Melanoma is currently divided on a genetic level according to mutational status. However, this classification does not optimally predict prognosis. In prior studies, we have defined gene expression phenotypes (high-immune, pigmentation, proliferative and normal-like), which are predictive of survival outcome as well as informative of biology. Herein, we employed a population-based metastatic melanoma cohort and external cohorts to determine the prognostic and predictive significance of the gene expression phenotypes. We performed expression profiling on 214 cutaneous melanoma tumors and found an increased risk of developing distant metastases in the pigmentation (HR, 1.9; 95% CI, 1.05-3.28; P=0.03) and proliferative (HR, 2.8; 95% CI, 1.43-5.57; P=0.003) groups as compared to the high-immune response group. Further genetic characterization of melanomas using targeted deep-sequencing revealed similar mutational patterns across these phenotypes. We also used publicly available expression profiling data from melanoma patients treated with targeted or vaccine therapy in order to determine if our signatures predicted therapeutic response. In patients receiving targeted therapy, melanomas resistant to targeted therapy were enriched in the MITF-low proliferative subtype as compared to pre-treatment biopsies (P=0.02). In summary, the melanoma gene expression phenotypes are highly predictive of survival outcome and can further help to discriminate patients responding to targeted therapy.

There is increasing evidence for the prognostic role of hypoxia in high-grade soft tissue sarcoma, and these data suggest that HIF-1α expression represents a candidate prognostic biomarker for clinical application in high-grade leiomyosarcoma and undifferentiated pleomorphic sarcoma.

Purpose: Androgen receptor (AR) status in breast cancer has received renewed interest over the last years especially in triple-negative disease (TNBC), but the prognostic value is still under debate. The aim of this study was to assess the distribution and prognostic value of AR in early breast cancer patients with or without adjuvant endocrine treatment. Patients and methods: AR was assessed on tissue microarray with the AR 441 antibody (Thermo Scientific) on a cohort consisting of 471 patients derived from two clinical studies: (1) 208 premenopausal node-negative patients of which 87% had received no adjuvant medical treatment and (2) 263 estrogen receptor (ER)+ and ER-, node-positive and –negative patients treated with 2 years of adjuvant tamoxifen. Nuclear AR was divided in 5 groups: 0-1%, 2-10%, 11-50%, 51-75%, and 76-100% positive cells, scored as 0-4. Cox proportional hazards regression, stratified by study, was used to model the impact of the prognostic factors on distant disease-free survival (DDFS), both using trend tests and a cut-off for positivity set at >10%, and log-rank tests to compare survival in different strata. Due to non-proportional hazards, the analysis was restricted to the first 5 years after diagnosis, a time period during which 95 patients developed distant recurrences. Results: 76% of all patients were AR+, and 89%, 48%, and 23% of the ER+, ER-, and TNBC, respectively. Positive associations were observed between AR, ER and progesterone receptor status (PgR), negative associations with Ki67, and histological grade, but no associations with tumour size, age or Human Epidermal Growth Factor Receptor 2 (HER2). In univariable analysis, when divided into 5 groups, AR was a prognostic factor for DDFS with a Hazard Ratio (HR) of 0.86 per step in fraction score (95% Confidence Interval (CI): 0.76-0.98, p=0.018), as was HER2, age, size, grade, node-status, PgR, and ER status. In the Kaplan-Meier curves for each study, a similar but weaker trend was found (log-rank test for trend p=0.14 and 0.057 for cohort 1 and 2, respectively). With a cut-off at 10%, a similar HR was found (HR=0.67, 95% CI:0.43-1.05, p=0.078). In multivariable analysis, adjusted for grade, tumour size, HER2, ER, node-status, and age, AR did not retain independent prognostic value (HR 1.04 95% CI:0.88-1.23, p=0.66). In the TNBC patients there were no significant differences in DDFS in the AR+ vs AR-patients, possibly due to few events and a small population (n=20/75). Conclusion: This study demonstrates that AR is a weak prognostic factor for recurrence in a cohort consisting of node-negative premenopausal patients without endocrine treatment and patients who have received adjuvant endocrine treatment. There was however no independent value in multivariable analyses. It is noteworthy that there were 23% AR positive TNBC patients, for whom there is currently no available targeted treatment. There are several ongoing studies with AR-targeted treatment in the metastatic setting, which if proven effective, may be transferred to studies in the adjuvant setting with the goal of improving long-term prognosis for TNBC. Taken together, AR may be clinically helpful for prognostic considerations and for selection of adjuvant treatment. Citation Format: Werner-Hartman L, Folkesson E, Nodin B, Malmström P, Fernö M, Nimeus E, Klintman M. Androgen receptor in early breast cancer: Distribution and prognostic value. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-08-30.

Background Proliferation is a key feature in breast cancer and also a clinical important factor for prognosis and treatment prediction. In the St Gallen Consensus of 2011, immunohistochemically analysed Ki67 was adopted as a surrogate marker to distinguish the “luminal A” from the “luminal B” subtype, in order to select patients with estrogen receptor positive disease expected to benefit from adjuvant chemotherapy. The mouse monoclonal antibody MIB1 is the generally accepted antibody for assessment of Ki67. However, other antibodies have been developed, e.g. the rabbit monoclonal antibody SP6. The assessment of Ki67 with MIB1 is unfortunately associated with a considerable lack of reproducibility. Rabbit monoclonal antibodies generally tend to have higher specificity without loss of sensitivity, compared to corresponding mouse monoclonal antibodies. SP6 has also been found to have reduced background staining compared to MIB1. According to these advantages, SP6 may be an alternative to MIB1 for routine staining of Ki67. Any methodological modification should, according to international recommendations, be compared against a clinically validated assay and demonstrate acceptable concordance before being introduced in clinical routine. The analysis of Ki67 with SP6 therefore needs to be evaluated and compared to MIB1 in a cohort of breast cancer patients with clinical follow-up. Aims To compare the antibodies SP6 and MIB1 for immunohistochemical assessment of Ki67 in primary breast cancer regarding prognostic strength and reproducibility of the evaluation. Methods Tissue microarray from a cohort of 237 premenopausal women with node-negative breast cancer was used for assessment of Ki67, with both SP6 and MIB1, by three different investigators. The 7th decile was applied for defining cut-off. Distant disease-free survival (DDFS) was used as endpoint and the follow-up was restricted to 5 years. Results Ninety per cent of the samples were classified into the same group, either high or low Ki67, irrespective of antibody used. Ki67 (high vs. low), analysed with both antibodies was associated to DDFS (34 events) in the univariable analyses (SP6: HR 2.6, 95% CI 1.3-5.2, p = 0.01 and MIB1: HR 2.8, 95% CI 1.4-5.7 p = 0.004) and showed borderline significance for DDFS in the multivariable analyses, also including HER2, age, and tumour size (SP6: HR 2.0, 95% CI 0.93-4.5, p = 0.074 and MIB1: HR 2.2, 95% CI 0.97-4.8, p = 0.058). The agreement between different assessors was somewhat higher for MIB1 than for SP6 (kappa-values 0.83-0.88 vs. 0.72-0.77). Conclusions SP6 was not superior to MIB1 and the two antibodies were comparable in the assessment of Ki67 for prognostic considerations in primary breast cancer. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-11-15.

Introduction Breast conserving surgery (BCS) with postoperative radiotherapy (RT) is recommended for the majority of patients with early breast cancer. Meta-analyses of randomized trials have shown reduction of breast cancer and overall mortality after RT but increased cardiovascular mortality Aim To evaluate side effects of RT after BCS Patients and Methods Trial SweBCG 91RT randomized 1187 women with stage 1-IIA breast cancer after BCS to RT or to no further treatment. RT was administered as tangential opposing beams to the breast to a target dose of 48-54 Gy. Three-dimensional dose planning on multiple CT slices 10 mm apartwas used for 69% of the patients and for the remainder two-dimensional planning. All hospital records and radiotherapy charts were monitored. Update of mortality, cause of death and morbidity was made using the Swedish personal identification numbers and the following Swedish national registers: the Cancer Register, the Population Register, the Cause of Death Register, the Inpatient and the SWEDEHEART registers. For analyses the diagnoses were grouped as follows: breast cancer, cardiovascular disease, cerebrovascular disease, lung cancer and benign pulmonary disease Statistics Overall mortality was calculated using the Kaplan-Meier method, whereas cumulative incidence functions with other causes of death as competing events were used for cause-specific mortality and morbidities. For all outcomes, log-rank tests were used to compare between treatment groups up to 20 years. We studied two populations: 1) patients according to intention to treat for overall and breast cancer mortality and 2) patients treated per protocol for side effects. Cause-specific death was allocated to a certain group when either the underlying or a contributing cause belonged to the disease group Results After 20 years, overall mortality was 42.9% after BCS and 42.5% after BCS+RT (p=0.8), and breast cancer mortality was 18.0% vs 15.8% (p=0.3). The cumulative incidence of mortality from heart disease was 12.4% after BCS and 13.0% after BCS+RT (p=0.8) with no difference for left or right side. Ischemic heart disease and congestive heart failure were the most common cardiac diagnoses. The cumulative incidence of cerebrovascular mortality was 3.4% among controls and 6.7% after RT (p=0.016). Of the patients with cerebrovascular death, 50% also had a cardiac cause of death. Other cause-specific mortalities investigated were similar regardless of RT: lung cancer 1.7% vs 1.9% (p=1.0), benign pulmonary disease 7.1% vs 6.4% (p=0.5) Morbidity Morbidity outcomes were also similar in control and irradiated patients: the cumulative incidence of hospital admissions with cardiac diagnoses 29.7% vs 31.0% (p=0.7), and for cerebrovascular morbidity 11.6% vs 13.7% (p= 0.33) respectively Radiation exposure The original dose plans were retrieved from 125 patients. Doses to organs at risk were recalculated. Median of mean heart dose was 3.0 Gy (1.1-8.2) for left and 1.0 Gy (0.5-2.5) for right-sided RT Conclusions After 20 years tangential RT in a randomized trial was not associated with increased cardiac mortality. A minor increase in cerebrovascular mortality was seen, but the causality is unclear. Citation Format: Malmström P, Karlsson P, Holmberg E, Lundstedt D, Holmberg L, Werner-Hartman L, Koul S, Kjellén E, Killander F. No increased cardiovascular mortality after twenty years in a randomized trial of radiotherapy after breast conserving surgery, SweBCG 91RT, from the Swedish breast cancer group [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 PD8-10.