Stage Breast Research Articles

Abstract PD2-03: Recurrent functionally diverse in-frame ESR1 gene fusions drive endocrine resistance in breast cancer

Abstract Background. We previously reported an alternative ESR1 somatic gain-of-function chromosomal translocation event in a patient presenting with aggressive, endocrine therapy resistant estrogen receptor (ER) positive disease, producing an in-frame fusion gene consisting of N-terminal ESR1 and the C-terminus of the Hippo pathway coactivator YAP1 (ESR1-YAP1). We recently identified another ESR1 fusion through RNA sequencing (RNA-seq) in advanced stage ER+ disease from a chest wall recurrence in a male patient that was refractory to multiple lines of treatment. Two examples of fusions discovered in primary breast cancer samples include ESR1 fused in-frame to C-terminal sequences from NOP2 (ESR1-NOP2), identified in a resistant cohort from a RNA-seq screen focused on 81 primary breast cancers from aromatase inhibitor clinical trials, and a second ESR1 fusion, fused in-frame to the entire coding sequence of POLH (ESR1-POLH), that was identified from RNA-seq analysis of 728 Cancer Genome Atlas breast samples. This current study extends our previous characterization of ESR1-YAP1 by comparing functional and pharmacological properties of these three additional ESR1 gene fusion events of both early stage and advanced breast cancers. Methods. In vitro and in vivo experiments were conducted to test ESR1 fusions to induce therapeutic resistance, and metastasis. The transcriptional and binding properties of each fusion was also examined. Pharmacological inhibition with Palbociclib, a cyclin-dependent kinase 4/6 inhibitor, was utilized to assess drug sensitivity in ESR1 fusion containing breast cancer cells and in a patient derived xenograft (PDX) model expressing ESR1-YAP1 (WHIM18). Results. The YAP1 and PCDH11x fusions conferred estrogen-independent and fulvestrant-resistant growth. Immunohistochemistry revealed significantly higher numbers of ER+ cells in lungs of mice xenografted with T47D cells expressing the YAP1 and PCDH11x fusions compared to YFP control, NOP2 and POLH fusions. Results from ChIP-seq and microarray studies suggest that these two fusions promote proliferation and metastasis through genomic action by binding estrogen response elements (ERE) and subsequent gene activation. We thereby define these fusions as “canonical” fusions compared to “non-canonical” NOP2 and POLH fusions, which demonstrated dramatically decreased genomic binding ability. The non-canonical fusions induced genes associated with basal-like breast cancer and promoted HER2, EGFR, and MAPK gene expression signatures in contrast to genes associated with cell cycle/proliferation induced by canonical fusions. The proliferative ability of canonical fusion-containing ER+ cells was inhibited by Palbociclib in a dose-dependent manner. In vivo WHIM18 tumors in mice fed with Palbociclib-containing chow demonstrated significantly reduced tumor volume, growth rate, and weight compared to tumors in mice on control chow. Conclusions. In-frame ERE activating canonical fusions occur in end-stage drug resistant advanced breast cancer and can be added to ESR1 point mutations as a class of recurrent somatic mutation that may cause acquired resistance. Growth induced by these fusions can be antagonized by Palbociclib and is potentially clinically helpful. Citation Format: Lei JT, Shao J, Zhang J, Iglesia M, Cao J, Chan DW, He X, Kosaka Y, Schmidt C, Matsunuma R, Haricharan S, Crowder R, Hoog J, Phommaly C, Goncalves R, Ramalho S, Lai W-C, Hampton O, Rogers A, Tobias E, Parikh P, Davies S, Ma C, Suman V, Hunt K, Watson M, Hoadley KA, Thompson A, Chen X, Perou CM, Creighton CJ, Maher C, Ellis MJ. Recurrent functionally diverse in-frame ESR1 gene fusions drive endocrine resistance in breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr PD2-03.

Abstract P2-01-06: Patterns of axillary evaluation in older patients (pts) with breast cancer and impact on adjuvant therapy

Abstract Background: Axillary lymph node status has traditionally been a key factor in informing adjuvant therapy recommendations for pts with breast cancer. With increased emphasis on tumor biology, this information may be less relevant, particularly in older populations where competing comorbidity frequently influences treatment decisions. We examined patterns of axillary surgery in older breast cancer pts and the impact axillary surgery has on treatment receipt. Methods: We identified women aged ≥65 with Stage I-III invasive breast cancer diagnosed during 2012-2013 from the National Cancer Data Base who did not have clinically positive nodes and underwent cancer-directed surgery. Nodal surgery type and receipt of adjuvant therapies were examined. Multivariable logistic regression was used to examine the associations of axillary surgery receipt with pt, clinical and facility factors. Results: Among 69,414 eligible women, 40% were aged 65-70, 42% aged 71-80 and 18% aged >80. 91% had axillary surgery (67% sentinel lymph node biopsy, 11% axillary lymph node dissection, 13% unspecified axillary surgery), and 24% of pts had pathologically positive nodes. 10% of pts (stage IIB-III) received adjuvant chemotherapy, 81% (hormone receptor positive) received adjuvant hormonal therapy, 67% (breast conservation or stage III postmastectomy) received radiation. In adjusted analyses, increasing age and neoadjuvant hormonal therapy were strongly associated with lower odds of axillary surgery. Region and mastectomy were strongly associated with higher odds of axillary surgery. The table shows variables associated with axillary surgery. VariableAdjusted OR (95% CI) for having any axillary surgery*Age (vs. 65-70) 71-75.64 (.58-.71)76-80.34 (.31-.37)>80.08 (.07-.09)Diagnosed in 2013 (vs. 2012)1.08 (1.02-1.15)Stage (vs. II) I1.25 (1.13-1.38)III.73 (.60-.89)Grade (vs. 1) 21.22 (1.14-1.31)31.24 (1.13-1.37)HER2 status (vs. positive) Negative.83 (.73-.93)Tumor size (vs. ≤2 cm) >2-5cm1 (.91-1.11)>5cm.56 (.47-.67)Comorbidity score (vs. 0) 1.85 (.79-.92)>/=2.62 (.56-.68)Region (vs. New England)Range 1.66-2.67 (1.42-3.12)Case volume (vs. high) Low.82 (.73-.93)Medium.98 (.91-1.06)Insurance (vs. private) Uninsured.69 (.44-1.09)Medicaid.67 (.52-.86)Medicare.84 (.77-.93)Median household income ($) (vs. >46K) <30,0001.06 (.95-1.18)30,000-34,9991.11 (1.01-1.21)35,000-45,9991.08 (1.01-1.16)Neoadjuvant hormonal therapy (vs. not).49 (.42-.59)Surgery (vs. BCS) Mastectomy, no recon3.37 (3.09-3.68)Mastectomy, +recon2.76 (2.16-3.51) *Adjusted for table variables plus race, hormone receptor status, and facility type (none significantly associated with axillary surgery) Axillary surgery and younger age were significantly associated with receipt of adjuvant chemotherapy, radiation, and hormonal therapy. Conclusion: Within the NCDB, 91% of pts age ≥65 with clinically node-negative breast cancer undergo surgical staging of the axilla, and axillary surgery was associated with adjuvant therapy receipt. The impact of routine node assessment on treatment and outcome has been questioned, and further study in this population of pts is warranted. Citation Format: Dominici LS, Sineshaw H, Jemal A, Lin A, King TA, Freedman RA. Patterns of axillary evaluation in older patients (pts) with breast cancer and impact on adjuvant therapy [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-01-06.

Immediate-term chemotherapy-related cognitive impairment (CRCI) following administration of intravenous (IV) chemotherapy.

146 Background: The acute impact of chemotherapy on cognition is unknown. This study utilized performance on the psychomotor vigilance task (PVT) and trail-making test B (TMT) to assess CRCI immediately following chemotherapy administration. Methods: Patients aged 18-80 years receiving first-line IV chemotherapy for any stage of breast or colorectal cancer were eligible. Patients with brain metastases, neurologic disorders or allergic reactions to chemotherapy were excluded. Patient symptoms, peripheral neuropathy and Stanford Sleepiness Scale were assessed. A five-minute PVT and TMT were completed on a tablet computer pre-chemotherapy and immediately post-chemotherapy. Paired Wilcoxon Rank Sum tests were used to assess changes in median PVT reaction time, TMT completion time, TMT errors and PVT lapses. A priori, increases of 20 ms or over in median PVT reaction times (approximating reaction time changes with blood alcohol concentrations of 0.04 to 0.05 g%) were considered clinically relevant. Results: 144 patients (74 breast, 70 colorectal, median age 55.5 years) were tested. Post-chemotherapy, median PVT reaction time slowed by an average of 12.4 ms (p = 0.01). Post-chemotherapy median PVT times slowed by over 20 ms in 59 patients (40.9%). TMT completion post-chemotherapy was faster by an average of 6.1 seconds (p < 0.001). No differences were seen in TMT errors (p = 0.417) or PVT lapses (p = 0.845). Change in median PVT reaction time was not associated with age, gender, number of prior chemotherapy cycles, use of paclitaxel (which contains alcohol), peripheral neuropathy grade, or self-reported anxiety, fatigue or depression. Conclusions: Median PVT reaction time was significantly slower immediately after chemotherapy compared to a pre-chemotherapy baseline, and impairment correlating to effects of alcohol was seen in 41% of patients. This effect appears independent of age, self-reported symptoms or prior chemotherapy cycles. Further studies assessing the functional implications of immediate-term CRCI are warranted.

Thymosin beta 10 is a key regulator of tumorigenesis and metastasis and a novel serum marker in breast cancer

BackgroundThymosin beta 10 (TMSB10) has been demonstrated to be involved in the malignant process of many cancers. The purpose of this study was to determine the biological roles and clinical significance of TMSB10 in breast cancer and to identify whether TMSB10 might be used as a serum marker for the diagnosis of breast cancer.MethodsTMSB10 expression was evaluated by immunohistochemical analysis (IHC) of 253 breast tumors and ELISA of serum from 80 patients with breast cancer. Statistical analysis was performed to explore the correlation between TMSB10 expression and clinicopathological features in breast cancer. Univariate and multivariate Cox regression analysis were performed to examine the association between TMSB10 expression and overall survival and metastatic status. In vitro and in vivo assays were performed to assess the biological roles of TMSB10 in breast cancer. Western blotting and luciferase assays were examined to identify the underlying pathway involved in the tumor-promoting role of TMSB10.ResultsWe found TMSB10 was upregulated in breast cancer cells and tissues. Univariate and multivariate analysis demonstrated that high TMSB10 expression significantly correlated with clinicopathological features, poor prognosis and distant metastases in patients with breast cancer. Overexpression of TMSB10 promotes, while silencing of TMSB10 inhibits, proliferation, invasion and migration of breast cancer cells in vitro and in vivo. Our results further reveal that TMSB10 promotes the proliferation, invasion and migration of breast cancer cells via AKT/FOXO signaling, which is antagonized by the AKT kinase inhibitor perifosine. Importantly, the expression of TMSB10 is significantly elevated in the serum of patients with breast cancer and is positively associated with clinical stages of breast cancer.ConclusionTMSB10 may hold promise as a minimally invasive serum cancer biomarker for the diagnosis of breast cancer and a potential therapeutic target which will facilitate the development of a novel therapeutic strategy against breast cancer.

Big Data, Predictive Analytics, and Quality Improvement in Kidney Transplantation: A Proof of Concept.

We sought proof of concept of a Big Data Solution incorporating longitudinal structured and unstructured patient-level data from electronic health records (EHR) to predict graft loss (GL) and mortality. For a quality improvement initiative, GL and mortality prediction models were constructed using baseline and follow-up data (0-90 days posttransplant; structured and unstructured for 1-year models; data up to 1 year for 3-year models) on adult solitary kidney transplant recipients transplanted during 2007-2015 as follows: Model 1: United Network for Organ Sharing (UNOS) data; Model 2: UNOS & Transplant Database (Tx Database) data; Model 3: UNOS, Tx Database & EHR comorbidity data; and Model 4: UNOS, Tx Database, EHR data, Posttransplant trajectory data, and unstructured data. A 10% 3-year GL rate was observed among 891 patients (2007-2015). Layering of data sources improved model performance; Model 1: area under the curve (AUC), 0.66; (95% confidence interval [CI]: 0.60, 0.72); Model 2: AUC, 0.68; (95% CI: 0.61-0.74); Model 3: AUC, 0.72; (95% CI: 0.66-077); Model 4: AUC, 0.84, (95 % CI: 0.79-0.89). One-year GL (AUC, 0.87; Model 4) and 3-year mortality (AUC, 0.84; Model 4) models performed similarly. A Big Data approach significantly adds efficacy to GL and mortality prediction models and is EHR deployable to optimize outcomes.

Thermal Patterns in Peripheral Regions of Breast during Different Stages of Development

Background: Mathematical modelling of bio thermal processes is widely used to enhance the quantitative understanding of thermoregulation system of human body organs. This quantitative knowledge of thermal information of various human body organs can be used for developing clinical applications. In the past the investigators have studied thermal distribution in hemispherical shaped human breast in presence of spherical shaped tumor. The shape and size of the breast as well as tumor may also affect thermal distribution which can have serious implications in thermography. In the present paper a model of thermal disturbances in peripheral regions of ellipsoidal shaped breast for two dimensional steady state case. The modelling study will provide biomedical scientist vital insights of thermal changes occurring due to shape and size of breast which can influence the development of protocols of thermography for diagnosis and treatment of tumors in women’s breast. Method: We have incorporated the significant parameters like blood flow, metabolic activity and thermal conductivity in the thermal model for normal and malignant tissues. The controlled metabolic activity has been incorporated for normal tissues. The peripheral regions of breast are divided into three major layers mainly epidermis, dermis and sub dermal tissues. The outer surface of the breast is assumed to be exposed to the environment and the heat loss takes place by conduction, convection, radiation and evaporation. The finite element approach is employed to obtain the solution. Results: By selecting appropriate model parameters we have shown the spatial thermal variation in different stages of breast according to their shapes which could be replicated by the proposed model. Conclusions: The proposed model was successfully employed to study the impact of different sizes and shapes in peripheral regions of elliptical shaped woman’s breast. The proposed model is more realistic in terms of shape and size of woman’s breast in comparison to earlier models reported in the literature. The changes in slope of the thermal curves at the junctions of various peripheral layers are due to the non homogenous nature of the region.

Serum butyrylcholinesterase and zinc in breast cancer.

Even though, a large number of serological, molecular markers have been proposed for breast cancer screening, most of them lack specificity, sensitivity, prognostic value, and cost effectiveness. Butyrylcholinesterase (BChE) and its genes are aberrantly expressed in a variety of human cancers. It has-been linked to tumorigenesis, cell proliferation, and cell differentiation. Zinc (Zn) is a cofactor for superoxide dismutase, an enzyme that protects cellular components against free radical-induced damage and carcinogenesis. Therefore, the aim of present study was to estimate and compare serum BChE and serum Zn levels in healthy controls and biopsy proven breast cancer patients before definitive therapy. To estimate and compare serum BChE and serum Zn levels in healthy controls and biopsy proven breast cancer patients before definitive therapy. Serum BChE and Zn were estimated in 46 newly diagnosed (preoperative) female patients with breast cancer and 50 healthy female volunteers. Serum BChE and Zn were estimated by spectrophotometric method. Data was expressed as median and inter quartile range. Comparisons between different stages of cancer were done using Kruskal-Wallis test. There was a significant increase in serum BChE and Zn in breast cancer patients compared to controls (P < 0.001). Serum BChE showed a significant increase and Zn was significantly decreased in different stages of breast cancer. Both BChE and Zn are inexpensive and can easily be analyzed and may play a role in the management of breast cancer.

Clinicopathological Features of Triple Negative Breast Carcinoma.

Breast carcinoma is one of the most common malignancies affecting women in developing countries. Molecular studies of breast carcinoma have classified the tumour based on the immunohistochemical staining into 4 subtypes, such as Luminal A, Luminal B, HER2/neu Positive and Triple Negative Breast Carcinoma (TNBC). TNBCs are reported to have an aggressive behaviour and wide metastasis, leading to selective treatment outcomes. The aim was to study the clinicopathological features such as age, site, tumour size, histopathological type, histologic grade, lymph node status, stage and treatment outcomes of triple negative breast carcinoma. A retrospective study was conducted on 108 cases of breast carcinoma received during the period of 2 years. The tumour was classified based on immunohistochemical staining into four subtypes. The clinicopathological details, histomorphological and immunohistochemical features of TNBC were studied. Of the 108 patients, 34 patients were diagnosed as TNBC. The average age at presentation was 48 years. Most of the cases showed Nottingham Modification of Scarff Bloom-Richardson (NMBR) grade 3 (55.9%) and stage II (67.6%). Ly-mph node metastasis was seen in 50% of cases. Infiltrating ductal carcinoma (not otherwise specified) type (91.2%) was the most common histological type. Among the other subtypes, Luminal A carcinoma was the most common (36.1%), followed by TNBC (31.5%) and HER2/neu positive carcinomas (28.7%). Compared to the other types of tumours, TNBC showed the most frequent distant lymph node metastasis (50%) when compared to luminal A (38.5%), luminal B (25%), HER2/neu positive (48.4%). Unlike the other types of tumours, TNBC were mostly high-grade. TNBC have an aggressive behaviour compared to other subtypes with higher NMBR grade, nuclear pleomorphism, high mitotic rate and lymph node metastasis.

ROLE OF SURGERY IN PATIENTS WITH ADVANCED CANCER BREAST WITH OR WITHOUT INITIAL CHEMO-HORMONO-RADIOTHERAPY

Introduction: Role of surgery has changed or changing globally in the form of down staging and surgery in locally advanced breast cancer to Modified Radical Mastectomy with axillary clearance in locally operable stage IV breast cancer with good performing status. Aim & Objective: To study the role of surgery in patients with advanced cancer breast with or without initial Chemo-Hormono-Radiotherapy. Material and methods: The study includes 100 cases of advanced breast cancer-stage IIIb and stage IV, undergoing surgery with or without initial Chemo-Hormono-Radiotherapy. Information regarding family history of similar complaints, operative detailed history, treatment received by the patients was recorded also recorded. Postoperatively patients were followed up every monthly for completion Chemotherapy and clinical examination for first 3 months than 3 monthly interval. Patients were thoroughly examined for local site or Axillary recurrence and progress of the disease in the form of supraclavicular lymph node, opposite breast, arm edema, bony tenderness, chest infection and liver enlargement. Result: The highest percentage of breast cancer is reported in age group 51-60 years of age. Mean age of presentation was 52 years. The minimum age of presentation was 30 years and maximum was 80 years. Our study reported infiltrating ductal carcinoma to be the most frequent of all carcinoma of breast, comprising 95% of cases. 7% of patients underwent Palliative Mastectomy while 73% of cases underwent Modified Radical Mastectomy with Axillary clearance. Axillary clearance (level III) was done in 80 patients. While Level I axillary clearance was done in 12 patients. 7 patients underwent palliative mastectomy. Only 58 patients received Neoadjuvent Chemotherapy. 75% of patients received Radiotherapy postoperatively. Conclusion: Even though there is definite role of surgery in breast cancer in the early stage of breast cancer, the incidence of patients presenting with advanced breast cancer in a rural population is alarming. The treatment of stage IIIb and stage IV breast cancer is multimodal, evolving and challenging which is further complicated by resistance or refusal of Neoadjuvent Chemotherapy or Radiotherapy by the rural population. Key words: Advanced cancer breast, Chemo-Hormono-Radiotherapy, Treatment modalities.

Pubertal Progression and Reproductive Hormones in Healthy Girls With Transient Thelarche.

Detailed evaluation of pubertal progression in girls from longitudinal studies is sparse, and the phenomenon of transient thelarche (TT), defined as the appearance, regression, and subsequent reappearance of breast buds, in healthy girls remains undescribed. To describe TT in terms of pubertal progression, growth, genotypes, and reproductive hormones and to apply new puberty nomograms for breast stages, pubic hair, and menarche. A prospective, longitudinal population-based study. Ninety-eight healthy Danish schoolchildren (Caucasian girls) followed longitudinally as part of the COPENHAGEN Puberty Study were included in the evaluation of TT. A total of 1466 girls from 2 cross-sectional studies were included in the creation of the puberty nomograms. None. Pubertal progression, specifically thelarche, reproductive hormones, genotype, and growth. Twelve of 98 (12%) girls experienced TT. A larger proportion of girls with TT entered puberty by the pubarche pathway (50%) compared with girls with normal progression (15.4%), P = 0.014. Girls with TT progressed through puberty normally when evaluated using puberty nomograms. Reproductive hormones and growth velocity were lower at the first (transient) thelarche than the second (permanent) thelarche. TT is a frequent phenomenon that appears to be a peripheral occurrence independent of central puberty. It does not appear to affect subsequent pubertal progression as evaluated by our new puberty nomograms.

Update on pubertal development among primary school students in Shanghai, 2014

Objective: To investigate the current prevalence of pubertal development in healthy Shanghai schoolchildren. Methods: This study was a cross-sectional investigation focused on current pubertal development conducted in healthy Shanghai schoolchildren by multi-stage cluster sampling. The sample included 17 571 children in grades 1-5 investigated in June 2014. The data were weighted by inverse probability weighting (IPW) to make them more representative. At examination, stages of breast and pubic hair development were rated according to the Tanner method. Testicular volume was determined. Data on menarche and spermatorrhea were collected by the status quo method. The rates of precocious puberty, breast, and pubic hair development of Tanner stage ≥Ⅱ in girls aged 6-7 years, menarche in girls aged 6-9 years, and testicular volume ≥4 ml and pubic hair development of Tanner stage ≥Ⅱ in boys aged 6-8 years were calculated. All the data were weighted by IPW. Results: After data processing, 16 197 children's data were analyzed. In girls aged 6-7 years, 17.2% and 2.5% showed evidence of breast and pubic hair development at Tanner stage ≥Ⅱ, respectively. In girls aged 6-9 years, 0.3% had experienced menarche. Schoolgirls' rate of menarche was 4.7%. In girls aged 6-7 years, 19.0% were diagnosed with precocious puberty according to the classic criteria. In boys aged 6-8 years, 1.7% had testicular volume ≥4 ml, and 0.6% showed evidence of pubic hair development at Tanner stage ≥Ⅱ. Schoolboys' incidence rate of spermatorrhea was 0.1%. In boys aged 6-8 years, 2.3% were diagnosed with precocious puberty according to the classic criteria. All the numbers above were weighted. Conclusion: Proper education on adolescence and sex is essential for Shanghai schoolchildren.

Initial experience of automated breast volume scanning (ABVS) and ultrasound elastography in predicting breast cancer subtypes and staging

ObjectivesBreast cancer is a heterogeneous disease consisting of distinct histopathological subtypes with different clinical outcomes. In this article, we identified the automated breast volume scanning (ABVS) and shear wave velocity (SWV) characteristics of different pathological types of breast carcinoma. MethodsA retrospective review of both ABVS and SWV imaging of 118 consecutive breast masses was performed. The imaging features of both techniques were assessed with reference to histopathological results. ResultsEcho heterogeneity with a smooth and lobulated margin was a significant feature more frequently found in mucinous carcinoma groups (100%, P < 0.05). Between different stages of ductal carcinoma, echo homogeneity was more likely in high-grade ductal carcinomas (P < 0.05). SWV differences existed between inside tumor areas and either tumor boundary or tissues outside the tumors (P < 0.05), and values differed between different breast carcinoma stages. The central and tumor margin areas of ductal carcinomas were much harder than in tubular carcinoma and micro-carcinoma, respectively (P < 0.05). SWV ROC curve analyses yielded a cut-off value of 3.015 m/s between ductal carcinoma in situ (DCIS) and invasive ductal carcinoma in the central part of lesions, with 83.5% sensitivity and 80% specificity for T0 vs T1-3 staging. ConclusionsSince some features were associated with different breast carcinoma types and stages, ABVS and SWV imaging has the potential to give clues about breast carcinoma differentiation in a non-invasive manner.

The Role of 18F-Sodium Fluoride PET/CT Bone Scans in the Diagnosis of Metastatic Bone Disease from Breast and Prostate Cancer.

We describe the role of 18F-sodium fluoride (18F-NaF) PET/CT bone scanning in the staging of breast and prostate cancer. 18F-NaF PET was initially utilized as a bone scanning agent in the 1960s and early 1970s, however, its use was restricted by the then-available γ-cameras. The advent of hybrid PET/CT cameras in the late 1990s has shown a resurgence of interest in its use and role. After a brief introduction, this paper describes the radiopharmaceutical properties, dosimetry, pharmacokinetics, and mechanism of uptake of 18F-NaF. The performance of 18F-NaF PET/CT is then compared with that of conventional bone scintigraphy using current evidence from the literature. Strengths and weaknesses of 18F-NaF PET/CT imaging are highlighted. Clinical examples of improved accuracy of diagnosis and impact on patient management are illustrated. Limitations of 18F-NaF PET/CT imaging are outlined.

“A study of Serum levels of LDH and ALP in carcinoma breast: An Observational study”

Background:- Breast Cancer is one of the major surgical problems in India particularly among the younger women. Unfortunately, most breast cancers do not produce any symptoms until the tumors are either too large to be removed surgically or cancerous cells have already spread to other tissues i.e. metastasis has taken place. Therefore, there is a need for noninvasive and sensitive methods to detect growths small in size which escape routine examination .There is, therefore, need of simple investigations, which can be easily assayed, are less expensive and can detect metastasis. Aims And Objectives: To study the serum levels of LDH and ALP in cases of breast lesions and to evaluate whether their levels are significantly higher in cases of breast malignancies as compared to cases of benign breast lesions Material And Methods:- A total number of 160 subjects were studied, comprising of 80 cases of benign breast lesions and 80 Breast cancer subjects. Breast cancer subjects were further divided depending on stage of the cancer. Serum levels of LDH and ALP were estimated in all the subjects. RESULT:- The present observational study has shown a significant elevation in serum levels of LDH and ALP in cases of carcinoma Breast as compared to benign breast lesions. These levels raised significantly with severity (stage) of carcinoma Breast. Conclusion:- Serum LDH and ALP may serve as biomarkers in carcinoma breast and may help in early detection of progression of the disease. These enzymes can be easily assayed in laboratories that do not have sophisticated technologies.

The challenge of measuring intra-individual change in fatigue during cancer treatment.

To evaluate how well three different patient-reported outcomes (PROs) measure individual change. Two hundred and fourteen patients (from two sites) initiating first or new chemotherapy for any stage of breast or gastrointestinal cancer participated. The 13-item FACIT Fatigue scale, a 7-item PROMIS® Fatigue Short Form (PROMIS 7a), and the PROMIS® Fatigue computer adaptive test (CAT) were administered monthly online for 6months. Reliability of measured change was defined, under a population mixed effects model, as the ratio of estimated systematic variance in rate of change to the estimated total variance of measured individual differences in rate of change. Precision of individual measured change, the standard error of measurement of change, was given by the square root of the rate-of-change sampling variance. Linear and quadratic models were examined up to 3 and up to 6months. A linear model for measured change showed the following by 6 and 3months, respectively: PROMIS CAT (0.363 and 0.342); PROMIS SF (0.408 and 0.533); FACIT (0.459 and 0.473). Quadratic models offered no noteworthy improvement over linear models. Both reliability and precision results demonstrate the need to improve the measurement of intra-individual change. These results illustrate the challenge of reliably measuring individual change in fatigue with a level of confidence required for intervention. Optimizing clinically useful measurement of intra-individual differences over time continues to pose a challenge for PROs.

Abstract 2160: Targeting SMC1 in combination therapy for triple negative breast cancer

Abstract Triple negative breast cancer (TNBC) defined primarily by lack of expression of estrogen and progesterone hormone receptors (HR) and lacking HER2 overexpression and/or gene amplification (HER2) is an aggressive subtype with poor prognosis. Presentation is frequently at an advanced stage, and TNBC is particularly prevalent in younger women, African Americans, and/or in women with BRCA1 and less frequently with BRCA2 gene mutations. Over the last several years, although great progress has been made in the treatment of early HR positive and HER2+ (as well as advanced stage) breast cancer, relapse/progression-free and overall survival of patients with TNBC remains a serious problem. Molecular subtyping identified subsets of TNBC, several of which are thought to be DNA-repair deficient (either due to germ-line mutations such as BRCA1 or 2, or somatic mutations affecting DNA-repair), may be amenable to treatment with DNA-damaging agents such as platinum compounds and PARP (poly(ADP-ribose) polymerase-1) inhibitors. This could be particularly useful in BRCA-mutated cancers. As for the other TNBC subtypes, development of novel therapeutic agents, represents a high priority. In response to this clinical need, we tested the efficacy of a novel combinatorial strategy which targets differentiated cells and TNBC cancer stem cell like population (CSCs). Structural maintenance of chromosome-1 (SMC1) was selected as a tumor antigen based on our published studies showing its surface localization and the role in cell proliferation, survival and metastasis in TNBC cells. Bioinformatics analyses using public available patient cohorts with clinicopathological information such as TCGA, NCBI and BRAVO showed statistically higher mRNA overexpression of SMC1 in TNBC compared to HR+ and HER2+ breast cancer. To target SMC1, we developed several monoclonal antibodies against the epitope of SMC1 which showed similarity to cell-adhesion peptides in order to facilitate biochemical and cellular assays. The impact of SMC1 inhibition in combination with veliparib, an inhibitor of PARP-1 was tested in BRCA1-wild-type (MDA-MB-231) vs. mutant (MDA-MB-436) TNBC cells and in CSCs (CD44+/CD24low-) sorted from these cells by MTT and colony forming assays. TUNEL and Annexin-V flipping assays were used to determine cell apoptosis. The role of intrinsic vs. extrinsic pathways of apoptosis was examined by measuring activation of Caspase 8 and 9 as well as measurements of cytochrome-c release from mitochondria. Our results showed that Mab-S15 recognizes SMC1 which is selectively expressed on the surface of cancer cells (including CSC’s) and differentiated TNBC’s, but has a minimal distribution in normal cells. The antibody distinctly inhibits the survival of TNBC in culture, including CSC’s, sorted from TNBC’s. The Mab-S15 and veliparib combination was more effective than the individual agents in inhibiting the growth and survival of TNBC cells including those with a functional BRCA1. Citation Format: Sushma Yadav, David Berz, George Somlo, Kimlin Ashing, Yate-Ching Yuan, Robert J. Hickey, Sailee Yadav, Arthur D. Riggs. Targeting SMC1 in combination therapy for triple negative breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2160.

Abstract 2130A: Epigenetic silencing of TGFβ1, BCL6, KILLIN and CTSZ is related to trastuzumab resistance in HER2-positive breast cancer models

Abstract Introduction: The major clinical problem for HER2 breast cancer target therapies is the acquisition of resistance. Trastuzumab has been used for the treatment of HER2-positive early stage and metastatic breast cancers. Notoriously, up to 62% patients that respond to the initial treatment develop resistance within a year. Different cellular and molecular alterations may contribute to the development of Trastuzumab resistance, including epigenetic changes. The aim of our study has been the evaluation of epigenetic differences between Trastuzumab-sensitive and -resistant breast cancer cell lines to determine whether they might be used as potential biomarkers to help in the treatment assessment and monitoring. Experimental procedures: To understand the epigenetic changes that may be associated with Trastuzumab resistance we compared the DNA methylation status of ∼450000 CpG sites from a DNA methylation microarray performed in Trastuzumab-sensitive and resistant cells. To identify the differentially methylated CpGs we considered differences between methylation groups of ≥60%. The functions associated to genes were analysed using the Gene Ontology and were used to select a small number of candidate genes. Their differences in promoter methylation status were validated by expression techniques (qPCR) and subsequently confirmed by demethylation agent (5-aza-DC) treatment followed by qPCR. The Trastuzumab resistant HER2-positive cell line (SKTR) had been previously developed in our laboratory by exposing SKBr3 cells to Trastuzumab, starting with 1μM for three months of exposure and increasing up to 2 μM for a 12 months period. Summary of the new, unpublished data: The DNA methylation microarray data showed that different 5’-CpG island sites had altered methylation profile when comparing Trastuzumab-resistant (SKTR) and -sensible SKBr3 cells. We selected 4 candidate genes with a hypermethylation profile in -resistant vs. -sensible cells: TGFβ1 (Transforming Growth Factor Beta 1), BCL6 (B-Cell CLL/Lymphoma 6), KILLIN (P53-Regulated DNA Replication Inhibitor) and CTSZ (Cathepsin Z). The hypermethylation of these four candidate genes in SKTR was in agreement with their expression downregulation observed by PCR analysis. SKTR treatment with the demethylation agent 5-aza-2’-deoxycytidine restored the expression levels of all four genes to the levels of Trastuzumab-sensitive cells. Although previously related to breast cancer, TGFβ1, BCL6, KILLIN and CTSZ have not earlier been associated with the development of Trastuzumab resistance in breast cancer or in other cancer types and may be further studied as potential biomarkers. Conclusions: These results provide a basis for future studies to validate the hypermethylation status of these genes as a predictive or monitoring biomarker of Trastuzumab resistance in HER2+ breast cancer patients. Citation Format: Sonia Palomeras, Angel Diaz-Lagares, Sarrats Ariadna, Crujeiras Ana Belen, Giro-Perafita Ariadna, Juan Sandoval, Marc Rabionet, Manel Esteller, Teresa Puig. Epigenetic silencing of TGFβ1, BCL6, KILLIN and CTSZ is related to trastuzumab resistance in HER2-positive breast cancer models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2130A.

Abstract SY20-03: Metastatic latency and immune evasion by cancer stem cells

Abstract Metastasis frequently develops, years after the removal of a primary tumor, from a minority of disseminated cancer cells that survived as latent entities. An improved understanding of the location and survival mechanisms of disseminated cancer cells, and their ability to enter a quiescent state, evade immune surveillance, and retain tumor reinitiating capacity is needed in order to better prevent and treat metastasis. To this end, we developed latency competent cancer cell (LCC) models from early stage human lung and breast carcinoma cell lines and defined the mechanisms that prevent the outgrowth, support the long-term survival, and maintain the tumor-initiating potential of these cells during latent metastasis. LCC cells show stem cell-like characteristics and express Sox transcription factors, which are essential for their survival in host organs under restrictive conditions and for metastatic outgrowth under permissive conditions. Through autocrine suppression of WNT signaling and heightened TGF-β signaling, LCC cells self-impose a slow-cycling state with broad downregulation of NK cell activators. By actively entering quiescence, metastatic stem cells can evade innate immunity to remain latent for extended periods. By persistently reentering the proliferative state, these cells may evolve organ-specific colonization traits to take over the host stroma. Citation Format: Joan Massague. Metastatic latency and immune evasion by cancer stem cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr SY20-03.

Efficacy and Safety of Gonadotropin-Releasing Hormone Agonist Treatment to Suppress Puberty in Gender Dysphoric Adolescents

IntroductionPuberty suppression using gonadotropin-releasing hormone agonists (GnRHas) is recommended by current guidelines as the treatment of choice for gender dysphoric adolescents. Although GnRHas have long been used to treat precocious puberty, there are few data on the efficacy and safety in gender dysphoric adolescents. Therefore, the Endocrine Society guideline recommends frequent monitoring of gonadotropins, sex steroids, and renal and liver function. AimTo evaluate the efficacy and safety of GnRHa treatment to suppress puberty in gender dysphoric adolescents. MethodsForty-nine male-to-female and 67 female-to-male gender dysphoric adolescents treated with triptorelin were included in the analysis. Main Outcome MeasuresPhysical examination, including assessment of Tanner stage, took place every 3 months and blood samples were drawn at 0, 3, and 6 months and then every 6 months. Body composition was evaluated using dual energy x-ray absorptiometry. ResultsGnRHa treatment caused a decrease in testicular volume in 43 of 49 male-to-female subjects. In one of four female-to-male subjects who presented at Tanner breast stage 2, breast development completely regressed. Gonadotropins and sex steroid levels were suppressed within 3 months. Treatment did not have to be adjusted because of insufficient suppression in any subject. No sustained abnormalities of liver enzymes or creatinine were encountered. Alkaline phosphatase decreased, probably related to a slower growth velocity, because height SD score decreased in boys and girls. Lean body mass percentage significantly decreased during the first year of treatment in girls and boys, whereas fat percentage significantly increased. ConclusionTriptorelin effectively suppresses puberty in gender dysphoric adolescents. These data suggest routine monitoring of gonadotropins, sex steroids, creatinine, and liver function is not necessary during treatment with triptorelin. Further studies should evaluate the extent to which changes in height SD score and body composition that occur during GnRHa treatment can be reversed during subsequent cross-sex hormone treatment.

Associations Between Maternal Pregravid Obesity and Gestational Diabetes and the Timing of Pubarche in Daughters.

We investigated whether in utero exposure to maternal pregravid obesity and/or gestational diabetes mellitus (GDM) was associated with early puberty in girls. We used data from a longitudinal study of 421 mother-daughter pairs enrolled in an integrated health services organization, Kaiser Permanente Northern California (2005-2012). Girls aged 6-8 years were followed annually through ages 12-14 years. Onset of puberty was assessed using study clinic-based Tanner staging. We examined associations of self-reported pregravid obesity and maternal GDM with timing of the daughter's transition to pubertal maturation stage 2 or above for development of breasts and pubic hair, using accelerated failure time regression models with interval censoring to estimate time ratios and hazard ratios and corresponding 95% confidence intervals. Maternal obesity (pregravid body mass index (BMI; weight (kg)/height (m)(2)) ≥30) was associated with a daughter's earlier transition to breast and pubic hair stage 2+ in comparison with girls whose mothers had pregravid BMI <25. These associations were attenuated and not statistically significant after adjustment for covariates. Girls whose mothers had both pregravid BMI ≥25 and GDM were at higher risk of an earlier transition to pubic hair stage 2+ than those whose mothers had neither condition (adjusted time ratio = 0.89, 95% confidence interval: 0.83, 0.96; hazard ratio = 2.97, 95% confidence interval: 1.52, 5.83). These findings suggest that exposure to maternal obesity and hyperglycemia places girls at higher risk of earlier pubarche.