Growth Factor Receptor 2-positive Tumors Research Articles

Healthy Eating and Mortality among Breast Cancer Survivors: A Systematic Review and Meta-Analysis of Cohort Studies.

This systematic review examined the effect of diet quality, defined as adherence to healthy dietary recommendations, on all-cause and breast cancer-specific mortality. Web of Science, Medline, CINAHL, and PsycINFO databases were searched to identify eligible studies published by May 2021. We used a random-effects model meta-analysis in two different approaches to estimate pooled hazard ratio (HR) and 95% confidence interval (CI) for highest and lowest categories of diet quality: (1) each dietary quality index as the unit of analysis and (2) cohort as the unit of analysis. Heterogeneity was examined using Cochran’s Q test and inconsistency I2 statistics. The risk of bias was assessed by the Newcastle–Ottawa Scale for cohort studies, and the quality of evidence was investigated by the GRADE tool. The analysis included 11 publications from eight cohorts, including data from 27,346 survivors and seven dietary indices. Both approaches yielded a similar effect size, but cohort-based analysis had a wider CI. Pre-diagnosis diet quality was not associated with both outcomes. However, better post-diagnosis diet quality significantly reduced all-cause mortality by 21% (HR = 0.79, 95% CI = 0.70, 0.89, I2 = 16.83%, n = 7) and marginally reduced breast cancer-specific mortality by 15% (HR = 0.85, 95% CI = 0.62, 1.18, I2 = 57.4%, n = 7). Subgroup analysis showed that adhering to the Diet Approaches to Stop Hypertension and Chinese Food Pagoda guidelines could reduce breast cancer-specific mortality. Such reduction could be larger for older people, physically fit individuals, and women with estrogen receptor-positive, progesterone receptor-negative, or human epidermal growth factor receptor 2-positive tumors. The risk of bias in the selected studies was low, and the quality of evidence for the identified associations was low or very low due to imprecision of effect estimation, inconsistent results, and publication bias. More research is needed to precisely estimate the effect of diet quality on mortality. Healthcare providers can encourage breast cancer survivors to comply with healthy dietary recommendations to improve overall health. (Funding: University of Central Florida Office of Undergraduate Research, Registration: PROSPERO-CRD42021260135).

Ultrasound Imaging Morphology is Associated with Biological Behavior in Invasive Ductal Carcinoma of the Breast.

Objectives:Ultrasound (US) is commonly used for diagnostic evaluation of breast lesions. The objective of this study was to investigate the association between US imaging morphology from routine radiologists’ interpretation and biological behavior such as receptor status and tumor grade determined from histopathology in invasive ductal carcinoma (IDC).Material and Methods:This retrospective study included 453 patients with pathology-verified diagnosis of IDC who had undergone US imaging and had surgery over a 5-year period. US and surgical pathology reports were reviewed and compiled. Correlation analyses and age-adjusted multivariable models were used to determine the association between US imaging morphology and receptor status, tumor grade, and germ line mutation of the breast cancer genes (BRCA1 and BRCA2). The odds ratio (OR), area under receiver operating characteristic curve (AUC), and 95% confidence intervals (CI) were obtained.Results:The likelihood for high-grade cancer increased with size (OR: 1.066; CI: 1.042–1.091) and hypo-echogenicity (OR: 2.044; CI: 1.337–3.126), and decreased with angular or spiculated margins (OR: 0.605; CI: 0.393–0.931) and posterior acoustic shadowing (OR: 0.352; CI: 0.238–0.523). These features achieved an AUC of 0.799 (CI: 0.752–0.845) for predicting high-grade tumors. The likelihood for Estrogen Receptor-positive tumors increased with posterior acoustic shadowing (OR: 3.818; CI: 2.206–6.607), angulated or spiculated margins (OR: 2.596; CI: 1.159–5.815) and decreased with US measured tumor size (OR: 0.959; CI: 0.933–0.986) and hypoechoic features (OR: 0.399; CI: 0.198– 0.801), and achieved an AUC of 0.787 (CI: 0.733–0.841). The likelihood for Progesterone Receptor-positive tumors increased with posterior acoustic shadowing (OR: 2.732; CI: 1.744–4.28) and angulated or spiculated margins (OR: 2.618; CI: 1.412–4.852), and decreased with US measured tumor size (OR: 0.961; CI: 0.937–0.985) and hypoechoic features (OR: 0.571; CI: 0.335–0.975), and achieved an AUC of 0.739 (CI: 0.689–0.790). The likelihood for Human epidermal growth factor receptor 2-positive tumors increased with heterogeneous echo texture (OR: 2.141; CI: 1.17– 3.919) and decreased with angulated or spiculated margins (OR: 0.408; CI: 0.177–0.944), and was marginally associated with hypoechoic features (OR: 2.101; CI: 0.98–4.505) and circumscribed margins (OR: 4.225; CI: 0.919–19.4). The model with the aforementioned four US morphological features and achieved an AUC of 0.686 (CI: 0.614–0.758). The likelihood for triple-negative breast cancers increased with hypo-echogenicity (OR: 2.671; CI: 1.249–5.712) and decreased with posterior acoustic shadowing (OR: 0.287; CI: 0.161–0.513), and achieved an AUC of 0.739 (CI: 0.671– 0.806). No statistical association was observed between US imaging morphology and BRCA mutation.Conclusion:In this study of over 450 IDCs, significant statistical associations between tumor grade and receptor status with US imaging morphology were observed and could serve as a surrogate imaging marker for the biological behavior of the tumor.

The cost impact of unselective vs selective MammaPrint testing in early-stage breast cancer in Southern Africa

BackgroundMammaPrint (MP) has been applied in South Africa (SA) for decision-making in early-stage hormone receptor-positive breast cancer since 2006. The cost-impact of MP in SA has not been assessed. AimTo assess different MP testing strategies for cost-minimization in early-stage breast carcinoma using a funder perspective. MethodsClinico-pathologic information was extracted from a prospectively collected database. Clinical risk stratification was done using Adjuvant Online! (AOL) and the Predict V2.1 algorithm (www.predict.nhs.uk). An unselected MP testing strategy was compared to a selective strategy, testing only clinically high risk (cHigh) patients. Excluding human epidermal growth factor receptor-2 positive tumours, the costs for chemotherapy treatment and MP using funding data were used to evaluate the financial impact of these strategies. ResultsIn 583 patients with 601 tumours, 52% were clinically low risk (cLow) (AOL) while the average Predict 10-year survival with chemotherapy was 2.9%. MP correlated strongly with Predict and 318 (60%) patients were MP low risk. Unselective testing allowed omission of chemotherapy in 44 (8.4%) patients but escalated cost by 57.7%. Using a selective testing strategy, only 251 would be tested, de-escalating treatment in 138 (55%) and reducing cost by 19.5%. Considering a Predict value up to 3.2% as cHigh, cost would be up to 7.3% (p = 0.0467) lower with a selective testing strategy. ConclusionMP allowed reduction in the use of adjuvant chemotherapy. Unselective use of MP increases overall costs. A selective testing strategy through clinical risk stratification using AOL/Predict results in substantial cost saving.

Combination of pegylated liposomal doxorubicin and docetaxel as neoadjuvant therapy for breast cancer with axillary lymph node metastasis.

ObjectiveTo evaluate the efficacy and safety of the combination of pegylated liposomal doxorubicin and docetaxel as neoadjuvant therapy for breast cancer (BC) in patients with axillary lymph node metastasis.MethodsIn this single-arm study, 91 patients with clinical stage IIA–IIIc breast cancer received six cycles of pegylated liposomal doxorubicin plus docetaxel as neoadjuvant chemotherapy (NAC). Trastuzumab was allowed for patients with human epidermal growth factor receptor 2-positive tumors. The primary endpoint was pathologic complete response (pCR) in the breast after surgery. The overall response rate (ORR), Miller–Payne (MP) score of the primary tumors, and incidence of adverse events were also evaluated.ResultsIn total, 88 patients completed all cycles of NAC. Fourteen patients (15.4%, 95% confidence interval [CI] = 7.8–22.9) achieved pCR. The ORR was 89% (95% CI = 82.5–95.6), and 72 lesions (79.1%) were rated as MP grade 3 or higher. The left ventricular ejection fraction (LVEF) was within the normal range, although four (4.4%) patients experienced an LVEF decline exceeding 10%. No symptomatic cardiac events were reported.ConclusionPreoperative NAC with pegylated liposomal doxorubicin and docetaxel appears effective and safe for treating BC with axillary lymph node metastasis.

Frequent methylation of genes encoding wnt pathway antagonists: Secreted frizzled-related protein 1 and dickkopf 3 in invasive breast cancer

Introduction: Wnt signaling pathway is often dysregulated in the pathogenesis of various malignancies, including breast cancer. This might be related to methylation of the genes encoding antagonists of this signaling pathway. Aim: The aim of the study was to analyze the methylation status of the promoter regions of Wnt antagonists-secreted frizzled-related protein 1 (sFRP1) and Dickkopf 3 (DKK3) and to determine their correlation with clinicopathological parameters and survival outcome in patients with primary invasive ductal breast cancer. Materials and Methods: The methylation status of sFRP1 and DKK3 was analyzed in 160 breast tumor samples using methylation-specific polymerase chain reaction. Statistical analysis was performed using SPSS software. P ≤ 0.05 was considered as statistically significant. Results: The promoter region of sFRP1 and DKK3 genes was found to be methylated in 76% and 64% of total invasive ductal breast cancer patients, respectively. The promoter methylation in sFRP1 and DKK3 genes was significantly associated with larger tumor size, positive lymph nodes, advanced stage, and perinodal extension of breast tumors. Further, sFRP1 methylation was associated with human epidermal growth factor receptor 2-positive tumors while DKK3 methylation was associated with Grade 3 tumors. Survival analysis demonstrated that sFRP1 methylation was correlated with reduced overall survival in breast cancer patients. Conclusion: Promoter methylation of Wnt pathway antagonists is frequent in breast cancer ultimately leading to probable upregulation of the pathway in these tumors. Hence, sFRP1 and DKK3 methylation may be used as a valuable biomarker in clinical breast cancer management.

Molecular subtypes are prognostic for N3 breast cancer patients in the modern therapeutic era.

Nodal (N) status and molecular subtypes are well-known prognostic factors for breast cancer patients. The aim of the present study was to evaluate whether there was a prognostic role of molecular subtypes for pN3a breast cancer patients in the modern therapeutic era. The present study retrospectively evaluated a total of 521 breast cancer patients who had 10 or more metastatic lymph nodes and received adjuvant systemic therapy at the Oncology Department of four different centers in Turkey between 2000-2015. Patients were divided into four molecular subtypes by immunohistochemical staining. There were no significant differences in relapse rates according to the molecular subtypes (P=0.07). The five year disease free survival rate was 62% for the whole study population, 67% for Luminal A tumors, 53% for Luminal B tumors, 64% for human epidermal growth factor receptor 2-positive tumors and 56% for triple negative tumors. Luminal A patients had a better progression free survival when compared with Luminal B (P=0.026) and triple negative (P=0.07) patients. pT stage (P<0.001), and breast cancer subtype (P<0.001), remained significant independent factors for disease free survival. Therefore, breast cancer subtypes are still prognostic for patients with pN3 breast cancer.

Optimal time for adjuvant therapy initiation in breast cancer patients: A single center experience

Introduction. This retrospective study evaluates the association between the time of chemotherapy initiation and disease free survival in regard to breast cancer subtypes and stage at diagnosis. Material and Methods. The study included a total of 1075 breast cancer patients, stages I - III, treated at Oncology Institute of Vojvodina, Serbia (from 2010 to 2012; n = 617). The gathered data included prognostic factors used in everyday practice. Patients were divided into three groups according to the interval between surgery and chemotherapy (? 30, 31 - 60, ? 61 days). Disease free survival was calculated. Results. Among the 617 patients, the 5-year disease free survival estimate was similar: 81.5%, 81.0%, 84.6% (log-rank test, p = 0.728) regarding the time of adjuvant chemotherapy initiation: ? 30 days, 31 - 60, and ? 61 days, respectively. The study showed that 85% of our breast cancer patients started adjuvant chemotherapy within 3 months after definitive surgery. In multivariate analysis, independent prognostic factors for disease free survival were nodal status and tumor size. The 5-year disease free survival estimate was 85.8% (p = 0.001) for patients with luminal- A subtype (Estrogen +, Progesterone high, human epidermal growth factor receptor 2-, Ki-67 &lt; 20%) with a median follow-up of 62,7 months; for patients with luminal-B (Luminal B (human epidermal growth factor receptor 2 -) Estrogen +, human epidermal growth factor receptor 2-, either Ki-67 high or Progesterone low, Luminal B (human epidermal growth factor receptor +) Estrogen +, human epidermal growth factor receptor 2 +, any Ki-67, any Progesterone) 78.3% (p = 0.534) with a median follow-up of 55.9 months; for patients with triple negative breast cancer it was 73.4% with a median follow-up of 58,1 months, and for patients with human epidermal growth factor receptor 2+ it was 77.1% (p = 0.448) with a median follow-up of 55.5 months. Conclusion. Early initiation of adjuvant chemotherapy is particularly important in patients with advanced- stage breast cancer at diagnosis, and those with trastuzumab- treated triple-negative breast cancer and human epidermal growth factor receptor 2-positive tumors.

Care patterns and changes in treatment for nonmetastatic breast cancer in 2013-2014 versus 2005: a population-based high-resolution study.

Studies on recent trends in patterns of care for breast cancer patients are scarce. This study aims to examine the patterns and trends in the treatment of women with nonmetastatic breast cancer according to major recommended treatment options. A population-based study was carried out in Navarra, Spain, including all women with a primary invasive nonmetastasized breast cancer, diagnosed in 2005 and in 2013-2014. We compared patients' characteristics and treatment patterns between periods. Factors associated with receipt of recommended treatment were examined by multivariate logistic regression. Of the 719 patients included, 90% received guideline-adherent locoregional treatment. Over the two periods, there was an increasing use of sentinel lymph node biopsy as opposed to axillary lymph node dissection as the first axillary procedure. Among women with oestrogen receptor-positive tumours, 96% received endocrine therapy. The proportion of high-risk patients who were treated with chemotherapy increased between the two periods from 65 to 74% (P=0.079) and, among patients with human epidermal growth factor receptor 2-positive tumours, the receipt of targeted treatment increased from 37 to 72% (P<0.001). The main factors associated independently with a lower probability of receiving recommended treatment were age 70 years or older for all treatment modalities and comorbidity for locoregional treatment and chemotherapy. The proportion of women with breast cancer who received treatment according to recent European guidelines in Navarra has increased from 2005 to 2013-2014, resulting in a high level of adherence to standard care. Most failures in adherence to these standards are related to older age or comorbidities.

Inhibitory effects of human epidermal growth factor receptor-2-single-chain antibody on human epidermal growth factor receptor-2 positive tumors

Objective To study the inhibitory effect of single-chain antibody (scFv) against human epidermal growth factor receptor-2 (Her-2) on tumor cell T6-17 proliferation. To investigate the potential of Her-2-scFv protein targeted therapy of Her-2 positive tumors. Methods In our study, Her-2-scFv gene fragment was amplified from plasmid pUC57-Her-2-scFv by polymerase chain reaction (PCR). Then Her-2-scFv gene fragment was cloned into the prokaryotic expression vector pET28a though double enzyme, and get the recombinant plasmid pET28a-Her-2-scFv. Plasmid pET28a-Her-2-scFv was transformed into Rosetta of E. coli. Her-2-scFv protein successful expression was detected by Western blotting. Her-2 positive tumor cell T6-17 proliferation inhibition were tested by methyl thiazol tetrazolium (MTT) assay. The tumor volume was measured by animal model of gastric cancer in vivo test. Results Her-2-scFv gene fragments were received by PCR successfully. Location of Her-2-scFv gene and its sequence in recombinant plasmid pET28a-Her-2-scFv is in a right direction by gene sequencing. MTT assay showed that the growth inhibition ratio of Her-2-positive tumor cell T6-17 in Her-2-scFv group was 33.52%, trastuzumab treatment group was 34.79%, with no statistic significance between them (P=0.429). Compared with the negative control group (PBS group), the differences were statistically significant (P=0.000). In vivo experiments: The inhibiting tumor rates of 100 ng/kg group, 250 ng/kg group, 500 ng/kg group were 33.7%, 27.2%, 63.3%, separately. Compared with the control group, the difference were statistically significant (P=0.027, 0.035, 0.000). In Hessaitin group, the inhibition rate was 70.2%, compared with 500 ng/kg group, there was no statistically significant difference (P=0.643). Conclusion Her-2-scFv protein can inhibit the proliferation of T6-17 positive tumor cells in vitro and in vivo. Key words: Human epidermal growth factor receptor-2; Single-chain antibody; Tumor cell T6-17

Niche-localized tumor cells are protected from HER2-targeted therapy via upregulation of an anti-apoptotic program in vivo

Several lines of evidence suggest that components of the tumor microenvironment, specifically basement membrane and extracellular matrix proteins, influence drug sensitivities. We previously reported differential drug sensitivity of tumor cells localized adjacent to laminin-rich extracellular matrix in three-dimensional tumor spheroid cultures. To evaluate whether differential intra-tumor responses to targeted therapy occur in vivo, we examined the sensitivity of human epidermal growth factor receptor 2-positive tumors to lapatinib using a previously described ductal carcinoma in situ-like model characterized by tumor cell confinement within ductal structures surrounded by an organized basement membrane. Here we show that tumor cells localized to a ‘niche’ in the outer layer of the intraductal tumors adjacent to myoepithelial cells and basement membrane are resistant to lapatinib. We found that the pro-survival protein BCL2 is selectively induced in the niche-protected tumor cells following lapatinib treatment, and combined inhibition of HER2 and BCL-2/XL enhanced targeting of these residual tumor cells. Elimination of the niche-protected tumor cells was achieved with the HER2 antibody–drug conjugate T-DM1, which delivers a chemotherapeutic payload. Thus, these studies provide evidence that subpopulations of tumor cells within specific microenvironmental niches can adapt to inhibition of critical oncogenic pathways, and furthermore reveal effective strategies to eliminate these resistant subpopulations.

Prognostic impact of total and tyrosine phosphorylated GIV/Girdin in breast cancers.

Gα-interacting vesicle-associated protein (GIV, aka Girdin) is a guanine exchange factor (GEF) for the trimeric G protein Gαi and a bona fide metastasis-related gene that serves as a platform for amplification of tyrosine-based signals via G-protein intermediates. Here we present the first exploratory biomarker study conducted on a cohort of 187 patients with breast cancer to evaluate the prognostic role of total GIV (tGIV) and tyrosine phosphorylated GIV (pYGIV) across the various molecular subtypes. A Kaplan-Meier analysis of recurrence-free survival showed that the presence of tGIV, either cytoplasmic or nuclear, carried poor prognosis, but that nuclear tGIV had a greater prognostic impact (P = 0.007 in early and P = 0.0048 in late clinical stages). Activated pYGIV in the cytoplasm had the greatest prognostic impact in late clinical stages (P = 0.006). Furthermore, we found that the prognostic impacts of cytoplasmic pYGIV and nuclear tGIV were additive (hazard ratio 19.0548; P = 0.0002). Surprisingly, this additive effect of nuclear tGIV/cytoplasmic pYGIV was observed in human epidermal growth factor receptor 2-positive tumors (hazard ratio 16.918; P = 0.0005) but not in triple-negative breast cancers. In triple-negative breast cancers, tGIV and cytoplasmic pYGIV had no prognostic impact; however, membrane-association of pYGIV carried a poor prognosis (P = 0.026). Both tGIV and pYGIV showed no correlation with clinical stage, tumor size, pathologic type, lymph node involvement, and BRCA1/2 status. We conclude that immunocytochemical detection of pYGIV and tGIV can serve as an effective prognosticator. On the basis of the differential prognostic impact of tGIV/pYGIV within each molecular subtype, we propose a diagnostic algorithm. Further studies on larger cohorts are essential to rigorously assess the effectiveness and robustness of this algorithm in prognosticating outcome among patients with breast cancer.-Dunkel, Y., Diao, K., Aznar, N., Swanson, L., Liu, L., Zhu, W., Mi, X.-Y., Ghosh, P. Prognostic impact of total and tyrosine phosphorylated GIV/Girdin in breast cancers.

Prognostic significance of inflammatory factors expression by stroma from breast carcinomas.

The aim of this work was to evaluate the expression and clinical relevance of some cytokines in breast carcinomas. An immunohistochemical study using tissue arrays and specific antibodies against interleukin 1β (IL-1β), IL-6, IL-10, IL-17, interferon β (IFNβ) and nuclear factor kappa B (NFκB) was performed in 108 breast carcinomas. Most studied cytokines were mainly expressed by cancer cells but also by stromal cells as cancer-associated fibroblasts (CAFs) or mononuclear inflammatory cells (MICs). Global expression (score) of IL-1β and IL-17 was positively associated with histological grade; human epidermal growth factor receptor 2-positive tumors showed a higher global expression of IFNβ but a lower global expression of NFκB; and node-negative tumors showed a higher global expression of IL-6. High score of IL-6 was significantly associated with both longer relapse free-survival (RFS) and overall survival (OS). Moreover, the expression of IL-1β by each stromal cells (CAFs and MICs) was significantly associated with both longer RFS and OS, whereas the expression of IL-10 by these cells was significantly associated with both shorter RFS and OS. However, the combination of IL-1β, IL-6 and IL-10 expression by MICs reached an important association with prognosis and improved our previously reported prognostic signification based on the matrix metalloprotease 11 status by MICs. The combination of IL-1β, IL-6 and IL-10 expression by MICs was significant and independently associated with distant RFS in a multivariate analysis. Therefore, the combination of the expression of IL-1β, IL-6 and IL-10 may serve as promising biomarkers of MICs with prognostic significance, contributing to a better characterization of breast carcinomas microenvironment.

Perinatal and childhood factors and risk of breast cancer subtypes in adulthood

BackgroundAccumulated exposure to hormones and growth factors during early life may influence the future risk of breast cancer (BC). This study examines the influence of childhood-related, socio-demographic and anthropometric variables on BC risk, overall and by specific pathologic subtypes. MethodsThis is a case-control study where 1539 histologically-confirmed BC cases (23–85 years) and 1621 population controls, frequency matched by age, were recruited in 10 Spanish provinces. Perinatal and childhood-related characteristics were directly surveyed by trained staff. The association with BC risk, globally and according to menopausal status and pathologic subtypes, was evaluated using logistic and multinomial regression models, adjusting for tumor specific risk factors. ResultsBirth characteristics were not related with BC risk. However, women with high socioeconomic level at birth presented a decreased BC risk (OR=0.45; 95% CI=0.29–0.70), while those whose mothers were aged over 39 years at their birth showed an almost significant excess risk of hormone receptor positive tumors (HR+) (OR=1.35; 95% CI=0.99–1.84). Women who were taller than their girl mates before puberty showed increased postmenopausal BC risk (OR=1.26; 95% CI=1.03–1.54) and increased HR+ BC risk (OR=1.26; 95% CI=1.04–1.52). Regarding prepubertal weight, while those women who were thinner than average showed higher postmenopausal BC risk (OR=1.46; 95% CI=1.20–1.78), associated with HR+ tumors (OR=1.34; 95% CI=1.12–1.61) and with triple negative tumors (OR=1.56; 95% CI=1.03–2.35), those who were heavier than average presented lower premenopausal BC risk (OR=0.64; 95% CI=0.46–0.90) and lower risk of epidermal growth factor receptor positive tumors (OR=0.61; 95% CI=0.40–0.93). ConclusionThese data reflect the importance of hormones and growth factors in the early stages of life, when the mammary gland is in development and therefore more vulnerable to proliferative stimuli.

Family history and outcome of young patients with breast cancer in the UK (POSH study).

Young patients presenting to surgical clinics with breast cancer are usually aware of their family history and frequently believe that a positive family history may adversely affect their prognosis. Tumour pathology and outcomes were compared in young British patients with breast cancer with and without a family history of breast cancer. Prospective Outcomes in Sporadic versus Hereditary breast cancer (POSH) is a large prospective cohort study of women aged less than 41 years with breast cancer diagnosed and treated in the UK using modern oncological management. Personal characteristics, tumour pathology, treatment and family history of breast/ovarian cancer were recorded. Follow-up data were collected annually. Family history data were available for 2850 patients. No family history was reported by 65·9 per cent, and 34·1 per cent reported breast/ovarian cancer in at least one first- or second-degree relative. Patients with a family history were more likely to have grade 3 tumours (63·3 versus 58·9 per cent) and less likely to have human epidermal growth factor receptor 2-positive tumours (24·7 versus 28·8 per cent) than those with no family history. In multivariable analyses, there were no significant differences in distant disease-free intervals for patients with versus those without a family history, either for the whole cohort (hazard ratio (HR) 0·89, 95 per cent c.i. 0·76 to 1·03; P = 0·120) or when stratified by oestrogen receptor (ER) status (ER-negative: HR 0·80, 0·62 to 1·04, P = 0·101; ER-positive: HR 0·95, 0·78 to 1·15, P = 0·589). Young British patients presenting to breast surgical clinics with a positive family history can be reassured that this is not a significant independent risk factor for breast cancer outcome.

Effects of Exercise during Adjuvant Chemotherapy on Breast Cancer Outcomes

Observational studies suggest that physical activity after a breast cancer diagnosis is associated with improved cancer outcomes; however, no randomized data are available. Here, we report an exploratory follow-up of cancer outcomes from the Supervised Trial of Aerobic versus Resistance Training (START). The START was a Canadian multicenter trial that randomized 242 breast cancer patients between 2003 and 2005 to usual care (n = 82), supervised aerobic (n = 78), or resistance (n = 82) exercise during chemotherapy. The primary end point for this exploratory analysis was disease-free survival (DFS). Secondary end points were overall survival, distant DFS, and recurrence-free interval. The two exercise arms were combined for analysis (n = 160), and selected subgroups were explored. After a median follow-up of 89 months, there were 25/160 (15.6%) DFS events in the exercise groups and 18/82 (22.0%) in the control group. Eight-year DFS was 82.7% for the exercise groups compared with 75.6% for the control group (HR, 0.68; 95% confidence interval (CI), 0.37-1.24; log-rank, P = 0.21). Slightly stronger effects were observed for overall survival (HR, 0.60; 95% CI, 0.27-1.33; log-rank, P = 0.21), distant DFS (HR, 0.62; 95% CI, 0.32-1.19; log-rank, P = 0.15), and recurrence-free interval (HR, 0.58; 95% CI, 0.30-1.11; Gray test, P = 0.095). Subgroup analyses suggested potentially stronger exercise effects on DFS for women who were overweight/obese (HR, 0.59; 95% CI, 0.27-1.27), had stage II/III cancer (HR, 0.61; 95% CI, 0.31-1.20), estrogen receptor-positive tumors (HR, 0.58; 95% CI, 0.26-1.29), human epidermal growth factor receptor 2-positive tumors (HR, 0.21; 95% CI, 0.04-1.02), received taxane-based chemotherapies (HR, 0.46; 95% CI, 0.19-1.15), and ≥85% of their planned chemotherapy (HR, 0.50; 95% CI, 0.25-1.01). This exploratory follow-up of the START provides the first randomized data to suggest that adding exercise to standard chemotherapy may improve breast cancer outcomes. A definitive phase III trial is warranted.

Imaging of triple-negative breast cancer

Although triple-negative breast cancer (TNBC) has been studied extensively in the oncology and pathology literature, there are few reports on imaging features. Emerging data suggest that imaging features of TNBC are substantially different from other primary breast cancer immunotypes. In this work, we reviewed multi-modality imaging features of primary TNBC with emphasis on the appropriate niche for each technology in diagnosis, staging and management. TNBC lacks the typical suspicious mammographic features of breast cancer; namely irregular mass shape, spiculated margins and associated suspicious calcifications. Therefore, mammography alone is usually a sub-optimal tool for its initial diagnostic evaluation. Ultrasound has a much higher sensitivity, although its diagnostic capability may be impaired by associated benign features encountered in 21%-41% of TNBC lesions. Magnetic resonance imaging (MRI) consistently demonstrates the presence of all TNBC with a higher level of accuracy compared with other tumour sub-types, and provides a reliable baseline for neoadjuvant chemotherapy (NAC) follow-up. Preliminary studies also suggest that MRI may predict complete NAC response in TNBC more sensitively than other methods. [(18)F]2-fluoro-2-deoxy-D-glucose positron emission tomography ((18)F-FDG-PET) has a higher sensitivity for TNBC than estrogen-positive and human epidermal growth factor receptor 2-positive tumours, and axillary lymph node metastases are detected with more accuracy compared with other tumour molecular sub-types, although low FDG uptake of breast cancers in general may limit the routine clinical use of (18)F-FDG-PET in the initial diagnosis of TNBC patients and NAC follow-up.

Evaluation of Four Affibody-Based Near-Infrared Fluorescent Probes for Optical Imaging of Epidermal Growth Factor Receptor Positive Tumors

The epidermal growth factor receptor 1 (EGFR) has become an attractive target for cancer molecular imaging and therapy. An Affibody protein with strong binding affinity for EGFR, ZEGFR:1907, has been reported. We are interested in translating Affibody molecules to potential clinical optical imaging of EGFR positive cancers. In this study, four anti-EGFR Affibody based near-infrared (NIR) fluorescent probes were thus prepared, and their in vivo performance was evaluated in the mice bearing EGFR positive subcutaneous A431 tumors. The Affibody analogue, Ac-Cys-ZEGFR:1907, was synthesized using solid-phase peptide synthesis method. The purified small protein was then site-specifically conjugated with four NIR fluorescent dyes, Cy5.5-monomaleimide, Alex-Fluor-680-maleimide, SRfluor680-maleimide, or IRDye-800CW-maleimide, to produce four optical probes-Cy5.5-ZEGFR:1907, Alexa680-ZEGFR:1907, SR680-ZEGFR:1907, and 800CW-ZEGFR:1907. The EGFR binding property and specificity of the four NIR fluorescent Affibody probes were studied by fluorescence microscopy using high EGFR expressing A431 cells and low expressing MCF7 cells. The binding affinities of the probes (KD) to EGFR were further determined by flow cytometry. In vivo optical imaging of the four probes was performed in the mice bearing subcutaneous A431 tumors. The four NIR optical probes were prepared in high purity. In vitro cell imaging studies demonstrated that all of them could specifically bind to EGFR positive A431 cells while showing minimum uptake in low EGFR expressing MCF7 cells. Flow cytometry showed that Cy5.5-ZEGFR:1907 and Alexa680-ZEGFR:1907 possessed high binding affinity in low nanomolar range (43.6 ± 8.4 and 28.3 ± 4.9, respectively). In vivo optical imaging of the four probes revealed that they all showed fast tumor targeting ability and good tumor-to-normal tissue contrast as early as 0.5 h postinjection (p.i.). The tumor-to-normal tissue ratio reached a peak at 2 to 4 h p.i. by regional of interest (ROI) analysis of images. Ex vivo studies further demonstrated that the four probes had high tumor uptakes. Particularly, Cy5.5-ZEGFR:1907 and Alex680-ZEGFR:1907 displayed higher tumor-to-normal tissue ratios than those of the other two probes. This work demonstrates that Affibody proteins can be modified with different NIR fluorescent dyes and used for imaging of EGFR expressing tumors. Different NIR fluorescent dyes have variable impact on the in vitro binding and in vivo performance of the resulting Affibody based probes. Therefore, selection of an appropriate NIRF label is important for optical probe development. The probes developed are promising for further tumor imaging applications and clinical translation. Particularly, Alex680-ZEGFR:1907 and Cy5.5-ZEGFR:1907 are excellent candidates as EGFR-targeted probes for optical imaging.

Cy5.5-labeled Affibody molecule for near-infrared fluorescent optical imaging of epidermal growth factor receptor positive tumors

Affibody protein is an engineered protein scaffold with a three-helical bundle structure. Affibody molecules of small size (7 kD) have great potential for targeting overexpressed cancer biomarkers in vivo. To develop an Affibody-based molecular probe for in vivo optical imaging of epidermal growth factor receptor (EGFR) positive tumors, an anti-EGFR Affibody molecule, Ac-Cys-Z(EGFR:1907) (7 kD), is site-specifically conjugated with a near-IR fluorescence dye, Cy5.5-mono-maleimide. Using fluorescent microscopy, the binding specificity of the probe Cy5.5-Z(EGFR:1907) is checked by a high-EGFR-expressing A431 cell and low-EGFR-expressing MCF7 cells. The binding affinity of Cy5.5-Z(EGFR:1907) (K(D)) to EGFR is 43.6+/-8.4 nM, as determined by flow cytometry. For an in vivo imaging study, the probe shows fast tumor targeting and good tumor contrast as early as 0.5 h postinjection (p.i.) for A431 tumors, while MCF7 tumors are barely visible. An ex vivo imaging study also demonstrates that Cy5.5-Z(EGFR:1907) has high tumor, liver, and kidney uptakes at 24 h p.i.. In conclusion, Cy5.5-Z(EGFR:1907) shows good affinity and high specificity to the EGFR. There is rapid achievement of good tumor-to-normal-tissue contrasts of Cy5.5-Z(EGFR:1907), thus demonstrating its potential for EGFR-targeted molecular imaging of cancers.