bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P3-04-08: A proteo-probe for nucleotide excision repair in breast cancers
Authors: Dreze M , Calkins AS , Echelman DJ , Schnorenberg MR , Fell GL , Fisher DE , Iglehart JD , Lazaro J-B
Published: August 2014

The response of tumor cells to DNA damaging agents is often compromised, a weakness readily exploited for therapeutic purposes. Despite the general use of genotoxic treatments, the nature and extent of DNA damage in tumor cells is generally under-investigated, or remains unknown. We postulate DNA repair deficient breast tumor cells exhibit significantly more DNA damage after genotoxic treatments, compared to repair proficient cells.

We present a technology to detect specific DNA damage and monitor repair as a basis to categorize cancers into homogenous groups of malignancies. We rely on the use of biochemically purified DNA damage recognition complexes as ready-to-use reagents, or “proteo-probes” for DNA damage.

We developed a proteo-probe based on a purified DDB2 protein complex. DDB2 is a DNA damage recognition protein part of the nucleotide excision repair (NER) that recognizes and repairs bulky DNA adducts caused for example by UV irradiation or cisplatin. We show the DDB2 proteo-probe binds to nuclei of UV-irradiated cells. We followed the DDB2 proteo-probe signal over time, and observed a decrease concordant with the completion of NER. We then successfully distinguished repair proficient cells from genetically characterized NER-deficient cells obtained from patients. We conclude the DDB2 proteo-probe can be used to detect activation of NER by UV-induced DNA damage, and subsequent repair in situ.

It has been proposed that some breast cancers carry NER deficiencies. We screened a panel of cancer cell lines using the DDB2 proteo-probe and discovered obvious NER deficient (NER-D) cell lines. In addition, we correlated NER activity with cisplatin sensitivity. We further determined whether NER deficiency in breast cancer cell lines was caused by loss-of-function of NER genes through complementation.

The classification of breast cancer cell lines based on NER activity presented here suggests more investigation is needed about NER in breast cancers. Ultimately, we envision NER deficiencies could be used to select breast cancer patients for DNA-repair targeted therapy.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-04-08.
Abstract P3-06-03: Oral bisphosphonates and survival of breast and colon malignancies
Authors: Rennert G , Rennert HS , Pinchev M , Flugelman A , Kershenbaum A , Landsman K , Hadad R , Shulman K , Gruber SB
Published: August 2014

Bisphosphonates are traditionally used for treatment of osteoporosis and more recently for treatment and prevention of bone metastases in various malignancies. The use of 2nd-generation oral bisphosphonates has been reported to reduce the risk of developing breast and colon cancer but their influence on cancer survival has not been studied.

Methods Two large cohorts of consecutively diagnosed cases with breast or colorectal cancer were studied for the association between use of 2nd-generation oral bisphosphonates and cancer survival. Using computerized prescription records, sustained use of alendronate/risedronate was assessed in postmenopausal women with newly diagnosed breast (n = 2,843) or colorectal cancer (n = 1,706). Overall survival and cancer-specific survival were evaluated using time dependent analysis.

Results Postmenopausal women with breast cancer previously unexposed to bisphosphonates who used 2nd-generation bisphosphonates after diagnosis for at least one year had a significantly better survival than non-users, adjusted for age, tumor stage and grade (Overall survival: HR = 0.53, 0.33-0.86, breast cancer-specific survival: HR = 0.26, 0.10-0.71, p = 0.009). A similar advantageous hazard ratio was found in users with ER positive, ER negative and HER2neu positive tumors. A similar significantly better survival was noted for colorectal cancer after adjustment for age, tumor stage and grade (Overall survival: HR = 0.53, 0.33-0.85, colorectal cancer-specific survival: HR = 0.44, 0.21-0.88, p = 0.02). Women who used bisphosphonates before diagnosis did not exhibit a significant survival benefit. Pharmacogenetic studies of F(D)PPS (Farnesyl Pyrophosphate Synthase), a gene coding a key step in the mevalonate pathway revealed direct correlation between a minor homozygous status and survival in bisphosphonate users.

Conclusions The use of 2nd-generation bisphosphonates initiated after diagnosis was associated with a significant improvement in overall and in cancer-specific survival of postmenopausal women with breast or colon cancers.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-06-03.
Abstract P3-06-15: Breast cancer molecular profile according to BMI and menopausal status
Authors: Arpino G , Crispo A , De Angelis C , Thomas R , De Laurentiis M , Forestieri V , Colao A , De Placido S
Published: August 2014

Introduction:

High body mass index (BMI) is considered a risk factor for breast cancer onset and is correlated with bad prognosis in breast cancer patients. However, very few studies have associated clinical and molecular tumor characteristics with the patients’ BMI at the time of breast cancer diagnosis. The present study analyses how BMI influences tumor biology and correlates with different molecular subtypes according to menopausal status in a large cohort of patients with new diagnoses of early breast cancer (EBC).

Methods:

Overall 967 patients with a new diagnosis of EBC from 2007 till 2012 were recruited for this study. Clinical and tumor characteristics such as age, menopausal status, weight, height, tumor size (T), grading, estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER2) status, were prospectively collected. BMI was categorized into two groups (low: ≤25; high: >25). Associations between BMI and clinicopathological variables were performed by χ2 test.

Results:

A total of 594 (61%) post-menopause and 373 (39%) pre-menopause women were enrolled. Regardless of menopausal status, patients with high BMI were older (p<0.0001) and had larger tumors (p<0.0001). However, among postmenopausal patients, high BMI correlates with increased ER expression (p = 0.015), lower HER2 expression (p = 0.05) and less aggressive tumor subtypes as luminal A (p = 0.043). In the same patient subset, women with low BMI were more likely to develop a Luminal B, HER2 like or TN breast cancer. No correlation was observed between tumor molecular phenotypes and BMI among premenopausal women.

Conclusion:

Among postmenopausal patient, high BMI is associated with less aggressive and more endocrine sensitive EBC. This is consistent with the hypotheses that higher estrogen exposure of breast tissue in women with higher BMI may drive growth of these cancers. If further confirmed, our data suggests that weight control in this subset of women may help to prevent cancer development.

Table 1Clinical and Molecular CharacteristicsBMI ≤25BMI>25   N%N%totalp valueER     .015Positive15376.132884.3481 Negative4823.96115.7109 PgR     .2Positive14471.629676.1440 Negative5728.49323.9150 HER2     .05Positive3819.55013.388 Negative15780.532786.7484 MOLECULAR SUBTYPES     .043Luminal A11560.225771.2372 Luminal B3015.74813.378 HER2147.3143.928 TN3216.84211.674 

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-06-15.
Abstract P3-07-13: Evaluation of epidemiological and molecular differences in African American and Caucasian women with triple negative breast cancer
Authors: Ellsworth RE , Sturtz LA , Melley J , Means M , Shriver CD
Published: August 2014

Background: Breast cancer's (BC) higher mortality rates in African American women (AAW) have been attributed to the higher frequency of triple negative breast cancer (TNBC), an aggressive tumor subtype, in young AAW. What is not known is whether TNBC represents a single disease entity or differs in etiology, molecular characteristics or outcomes by population.

Methods: Demographic, pathological and survival data from AAW (n = 62) and CW (n = 98) with TNBC were analyzed using chi-square analysis, Student's t-tests, and log-rank tests. RNA from laser microdissected TNBC from a subset of patients was hybridized to HG U133A 2.0 microarrays and data analyzed with a FDR <0.05, >2-fold change to define significance.

Results: The frequency of TNBC compared to all BC was significantly higher in AAW (26%) compared to CW (8%), however, significant survival and pathological differences were not detected. The expression of CRYBB2 was ∼4-fold higher in tumors from AAW; these expression levels were able to classify just 64% of AAW patients correctly. Among demographic characteristics, AAW consumed significantly lower amounts of caffeine and alcohol, were less likely to breastfeed and more likely to be obese.

Conclusions: These data suggest that TNBC in AAW is not a unique disease compared to TNBC in CW. Rather, higher frequency of TNBC in AAW may, in part, be attributable to the effects of lifestyle choices. Because these risk factors are modifiable, they provide new opportunities for the development of risk reduction strategies that may effectively decrease survival disparities by preventing the development of TNBC in AAW.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-07-13.
Abstract P3-08-08: Pattern of use of an online primary health care information resource for breast cancer survivorship education: The pink ribbon survivors network
Authors: Fisher RE , Jensen L , Adams KA
Published: August 2014

Introduction: The Pink Ribbon Survivors Network, a 501(c)(3) nonprofit organization founded by cancer professionals, developed an online educational resource for breast cancer survivors, cancer care professionals, and primary health care providers. The organization created a separate online educational library for each of these three separate audiences. The organization's website launched in July, 2012.

Methods: The Primary Health Care Library for breast cancer survivorship education houses 15 unique categories. Utilizing the organization's administrative website, numerical pattern of access of each separate listing in the 15 categories can be obtained. In addition, Google Analytics data was utilized to determine total website activity for all three educational libraries.

Results: As of abstract submission, the Primary Healthcare Breast Cancer Survivorship Education Library contains 141 clinical articles in 15 categories. The website has been utilized in all 50 of the United States, and in 66 countries and 890 cities. The five most viewed categories of the 15 total categories are (in descending order): 1.) Lifestyle Modifications, 2.) Creating a Link between Cancer Care and Primary Care: Clinical Literature on Survivorship, 3.) Depression/Mood Disturbance, 4.) Consensus Panel Guidelines for Breast Cancer Follow up, and 5.) Gynecologic Issues: Sexuality/Fertility.

Conclusion: Since it's creation in mid- 2012, an online breast cancer survivorship resource is being utilized for educational review. Users of the website are preferentially interested in QOL, healthcare system, and Consensus guidance in their educational viewing. This system can be utilized to disseminate nationally recognized clinical advice for Primary Healthcare Providers as they care for breast cancer survivors in their routine healthcare practices.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-08-08.
Abstract P3-09-12: Effects of acupuncture on rehabilitation after breast cancer surgery - Preliminary results
Authors: Nazario AP , Giron PS , Simão CA , Facina G
Published: August 2014

Background: The increased incidence of breast cancer and successful treatment result in greater survival of these patients and consequently a greater number of survivors. These patients may have chronic condition of pain in the shoulder and upper limb, including cervical and thoracic spine. This paper seeks, through acupuncture, a new approach in the rehabilitation of these patients.

Method: This study was conducted in the Gynecological Department of São Paulo Federal University. The study included women who underwent surgical treatment of breast cancer, radical or conservative, and present pain in the region of the shoulder and upper limb, including cervical and thoracic spine after three months of surgery. Patients older than 18 years and pain level ≥ 3 on the Visual Analog Scale (VAS). We excluded patients with bilateral breast surgery, metastatic disease, vascular disorders and tactile sensitivity, with diabetes mellitus type I and II uncompensated and education level less than four years.

The patients were randomly divided into two groups, with 15 patients per group who were treated weekly for 10 weeks, one group treated with kinesiotherapy standard, pre-defined, based on stretching the neck muscles, scapular exercises and upper limb strength lasting 30 minutes and the other group was performed the same protocol described above kinesiotherapy followed by another 30 minutes of Acupuncture using pre-defined points.

Acupuncture points used were as follows: CV 3, SP 9, ST 36, KD 7, LV 3 - edema and treat gynecological disorders; VB 21, LI 15, TH 14, LU 5, LI 4, ST 38 and UB 60 - treat pain and difficulty of movement in the upper limb.

Patients were evaluated at baseline, after five weeks and at the end of 10 weeks. We used the Visual Analog Scale (VAS) for pain intensity ratings and the DASH questionnaire (Disabilities of the Arm, Shoulder and Hand Questionnaire) assessing functional capacity in diseases of the upper limb, measuring the skills to do certain activities as well as the symptoms.

Results: The ANOVA test was used for statistical analysis of pain relief by VAS scale in both groups. Both groups had significant improvement with p> 0.001 and no difference between groups, with p = 0.756. The same analysis of variance was used in the DASH questionnaire, with similar results, both groups improved significantly with p> 0.001 and no difference between groups with p = 0.200. In multiple comparisons all reviews are different from each other showing a gradual improvement.

Conclusion: We found that both types of treatment (kinesiotherapy or kinesiotherapy and acupuncture) are effective, and the evolution of the two groups was the same with a gradual improvement.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-09-12.
Abstract P3-10-04: Prevalence of pain and barriers to pain management in breast cancer survivors in Vietnam
Authors: Nguyen J , Tran TT , Ta TV , Ginsburg O
Published: August 2014

Introduction: Breast cancer-related pain significantly affects quality of life and is well-documented in developed countries. However, little is known about the extent of breast cancer-related pain in developing countries. A study at the National Cancer Institute of Vietnam reported that 44% of breast cancer patients experienced moderate to severe pain. However, this study did not include breast cancer survivors, and little is known about the prevalence of pain and its management in this patient population.

Objectives: Determine the prevalence of pain, adequacy of pain management, and barriers to care in breast cancer survivors in Vietnam.

Methods: We administered previously validated and published symptom assessment tools (the BPI and MDASI) to 160 unselected Vietnamese women diagnosed and treated for breast cancer in 2007. Here we present a preliminary description of the results for the first 40 breast cancer survivors enrolled in this study.

Results: The most frequently reported symptoms were pain (62.5%, with 40% reporting the pain as severe), memory loss (50%), numbness/tingling (50%), sleep disturbance (45%), and appetite loss (32.5%). Only 23% reported adequate symptom management, with the costs of treatment and the false belief that no treatments existed for the pain being frequently cited as barriers to care.

Discussion: Cancer-related pain is highly prevalent in breast cancer survivors treated at the largest cancer hospital in Vietnam. The majority of patients report inadequate management of pain, often attributable to costs and false beliefs regarding pain management. Initiatives aimed at patient education and improving accessibility to pain management could significantly improve the quality of life in this patient population.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-10-04.
Abstract P3-14-17: Results of a novel neoadjuvant chemotherapy (NAC) for breast cancer
Authors: Rivera-Rodriguez N , Cabanillas F , Lawrenson L , Negron V , Pavia O , Bruno M , Echenique M , Pardo W , Santiago K
Published: August 2014

Background: Achieving a pathologic complete response (pCR) after neoadjuvant chemotherapy (NAC) has been associated with improved disease free survival (DFS) and overall survival (OS). The Residual Cancer Burden Score (RCB) method is a useful tool that predicts DFS and OS after NAC. We present the results of pts with either triple negative or HER2 positive breast cancer treated with a novel NAC. Methods: 34 pts with localized breast cancer >1 cm with HER2+ (N = 19) or triple negative breast cancer (TNBC) (N = 15) were treated with this novel regimen consisting first of TEC (docetaxel 75 mg/m2, epirubicin 80 mg/m2, and cyclophosphamide 500 mg/m2) + PEG Filgrastim x 4 cycles. Following the 4th course, TNBC patients received 4 additional TEC cycles if they achieved CR by MRI, or were switched to a non cross-resistant regimen (vinorelbine, bevacizumab, capecitabine) if they had < CR. HER2+ pts received TEC x4 followed by docetaxel + trastuzumab x 4. RCB score was used to measure pathologic response. Pretreament PET scan was done and repeated after course 1 in order to correlate with RCB. Results: Median age was 56 (58 for Her2+ and 49 for TNBC). RCB = 0 (pCR) was achieved in 76%, while only 1 responded poorly (RCB = 3). There was no significant difference in the pCR rate between Her2+ and TNBC patients (74% vs 80% respectively), but there was a difference in the rate of pCR without DCIS and invasive cancer between these two (see table, p = 0.034). Pts with SUV drop > 5% after 1st TEC had 84% pCR while none with < 5% achieved pCR (p = 0.001). Comparison of our results with other NAC regimens reported in the literature is summarized in the table below:

ProtocolpCR (no invasive)pCR(no invasive and no DCIS)RCB 0-1Auxilio Cancer Center HER2+/TNBC74%/80%37%/73%79%/80%I-Spy trial 1 HER2+/TNBC45%/35%N/A61%/40%GeparQuinto HER2+/TNBCN/A45%/38.4%N/ANSABP B-40 TNBC51%N/AN/ANeoAltto HER2+47%N/AN/ANeoSphere HER2+39%N/AN/AMDACC FEC+trastuzumab HER2+60%N/AN/A

Conclusions: This novel chemotherapy approach results in a high pCR rate and RCB 0-1, which have been associated with improved clinical outcomes. Early PET can predict pCR. Although sample size is modest, results are encouraging and deserve further evaluation.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-14-17.
Abstract P4-01-03: HDACi (vorinostat) in metastatic breast cancer to restore sensitivity to ER-directed (AI) therapy: A phase II clinical trial with FES imaging correlates
Authors: Linden HM , Kurland BF , Link JM , Novakova A , Chai X , Specht JM , Gadi VK , Gralow JR , Schubert EK , Peterson LM , Eary J , Shields A , Mankoff DA , Krohn KA
Published: August 2014

Background: Histone deacetylase inhibitors (HDACi) have shown pre-clinical promise in estrogen receptor(ER)-modulation and restoring sensitivity to endocrine manipulation, suggesting potential clinical benefit (Sabnis 2011) (Huang 2000) in ER+ breast cancer. Vorinostat is an FDA-approved HDACi for CTCL, and could have a beneficial role in restoring ER-signaling in endocrine-resistant tumors (Munster 2011) (Yardley 2011). [F-18]fluoroestradiol (FES) PET imaging may be used to monitor regional tumor ER expression in patients with breast cancer (Linden 2011).

Methods: Patients with metastatic breast cancer with prior clinical benefit from endocrine manipulation who progressed on an AI therapy are eligible for this ongoing trial. In part A, patients were given vorinostat for 2 weeks, then resumed AI for 6 W. In part B (reflecting results of prior HDACi trials) patients are given vorinostat 400mg po daily 5/7 days 3/4 weeks while AI is given continuously. Paired FES and FDG PET are performed at baseline, week 2 and 8; clinical/radiologic assessment of disease is also performed at week 8. Patients with clinical benefit (response or stable disease) may continue on treatment until progressive disease or study withdrawal. Lesion-level analysis of the association between baseline FES uptake (logged) and FES/FDG ratio used generalized estimating equations (GEE) with small-sample adjustments to standard errors.

Results: 12/ 20 planned patients have accrued, and the treatment is well tolerated. Enrolled women were postmenopausal, the majority with primary infiltrating ductal tumors, bone/soft tissue dominant with longstanding metastatic disease, exposed to multiple endocrine and chemotherapy regimens. Five patients have had clinical benefit (2/4 on part B with greater HDACi exposure). One patient withdrew from the study due to toxicity. FES and FDG uptake was analyzed in 42 lesions in 11 patients. Average FES uptake was 2.0 (SULmean) for patients with clinical benefit, and 1.2 in patients with progressive disease by 8 weeks (p = 0.09). FES/FDG ratio at baseline was also associated with response (p = 0.04).

Conclusions: HDACi therapy is promising in relapsed ER+ breast cancer. Imaging of metabolic pathways in parallel with clinical trials may accelerate understanding of the underlying tumor biology and refine treatment selection.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-03.
Abstract P4-01-04: Role of FDG-PET/CT in evaluating surgical outcomes of operable breast cancer: Usefulness for malignant grade of triple-negative breast cancer
Authors: Ohara M , Hideo S , Akiko E , Masumoto N , Ozaki S , Kadoya T , Okada M
Published: August 2014

Background: The aim of this study was to evaluate the significance of 18F-fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) for speculating the malignant level and prognostic value of operable breast cancers.

Methods: Of 578 consecutive patients with primary invasive breast cancer who underwent curative surgery between 2005 and 2010, 311 patients (53.8%) who received FDG PET/CT before initial therapy were examined.

Results: Receiver operating characteristics (ROC) curve analysis showed the cutoff value of the maximum standardized uptake value (SUVmax) to predict cancer recurrence was 3.8 in all patients and 8.6 in patients with the triple-negative subtype, respectively. In all patients, 3-year DFS rates were 98.8% for patients with a tumor of SUVmax ≤ 3.8 and 91.6% for patients with a tumor of SUVmax > 3.8 (p < 0.001). High value of SUVmax was significantly associated with large tumor size (p < 0.001), lymph node metastasis (p = 0.040), high nuclear grade (p < 0.001), lymphovascular invasion (p = 0.032), negative hormone receptor status (p < 0.001), and positive HER2 status (p = 0.014). Based on the results of multivariate Cox analysis in all patients, high SUVmax (p = 0.001) and negative hormone receptor status (p = 0.005) were significantly associated with poor prognosis. In patients with triple-negative subtype, 3-year DFS rates were 90.9% for patients with a tumor of SUVmax ≤ 8.6 and 42.9% for patients with a tumor of SUVmax > 8.6 (p = 0.002), and high SUVmax was the only significant independent prognostic factor (p = 0.047).

Conclusion: FDG-PET/CT is useful for predicting malignant behavior and prognosis in patients with operable breast cancer, especially the triple-negative subtype.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-04.
Abstract P4-01-13: Fusion imaging using opto-acoustics and ultrasound to differentiate breast mass histopathology
Authors: Lavin P , Stavros AT , Tucker FL
Published: August 2014

Introduction: Opto-acoustics (OA) is a true fusion imaging technology that combines dual energy laser optic functional imaging with real time gray scale ultrasound morphologic imaging. The laser optic component uses two wavelengths that have been chosen to optimally demonstrate oxygenated and deoxygenated hemoglobin. The color-coded laser optic images are co-registered and temporally interleaved with real time gray scale images. OA fusion imaging benefits from the combination of the high contrast resolution of dual wavelength laser optics and the higher spatial resolution and penetration of gray scale ultrasound. OA can demonstrate both tumor neovascularity and the relative degrees of oxygenation-deoxygenation within breast mass and within the surrounding tissues.

Methodology: A series of 68 prospective patients with undiagnosed breast masses going to biopsy were evaluated before biopsy with a handheld device combining ultrasound co-registered with OA for the visualization of pre-defined internal and external features. The internal features included vascularity (V), blush (BU), and blood (BL) accumulation while external features included boundary zone vascularity and blush (BZ) and peri-tumoral radiating vessels (RV); each were scored on a 0-5 ordinal scale and were summed to get a total internal score, total external score, and a total score. A two-sided exact Jonckheere-Terpstra test was used to test the relationship between increasing scores and higher cancer grade.

Results: The study population consisted of 29 fibroadenomas (FAs) as well as 10 invasive ductal carcinoma (IDC), grade 1, 11 IDC grade 2, and 18 IDC grade 3 cases. OA Total (two-sided p = 3×10−8) and OA Total Internal (two-sided p = 6×10−9) were highly significant but OA Total External was not statistically significant; OA Total and OA Total Internal Features were highly related to histology with lower scores for FAs increasing progressively to highest scores for IDC 3. Moreover, all three internal OA features were highly significant: (1) V (two-sided p = 10−7), BU (two-sided p = 3×10−8), and BL (two-sided p = 5×10−6).

Cumulative contribution of each internal feature to the total internal score was also assessed for each histopathology subgroup to confirm independent contribution to the OA Total Internal Score. Pairwise correlations explained no more than 56% of the total variance.

Conclusions: We have established the prognostic value of OA Total and OA Internal Features as well as the validation of the contributions of each OA Internal Feature. Using the combination of individual internal and external OA features not only helps distinguish benign from malignant masses, but helps stratify malignant lesions by histologic grade. These abilities could be useful both for diagnosis and for monitoring non-surgical treatment of malignant masses.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-13.
Abstract P4-03-06: Initial ex-vivo study of the ability of an in situ fluorescence spectroscopy probe to distinguish benign from malignant breast tissues
Authors: Mathieu M-C , Moniz-Koum H , Vincent Y , Toullec A , Balleyguier C , Eyrignoux T , Vielh P , Chatellier F , Al Chab L , Fontaine-Aupart M-P , Farcy R , Delaloge S
Published: August 2014

Background:

The native fluorescence spectra of human cancerous and normal breast tissues under the selected excitation of a 405 nm light should differ according to their different content of NAD(P)H, FAD, collagen or retinol and derived fluorophores. Nodea Medical has developed an in situ probe, Probea®, installed in a 25-Gauge needle, which is able to measure endogenous tissue fluorescence under 405 nm specific wavelength. Tissues are excited by this blue light; their back-production of fluorescence (around 480 nm) is caught and immediately analyzed.

The objectives of the present preclinical study are to evaluate the ability of the Probea® probe to distinguish benign versus malignant breast lesions among freshly harvested ex-vivo surgical samples, and to evaluate the fluorescence behaviour of adjacent and remote normal tissues among cancer specimens.

Methods:

Fresh breast samples coming out of the operating room were harvested and analyzed prior to pathological fixation. Surgical specimens included mastectomies, lumpectomies, and plastic surgeries in different malignant or non-malignant situations. For each of them, the probe was placed within the main target surgical lesion, as well as within surrounding normal epithelial and fat tissues, and remote normal tissues where available (mastectomies or reduction mammoplasties).

We aimed at measuring different parameters of the endogenous fluorescence at each measurement point: maximum intensity, peak wavelength, level of signal stability, growth or spontaneous decay in a fixed position. Fluorescence signals harvested were compared to the detailed pathological analyses of each area explored within the surgical samples.

Results :

49 fresh breast samples were harvested and analyzed, of which 40 were from patients with cancer and 9 from patients without cancer.

The presence of a fluorescence signal within the main lesion was overall strongly associated with a cancer diagnosis (N = 40/49 malignant lesions versus 0/9 benign lesions, p = 0.001). The positive predictive value of the test in this preliminary series is 98%, whereas the negative predictive value is 100% analyzing the maximum fluorescence intensity with a cut-off of 1000. Interestingly, normal adjacent fat tissues surrounding and remote from breast cancer lesions also produced fluorescence which could also be considered as criteria to define malignancy. Complete analyses of multiple parameters are ongoing.

Conclusion:

In this initial preclinical study, the Probea® probe has shown an elective ability to distinguish benign and malignant lesions. Particularly, its negative predictive value appears very promising. A clinical study to confirm these findings is planned.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-03-06.
Abstract P4-04-01: Understanding estrogen receptor transcription in breast cancer
Authors: Carroll JS
Published: August 2014

Estrogen Receptor (ER) is the defining feature of luminal breast cancers, where is functions as a transcription factor. The traditional view of ER getting recruited to promoters of target genes is too simplistic. The recent discovery of ER-DNA interaction regions from ER+ breast cancer cell lines has revealed that ER rarely associates with promoter regions of target genes and instead associates with enhancer elements significant distances from the target genes. The genomic mapping of ER binding events also revealed the enrichment of DNA motifs for Forkhead factors. The Forkhead protein FOXA1 (HNF3a) was subsequently shown to bind to half of the ER binding events in the genome and was required for ER to maintain interaction with DNA. We have extended on these findings to explore ER and FoxA1 functional interactions in breast cancer, with a specific focus on changes in binding dynamics that occur during endocrine resistance. To this end, we have established transcription factor mapping approaches (ChIP-seq) in primary tumor material, to investigate differential ER/FoxA1 binding that occurs during tumor progression. We have functionally explored underlying factors that contribute to altered transcription factor binding and activity. In addition, we have recently established a method for rapid unbiased discovery of protein interactomes, which we have applied to discover ER and FoxA1 associated proteins. We find unexpected interactions between ER and additional nuclear receptors in ER+ breast cancer, which appear to be important modulators of cellular growth and endocrine response. These findings help delineate the complexes that influence ER transcriptional activity and ultimately impinge on tumor progression and drug sensitivity.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-04-01.
Abstract P4-04-06: Systematic analysis of molecular subgroups and age in patients with breast cancer
Authors: Liedtke C , Karn T , Gluz O , Becker S , Pusztai L , Holtrich U , Rody A
Published: August 2014

Background: Breast cancer (BC) molecular subgroups show significant differences with regard to prognosis. Similarly, young age at diagnosis is significantly associated with an unfavorable prognosis.

Methods: Our aim was to analyze the association between BC molecular subtypes and age at diagnosis. Publically available gene expression data (Affymetrix U133A) of 3488 BC patients was analyzed using MAS5.0 and pooled into a single database. Age information was available for 2604 of the patients (74.7%). Of a total of 579 cases with TNBC, 394 samples with most comparable array data were selected to avoid batch effects as previosly described. Age information was available for 309 of these 394 patients (78.4%). Significance Analysis of Microarrays (SAM) was performed to identify genes with differential expression according to age at diagnosis in these 309 patients with TNBC.

Results: Patients <40 years significantly more often presented with triple negative BC compared to patients 40-50 and >50 (34.8 vs. 25.4 vs. 17.3%, respectively). In contrast, a luminal A subtype was diagnosed less often in patients < 40 yrs (20.6 vs. 31.9 vs. 41.1%, respectively). Although we observed significant differences for EFS according to i) age at diagnosis and ii) molecular subtype, EFS differences between patients of distinct age were only modest after stratification for molecular subtype. Among patients with TNBC, 134 genes were upregulated and 627 genes were downregulated in patients < 40 yrs compared to patients ≥ 40 yrs (median FDR < 4%). Several of the downregulated genes in young TNBC patients belong to a metagene for the molecular-apocrine subtype of TNBC and several upregulated genes represent cell cycle genes. However, the effects for individual genes were rather small. Conclusion: The frequency of molecular subgroups differs with age at diagnosis. Young age at diagnosis is significantly associated with adverse prognosis. Within the group of TNBC patients young age is associated with high proliferation and a basal-like phenotype and inversely correlated with the molecular-apocrine subtype of TNBC.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-04-06.
Abstract P4-04-11: Correlation study of gene polymorphisms on 19p13.1 with triple negative breast cancer
Authors: Xu Z , Xing H , Wang L , Zhou H , Gu Y
Published: August 2014

Objective:The aim of this study was to investigate the association between triple negative breast cancer (TNBC) susceptibility and single nucleotide polymorphisms (SNPs) of MERIT40(c19orf62)and ANKLE1 genes located in 19p13.1, so as to find the clinical helpful molecular markers and therapeutic targets to the early detection and to guide individualized treatment.

Materials and Methods:Totally 252 patients with breast cancer diagnosed and treated in Breast Surgery Department of China-Japan Union Hospital affiliated to Jilin University were taken into the study, Among them,there were 124 cases identified as TNBC by using IHC technique, and others there were 128 cases as non-TNBC. Using common PCR assay and DNA sequencing to detect the SNPs of candidate genes. Finally, the association was analyzed between triple negative breast cancer group and non-triple negative breast cancer group with incidence and impact factors.

Results:Three genotypes were detected in rs8170,a SNP of MERIT40 gene located in 19p13.1.The frequencies of CC, CT and TT genotype in the triple negative breast cancer group and non-triple negative breast cancer group were 52.9% vs 93.5%, 41.2% vs 6.5% and 5.9% vs 0.0%.The difference of the distribution of genotypes between two groups was statistically significant (P = 0.004).Three genotypes were detected in rs2363956,a SNP of ANKLE1 gene also located in 19p13.1. The frequencies of TT, GT and GG genotype in the triple negative breast cancer group and non-triple negative breast cancer group were 35.7% vs 38.5%, 42.9% vs 53.8%and 21.4%vs7.7%.The difference of the distribution of genotypes between tow groups was no significant correlation (P = 0.464).Rs2363956 genotypes had obvious correlation with clinical stage (P = 0.005) and lymph node metastasis (P = 0.029) in non-triple negative breast cancer group, but not in triple negative breast cancer group(P = 0.323, P = 0.510).Neither rs8170 nor rs2363956 genotypes had obvious correlation with breast cancer-related immune group, such as EGFR,P53(P>0.05). Logistic regression analysis showed that TT genotype of rs8170 would increase the risk of in TNBC patients (OR = 1.426,95% CI = 1.266-2.872 P = 0.024).There was no significant association between the risk of incidence and the SNPs of ANKLE1 genes. Multivariate Logistic regression analysis showed that the polymorphism of ANKLE1 rs2363956 was an independent prognostic factor with lymph node metastasis and high clinical installments in for non-triple negative breast cancer (OR = 1.228, 95%CI = 1. 104-1.908 P = 0.045), but not for triple negative breast cancer.

Conclusions :The rate of incidence is comparatively higher in TNBC patients with TT genotype of MERIT40 rs8170, which may be an important molecular markers to predict the incidence of triple negative breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-04-11.
Abstract P4-05-03: Unique genetic, epigenetic, and transcriptomic changes in premenopausal breast cancer suggest novel strategies for therapy
Authors: Liao G , Hartmaier RJ , Luthra S , Chandran U , McGuire KP , Puhalla SL , Lee AV , Tseng GC , Oesterreich S
Published: August 2014

Breast cancer in premenopausal women (preM) is frequently associated with worse prognosis compared to that in postmenopausal women (postM) even when controlling for prognostic variables. In particular, preM ER+ tumors have a poor prognosis on endocrine therapy. There is, however, a paucity of studies characterizing molecular alterations in premenopausal tumors, a potential avenue for finding personalized therapies for this group of women.

We analyzed gene expression, CNV, methylation, and somatic mutations in tumors from preM (≤45; ER+ n = 110, and ER- n = 39) and postM (≥55, ER+ n = 392, and ER- n = 102) women in The Cancer Genome Atlas (TCGA). Unbiased hierarchical clustering of 2,900 most variably expressed genes (using both RNA-seq and Agilent expression array data) in the whole dataset (n = 643) identified four major subtypes which correlated highly with the PAM50 defined subtypes LumA, LumB, Basal and HER2; however, there wasn't any separation between preM and postM samples. Similarly, principal component analysis using 10,000 genes with the highest inter-quartile range (IQR) demonstrated high similarity across preM and postM samples. Direct examination of gene expression differences between PreM and PostM ER+ tumors using unpaired t-test (5% FDR) identified 3,044 differentially expressed genes. The genes most upregulated in premenopausal tumors included AREG, TFPI2, MSMB, TCN1, and GLRA3. Ingenuity Pathway Analysis revealed a highly significant enrichment for TGFb (p<1.9E-16) pathway activity in preM tumors. Intriguingly, no significant gene expression differences between preM and postM ER- tumors were identified. We thus then focused on genetic and epigenetic alterations that may underlie these transcriptomic changes in ER+ preM tumors.

Comparison of methylation (450K Illumina array) between preM and postM ER+ tumors showed a difference in 1% (n = 1,738) of the probes. Genes with the largest difference included ESR1, SIM2, and KLF6. Significant differences in DNA copy number variation (Affymetrix SNP 6.0 array) were also identified in ER+ preM tumors. A number of somatic mutations were significantly enriched in preM ER+ tumors including DSPP and GATA3. Integrated analysis also showed that approximately half of the observed differences in gene expression are driven by CNVs.

Conclusion: Our in silico study has identified a number of genes and pathways which are significantly altered between preM and postM ER+ breast cancer. Distinct genetic and epigenetic differences suggest unique etiology for some preM tumors. Currently ongoing Paradigm analysis, and confirmatory studies using METABRIC data are expected to further identify pathways that could specifically be targeted in premenopausal breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-03.
Abstract P4-05-05: Molecular effects of initial core biopsies in neoadjuvant window studies
Authors: Arthur LM , Turnbull AK , Renshaw L , Sabine VS , Bartlett JM , Sims AH , Dixon JM
Published: August 2014

Background

A growing number of studies are assessing the effects of drugs on breast cancers by investigating the molecular changes in tumours between the initial core biopsy taken at diagnosis and the excision specimen removed at surgery 2-3 weeks later. It is imperative to know if molecular profiles of tumours change over time without treatment and to assess whether the core biopsy itself influences the molecular profile. This study aimed to investigate changes in molecular profiles of breast cancer between the initial core biopsy and the excision specimen in patients who had no intervening treatment.

Methods

83 patients with paired fresh frozen tissue specimens from an initial diagnostic core biopsy and the later surgical excision biopsy have been studied. Following RNA extraction and processing, Illumina HT-12 BeadArrays were used for gene expression profiling. Samples were analysed for significant changes in gene expression and compared with a large dataset of patients treated with neoadjuvant letrozole.

Results

Analysis is on-going and results from all patients will be available by December 2013. Pairwise Rank Product analysis (false discovery rate 0.01) of the first 12 patients representing all subtypes of breast cancer (6 ER+, 4 HER2+, 2 triple negative) with a mean interval of 22.9 days (range 15 - 33 days) between specimens, identified 346 down regulated genes. Functional processes represented by these genes were; ribosomes, regulation of cell death and apoptosis, and protein complex biogenesis. Up-regulated processes (236 genes) included functions relating to regulation of transcription, mRNA processing/splicing and ECM remodelling.

No consistent changes in a 60 gene immune signature were seen across these 12 patients. Integration of this dataset with a large letrozole-treated dataset, demonstrated that less than 1% of genes that changed on-treatment with letrozole, changed without treatment.

Conclusions

This is the largest study investigating molecular profiles of the same breast cancer sampled at different times. It has shown:

· No consistent changes in expression of genes associated with wound healing and immune response 15-33 days after core biopsy

· Stability of the genetic profile of individual tumours despite multiple biopsies

· Stability in the genes that are changed by two weeks of treatment with daily letrozole.

This study provides definitive evidence of the validity of preoperative or window of opportunity molecular studies and has major implications for investigating the effects of new therapeutic drugs.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-05.
Abstract P4-05-06: Host inflammation and breast cancer molecular subtypes: Updated results from a TCGA sub-analysis
Authors: Allison KH , Chen Y-Y , Jensen KC , Sanders ME , Reis-Filho J , Lester SC , Johnson NB , Factor RE , Tse GMK , Shin SJ , Eberhard DA , Tan PH , Collins LC , Korski K , Waldman FM , Perou CM , Beck AH
Published: August 2014

Background: There is increasing evidence that the presence of a host inflammatory response to breast cancer may influence outcomes. Utilizing inflammation scores on the histology of breast cancer samples submitted for comprehensive molecular analyses for The Cancer Genome Atlas (TCGA), we provide an updated look at associations between the presence of host inflammation and breast cancer molecular and pathologic features.

Design: Experts in breast pathology reviewed the digital slides of breast cancer samples submitted for comprehensive molecular profiling to the TCGA and scored each case for the level of inflammation present (high/moderate vs mild/minimal). We tested pairwise associations between host inflammation and molecular subtypes (DNA copy-number, RNA expression, RPPA defined subtypes, miRNA subtypes, methylation subtypes) and pathological features by performing Chi-Square analyses. Multiple hypothesis testing correction was performed using the Bonferroni method.

Results: 598 breast cancer cases with TCGA molecular profiling data were scored by the expert breast pathologists for morphological features (including inflammation). 195 (33%) of these were scored as high/moderate inflammation. Cases with inflammation had a significantly higher rate of TP53 mutations (p = 9.0e-8) with 64 of 118 (54.2%) p53 mutant cases with inflammation. Inflammation was also significantly associated with PAM50 molecular subtypes (p = 2.2e-11), with the greatest enrichment among basal-like (64.5% of 70 basal-like cases had inflammation) and the greatest depletion among Luminal A (18.1% of 166 Luminal A cases had inflammation). Cases with inflammation were significantly less likely to be lobular (p = 1.5e-7), had less tubule formation (p = 0.0006), increased mitoses (p <2.2 e-16), increased nuclear pleomorphism (p = 2.9e-15), and increased necrosis (p = 2.9e-14).

Conclusions: There are strong associations between breast cancer molecular and pathological features and the host inflammatory response.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-06.