Background: Carboplatin (Cb) improves treatment efficacy among patients with triple negative breast cancer (TNBC). However, not all patients need additional chemotherapy and the benefit for patients with hormone receptor positive breast cancer (HR+BC) has not been extensively investigated. Tumor-specific copy number alterations may predict sensitivity to DNA damaging agents such as Cb. The I-BCT-1 (Improved Breast Cancer Therapy – 1) study was designed to discover biomarkers associated with treatment response, in patients with high-risk HR+BC and TNBC. Methods: Patients with HER2 negative high risk (by Ki67, ER/PgR and/or grade) cT2-4 tumors and no distant metastases were randomized 1:1 to paclitaxel (80 mg/m2, Q1W) alone or in combination with Cb (AUC6 Q3W), for 12 weeks, followed by four cycles of EC90 (epirubicin 90 mg/m2 and cyclophosphamide 600 mg/m2; Q3W). Tumor sampling was performed before treatment start, along the treatment, and at surgery, but only data related to initial sampling is reported here. Complex biomarkers based on somatic copy number alterations (the complex arm-wise aberration index (CAAI), loss of heterozygosity (LOH) and homologous recombination deficiency (HRD)) status were determined based on exome sequencing data. The influence of Cb on chemotherapy delivery and toxicities was analysed. Results: In the 187 patients randomized, the median follow-up was 4.7 years (range 1.0 – 7.9 years). Among the TNBC patients (n=72), the pCR rate was 69% in the Cb group (n=35) vs. 43% in the control group (n=37; p = 0.036). For HR+BC patients (n=115), the pCR rate was 15% in the Cb group vs. 11% in the control group (n.s.), but the fraction of these patients with residual cancer burden (RCB) 0 or 1 increased from 16 to 35% by Cb treatment (p = 0.033). For TNBC patients, event-free survival (EFS) was significantly improved in the Cb group compared to the control group (p=0.017, HR 0.24, 95% CI 0.07-0.86). For HR+BC patients, no improvement in EFS was observed (p=0.684, HR 1.19, 95% CI 0.52-2.71). The proportion of patients with serious adverse events was 42.1% in the Cb group vs. 18.5% in the control group (p<0.001). In TNBC patients receiving Cb, high CAAI was associated with an improved pCR rate (p=0.029). HRD has earlier been investigated as a marker for several DNA targeted therapies, including Cb, but no significant difference in pCR was found in the TNBC patients according to HRD status. In HR+BC patients receiving Cb, the pCR rate was significantly higher in patients with HRD tumors vs. patients with non HRD tumors (p=0.002; not associated with germline BRCA status). In TNBC patients with a low genomic fraction of LOH, the treatment outcome was better in the Cb group than in the control group (p=0.005). In TNBC patients who did not receive Cb, high genomic fraction of LOH was associated with a good prognosis. This was validated in an independent dataset of TNBC patients (n=144, p=0.034), indicating a limited additional effect of Cb for patients with tumor harbouring a high fraction of LOH. Conclusion: In TNBC patients, Cb improves the pCR rate and EFS. In HRBC patients, Cb improves the favorable outcome with RCB 0/1, but not EFS. The results suggest that different complex DNA based biomarkers predict response in hormone receptor positive and triple negative breast cancer: HRD should be further investigated as a predictive marker for Cb benefit in hormone receptor positive patients, while genomic fraction of LOH may be an important marker for treatment selection in triple negative breast cancer patients. Clinical trial identification: EudraCT number: 2013 – 004418 – 17 / ClinicalTrials.gov ID NCT02546232 Citation Format: Olav Engebraaten, Marianne Lislerud Smebye, Arne Valebjørg Pladsen, Øystein Garred, Elin Borgen, Eivind Valen Egeland, Mads Haugland Haugen, Maria Aanesland Dahle, Gunhild Mari Mælandsmo, Eivind Hovig, Sigve Nakken, Anne Alexandra Østgaard, Inger Riise Bergheim, Ellen Schlichting, Helle Skjerven, Anne-Lise Børresen Dale, Erik Wist, Bjørn Naume, Hege Russnes, Ole Christian Lingjærde. Carboplatin added to neoadjuvant chemotherapy in a randomized phase II study: Complex DNA based biomarkers predict response in hormone receptor positive and triple negative breast cancer [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P2-01-26.

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

Background: Early integration of oncology and patient-centered palliative care is the recommended clinical practice model for patients with advanced cancer. General and specific communication skills are necessary to achieve integrated patient-centered care, but require organized training to be adequately mastered. Challenges and barriers on several levels, i.e. organizational, professional and individual may, however, hamper implementation. The development, implementation, and evaluation of such an educational program focusing on communication skills contain many steps, considerations and lessons learned, which are described in this article.Methods: A multi-professional faculty developed, implemented, and evaluated an educational program through a 5-step approach. The program was part of a Norwegian cluster-randomized controlled trial aiming to test the effect of early integration of oncology and palliative care for patients with advanced cancer.Results: The result is the PALLiON educational program; a multi-faceted, evidence-based, and learner-centered program with a specific focus on physicians' communication skills. Four modules were developed: lectures, discussion groups, skills training, and coaching. These were implemented at the six intervention hospitals using different teaching strategies. Evaluation in a subgroup of participants showed a positive appraisal of the group discussions and skills training.Conclusion:We present our experiences and reflections regarding implementation and lessons learned, which should be considered in future developments and implementations; (1) Include experienced faculty with various backgrounds, (2) Be both evidence-based and learner-centered, (3) Choose teaching strategies wisely, (4) Expect resistance and skepticism, (5) Team up with management and gatekeepers, (6) Expect time to fly, and (7) Plan thorough assessment of the evaluation and effect.Trial registration: ClinicalTrials.gov identifier: NCT03088202.

Supplementary figures as referred to in the manuscript

Background: Migrant studies have shown an increase in breast cancer incidence rates among immigrants moving from a breast cancer low-incidence to a high-incidence country. However, 30 years after immigration, it remains equivocal to what degree metabolic factors and ethnic disparities affect breast cancer development and treatment. Methods: Using Cox regression models, we examined the association between ethnicity and breast cancer development, and whether this association varied by pre-diagnostic metabolic profiles among 13 802 women, aged 20-75 years, participating in the population-based Oslo Ethnic Breast Cancer Study. Ethnicity was categorized into: women of Western European descent (reference population) and women of non-western ethnicity (ethnic minority). The ethnic minority women were further subclassified into three groups: 1) South Asian, 2) Middle East and North African, and 3) all other non-western origin women. We defined four pre-diagnostic unfavorable metabolic factors (above median body mass index (>24.6 kg/m2), waist:hip ratio (>0.79), triglyceride:HDL-cholesterol ratio (>0.73), and blood pressure (>96.5 mmHg)), which were combined to define three metabolic profiles: (0-2, 3, and 4 unfavorable metabolic factors). A total of 557 women developed invasive breast cancer during a mean 16.5 years of follow-up. Detailed medical records were obtained. Results: Among women with an unfavorable metabolic profile, South Asian women, compared with Western European women, had a 2.3 times higher breast cancer risk (HR 2.30, 95% CI 1.18-4.49). Furthermore, the ethnic minority women, compared with the Western European women, were suggestively more likely to present with triple-negative breast cancer (OR 2.11, 95% CI 0.97-4.61), and less likely to complete all courses of planned taxane treatment (OR 0.26, 95% CI 0.08-0.82). No differences by ethnicity were observed in physicians’ decisions of planned breast cancer treatment, Conclusions: Our results support that metabolic factors, including body composition, serum lipids and blood pressure, are important when balancing breast cancer prevention and disease management among non-western women migrating from a breast cancer low-incidence to a high-incidence country. However, larger studies are needed. Citation Format: Trygve Lofterød, Hanne Frydenberg, Marit Veierød, Anne Karen Jenum, Jon B Reitan, Erik Wist, Inger Thune. The influence of metabolic factors, migration, and ethnic disparities on breast cancer risk and treatment [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-14-08.

Background Breast cancer risk remains higher in high-income compared with low-income countries. However, it is unclear to what degree metabolic factors influence breast cancer development in women 30 years after immigration from low- to a high-incidence country. Methods Using Cox regression models, we studied the association between pre-diagnostic metabolic factors and breast cancer development, and whether this association varied by ethnicity among 13,802 women participating in the population-based Oslo Ethnic Breast Cancer Study. Ethnic background was assessed and pre-diagnostic metabolic factors (body mass index, waist:hip ratio, serum lipids and blood pressure) were measured. A total of 557 women developed invasive breast cancer, and these women were followed for an additional 7.7 years. Results Among women with an unfavorable metabolic profile, women from south Asia, compared with western European women, had a 2.3 times higher breast cancer risk (HR 2.30, 95% CI 1.18-4.49). Compared with the western European women, the ethnic minority women were more likely to present with triple-negative breast cancer (TNBC) (OR 2.11, 95% CI 0.97-4.61), and less likely to complete all courses of planned taxane treatment (OR 0.26, 95% CI 0.08-0.82). Among TNBC women, above-median triglycerides:HDL-cholesterol (>0.73) levels, compared with below-median triglycerides:HDL-cholesterol (<= 0.73) levels, was associated with 2.9 times higher overall mortality (HR 2.88, 95% CI 1.02-8.11). Conclusions Our results support the importance of metabolic factors when balancing breast cancer prevention and disease management among all women, and in particular among non-western women migrating from a breast cancer low-incidence to a high-incidence country.

Abstract Background: Adjuvant breast cancer treatment may cause metabolic perturbations, such as dyslipidaemia, potentially exacerbating risk of cardiometabolic disease as well as risk of breast cancer recurrence. Physical exercise may have beneficial metabolic effects, but it’s effect on serum lipoprotein- and metabolite profiles during adjuvant breast cancer treatment including chemotherapy is not yet well established. Methods: The women participating in this pilot study of Energy Balance and Breast Cancer Aspects (EBBA)-II, were aged 38-69 years and diagnosed with stage I-II breast cancer. 60 breast cancer patients were randomized after surgery to a control group (n = 29, usual care) or an intervention group (n = 31, intervention), stratified by menopausal status. The patients in the intervention group received a detailed exercise program and met for supervised training sessions in groups of 10-12 women for 60 minutes twice a week during a 12 month period, and were in addition asked to perform at least 60 minutes of exercise at home (a total of 180 minutes of exercise weekly). Fasting serum samples were collected pre-surgery and after six months, and analysed by nuclear magnetic resonance (NMR)-spectroscopy and mass spectrometry. 170 metabolites and 109 lipoprotein subclass variables were quantified and analysed using orthogonalized partial least squares discriminant analysis. Statistical significance was assessed by permutation testing. Single variables were tested with Mann Whitney U-tests or multiple linear regression (NCT02240836). Results: The breast cancer patients (n = 60) had at pre-surgery the following means: Age at diagnosis of 55.4 years (38-69 years), low density lipoprotein (LDL)-cholesterol 145.4 mg/dl (3.76 mmol/L), high density lipoprotein (HDL)-cholesterol 70.4 mg/dl (1.82 mmol/L), and triglycerides 101.9 mg/dl (1.15 mmol/L), and 58.3 % of the patients underwent chemotherapy (paclitaxel/docetaxel/5-FU/epirubicin/cyclophosphamide based adjuvant chemotherapy). Physical exercise ameliorated chemotherapy-induced increases in very low density lipoprotein (VLDL)- and intermediate density lipoprotein (IDL)-associated lipids, and reduced triglyceride enrichment in LDL and HDL compared with chemotherapy controls (p = 0.003). Physical exercise also significantly increased apoA1 (4.6 % increase vs 11.3 % decrease, q = 0.02) and apoA2 (5.2 % increase vs 13.0 % decrease, q = 0.01) compared with chemotherapy control patients. The NMR-measured lipid signal at 1.55-1.60 ppm increased after six months in chemotherapy recipients, but this was attenuated among chemotherapy recipients in the intervention group. No statistically significant effect of physical exercise on serum levels of small-molecular metabolites was detected. Conclusion: Our findings suggest that physical exercise may prevent atherogenic alterations in lipoprotein profile induced by chemotherapy. The results indicate increased HDL particle number- and function, as well as increased triglyceride clearance in the intervention group. Thus, atherogenic alterations in lipoprotein profile may play a role in evaluating breast cancer treatment, and could potentially be biomarkers of importance for breast cancer prognosis and co-morbidity. Citation Format: Torfinn Støve Madssen, Vidar Gordon Flote, Inger Thune, Gro Falkener Bertheussen, Anders Husøy, Steinar Lundgren, Hanne Frydenberg, Erik Wist, Ellen Schlichting, Jon Lømo, Anne McTiernan, Tone Frost Bathen, Guro Fanneløb Giskeødegård. Lipoprotein and metabolite responses to physical exercise during adjuvant breast cancer treatment [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P1-13-01.

Abstract Background: Bevacizumab added to conventional neoadjuvant chemotherapy increase the proportion of patients achieving a pathological complete response (pCR). Identifying patients responding to antiangiogenic therapy have been challenging. The primary objective of this study was to determine the molecular characteristics and treatment induced changes of the primary tumors with reference to treatment response. Clinical outcome measurements according to treatment were exploratory endpoints. Recent updated clinical results, in addition to extended molecular analyses are presented. Methods: A phase II randomized clinical trial of HER2 negative primary tumors of ≥25 mm (n=132) was conducted, treated with neoadjuvant chemotherapy (4xFEC100 followed by taxane-based therapy) with or without the addition of bevacizumab. Biopsies were obtained at the time of diagnosis, after 12 weeks of treatment, and after 25 weeks at surgery. The response was evaluated using the criteria for determining the residual cancer burden (RCB). We derived a mean immune score per patient by calculating the average score from the 770 genes in the nCounter PanCancer immune panel to detect an association between immune activity and response to antiangiogenic therapy. In addition, the median five-year follow-up for disease recurrence are reported. Results: The addition of bevacizumab increased the RCB class 0 (pCR) rate in the study population from 12% to 17% and the rate of “good responders” (RCB class 0 and 1) from 24% to 33%, without reaching statistical significance. A pronounced effect of bevacizumab combination therapy was observed in the hormone receptor (HR) positive tumors, were the percentage of patients achieving RCB class 0 increased from 5% to 20% (Fisher's Exact test, p=0.02). More HR positive patients achieved a good response and fewer patients were poor responders (RCB class 3) in the combination treatment arm (Wilcoxon, p=0.035). Previously, our unsupervised analyses demonstrated an enrichment of immune related genes in pretreatment samples from patients responding to combination therapy. A significantly higher mean immune score (p<0.001) was detected among the HR positive patients who received bevacizumab and achieved RCB class 0 after neoadjuvant treatment (n=11, 20%) . Five-year follow-up data revealed a total of 21 events in the study population; 9 relapses in patients treated with combination therapy, and 12 relapses in patients treated with chemotherapy only. DFS was not statistically different between the treatment groups (log rank, p=0.4257). However, among the patients achieving a good response an improved DFS was observed for those treated with combination therapy (1/22 vs. 5/16, log rank, p=0.0254). Conclusion: Among locally advanced HER2-negative HR positive breast cancer patients, the addition of bevacizumab to neoadjuvant chemotherapy increased the rate of good responders and improved the DFS among these patients. An increased primary tumor immune score may predict good response to neoadjuvant antiangiogenic therapy in HR positive disease. Further studies are needed to validate the use of such immune panels for selection of patients most likely to benefit from antiangiogenic therapy. Citation Format: Gythfeldt HvdL, Engebråten O, Naume B, Wist E, Borgen E, Lien T, Lindgjærde OC, Garred O, Schlichting E, Silwal-Pandit L, Borresen-Dale AL. A translational and five-year clinical update in patients treated with neoadjuvant chemotherapy randomized to bevacizumab or control in HER2 negative breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-07-01.

Chemotherapeutic agents such as anthracyclines and taxanes are commonly used in the neoadjuvant setting. Bevacizumab is an antibody which binds to vascular endothelial growth factor A (VEGFA) and inhibits its receptor interaction, thus obstructing the formation of new blood vessels.

Background Breast cancer treatment has metabolic side effects, potentially affecting risk of cardiovascular disease (CVD) and recurrence. We aimed to compare alterations in serum metabolites and lipoproteins during treatment between recipients and non-recipients of chemotherapy, and describe metabolite profiles associated with treatment-related weight gain. Methods This pilot study includes 60 stage I/II breast cancer patients who underwent surgery and were treated according to national guidelines. Serum sampled pre-surgery and after 6 and 12 months was analysed by MR spectroscopy and mass spectrometry. In all, 170 metabolites and 105 lipoprotein subfractions were quantified. Results The metabolite and lipoprotein profiles of chemotherapy recipients and non-recipients changed significantly 6 months after surgery ( p < 0.001). Kynurenine, the lipid signal at 1.55–1.60 ppm, ADMA, 2 phosphatidylcholines (PC aa C38:3, PC ae C42:1), alpha-aminoadipic acid, hexoses and sphingolipids were increased in chemotherapy recipients after 6 months. VLDL and small dense LDL increased after 6 months, while HDL decreased, with triglyceride enrichment in HDL and LDL. At baseline, weight gainers had less acylcarnitines, phosphatidylcholines, lyso-phosphatidylcholines and sphingolipids, and showed an inflammatory lipid profile. Conclusion Chemotherapy recipients exhibit metabolic changes associated with inflammation, altered immune response and increased risk of CVD. Altered lipid metabolism may predispose for treatment-related weight gain.

Table S4. Aberrant genes in > 30% of the samples at week 25 within the Combination arm. (XLSX 25 kb)

High triglycerides and low levels of high density lipoprotein (HDL)-cholesterol are observed to promote tumor growth. However, whether breast cancer heterogeneity may explain the contradictory influence of triglycerides and cholesterol observed on breast cancer prognosis remains unclear.

Table S5. Aberrant genes in > 30% of the samples at week 12 within the Chemotherapy arm. (XLSX 71 kb)

Abstract 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

Methods. Immunohistochemical analysis for Ki-67. Figure S1. Distribution of PAM50 subtypes within subgroups based on HR and HER2 status. Bars represent percentage of total in each HR/HER2− group. Number is displayed on top of the bar. Figure S2. ROR score within each of the PAM50 subtypes for all patients (R statistical software package). Figure S3. Kaplan-Meier plots of BCSS (S3a) and DDFS (S3b) according to HR/HER2 subtypes in all 653 patients (a) and according to PAM50 subtypes within different HR/HER2 (b–e) subgroups. p Values were derived from log-rank tests. Figure S4. Kaplan-Meier plots of BCSS according to ROR categories for node-negative (a) and node-positive (b) HR+/HER2− patients. p Values were derived from log-rank tests. Figure S5. Kaplan-Meier plots of BCSS (a and b) and DDFS (c and d) according to ROR categories for node-negative luminal A HR+/HER2− patients with no adjuvant treatment (a, c) or treated with tamoxifen only (b, d). p Values were derived from log-rank tests. Figure S6. Correlation between Ki-67 expression and ROR score for the HR+/HER2− patients. ρ = 0.62, p