No validated biomarkers are available to monitor neoadjuvant treatment effects for breast cancer. Natural killer cell activity (NKA) has shown prognostic potential in other cancers. This study examined the association between NKA and treatment response. Patients had blood samples collected at baseline, before each treatment, and pre- and postoperatively. Plasma IFNγ levels were measured by ELISA as a surrogate marker of NKA, with 250 pg/mL as the cutoff for normal versus low NKA. Study endpoints were residual cancer burden (RCB) class, overall survival (OS), and invasive disease-free survival (IDFS). Seventy-eight patients were included. The five-year IDFS was 88.1% (95% confidence interval (CI) 73.7–94.9%) for patients with normal NKA versus 71.5% (95% CI 40.6–88.2%) for patients with low NKA (p = 0.049) preoperatively. At the fifth treatment cycle, the median IFNγ was 11 pg/mL (interquartile range (IQR) 0.5–124 pg/mL) in patients receiving supportive prednisolone and 753 pg/mL (IQR 192–1580 pg/mL) in patients not receiving supportive prednisolone. At the sixth treatment cycle, the corresponding values were 7 pg/mL (IQR 0–15 pg/mL) and 806 pg/mL (IQR 180–1631 pg/mL) (p < 0.0001). In conclusion, NKA may have prognostic potential as a biomarker. This study is the first to demonstrate that prednisolone impairs NKA measurement in breast cancer patients.

IntroductionShared decision making (SDM) has become a crucial element on the political agenda and represents a vital aspect of modern healthcare. However, successful implementation of SDM highly depends on the attitude of clinicians towards SDM. The overall aim of our study was to explore the experience of oncologists and nurses with SDM using the Decision Helper, an in-consultation decision aid, at four Danish radiotherapy departments.MethodsSemi-structured interviews were conducted with 20 clinicians. The participants were selected using purposive sampling to include nurses and oncologists, male and female, with different levels of experience with SDM and clinical work. The analysis was a data-driven, iterative process with inductive coding of all interviews and meaning condensation.ResultsTwo main themes emerged: “Using the Decision Helper changes the consultation” and “Change of attitude among Danish oncologists”. Each of the two themes included four elaborative subthemes, which are reported with supporting citations in this paper. In brief, the use of SDM and the Decision Helper should ideally be adjusted to the individual patient and depends highly on the oncologist. The participants described ambitions towards "making the right decision for this patient at this time”. The healthcare system, however, has pitfalls that may hinder SDM, e.g. rigid interpretation of guideline-based recommendations.ConclusionUsing an in-consultation Decision Helper has the potential for individualized, structured patient engagement in decision making. There is a need for patient decision aids in clinical guidelines to ensure patient engagement in decision making.Micro AbstractClinicians’ experience of shared decision making and their use of patient decision aids are crucial for the implementation of shared decision making. Fourteen oncologists and six nurses share their experiences in this paper. Overall, while our findings indicate that the use of an in-consultation patient decision aid (the Decision Helper) requires a change in their practice, the clinicians report that using decision aids has the potential for individualized, structured patient involvement in decision making.

e13106 Background: The Homeobox A9 (HOXA9) gene may act as a tumor suppressor in carcinomas and induce breast cancer progression. The clinical potential of methylated HOXA9 (meth-HOXA9) as a biomarker is unclear. Therefore, this study aims to examine the association between meth-HOXA9 in tumor tissue, meth-HOXA9 in plasma, and mortality. Additionally, the study examines the association between meth-HOXA9 in metastatic tissue and plasma at the time of recurrence. Methods: The study enrolled 51 patients with recurrent breast cancer between 2011 and 2015 recruited at the Department of Oncology, Vejle Hospital, University Hospital of Southern Denmark. Tissue samples were obtained from the primary tumor at the time of primary surgery between 1993 and 2013. Tissue samples from the metastases and plasma samples were obtained at the time of recurrence. The study follow-up period ran until death or January 2019. All samples were analyzed for meth-HOXA9 using droplet digital PCR. The optimal cut-off for meth-HOXA9 in tissue was established at ≥7.4% using tissue specimens from independent breast cancer patients and healthy controls. In plasma samples, the detection of ≥5 meth-HOXA9-containing droplets resulted in a positive test. Using Cox regression, we estimated crude and age-adjusted hazard ratios (HR) for mortality with 95% confidence intervals (CI) comparing patients with detectable and undetectable meth-HOXA9 in both tumor tissue and plasma. Fischer’s exact test was used to compare plasma and metastasis meth-HOXA9 status. Results: Among the 50 patients with data on tumor tissue meth-HOXA9, 80% had detectable meth-HOXA9 in the primary tumor, but there was no increased mortality when comparing patients with detectable and undetectable meth-HOXA9 (HR 1.09; CI 0.47-2.52). Only 34 patients had data on plasma meth-HOXA9 and 62% of these had detectable meth-HOXA9 at the time of recurrence. Patients with detectable meth-HOXA9 in plasma had higher mortality than those without (HR 3.95; CI 1.50-10.37). Among the 20 patients with data on plasma and metastatic tissue meth-HOXA9, there was no significant association between meth-HOXA9 in plasma and metastatic tissue (p = 1.00). Conclusions: These results indicate that detectable plasma meth-HOXA9 at the time of breast cancer recurrence is associated with increased mortality. However, there is no association between tumor tissue meth-HOXA9 and mortality. Metastatic tissue and plasma meth-HOXA9 are not associated. Our data support the potential relevance of plasma meth-HOXA9 as a biomarker for prognosis and mortality in recurrent breast cancer, which calls for further investigation.

ImportanceValidation of a new method for prognostication of de novo metastatic breast cancer (dnMBC) to better reflect the heterogenecity of the disease.ObjectiveTo perform external methodological validation of the Plichta staging system, a novel prognostic system for de novo metastatic breast cancer (dnMBC).Design, Setting, and ParticipantsThis retrospective cohort study used a multicenter, nationwide, population-based Danish Breast Cancer Group database to validate the new method. Participants were patients with dnMBC diagnosed between 2010 and 2019. Data were analyzed from April to June 2023.Main outcomes and measuresA recursive partitioning analysis (RPA) was performed, as demonstrated by Plichta and colleagues, to group patients with similar overall survival (OS) based on clinical factors. The main outcome was to group patients into 4 prognostic groups based on 3-year OS as stage IVa, greater than 70%; stage IVb, 50% to 70%; stage IVc, 25% to less than 50%; or stage IVd, less than 25%. Bootstrapping was applied for 1000 iterations, with final stage assignments based on the most commonly occurring assignment.ResultsA total of 1859 women were included with a median (IQR) age of 69 (57-77) years. With a median potential follow-up of 89.9 (95% CI, 86.4-95.1) months and a median OS of 31.7 (95% CI, 29.5-34.1) months, the RPA stratified patients into 10 groups, with organ sites, estrogen receptor status, and human epidermal growth factor receptor 2 status as the key clinical factors. Three-year survival rates ranged from 62% (95% CI, 56%-69%) to 8% (95% CI, 3%-21%), which were further combined into 3 stage groups: IVb, 59.4% (95% CI, 56.2%-62.8%); IVc, 39.4% (95% CI, 36.2%-43.0%); and IVd, 15.4% (95% CI, 11.2%-21.3%) (P < .001). Following bootstrapping, an IVa group emerged, resulting in 4 stage groups with separate 3-year OS rates identified as IVa, 75.8% (95% CI, 67.8%-84.7%); IVb, 58.8% (95% CI, 55.5%-62.3%); IVc, 39.2% (95% CI, 35.8%-43.0%); and IVd, 14.4% (95% CI, 10.8%-19.4%) (P < .001).Conclusions and relevanceThese findings provide external and independent validation of the methods applied in the novel Plichta staging system for dnMBC. This could guide future revisions of the current American Joint Committee on Cancer staging guidelines and may be incorporated as a stratification factor in clinical trials.

Background and purposeShared decision making (SDM) is a patient engaging process advocated especially for preference-sensitive decisions, such as adjuvant treatment after breast cancer. An increasing call for patient engagement in decision making highlights the need for a systematic SDM approach. The objective of this trial was to investigate whether the Decision Helper (DH), an in-consultation patient decision aid, increases patient engagement in decisions regarding adjuvant whole breast irradiation.Material and methodsOncologists at four radiotherapy units were randomized to practice SDM using the DH versus usual practice. Patient candidates for adjuvant whole breast irradiation after breast conserving surgery for node-negative breast cancer were eligible. The primary endpoint was patient-reported engagement in the decision process assessed with the Shared Decision Making Questionnaire (SDM-Q-9) (range 0-100, 4 points difference considered clinical relevant). Other endpoints included oncologist-reported patient engagement, decisional conflict, fear of cancer recurrence, and decision regret after 6 months.ResultsOf the 674 included patients, 635 (94.2%) completed the SDM-Q-9. Patients in the intervention group reported higher level of engagement (median 80; IQR 68.9 to 94.4) than the control group (71.1; IQR 55.6 to 82.2; p< 0.0001). Oncologist-reported patient engagement was higher in the invention group (93.3; IQR 82.2 to 100) compared to control group (73.3; IQR 60.0 to 84.4) (p < 0.0001).ConclusionPatient engagement in medical decision making was significantly improved with the use of an in-consultation patient decision aid compared to standard. The DH on adjuvant whole breast irradiation is now recommended as standard of care in the Danish guideline.

Background Clinical trials have shown zoledronic acid as a potent bisphosphonate in preventing bone loss, but with varying potency between patients. Human osteoclasts ex vivo reportedly displayed a variable sensitivity to zoledronic acid > 200-fold, determined by the half-maximal inhibitory concentration (IC50), with cigarette smoking as one of the reported contributors to this variation. To reveal the molecular basis of the smoking-mediated variation on treatment sensitivity, we performed a DNA methylome profiling on whole blood cells from 34 healthy female blood donors. Multiple regression models were fitted to associate DNA methylation with ex vivo determined IC50 values, smoking, and their interaction adjusting for age and cell compositions. Results We identified 59 CpGs displaying genome-wide significance ( p < 1e−08) with a false discovery rate (FDR) < 0.05 for the smoking-dependent association with IC50. Among them, 3 CpGs have p < 1e−08 and FDR < 2e−03. By comparing with genome-wide association studies, 15 significant CpGs were locally enriched (within < 50,000 bp) by SNPs associated with bone and body size measures. Furthermore, through a replication analysis using data from a published multi-omics association study on bone mineral density (BMD), we could validate that 29 out of the 59 CpGs were in close vicinity of genomic sites significantly associated with BMD. Gene Ontology (GO) analysis on genes linked to the 59 CpGs displaying smoking-dependent association with IC50, detected 18 significant GO terms including cation:cation antiporter activity, extracellular matrix conferring tensile strength, ligand–gated ion channel activity, etc. Conclusions Our results suggest that smoking mediates individual sensitivity to zoledronic acid treatment through epigenetic regulation. Our novel findings could have important clinical implications since DNA methylation analysis may enable personalized zoledronic acid treatment.

Neoadjuvant treatment of breast cancer is applied to an increasing extent, but treatment response varies and side effects pose a challenge. The vitamin E isoform delta-tocotrienol might enhance the efficacy of chemotherapy and reduce the risk of side effects. The aim of this study was to investigate the clinical effect of delta-tocotrienol combined with standard neoadjuvant treatment and the possible association between detectable circulating tumor DNA (ctDNA) during and after neoadjuvant treatment with pathological treatment response. This open-label, randomized phase II trial included 80 women with newly diagnosed, histologically verified breast cancer randomized to standard neoadjuvant treatment alone or in combination with delta-tocotrienol. There was no difference in the response rate or frequency of serious adverse events between the two arms. We developed a multiplex digital droplet polymerase chain reaction (ddPCR) assay for the detection of ctDNA in breast cancer patients that targets a combination of two methylations specific for breast tissue (LMX1B and ZNF296) and one cancer specific methylation (HOXA9). The sensitivity of the assay increased when the cancer specific marker was combined with the ones specific to breast tissue (p < 0.001). The results did not show any association between ctDNA status and pathological treatment response, neither at midterm nor before surgery.

Shared decision making (SDM) is on the international healthcare agenda among patient advocacy groups and decision makers in healthcare worldwide [1–5]. In Denmark, implementation of SDM with all ca...

Objective: Concerns of increased time consumption and of the impact on clinical decisions may restrain doctors from shared decision making (SDM). This paper evaluates consultation length and decisions made when using an in-consult patient decision aid (PtDA). Methods: This prospective cohort study compared an unexposed cohort with a cohort exposed to SDM and a PtDA in two preference-sensitive decision situations: invasive lung cancer diagnostics and adjuvant treatment for early breast cancer. Outcome measures were consultation length and decisions made. Results: The study included 261 consultations, 115 were in the SDM-exposed cohort. Consultations were inconsiderably longer in the SDM cohort; 2 min, 11 s (p = 0.2217) for lung cancer diagnostics and 3 min, 57 s (p = 0.1128) for adjuvant breast cancer treatment. In lung cancer diagnostics, consultation length became more uniform and decisions tended to become conservative after introduction of SDM. For adjuvant breast cancer, slightly more patients in the SDM cohort chose to decline treatment. Conclusion: Shared decision making did not take significantly longer time and led to slightly more conservative decisions. Practice implications: SDM may be implemented without considerable impact on consultation length. The impact on clinical decisions depends mainly on the clinical situation. (c) 2020 Elsevier B.V. All rights reserved.

Brain metastases are frequent in patients with lung cancer and a major cause of morbidity and mortality. Finding a biomarker predicting brain metastases could facilitate early start of treatment and thereby reduce morbidity and possibly improve overall survival. Previous studies suggest S100B as a possible biomarker for this purpose. This prospective study enrolled 185 patients with newly diagnosed stage IV non-small cell lung cancer (NSCLC). A total of 22 patients had brain metastases verified by magnetic resonance imaging or computed tomography at the time of enrollment. Serum S100B levels were measured in blood samples collected prior to any treatment from 22 patients who had brain metastases at enrollment and from 50 patients randomly selected from the remaining 163 patients without brain metastases at enrollment. No statistically significant difference was found in the levels of serum S100B between patients with and without brain metastases [range 0.018-0.209 μg/l, mean 0.049 μg/l, 95% confidence interval (CI), 0.032‐0.061 μg/l] and (range 0.016-0.130 μg/i, mean 0.044 μg/l, 95% CI, 0.037-0.051 μg/l), respectively, (P=0.852). Univariate analysis of prognostic factors for S100B indicated a correlation (P<0.2) with sex (P=0.088) and histology (adenocarcinoma vs. squamous cell carcinoma/others) (P=0.028). In the multivariate analysis only histology (P=0.029) remained statistically significant. Conclusion: The present study found no significant correlation between the level of serum S100B and the presence of brain metastases in patients with advanced NSCLC. The clear cut-off of S100B in patients with and without brain metastases reported in other studies could not be verified in this study. Further studies investigating the role of S100B as a biomarker for brain metastases in non-small cell lung cancer are warranted.

Background and purpose: This study presents Danish consensus guidelines for delineation of the heart and cardiac substructures across relevant Danish Multidisciplinary Cancer Groups. Material and methods: Consensus guidelines for the heart and cardiac substructures were reached among 15 observers representing the radiotherapy (RT) committees of four Danish Multidisciplinary Cancer Groups. The guidelines were validated on CT scans of 12 patients, each with five independent contour sets. The Sorensen-Dice similarity coefficient (DSC), the distance between the centers of the arteries and the mean surface distance were used to evaluate the inter-observer variation. Results: National guidelines for contouring the heart and cardiac substructures were achieved. The median DSC was 0.78-0.96 for the heart and the four cardiac chambers. For the four substructures of the left ventricle, the median DSC was 0.35-0.57. The coronary arteries were contoured in ten segments, with the best agreement for the left anterior descending coronary artery segments, with a median distance between the arteries ranging from 2.4-4.4 mm. The median variation was 3.7-12.8 mm for the right coronary artery segments and 3.7-6.2 mm for the left circumflex coronary artery segments, with the most pronounced inter-observer variation in the distal segment for all three coronary arteries. Conclusion: National guidelines for contouring the heart and cardiac substructures were developed across relevant Danish Multidisciplinary Cancer Groups, where RT dose to the heart is of concern. The inter-observer contour overlap was best for the heart and chambers and decreased for smaller structures. (C) 2020 Elsevier B.V. All rights reserved. Radiotherapy and Oncology 150 (2020) 121-127

The epidermal growth factor receptor (EGFR) and its ligands are involved in cancer pathogenesis and they might serve as circulating biomarkers. The current study aims to investigate if abnormal pre-treatment serum levels of EGFR and EGFR ligands are present in women with early-stage breast cancer and if up- or downregulation of EGFR and EGFR ligands occur in defined patient subgroups. Pre-treatment serum samples were obtained from 311 women with newly diagnosed early-stage breast cancer and from 419 healthy women and analysed for EGFR and the ligands: Epidermal growth factor (EGF), heparin-binding epidermal growth factor (HBEGF), betacellulin (BTC), amphiregulin (AREG), and transforming growth factor α (TGF-α). Previously, age-dependent 95% reference intervals for EGFR and the EGFR ligands have been established based on the healthy women population. S-EGFR, S-EGF, S-HBEGF, S-AREG, and S-TGFα were all significantly different in women with breast cancer compared to healthy women (p < 0.05). Elevated S-EGFR, according to the reference intervals, was present in 11.3% of breast cancer patients, whereas decreased S-EGF was found in 11.6%. Elevated S-EGFR was associated with estrogen receptor positivity of tumor (ER+) and a subgroup of ER + breast cancer patients showed markedly elevated S-EGFR (>120 ng/mL).

Brain metastases are frequent in patients with lung cancer and a major cause of morbidity and mortality. Finding a biomarker predicting brain metastases could facilitate early start of treatment and thereby reduce morbidity and possibly improve overall survival. Previous studies suggest S100B as a possible biomarker for this purpose. This prospective study enrolled 185 patients with newly diagnosed stage IV non-small cell lung cancer (NSCLC). A total of 22 patients had brain metastases verified by magnetic resonance imaging or computed tomography at the time of enrollment. Serum S100B levels were measured in blood samples collected prior to any treatment from 22 patients who had brain metastases at enrollment and from 50 patients randomly selected from the remaining 163 patients without brain metastases at enrollment. No statistically significant difference was found in the levels of serum S100B between patients with and without brain metastases [range 0.018-0.209 µg/l, mean 0.049 µg/l, 95% confidence interval (CI), 0.032-0.061 µg/l] and (range 0.016-0.130 µg/i, mean 0.044 µg/l, 95% CI, 0.037-0.051 µg/l), respectively, (P=0.852). Univariate analysis of prognostic factors for S100B indicated a correlation (P<0.2) with sex (P=0.088) and histology (adenocarcinoma vs. squamous cell carcinoma/others) (P=0.028). In the multivariate analysis only histology (P=0.029) remained statistically significant. Conclusion: The present study found no significant correlation between the level of serum S100B and the presence of brain metastases in patients with advanced NSCLC. The clear cut-off of S100B in patients with and without brain metastases reported in other studies could not be verified in this study. Further studies investigating the role of S100B as a biomarker for brain metastases in non-small cell lung cancer are warranted.

Epidermal growth factor receptor (EGFR) and its ligands are involved in cancer pathogenesis. The emerging role of treatments co-targeting the EGFR system in breast cancer has increased the need to identify companion biomarkers. The aim of this study is to investigate whether pretreatment serum levels of EGFR and EGFR ligands in early-stage breast cancer patients might provide prognostic information as a stepping stone for further investigation. The study, which included 311 early-stage breast cancer patients, investigated associations between preoperative serum levels of EGFR and EGFR ligands (epidermal growth factor, heparin-binding epidermal growth factor (HBEGF), amphiregulin, transforming growth factor-α and betacellulin) and survival. Cutoffs were determined using Youden’s method, and overall survival (OS) and invasive disease-free survival (IDFS) were evaluated using Cox regression. Preoperative S-EGFR < 60.3 ng/mL was associated with shorter OS and IDFS in both univariate analyses and when adjusting for standard prognostic factors (p < 0.05). Preoperative S-HBEGF < 21.4 pg/mL was associated with shorter OS in both univariate and multivariate analyses, whereas association with shorter IDFS could only be demonstrated in the univariate analysis. In conclusion, our study demonstrated shorter survival in early-stage breast cancer patients who had low pretreatment levels of either S-EGFR or S-HBEGF.