects the wages.

BACKGROUND: Patients with HER2 negative primary tumors of ≥25 mm were treated with neoadjuvant chemotherapy (4 x FEC100 + 12 weeks of taxane-based therapy) and randomized (1:1) to receive bevacizumab or not. Mammography, ultrasound and MR imaging were used for response evaluation, in addition to the final pathology assessment after surgery. HYPOTHESIS: RPPA proteomic analyses support identification of molecular mechanisms associated with clinical response to bevacizumab treatment. METHODS: Tumor responses were evaluable in 132 patients; of which 66 received bevacizumab. Ratio of the tumor size at final pathology assessment, and at inclusion was calculated to obtain a continuous scale of response reflecting the percentage of tumor shrinkage in response to therapy. Tumor material was obtained at screening, 12 weeks into treatment and at surgical removal of tumors at 25 weeks. Lysates from each sample was analyzed on reverse phase protein arrays (RPPA) for expression levels of 210 proteins of which 54 were phospho-specific. RESULTS: Several proteins were found for which expression prior to treatment reflected a better response on tumor shrinkage in the combination treatment arm (chemotherapy+bevacizumab). The proteomic response from week 0 to 12 in both treatment arms had an overall similar profile regarding up- and down-regulated proteins; however, the combination treatment (FEC100 + bevacizumab) induced a more pronounced effect on regulation of each protein. This might reflect the capability of bevacizumab therapy to potentiate the effects of the anthracyclin based chemotherapy from week 0 to 12. Conversely, from week 12-25 (taxane-based therapy + bevacizumab) this effect was lost or even reversed, except for certain phosphoproteins where potentiation imposed by bevacizumab was sustained throughout the whole treatment period. We are in the process of analyzing the impact of phosphorylation and thus protein activation states on treatment response. Furthermore, tumors with low hormone receptor pathway score demonstrated a better response in the combination treatment (chemotherapy+bevacizumab). Additionally, in these good responders the hormone signaling pathway was significantly upregulated during treatment. Further investigations are conducted to determine if this was due to selective ablation of hormone receptor negative tumor cells, or a re-programming of the molecular phenotype of cells present prior to treatment. The above mentioned results have potentially important clinical relevance and will be further investigated with respect to subtypes and the biological pathways affected by antiangiogenic therapy. Citation Format: Haugen MH, Lindgjærde OC, Krohn M, Zhao W, Lindholm EM, Silwal-Pandit L, Borgen E, Garred Ø, Fangberget A, Holmen MM, Schlichting E, Skjerven H, Lundgren S, Wist E, Naume B, Mælandsmo GM, Lu Y, Børresen-Dale A-L, Mills GB, Engebråten O. Proteomic response in breast cancer treated with neoadjuvant chemotherapy with and without bevacizumab: Reverse phase protein array (RPPA) results from NeoAva - A randomized phase II study [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 P6-13-01.

The NeoAva study is a phase II clinical trial of patients with HER2 negative primary tumors of ≥25 mm treated with neoadjuvant chemotherapy (4 x FEC100 + 12 weeks of taxane-based therapy) and randomized (1:1) to receive bevacizumab or no bevacizumab. Mammography, ultrasound and MR imaging were used for response evaluation, in addition to final pathology assessment. Tumor response were evaluable in 131 patients; of which 66 received bevacizumab in addition to chemotherapy. Tumor material was obtained at screening, 12 weeks into treatment and at surgical removal of tumors at 25 weeks. mRNA expression profiling was performed on Agilent 8x60K platform and the tumors were classified into LuminalA, LuminalB, Her2-enriched, Basal and Normal-like subtypes using the PAM50 classifier. Ratio of the tumor size at final pathology assessment, and at inclusion (by radiology assessment) was calculated to obtain a continuous scale of response reflecting the percentage of tumor shrinkage in response to therapy. Genomic Grade Index (GGI scores) based on expression profiles of 97 genes (including cell-cycle and proliferation genes) were calculated. There were no significant differences in the tumor size, lymph node, hormone receptor status or PAM50 subtypes between the treatment arms. pCR in breast and axilla were obtained in 14 (21.1%) patients in the chemo+bev arm, and in 7 (10.6%) patients in the chemo-only arm. Tumors that obtained pCR were in higher number ER negative and TP53 mutated and exhibited Basal-like phenotype. The overall pCR rates were higher in the ER negative tumors compared to ER positive tumors {39.1% (9 of 23) vs 11.1% (12 of 108)}. However, addition of bevacizumab seemed to improve pCR in the ER positive patient group (9 vs 3) and not in ER negative patient group (5 vs 4). On evaluating the continuous response variable, ER status, TP53 mutation status and PAM50 subtypes were significantly associated to response (p < 0.001). GGI scores were highly correlated to response (p< 0.001), i. e tumors with higher GGI scores showed better response. Importantly, when the chemo+bev and the chemo-only arms were evaluated separately, although similar trend of associations was observed in both arms, the associations were found to be enhanced in the chemo+bev arm. Next, we evaluated a shift in PAM50 subtypes across the timepoints. A shift towards a better prognosis group, i.e Luminal A or Normal-like profile was observed in response to therapy. Distribution of Luminal A and Normal-like tumors at week 25, (and not at screening or week 12) was significantly different in the chemo+bev vs chemo-only group (p = 0.026, Fisher’s exact test). GGI scores regressed across timepoints reflecting the loss of aggressive and proliferating component of the tumors in response to therapy. GGI scores in the chemo+bev group became significantly lower (p < 0.01) already at week 12. This suggests that the removal of the proliferating component of the tumors by chemotherapy is accelerated and improved by addition of bevacizumab. These results, with potentially important clinical relevance will be further investigated with respect to subtypes and the molecular changes induced by antiangiogenic therapy. Citation Format: Olav Engebraaten, Laxmi Silwal-Pandit, Marit Krohn, Elen K Møller, Silje Nord, Thomas Fleischer, Hedda von der Lippe Gythfelt, Elin Borgen, Øystein Garred, Anne Fangberget, Marit Muri Holmen, Ellen Schlichting, Helle Skjerven, Steinar Lundgren, Vessela N Kristensen, Ole Christian Lingjaerde, Erik Wist, Bjørn Naume, Anne-Lise Børresen-Dale. Molecular response in breast cancer treated with neoadjuvant chemotherapy with and without bevacizumab: Results from NeoAva - a randomized phase II study [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-11-14.

Purpose To explore possible associations between in vivo pharmacokinetic dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters and the presence of disseminated tumor cells (DTCs) in bone marrow in breast cancer patients at the time of diagnosis. Materials and Methods Thirty-seven women with breast cancer (stage T2–4N0–1M0) were included. Patients were classified as DTC+ if one or more DTCs were detected by immunocytochemistry. DCE-MRI was acquired with a radial 3D T1-weighted spoiled gradient echo sequence with k-space weighted image contrast. Ktrans, kep, and ve were calculated using the extended Tofts model and a population-derived arterial input function. The nonparametric Mann–Whitney U-test was used to compare the histogram distributions of the pharmacokinetic parameters for the DTC+ and the DTC− patients. Results DTCs were detected in 7 of the 37 patients (19%). In DTC+ patients, the distribution of tumor Ktrans and kep were significantly (P < 0.01) more shifted towards lower values than in DTC− patients. Conclusion An association between vascular dependent pharmacokinetic DCE-MRI parameters and the presence of DTCs were found. Compared to DTC− patients, DTC+ patients had poorer perfusion and permeability, indicative of hypoxia. Thus, pharmacokinetic parameters might be surrogate biomarkers of metastatic potential and future relapse.J. Magn. Reson. Imaging 2014;40:1382–1391. © 2014 Wiley Periodicals, Inc.

Tumor heterogeneity is an area of intense research, revealing tumors with high complexity consisting of different subclones and infiltrating cells. Identification of subclones that are resistant to therapy may be critical to improve treatment outcome. The NeoAva study is a randomized phase II, clinical trial of Her2 negative breast cancer patients treated in a neoadjuvant setting with chemotherapy (FEC and taxane) +/- bevacizumab. Core needle biopsies were obtained at screening and after 12 weeks, and the tumor was surgically removed after 25 weeks. DNA copy number changes in the tumors were analyzed using Affymetrix SNP Array 6.0. Allele specific copy number changes were assessed using the Allele-Specific Copy number Analysis of Tumors (ASCAT) algorithm (Van Loo, Norgard et al., PNAS 2010) and allele-specific Piecewise Constant Fitting (asPCF) algorithms (Nilsen, Liestol et al., BMC Genomics 2012). Measures of genomic instability were obtained through the complex arm-wise aberration index (CAAI) that captures local rearrangements (‘firestorms’) (Russnes, Vollan et al., Sci Transl Med 2010). Changes in copy number aberrations between the three different time points were observed in almost all tumors. Some tumors showed a decrease in tumor percentage and aberrations after just 12 weeks of treatment, where others showed loss of aberrations only at the time of surgery (25 weeks). Most of the tumors that did retain aberrations at all time points during treatment, did not demonstrate any decrease in tumor size. Other profiles indicated subclonal reduction, where some aberrations are kept throughout treatment and others disappear. Many of the tumors shrinking in size showed fewer whole arm aberrations than before treatment, but retained their focal amplicons. Some of the tumor aberrations seem to disappear after 12 weeks, but to reappear after 25 weeks, but with the addition of novel aberration. Complex rearrangements were identified in 67% of tumors before treatment. The most frequent ‘firestorms’ were found on 20p, 11q and 8p. Some events were persistent through therapy, but the majority changed. An association between complex tumor genomes and patients having progressive disease/non-responders were observed. These results show the complex structure of a tumor and suggest that heterogeneity will influence the response to treatment. The subclonal patterns of tumors may be of great importance for clinical decision-making, as well as for monitoring treatment efficacy. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-14-01.

Background: Results from studies with bevacizumab in addition to traditional neoadjuvant therapy (NAT) indicate a need for predictive biomarkers. As a sub-study of the NeoAva study (a neoadjuvant study), the aim was to investigate the potential association between the presence of circulating endothelial cells (CECs) in peripheral blood (PB), circulating tumor cells (CTCs) in PB and disseminated tumor cells (DTCs) in bone marrow (BM) at different time points during NAT +/- bevacizumab and at one year follow-up, and therapy response. Patients and methods: A total of 150 HER2-negative patients with cT2-4 (≥ 2.5cm) N0-3 M0 status were randomized to receive NAT with or without bevacizumab. In this sub-study, 90 patients have so far been analyzed. Of these, 82 received chemotherapy (FEC100→taxane) and 8 received endocrine therapy (letrozole) + / - bevacizumab. Therapy response was evaluated according to the RECIST criteria and achievement of pathological complete response (pCR). Number of CECs, CTCs and DTCs were assessed at baseline after 12 weeks, at surgery (after 24 weeks) and at one year follow-up. Blood samples were analyzed by CellSearch® to assess CEC and CTC counts. The detection of DTCs was performed by immunocytochemical analysis of 2 × 106 BM mononuclear cells. Results: The pathological complete response rate was 10 out of 90 (11.1%), eight of these patients received bevacizumab. For bevacizumab-treated patients with a change in CEC counts from baseline to time of surgery below median change (27 CECs), 35% (6/17) achieved pCR compared to 6% (1/18) in the group with a CEC count-increase above median change (p = 0.035). The corresponding pCR rates for patients not receiving bevacizumab (median CEC change 131 CECs) were 0% (0/15) and 13% (2/14), respectively. Stepwise testing of thresholds for CEC changes in the bevacizumab-arm revealed significant associations to pCR for change-values between 20 and 40. CTC- and DTC-status or -changes were not associated with tumor response or CEC changes. Conclusion: The presented results indicate that the level of change in the number of circulating endothelial cells during neoadjuvant therapy including bevacizumab is associated with the pathological complete response rate in breast cancer patients. This supports additional testing of CECs as a surrogate marker for response to this treatment. The analyses will be up-dated with results from the rest of the included patients. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-21.

Patients treated with bevacizumab in addition to regular neoadjuvant chemotherapy achieve an increased rate of pathological complete response (pCR). The molecular characteristics of responding and non-responding tumors, including how treatment combinations influence the gene expression profiles and the signaling pathways, may be useful predictors of antiangiogenic response. The NeoAva study included patients with HER2 negative primary tumors of ≥25 mm that were randomized (1:1) to receive neoadjuvant chemotherapy (4 x FEC100 + 12 weeks of taxane-based therapy) with or without bevacizumab. Mammography, ultrasound and MR imaging were used for response evaluation, in addition to final pathology assessment. In the first part of the study 74 patients were evaluable for tumor response. The tumor size at time of inclusion was T2, T3 and T4 in 24.3%, 67.6% and 8.1% of the patients, respectively. Lymph node metastases were detected in 56.7% of the patients at inclusion and 82.4% were hormone receptor positive. There were no significant differences in the tumor size, lymph node or hormone receptor status between the treatment arms. The patients were randomized with bevacizumab + chemotherapy (n = 37) and treatment with chemotherapy alone (n = 37). Of the nine patients who achieved pCR in breast and axilla (12.2%), seven patients received bevacizumab (7/37), while two were treated with chemotherapy alone (2/37). Four of the patients with pCR were hormone receptor negative, of which three received bevacizumab. Of the remaining five hormone receptor positive tumors that achieved complete response, four received bevacizumab. In the second part of the study we evaluated gene expression signatures by RNA microarray and the time-response of pathways to treatment, using pathway analysis that integrates copy number and gene expression (Paradigm). Biopsies for molecular analyses were collected before therapy, after 12 weeks, and at surgery. Treatment associated gene expression changes to chemotherapy were subtracted, and bevacizumab associated differential expression was observed for 1069 genes. Furthermore, molecular profiling of the tumor tissue was performed at DNA level by copy number analysis (Affymetrix, SNP6.0) and mRNA level by gene expression arrays(Agilent 60K). At the screening time point, we found high proliferation through the activity of cyclin E and B and the transcription factors E2F1 and FOXM1. At 12 weeks, there was a strong increase in predicted p53 signaling, due to increased activity of downstream target genes. The 12 week timepoint was also characterized by an increase of Calmodulin 1, MAPK3, as well as Peroxisome proliferator-activated receptor alpha (PPAR-alpha), and both trends continued to the 25 week time point. At 25 weeks, there were broad increases in ERK1/2, JUN, and FOS signaling. The 25 week timepoint also showed a T-cell response signature that from increased activity of GATA3, IL6/IL6R, IL4, and NFATC1 and NFATC2. These results suggest that there are measurable and strongly significant aberrations in molecular activity during treatment, which may be useful to monitor treatment response. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-14-02.

Withdrawn by Author Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-17-02.

Objective To explore the predictive value of MRI parameters and tumour characteristics before neoadjuvant chemotherapy (NAC) and to compare changes in tumour size and tumour apparent diffusion coefficient (ADC) during treatment, between patients who achieved pathological complete response (pCR) and those who did not. Methods Approval by the Regional Ethics Committee and written informed consent were obtained. Thirty-one patients with invasive breast carcinoma scheduled for NAC were enrolled (mean age, 50.7; range, 37–72). Study design included MRI before treatment (Tp0), after four cycles of NAC (Tp1) and before surgery (Tp2). Data in pCR versus non-pCR groups were compared and cut-off values for pCR prediction were evaluated. Results Before NAC, HER2 overexpression was the single significant predictor of pCR (p=0.006). At Tp1 ADC, tumour size and changes in tumour size were all significantly different in the pCR and non-pCR groups. Using 1.42×10−3 mm2/s as the cut-off value for ADC, pCR was predicted with sensitivity and specificity of 88% and 80%, respectively. Using a cut-off value of 83% for tumour volume reduction, sensitivity and specificity for pCR were 91% and 80%. Conclusion ADC, tumour size and tumour size reduction at Tp1 were strong independent predictors of pCR. publishedVersion

Background . For patients with locally advanced breast cancer (LABC) undergoing neoadjuvant chemotherapy (NACT), the European Guidelines for Breast Imaging recommends magnetic resonance imaging (MRI) to be performed before start of NACT, when half of the NACT has been administered and prior to surgery. This is the fi rst study addressing the value of flapparent diffusion coeffi cients (ADCs) obtained from diffusion-weighted (DW) MRI at the recommended time points for pretreatment prediction and monitoring of treatment response. Materials and methods . Twenty-fi ve LABC patients were included in this prospective study. DW MRI was performed using single-shot spin-echo echo-planar imaging with b-values of 100, 250 and 800 s/mm 2 prior to NACT, after four cycles of NACT and at the conclusion of therapy using a 1.5 T MR scanner. ADC in the breast tumor was calculated from each assessment. The strength of correlation between pretreatment ADC, ADC changes and tumor volume changes were examined using Spearman’s rho correlation test. Results . Mean pretreatment ADC was 1.11 � 0.21 � 10 � 3 mm 2 /s. After 4 cycles of NACT, ADC was signifi cantly increased (1.39 � 0.36 � 10 � 3 mm 2 /s; p� 0.018). There was no correlation between individual pretreatment breast tumor ADC and MR response measured after four cycles of NACT (p� 0.816) or prior to surgery (p� 0.620). Conclusion . Pretreatment tumor ADC does not predict treatment response for patients with LABC undergoing NACT. Furthermore, ADC increase observed mid-way in the course of NACT does not correlate with tumor volume changes.

Antiangiogenic therapy using bevacizumab has proven effective for a number of cancers; however, in breast cancer there is an unmet need to identify patients that benefit from such treatment. Sampling of tumor biopsies before and during treatment, as well as at the time of surgery enables the assessment of response at multiple molecular levels. At the proteomic level reverse phase protein analysis (RPPA) support expression of numerous cancer associated proteins simultaneously, which can further be used to unravel molecular mechanisms associated with clinical response to bevacizumab treatment. In this phase II clinical trial, patients with HER2 negative primary tumors of ≥25 mm were treated with neoadjuvant chemotherapy (4 x FEC100 + 12 weeks of taxane-based therapy) and randomized (1:1) to receive bevacizumab or not. Mammography, ultrasound and MR imaging were used for response evaluation, in addition to final pathology assessment. Tumor responses were evaluable in 132 patients; of which 66 received bevacizumab. Ratio of the tumor size at final pathology assessment, and at inclusion was calculated to obtain a continuous scale of response reflecting the percentage of tumor shrinkage in response to therapy. Tumor biopsies were removed before start of treatment, at week 12 at the start of taxane-based tharapy and at the time of surgery. Lysates from each sample was analyzed on reverse phase protein arrays (RPPA) for expression levels of 210 proteins of which 54 were phospho-specific. The addition of bevacizumab to the chemotherapy do not alter proteomic response from week 0 to 25 to such extent that this patient group cluster naturally together. While the proteomic response from week 0 to 12 in both treatment arms had an overall similar profile regarding up- and down-regulated proteins, the combination treatment (FEC100 + bevacizumab) induced substantially more effect on the regulation of each protein. This suggests that bevacizumab treatment have the capability to potentiate the effects of the anthracyclin based chemotherapy from week 0 to 12. Conversely, from week 12-25 (taxane-based therapy + bevacizumab) this effect was lost or even reversed, possibly due to a de-vascularized and less accessible tumor. An exception to this observation was a few phospho-proteins that do seem to have sustained stronger regulation over the whole treatment period. We are in the process of analyzing in more detail the impact of phosphorylation and thus protein activation states on treatment response. Deciphering molecular response and activity regulation at the proteomic level is a promising approach and may reveal novel knowledge with potential important clinical relevance. Citation Format: Mads H. Haugen, Ole Christian Lingjaerde, Marit Krohn, Wei Zhao, Evita M. Lindholm, Laxmi Silwal-Pandit, Elin Borgen, Øystein Garred, Anne Fangberget, Marit M. Holmen, Ellen Schlichting, Helle K. Skjerven, Steinar Lundgren, Erik Wist, Bjørn Naume, Gunhild M. Maelandsmo, Yiling Lu, Anne-Lise Boerresen-Dale, Gordon B. Mills, Olav Engebraaten. Bevacizumab potentiates the proteomic response to neoadjuvant chemotherapy in breast cancer patients: Rppa exploration of consecutive tumor samples in the NeoAva randomized phase II trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1813. doi:10.1158/1538-7445.AM2017-1813

To explore possible associations between in vivo pharmacokinetic dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters and the presence of disseminated tumor cells (DTCs) in bone marrow in breast cancer patients at the time of diagnosis.Thirty-seven women with breast cancer (stage T2-4N0-1M0) were included. Patients were classified as DTC+ if one or more DTCs were detected by immunocytochemistry. DCE-MRI was acquired with a radial 3D T1 -weighted spoiled gradient echo sequence with k-space weighted image contrast. K(trans), kep, and ve were calculated using the extended Tofts model and a population-derived arterial input function. The nonparametric Mann-Whitney U-test was used to compare the histogram distributions of the pharmacokinetic parameters for the DTC+ and the DTC- patients.DTCs were detected in 7 of the 37 patients (19%). In DTC+ patients, the distribution of tumor K(trans) and kep were significantly (P0.01) more shifted towards lower values than in DTC- patients.An association between vascular dependent pharmacokinetic DCE-MRI parameters and the presence of DTCs were found. Compared to DTC- patients, DTC+ patients had poorer perfusion and permeability, indicative of hypoxia. Thus, pharmacokinetic parameters might be surrogate biomarkers of metastatic potential and future relapse.

To explore how apparent diffusion coefficients (ADCs) in malignant breast lesions are affected by selection of b values in the monoexponential model and to compare ADCs with diffusion coefficients (Ds) obtained from the biexponential model. Twenty-four women (mean age 51.3 years) with locally advanced breast cancer were included in this study. Pre-treatment diffusion-weighted magnetic resonance imaging was performed using a 1.5-T system with b values of 0, 50, 100, 250 and 800 s/mm2. Thirteen different b value combinations were used to derive individual monoexponential ADC maps. All b values were used in the biexponential model. Median ADC (including all b values) and D were 1.04 × 10-3 mm2/s (range 0.82–1.61 × 10-3 mm2/s) and 0.84 × 10-3 mm2/s (range 0.17–1.56 × 10-3 mm2/s), respectively. There was a strong positive correlation between ADCs and Ds. For clinically relevant b value combinations, maximum deviation between ADCs including and excluding low b values (

2523 Background: The molecular characteristics of responding and non-responding breast cancers when treated with antiangiogenic therapy are largely unknown. Methods: To investigate molecular altera...

The present study was a proof-of-concept study to provide an initial indication of the efficacy and safety of imaging malignant breast tumors using (99m)Tc-NC100692. The agent is a small peptide with high affinity for integrin receptors that are upregulated and expressed preferentially on proliferating endothelial cells.Sixteen patients with suggestive mammographic findings and 4 patients with benign lesions were included. The "standard of truth" was based on the histopathologic diagnosis of the recruited patients. All subjects received up to 75 microg of (99m)Tc-NC100692 with an average (99m)Tc activity of 694 MBq (range, 561-747 MBq). Safety endpoints were treatment-emergent adverse events (AEs) and changes in a limited physical examination, electrocardiogram (ECG) recordings, blood biochemistry, hematology, coagulation, vital signs, and urine analysis after administration of (99m)Tc-NC100692 and throughout the 24-h follow-up. Static images and SPECT were acquired between 40 min and 2.5 h after injection of the agent. Two experienced nuclear medicine physicians read the images in a nonblinded fashion.Nineteen of 22 malignant lesions were detected using (99m)Tc-NC100692 scintigraphy. Twenty lesions confirmed as malignant by histopathology were seen on mammography or ultrasound. Two additional lesions were identified from histopathology alone. Safety parameters evaluated through the follow-up period of 2.5 h included clinical laboratory tests, vital signs, and ECG. Five of 20 subjects experienced nonserious AEs, and all AEs were classified as mild. One subject experienced an AE (dysgeusia) possibly related to administration of (99m)Tc-NC100692. This AE was mild and lasted only for a few minutes. No deaths, serious AEs, or withdrawals due to AEs occurred during the study.Nineteen of 22 malignant lesions (86%) were clearly detected via scintigraphic imaging after administration of (99m)Tc-NC100692. Overall, the efficacy data in subjects with suspected breast lesions suggest that (99m)Tc-NC100692 scintigraphy may be effective in detecting malignant lesions. The use of (99m)Tc-NC100692 in subjects with breast cancer is safe and well tolerated. Further studies are warranted to assess the clinical potential of (99m)Tc-NC100692.

Tumor heterogeneity is an area of intense research, revealing tumors with high complexity consisting of different subclones and infiltrating cells. Identification of subclones that are resistant to therapy may be critical to improve treatment outcome. The NeoAva study is a randomized phase II, clinical trial of Her2 negative breast cancer patients treated in a neoadjuvant setting with chemotherapy (FEC and taxane) +/- bevacizumab. Core needle biopsies were obtained at screening and after 12 weeks, and the tumor was surgically removed after 25 weeks. DNA copy number changes in the tumors were analyzed using Affymetrix SNP Array 6.0. Allele specific copy number changes were assessed using the Allele-Specific Copy number Analysis of Tumors (ASCAT) algorithm (Van Loo, Norgard et al., PNAS 2010) and allele-specific Piecewise Constant Fitting (asPCF) algorithms (Nilsen, Liestol et al., BMC Genomics 2012). Measures of genomic instability were obtained through the complex arm-wise aberration index (CAAI) that captures local rearrangements (‘firestorms’) (Russnes, Vollan et al., Sci Transl Med 2010). Changes in copy number aberrations between the three different time points were observed in almost all tumors. Some tumors showed a decrease in tumor percentage and aberrations after just 12 weeks of treatment, where others showed loss of aberrations only at the time of surgery (25 weeks). Most of the tumors that did retain aberrations at all time points during treatment, did not demonstrate any decrease in tumor size. Other profiles indicated subclonal reduction, where some aberrations are kept throughout treatment and others disappear. Many of the tumors shrinking in size showed fewer whole arm aberrations than before treatment, but retained their focal amplicons. Some of the tumor aberrations seem to disappear after 12 weeks, but to reappear after 25 weeks, but with the addition of novel aberration. Complex rearrangements were identified in 67% of tumors before treatment. The most frequent ‘firestorms’ were found on 20p, 11q and 8p. Some events were persistent through therapy, but the majority changed. An association between complex tumor genomes and patients having progressive disease/non-responders were observed. These results show the complex structure of a tumor and suggest that heterogeneity will influence the response to treatment. The subclonal patterns of tumors may be of great importance for clinical decision-making, as well as for monitoring treatment efficacy. Citation Format: Elen K. Møller, Silje Nord, Hans Kristian Moen Vollan, Hedda von der Lippe Gythfeldt, Hege Edvardsen, Laxmi Silwal- Pandit, Marit Krohn, Thomas Fleischer, Ellen Schlitchting, Elin Borgen, Øystein Garred, Anne Fangberget, Marit M. Holmen, Helle Skjerven, Steinar Lundgren, Erik Wist, Bjørn Naume, Anne-Lise Børresen-Dale, Ole Christian Lingjærde, Olav Engebråten, Vessela N. Kristensen. A time course study of genomic instability in breast cancer patients receiving neoadjuvant therapy with or without bevacizumab. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1533. doi:10.1158/1538-7445.AM2014-1533