bluuebunny/crossref_metadata_2025 huggingface dataset:
Epidermal growth factor receptor (EGFR) as a prognostic factor over 10 years in non-metastasic colon carcinoma (NMCC)
Authors: Pericay C. , Santos-Vivas C. , Pous-Saltor E. , Bonfill-Abella T. , Dotor-Navarro E. , Fernández-Morales L. , Dalmau- Pórtulas E. , Casalots-Casado A. , Rey-Ruhí M. , Saigí-Grau E.
Published: March 2020

21062

Background: EGFR is a protooncogen that facilitates metastasis in cancer. It is overexpressed in colorectal carcinoma in 40% to 80%, and correlates a worse prognosis. Aim: To find patterns of EGFR overexpression in NMCC and to correlate these patterns with the follow-up. Methods: We analized NMCC with a selection criteria: 1. primary adenocarcinoma of the colon, without metastasis and with curative surgery, 2. pT3 pN0–2 pM0, progression-free in the first 6 months postsurgery, 3. initial follow-up began 6 months postsurgery, 4. minimum follow-up over 5 years. Dako PharmaDx kit (Glostrup, Denmark) was used for immunohistochemical study for EGFR. Dako staining and A431-AAM cells were used as positive control. Immunoreactivity for membrane (1(+), 2(+) and 3(+) intensity) and for cytoplasma was evaluated. Proportions of immunostained cells were also assessed. Results: 195 patients (p) were eligible. Year of diagnosis: 1994: 1p; 1995: 4p; 1996: 20p; 1997: 29p; 1998: 33p; 1999: 36p; 2000: 34p; 2001: 38p. Male/female: 93/102; Mean age: 71 years (28- 94). Location: 116/79 left/right. Stage II/III: 125/70. EGFR membrane positivity: 84/195 cases (43%), 49/125 (39%) in Stage II and 35/70 (50%) in Stage III. With a median follow-up of 7 years: Stage II: 29/125 relapsed (EGFR±: 12(24%)/17(22%)). Stage III: 33/70 relapsed (EGFR± : 21(60%)/12(34%)). EGFR membrane positivity was a poor prognostic factor with a significant progression-free survival (PFS) (p=0.042) and a non-significant overall survival (OS) (p=0.066). When follow-up period was increased over 10 years (patients from 1994 to 1998), EGFR membrane positivity maintained its poor prognostic factor in PFS (p=0.009) and presented a significant OS (p=0.02). Non-significant differences were found in immunostaining ratios. Conclusions: In patients with non-metastatic colon cancer, EGFR membrane positivity was a tumor progression prognostic factor, and this poor prognosis was maintained over time. Study supported by a grant from C.I.R. Hospital de Sabadell. Consorci Sanitari Parc Taulí. Sabadell and a grant from Merck Laboratory.

No significant financial relationships to disclose.
Clinical validation of a self-organizing map (SOM)-based quantitative prediction algorithm to determine the site of origin of cancers of unknown primary (CUP)
Authors: Potts S. J. , Chen R. , Qu K. Z. , Crossley B. , Bender R. A.
Published: March 2020

21064

Background: As many as 100,000 new cases of CUP are diagnosed each year in the United States. Identifying the site of origin of the CUP is critical for effective cancer management, and several genomic assays have been developed to facilitate this process. The genomic profile generated does not always fit into a discrete classification, however, and communicating results in a simple, easily understood format is important to clinical practice. Methods: We created a novel algorithm that combines SOMs for qualitative visualization with a quantitative clustering algorithm to predict the site of origin of metastatic cancers. We trained the algorithm on a dataset of 231 tumor-bank samples from 9 primary sites. After fixing all thresholds and algorithm settings, we evaluated the algorithm's performance in making a correct first or second prediction on a second blinded and independent dataset of 63 samples. Results: The algorithm correctly predicted the primary site in 82% of cases with the first prediction, and 89% with either the first or second prediction. The map visualization component helped to reduce the biological and mathematical complexity of the test into an easy to understand format for practicing physicians. The map- based approach allowed a fuzzy prediction that complements a class-based prediction and provides substantial additional information on the relationship between an unknown specimen and other related specimens of known provenance. Conclusion: CUP is a formidable clinical problem; correctly identifying the primary site and rendering specific treatment translates into improved patient outcomes. The use of an SOM to depict the genomic profile of the tumor specimen makes it easier for the clinician or pathologist to understand what tumor type the CUP most closely resembles and facilitates further diagnostic or therapeutic decisions.

No significant financial relationships to disclose.
Expression of the glutathione-S-transferase π (GST-π) protein correlates with survival in patients with stage I non-small cell lung cancer (NSCLC)
Authors: Gajra A. , Vajpayee N. , Wade M. , Gamble G. P. , Rajan A. , Graziano S. L.
Published: March 2020

21065

Background: It is important to identify prognostic markers in stage I NSCLC given the 30%-40% risk of recurrence and death despite resection. The role of platinum-based adjuvant chemotherapy remains unclear in stage I NSCLC. The glutathione pathway prevents cellular DNA damage by oxygen free radicals and cytotoxic compounds. GST-p catalyzes conjugation of glutathione to cytotoxic agents and high expression may predict for platinum resistance. The role of GST-p as a prognostic factor in early stage lung cancer is not well evaluated. Methods: In a retrospective cohort of 93 patients with resected stage I NSCLC we used immunohistochemistry (IHC) to study the association of GST-p expression with risk of tumor recurrence and death. None of the patients received adjuvant therapy. IHC was quantified by a visual grading intensity of cytoplasm staining (0= no immunoreactivity; 1=weak; 2= clear reactivity in >50% cancer cells and 3= strong immunoreactivity). Grades 2 and 3 were classified as high and grades 0 and 1 as low GST-p respectively. Results: The mean age of the patients was 67 years. Of the 93 patients, there were 45 (48%) men, 71 (76%) had adenocarcinoma and 66 (71%) had stage IA disease. The median follow-up time was 5.4 years. High GST-p expression was noted in 40 (43%) and 53 had low GST expression. The overall survival at 5 years was 67% with 31 deaths. Patients with high GST expression had a higher overall survival at 5 years- 32/40 (75%) vs. 30/53 (57%) in the low GST group (p=0.018). There was no significant difference in terms of tumor recurrence or recurrence free survival. After adjusting for age, gender, histology, and T stage, patients with low GST-p expression remained at an increased risk of death with a hazard ratio of 2.86 (1.16–7.61, p= 0.015). Conclusions: In this cohort, high expression of GST-p served as an independent prognostic factor in resected stage I NSCLC. These early findings with GST-p are similar to those from the IALT trial wherein the high expression of ERCC1 was associated with chemoresistance but conferred a survival advantage in the absence of chemotherapy (Olaussen, NEJM 2006). These findings merit evaluation in larger prospective series given the small, exploratory and retrospective design of the present study.

No significant financial relationships to disclose.
A de novo deletional germline mutation in the MLH1 gene resulting in early onset colorectal cancer in a woman with a negative family history: A case report
Authors: Zakalik D. , Goldstein N. S. , Ducaine W. L.
Published: March 2020

21067

Background: Hereditary non-polyposis colorectal cancer (HNPCC) is a genetic disorder that results in an increased risk of early onset colorectal cancer (CRC). HNPCC is caused by germline mutations in DNA mismatch repair (MMR) genes, including MLH1 and MSH2, and is transmitted in an autosomal dominant manner. De novo germline mutations in MMR genes are exceedingly rare. We now describe a case of a de novo germline mutation in MLH1 associated with early onset CRC in a young woman with a negative family history. Methods: We present a case of a 31-year-old Caucasian female who presented with abdominal pain. Colonoscopy revealed a moderately differentiated adenocarcinoma with focal mucin production involving the sigmoid colon, with a final staging of T4N0M0. Genetic testing was offered because of her young age at diagnosis, having fulfilled the Bethesda guidelines. Microsatellite instability (MSI) testing using a panel of six microsatellite loci was performed on the tumor sample from the affected patient. Immunohistochemistry (IHC) testing for MLH1 and MSH2 was performed. Following counseling and with informed consent, DNA was isolated and sequenced using bi-directional PCR of the MLH1 gene. All exons of MLH1 and MSH2 were analyzed by standard Southern blot methods. Paternity was established using eight genetic loci. Results: The patient's tumor revealed high MSI and complete absence of MLH1 immunoreactivity. MSH2 IHC staining was normal. A large deletional mutation involving exons 5–12 of MLH1 was identified by Southern blot analysis. The patient's parents and siblings were tested and found to have wild type MLH1. Paternity was confirmed with greater than 99.9% certainty. Conclusions: De novo mutations in MMR genes are a rare cause of HNPCC. We report the first case of a large de novo deletion in the MLH1 gene accounting for early onset CRC. Such de novo mutations, albeit rare, must be considered in patients who present with early onset CRC and a negative family history. These results support the use of the Bethesda guidelines to identify individuals who may carry mutations in the MMR genes.

No significant financial relationships to disclose.
Predicting nonsentinel node status in breast cancer preoperatively by molecular characterization of core needle biopsies with multiparametric protein analyses
Authors: Sauer G. W. , Schneiderhahn-Marra N. , Kurzeder C. , Koretz K. , Kreienberg R. , Joos T. , Deissler H.
Published: March 2020

21068

Background: Axillary lymph node dissection (ALND) still remains the standard treatment for breast cancer patients with sentinel lymph node (SLN) metastases. However, since only 40% to 60% of patients show additional lymph involvement, complete ALND offers no additional benefit for almost 50% of patients but carries a significant risk of axillary morbidity. In an attempt to achieve a more precise prediction for the individual patient a multivariable logistic-regression analysis of a large data set of eight histopathological variables has been published by the Memorial Sloan-Kettering Cancer Center (MSKCC) that predicts the likelihood of metastases in Non-SLNs with a ROC of 0,71. To achieve a more precise estimation of additional lymph node involvement preoperatively we analysed molecular markers of potential predictive value. Methods: Beside histopathological variables, fifty proteins of potential prognostic and predictive value were preoperatively quantified in lysates from 120 core needle biopsies with multiplex sandwich immunoassays. Biopsies, taken at the Breast Cancer Center of the University of Ulm Medical School, were classified as invasive mammary carcinomas. Variables which could be used to improve the accuracy of prediction of non-sentinel lymph node (NSLN) involvement were defined. Results: We demonstrated appropriate sensitivity, reproducibility, and robustness for this protein microarray technology to characterize proteins in core needle biopsies and generate reliable data sets. In an initial univariate data analysis (Mann Whitney test) six of the analysed parameters (TIMP2, p= 0,027; FasR, p = 0,0025; MIF, p= 0,030; FGF-2, p= 0,0020, PDGF AA/BB p= 0,027; RANTES p= 0,024) correlated with NSLN involvement beside known histopathological variables (i.e. tumor size, p= 0,006; grading, p = 0,0009). Conclusions: We defined molecular markers of diagnostic value to predict NSLN involvement. This might be an acceptable substitute to the known histopathological variables by the MSKCC nomogram to predict potential NSLN involvement preoperatively.

No significant financial relationships to disclose.
The prognostic value of nodal ratios in node-positive breast cancer: AUBMC experience
Authors: Shamseddine A. , Hatoum H. , Salem Z. , Abdel Khalek Z. , El Saghir N. , Taher A.
Published: March 2020

21072

Background: Axillary lymph node metastasis has proven to be the most important factor affecting overall survival (OS) and disease free survival (DFS) in patients with breast cancer. Recent evidence suggests that axillary lymph node ratio (LNR) may be at least as important as absolute number of involved lymph nodes in predicting OS and DFS. The aim of this retrospective study is to evaluate the impact of axillary nodal ratios in node-positive breast cancer as a prognostic factor for survival. Methods: Data from 1181 patients with stage I, II and III breast cancer diagnosed at AUBMC between 1990 and 2001 were studied. The median age at diagnosis was 50 years (23 - 88); the median number of lymph nodes dissected was 17 (0 - 49). Survival was compared in 737 patients with node-positive disease according to a LNR below or more than 0.25 (defined as number of involved lymph nodes divided by total dissected axillary lymph nodes). Results: Patients with LNR = 0.25 had a median follow-up of 30 months (1.2–156) and a median DFS of 26 months (1–156). The 5-year survival was 26.2% (94/358) and the 5-year DFS was 22.9% (82/358). Patients with LNR <0.25 had a median follow-up of 36 months (1.2–157) and a median DFS of 36 months (1–157). The 5-year survival of 33.2% (245/737) and the 5-year DFS was 29.8 % (220/737). LNR showed significance as a continuous variable and a categorical variable (0, < 0.25, and = 0.25) with a p < 0.001 Conclusions: LNR significantly predicts OS and DFS in node-positive primary breast cancer.

No significant financial relationships to disclose.
EGFR/K-ras mutations in lung adenocarcinoma and atypical adenomatous hyperplasia (AAH)
Authors: Matsumoto S. , Takuwa T. , Kondo N. , Okumura Y. , Fukuoka K. , Nakano T. , Hasegawa S. , Tanaka F.
Published: March 2020

21074

Background: Mutations in the epidermal growth factor receptor (EGFR) gene and k-ras gene are found in 10–30% of non-small cell lung cancer, and are useful to predict the efficacy of EGFR-tyrosine kinase inhibitors. Such mutations are also onco- developmentally important especially in lung adenocarcinoma, but the exact incidences of such mutations in adenocarcinoma/AAH and surrounding normal lung. Methods: A total of 45 consecutive patients with lung adenocarcinoma and/or AAH (18 females; mean age, 65 years) who received thoracotomy from August 2005 through December 2006. In each case, EGFR/k-ras mutations were examined in specimens cut from adenocarcinoma/AAH and normal lung; in the present study, pure bronchioalveolar carcinoma (BAC) was reviewed separately from adenocarcinoma. Results: As 12 (27%) of all 45 patients had multiple adenocarcinoma/BAC/AAH lesions, mutations were examined in 59 lesions (41 adenocarcinomas, 7 BACs, 11 AAHs) and 45 normal lung tissues. EGFR and k-ras mutations were documented in 37% (15/41) and 2% (1/41) of adenocarcinomas, and no EGFR/k-ras mutation was documented in any BAC, AAH, or normal lung tissue. Next, clinical characteristics of patients were compared between patients with solitary adenocarcionma/BAC/AAH lesion and those with multiple lesions; non-smoker were more frequently documented in multiple-lesions group (42%) than in solitary-lesion group (15%), and there was no significant difference in other characteristics such as sex and age. Conclusions: EGFR mutation was not documented in non-invasive (BAC) or pre-neoplastic (AAH) lesion in this series, which may suggest that EGFR mutation play a critical role in the development of invasive adenocarcinoma.

No significant financial relationships to disclose.
Expression of vascular endothelial growth factor (VEGF), p53 in Molecular breast cancer subtypes of pre-menopausal African-American women
Authors: Ihemelandu C. U. , Leffall L. D. , Naab T. J. , Frederick W. A.
Published: March 2020

21084

Background: Tumor growth and metastasis have been shown to be dependent on angiogenesis. With the current classification of breast tumors into molecular subtypes with distinct prognosis and response to treatment, we sort to analyze the expression of the angiogenesis markers in molecular subtypes and determine their association with clinicopathologic variables of prognostic significance. Methods: A retrospective analysis of women diagnosed with breast cancer from 1998–2005, who had assessable data for ER, PR, and Her-2/neu status. The molecular subtypes were defined as: luminal A, luminal B, basal-like , and Her-2/neu. Results: All molecular breast cancer subtypes overexpressed VEGF, with no statistically significant difference noted between the subtypes: - luminal A (69.8%) basal-like (71.1%), luminal B (70.0%), Her-2/neu (71.0%) (p=.99). Subtypes differed significantly in expression of p53 (p<.000), with the basal-like and Her-2/neu subtypes more likely to be associated with p53 mutations (51.7%) and (54.1%) respectively. No statistically significant association between p53 protein and increased VEGF expression was noted (p=.176) Statistically significant associations between p53 protein and prognostic factors ER (p<.000), PR (p>.000), histologic grade (p<.000), S-phase fraction (p<.001) were noted. A significant inverse correlation was noted between p53 expression and thrombospodin for the age-group <35 years (rho -.810; p=.003). VEGF showed no significant association with the prognostic factors ER, PR, histologic grade and S-phase fraction. A tendency not reaching statistical significance was found between VEGF and angiogenesis (p=.09). A direct correlation between VEGF and thrombospodin was noted in the age- group < 35 years (rho .800; p=.01). Expression of VEGF and thrombospodin did not correlate with survival outcome; however angiogenesis seemed to correlate with survival outcome. Survival outcome was influenced by molecular subtypes with the basal-like and Her-2/neu subtypes having a poorer outcome (p=.01). Conclusions: VEGF expression is not related to p53 status or survival outcome in molecular breast cancer subtypes of pre-menopausal African-American women.

No significant financial relationships to disclose.
VEGF-Trap (V-T) overcomes defects in dendritic cell (DC) differentiation without improvement in antigen-specific immune responses
Authors: Sosman J. A. , Fricke I. , Mirza N. , Dupont J. , Lockhart A. C. , Jackson A. , LaFleur B. , Gabrilovich D.
Published: March 2020

21087

Background: Induction of antitumor immune responses requires adequate DC function. Well-documented DC defects in cancer mediated through tumor-derived vascular endothelial growth factor (VEGF) may lead to tumor escape and limit efficacy of cancer vaccines. V-T with extracellular domains of VEGFR 1/ 2 coupled to Fc IgG1 binds all VEGF-A isoforms. Methods: To determine if inhibition of VEGF signaling improves immune responses, we evaluated 15 cancer pts treated with V-T (2.0–7.0 mg/kg) IV over 1 hr Q 2 wks on a phase I trial. To evaluate immune parameters, PBMC were collected prior to, days 15, 29, and 57 of V-T treatment. Ligand blockade was achieved in all patients. Mature DCs (myeloid and plasmacytoid subsets), and regulatory T cells (T regs) were characterized by expression pattern of CD11c, CD86, CD40, CCR7, CD83, CD123, CD4, CD25, and GITR. T cell function was assessed by allogeneic mixed leukocyte reaction (MLR) and proliferation to tetanus toxoid, influenza, or PHA. Results: V-T treatment did not affect the presence of the total population of DCs, myeloid or plasmacytoid subsets, or myeloid-derived suppressor cells (MDSC). It significantly increased the proportion of mature DCs expressing CD86, CD40, CD83, and CCR7R (p<0.05) without overall improvement in T cell immunity. At 8 wks, 11 pts had either stable disease (SD) or partial response (PR) to the therapy and 4 pts had progressive disease (PD), both with no significant differences in the proportion of MDSC or total DCs. The proportion of mature DCs was significantly increased in pts with SD and PR (p<0.025), but remained unchanged in PD pts. However, functional tests were similar in both groups. Another subset analysis revealed significant improvement in immune responses in 6 pts who had stable or decrease in the MDSC proportion, but not in 9 pts with increases in MDSC. During V-T treatment, Tregs further increased above baseline in pts (p=0.048). Conclusions: These data demonstrate that inhibition of VEGF signaling improves differentiation of DCs in cancer patients, while alone it was not sufficient to improve immune responses. These data illustrates the multifaceted nature of immune deficiency in cancer pts and points to a need for complex approaches to modulation of immune reactivity.

No significant financial relationships to disclose.
Does triple-negative breast cancer (TNBC) have distinct clinicopathologic characteristics and prognostic significance?
Authors: Rhee J. , Oh S. , Oh D. , Im S. , Lee S. , Kim D. , Heo D. , Park I. , Bang Y. , Kim T.
Published: March 2020

21088

Background: Studies have suggested that TNBC, defined by estrogen receptor-negative, progesterone receptor-negative, and HER2-negative, may represent the subset of breast cancer(BC) with different biologic behavior. Here we investigated the clinicopathologic characteristics of TNBC and its prognostic significance in Korean BC patients. Methods: Patients diagnosed as invasive BC and underwent curative surgery at Seoul National University Hospital between Jan. 2000 and Jun. 2003, were reviewed, retrospectively. We excluded the patients whose immunohistochemistry for hormone receptor nor HER2 status had not been evaluated, and who had been treated with adjuvant trastuzumab or neoadjuvant chemotherapy (CT). Clinicopathologic variables (age, T and N stage, endovascular or lymphatic tumor emboli, nuclear and histologic grade, p53, bcl2, Ki67) and 3 year relapse free survival (3YRFS) rate of TNBC were compared with those of non- TNBC. Results: 1,136 patients were eligible for analysis. The median follow-up was 48.7 months. 341 patients underwent breast conserving surgery followed by adjuvant radiotherapy. 249 patients were TNBC and 62.1% of those were node negative. 86.4% of node negative TNBC, 88.3% of node positive TNBC, 53.9% of node negative non-TNBC, and 90.2% of node positive non-TNBC received adjuvant CT. Compared with non-TNBC, TNBC was correlated with younger age (age<35,14.1% vs. 8.2%, p=0.013), higher nuclear and histologic grade(62.2% vs. 23.6%, p=0.001;60.2% vs. 24.6%, p=0.001, respectively); positive staining for p53 (p=0.001) and higher positivity for Ki67 (p=0.001), suggesting the biologic aggressiveness of TNBC. During the follow-up periods, 17.3% of TNBC were relapsed. In particular, 3YRFS in node negative TNBC and non-TNBC were 86% and 96%, respectively (p<0.001). But, in node positive BC, 3YRFS was not different between TNBC and non-TNBC (80.6% vs. 83%, p=0.99). Conclusions: We confirm that TNBC shows more aggressive clinicopathologic characteristics and in particular, higher relapse in node negative BC. Thus, triple-negativity(TN) may be integrated into risk factor analysis in node negative BC. Final results of more detailed molecular analysis for TNBC would be available in the meeting.

No significant financial relationships to disclose.
Null allele polymorphisms in the GSTT1 and GSTM1 genes in 705 women from a mammographic breast cancer screening program (NMPOA) in southern Brazil
Authors: Aguiar E. , Giacomazzi J. , Palmero E. I. , Kalakun L. , Schuler-Faccini L. , Duarte Filho D. , Skonieski G. , Caleffi M. , Giugliani R. , Ashton-Prolla P.
Published: March 2020

21091

Background: Several genetic polymorphisms in genes related with metabolism have been associated with breast cancer (BC) risk. Among these, the gluthatione-S-transferase M1 and T1 null genotypes have been associated with slightly increased BC risk in some populations. In Brazil, BC is a significant public health problem, due to its high incidence and mortality rates. In Porto Alegre, Brazil`s southernmost capital, a multidisciplinary BC Prevention Project - the Nucleo Mama Porto Alegre Cohort (NMPOA)- was started in 2004 and includes a mammographic screening program for women ages 40–69 years. Goal:Determine the allelic and genotypic frequencies of GSTM1 and GSTT1 null alleles in women undergoing annual mammographic screening and correlate its presence with mammography results and presence of additional BC risk factors at baseline and after 10 years. Methods: A sample of 705 women from the NMPOA BC screening program was consecutively enrolled from November/2005 until March/2006. Mammography results (BIRADS categories) and BC risk information (5-yr and vital estimates using the Gail model, family history of BC, body mass index) were obtained by chart review. Genotyping was performed by multiplex polymerase chain reaction (PCR). Results: Of the 705 patients studied, 145 (20.6%) and 314 (44.5%) had the GSTT1 and GSTM1 null alleles, respectively. Genotypically, 67 (9,5%) were null homozygous for both genes (GSTM1- and GSTT1- ); 78 (11,1%) were GSTT1- and non-null for GSTM1 (GSTT1- and GSTM1+); 247 (35%) were GSTT1+ and GSTM1- and 313(44,4%) were GSTM+ and GSTT+. There was a statistically significant association of the GSTT1+ allele with low-risk mammographic findings (category BIRADS 1; p<0,05). Conclusions: The genotypic and allelic frequencies of the GSTT1 and GSTM1 null alleles were not significantly different from those reported previously for other populations. The non-null GSTT1 allele was associated with lower category mammographic findings. Prospective clinical evaluation of the women followed in this program and correlation of genotype with clinical findings may elucidate additional risks associated with the presence of these polymorphisms.

No significant financial relationships to disclose.
In vivo and in vitro efficacy of capecitabine (X) + tamoxifen (TAM) in breast cancer (BC)
Authors: Mori K. , Yamaguchi Y. , Sawada N. , Kondoh K. , Hayashi S.
Published: March 2020

21092

Background: In vitro studies in BC cell lines suggested antagonism between TAM and 5-FU. Thymidine phosphorylase (TP) activates X to 5-FU in tumors. X activity correlates with tumor TP concentrations in vivo. Methods: We studied antitumor efficacy of X + TAM in vivo and in vitro in human BC models. Nude mice were inoculated s.c. with estradiol, then MCF-7 cells 1 day later. When tumors were ∼300 mm3, mice received 6 weeks’ oral vehicle (control), X (d1–14 q21d) at MTD (539 mg/kg) or 2/3 MTD, and/or TAM at 100 or 30 mg/kg/d. We also analyzed impact of 5-FU and doxifluridine (5'-DFUR, an intermediate of X) + TAM on estrogen receptor (ER) signals in an in-vitro culture system. ER signals were monitored by expression of green fluorescent protein (GFP) in MCF-7 BC cells transfected with the estrogen-responsive element (ERE)-GFP gene (MCF-7-E10). GFP expression was induced in MCF-7-E10 cells in the presence of estradiol at 3 pM or BC tissue supernatant. Results: X at 2/3 MTD + TAM 30 mg/kg were significantly more active than the highest dose of X or TAM alone. Tumor TP concentrations were significantly higher in TAM- than vehicle-treated mice. In the ER signal system, GFP expression of MCF-7-E10 was reduced by 4-hydroxytamoxifen (4-OHT, active form of TAM) at 0.01 and 0.1 nM. When added to 4-OHT, 5-FU 0.3–30 μM or 5'-DFUR 3–10 μM reduced GFP expression more than either agent alone. In vitro, 5-FU and 5'-DFUR inhibited proliferation of MCF-7-E10 cells regardless of 4-OHT. Additive effects could not be confirmed as 4-OHT alone showed only marginal anti- proliferative activity at 0.01–0.1 nM. Conclusion: X and TAM are not antagonistic in this model. TAM may augment X activity via TP upregulation in BC tissues. TAM and X intermediates showed no clear antagonism in vitro in an ER signal system. All-oral X + TAM merits evaluation as combination therapy in breast cancer.

[Table: see text]
Genetic testing in young women with breast cancer: Results from a web-based survey
Authors: Shin J. A. , Gelber S. , Garber J. , Rosenberg R. , Przypyszny M. , Winer E. , Partridge A.
Published: March 2020

21093

Background: Young women with breast cancer have an increased risk of harboring a BRCA1/2 mutation. The frequency of genetic testing in this population is not well described. We evaluated the reported frequency and factors associated with genetic testing among young breast cancer survivors identified through the Young Survival Coalition (YSC), an international advocacy group for young women with breast cancer. Methods: Items regarding family history and genetic testing were included in a large web-based survey addressing quality of life and fertility issues for young women with breast cancer. All YSC members were invited by email in March 2003 (N= 1,703 women) to participate in this cross-sectional survey. Results: 657 women completed the on-line survey; 622 were eligible for this analysis (age <40, no metastatic or recurrent disease). Mean age at breast cancer diagnosis was 33 years; mean age when surveyed 35.5 years. Stages included: 0 (10%), I (27%), II (49%), III (12%), missing (3%). 90% of women were white; 64% married; 49% with children; 78% had at least a college education; 42% of women reported a 1st or 2nd degree relative with breast or ovarian cancer, and 13% considered themselves high-risk for harboring a genetic mutation at the time of diagnosis. At the time of the survey, 23% of women had undergone genetic testing, and 26% of those tested reported that a mutation was found. In a multivariate model, women who were younger (age 36–40 vs. age =30, O.R. 2.26, p=0.004), more educated (< college vs. > college education, O.R. 2.62, p=0.0009), had a family history of breast or ovarian cancer (O.R. 3.15, p<0.0001), and had had a mastectomy (O.R. 1.99, p=0.001) were more likely to have undergone genetic testing. Non-significant covariates included: age at survey, stage, time since diagnosis, race, marital status, employment, finances, insurance, number of children, comorbidities, baseline anxiety and depression, and fear of recurrence. Conclusion: The majority of women diagnosed with breast cancer age 40 and younger do not undergo genetic testing. Younger, more educated women with a family history of breast or ovarian cancer are more likely to get tested. Further research to define the appropriateness of genetic testing in this relatively high-risk population is warranted.

No significant financial relationships to disclose.
Topoisomerase-2alpha (T-2a), Ki67, Her-2 and response to neoadjuvant anthracycline-containing chemotherapy in breast cancer. A prospective, correlation study
Authors: Bari M. , Sartori D. , Zambenedetti P. , Tacchetti G. , Sicari U. , Rosetti F. , Iop A. , Pappagallo G. L. , Vinante O.
Published: March 2020

21094

Background: T-2a creates a reversible double-strand DNA break allowing DNA doubling. Anthracyclines (A) stabilize the DNA double-strand breaks, and T-2a is probably the primary molecular target of A. Due to the close location of T-2a and Her-2 genes on chromosome 17, T-2a gene aberrations are mainly associated with Her-2 gene amplification; while a correlation exists between Her-2 amplification and protein overexpression this is not true for T-2a. A linear correlation between T-2a and Ki67 labeling indices was found, suggesting that both essentially reflect cellular proliferation. The correlation between T-2a overexpression and both Her-2 and Ki67 was investigated in a series of consecutive patients undergone neoadjuvant A-containing chemotherapy for locally-advanced breast cancer. Material and Methods: T-2a expression was measured by means of monoclonal antibody Ki-S1; thresholds (ts) for immunopositivity were tested at 10%, 15% and 20%, respectively. Both the anti-c-erb-B2 primary antibody (clone CB11) and the Dako test were employed to recognize c-erb-B2 protein. Ki67 was measured using the MIB-1 antibody, with ts for positivity at 10%. Patients were required to have a cT>2cm breast cancer. The neoadjuvant chemotherapy included Adriamycin 60mg/m2 or Epirubicin 75mg/m2, in combination with Paclitaxel (175 mg/m2), every 3 weeks for 4 cycles. Bivariate correlations were performed according to Pearson. Results: 38 patients were enrolled until August, 2006. A significant correlation between T-2a and Ki67 was found (r=.598; P<.000); the T-2a positivity rate within Ki67 positive patients was of 90% (10% ts), 86% (15% ts), and 67% (20% ts). No correlation appeared between T-2a and Her-2 labeling indices (r=.150; P=.391); the T-2a positivity rate within Dako +++ patients was 75% (for both 10% and 15% ts) and 50% (for 20% ts). The overall response rate by T-2a overexpression was 61% (10% ts), 70% (15% ts), and 72% (20% ts). Conclusions: We provide a further evidence of the correlation between T-2a and Ki67. We can also generate the hypothesis that a 20% ts for T-2a correlate with a better prediction of response to A-containing chemotherapy; at the same ts, no correlation with Her-2 status was found.

No significant financial relationships to disclose.
A comparison of tumor characteristics between ER+/PR+ and ER+/PR- breast tumors in Chinese women
Authors: Zhou L. , Yin W. , Lu J. , Shi D. , Liu G. , Di G. , Wu J. , Shen K. , Lu H. , Shen Z. , Shao Z.
Published: March 2020

21097

Background: Although breast caner patients with any one of the two sex hormone receptors positive can be treated with endocrine therapy, many clinical data showed that there was different response to endocrine therapy for patients with ER+/PR+ and with ER+/PR- tumors. The aim of this study was to find out the factors related to PR expression by comparing the ER+/PR+ tumors and ER+/PR- tumors clinically and biologically. Methods: Between January 1990 to August 2006, 5,191 female breast cancer patients with known ER/PR expression status who received operation in our hospital were enrolled onto this retrospective study. Clinical and biological features of 2,227 patients with ER+/PR+ tumors were compared with those of 909 patients with ER+/PR- tumors. χ2 test was used for univariate analysis and logistic regression for multivariate analysis. Disease-free survival (DFS) and overall survival (OS) was calculated using Kaplan-Meier analyses, and all statistical tests were two-sided. Results: The peak onset age of patients with ER+/PR+ tumors and ER+/PR- tumors was 50, and it was significantly higher than that of patients with ER- tumors, which is 48(P=0.001). Univariate analysis showed that ER+/PR- tumors were larger in size, had more lymph nodes of metastasis, were higher in tumor grade than ER+/PR+ tumors. Furthermore, the expression of ER and CathepsinD was significantly lower, and CerbB-2 expression was higher in ER+/PR- tumors than in ER+/PR+ tumors. Multivariate analysis indicated that positive PR expression was associated with the level of ER(OR=1.792, P=0.000), CathepsinD(OR=1.380, P=0.035)and CerbB-2(OR=0.639, P=0.007). DFS(P=0.004) and OS(P=0.009) were higher among patients with PR-expressing tumors than with PR- negative tumors. Conclusions: ER+/PR+ tumors and ER+/PR- tumors may have the same etiology which is different from that of ER- tumors. Because of low ER level and changes of the expression of CerbB-2 and CathepsinD, the tumors that lacked PR expression display more aggressive features and have worse prognosis. According to these differences, new target of therapy and endocrine regimen may provide the possibility of improving the response and prognosis of endocrine therapy for patients with ER+/PR- tumors.

No significant financial relationships to disclose.
The rule of mortality risk in young women (≤35 years) with primary breast cancer
Authors: Yin W. , Lu J. , Liu G. , Di G. , Wu J. , Shen K. , Shen Z. , Shao Z.
Published: March 2020

21098

Background: The prognosis of young women (=35 years) is poorer than that of women over 35 years, which may indicate that they bear different biologic characteristics. However, such data is still not available at home and abroad as the rule of mortality risk in young women and its distinction from that in women over 35 years. We sought to investigate the clinical characteristics, prognosis and the rule of mortality risk in young women (=35 years) with primary breast cancer. Methods: We performed a retrospective study of 2331 female unilateral breast cancer patients undergoing surgery in our hospital from January 1990 to October 2005.They were divided into two groups with the cut-off of 35 years. Selection criteria included complete data on the following: age, tumor size, axillary nodal status, estrogen receptor (ER) status, menopausal status.Overall survival curve was estimated by Kaplan-Meier method and annual mortality hazard by hazard function. Their clinicopathologic indices, overall survival curve and annual mortality hazard were analyzed. Results: There was statistical significance between =35 years and >35 years group in terms of tumor size( P =0.0231), axillary nodal status( P <0.001) and estrogen receptor status( P =0.064). 10-year overall survival was 72% and 79% ( P = 0.0081), respectively, for =35 years and >35 years group. Although annual mortality hazard curve for both groups showed double-peaked pattern, there also exist some differences. Mortality surges for =35 years patients appeared earlier than those for >35 years counterparts. Moreover, =35 years group showed a minor increase between double peaks, which cannot be found in >35 years group. Conclusions: Between the two groups, different tumor biology leads to the discrepancy of overall survival and mortality pattern, which might indicate distinct principles and measures in the follow-up and treatment for young patients to provide the possibility of improving the therapeutic effect and reducing the mortality hazard.

No significant financial relationships to disclose.
A comparative analysis of angiogenesis between male and female breast cancers
Authors: Bakkar R. , Nahleh Z. , Bui H. , Samaan S. , Sanders J. , Namakydoust A. , Komrokji R.
Published: March 2020

21101

Background: Tumor angiogenesis and vascularization are essential for the growth and metastasis of tumors. VEGF-C expression and peritumoral lymphocyte density (PLD) are markers of angiogenesis. They have been correlated with poor prognosis in female breast cancer (FBC). The purpose of this study is to characterize VEGF-C expression and PLD in MBC and correlate with FBC specimens. Method: We reviewed records of patients diagnosed with MBC and FBC at the Cincinnati VAMC, from 1989 to 2006. Pathology slides were retrieved. We used VEGF-C (Host Rabbit, PAD: Z-CVC7) . Imunohistochemical stains of VEGF-C were given scores of 0 to 3+ based on nuclear stains. PLD was analyzed based on the number of lymphocyte cells surrounding the tumor; score of 0 to 3+. Slides were reviewed independently by two pathologists. Results: We found nine MBC cases and selected 9 FBC cases. Mean age was 72 in the male patients and 62 in the females. Stages of disease were distributed as follows in MBC versus FBC, 11% versus 22% stage 0, 23% versus 23% stage I, 44% versus 44% stage II, and 22% versus 11% stage IV. Ductal carcinoma was the predominant histology in 88% of FBC and 88% MBC. Other histological types included papillary (1 MBC) and lobular (1 FBC). Among the invasive MBC tumors , 75 % were ER+/PR +, 13% ER+/PR-, and 12% ER -/PR- , compared to 72 % ER+/PR+, 14% ER+/PR-, and 14% ER-/PR- in FBC. Eight out of the 9 MBC cases (89%) stained positive for VEGF-C expression, compared to one FBC case (11%). The 1 male breast intraductal carcinoma was positive for VEGF-C expression, compared to none of the two intraductal FBC. PLD was more intense in male than female tumors : score 0 or 1+: 44% in MBC versus 67% in FBC , score 2+: 22% in MBC Versus 22% in FBC, and score 3+: 22% in MBC versus 11% in FBC. VEGF-C expression did not seem to correlate with ER/PR status. The median survival for patients with MBC was 4.5 years and for patients with FBC 6.9 years. Conclusion: VEGF-C expression and PLD were more pronounced in MBC versus FBC. This finding may correlate with more aggressive behavior of breast tumor cells in male patients, more intense angiogenic reaction and lower median survival. Further studies are warranted to further elucidate the role of angiogenesis in male breast cancer and explore potential antiangiogenic therapeutic modalities.

No significant financial relationships to disclose.
Prognostic impact of MKI67 and MMP1 in node-negative invasive ductal and invasive lobular carcinoma of the breast
Authors: Schmidt M. , Boehm D. , von Toerne C. , Lehr H. A. , Hengstler J. G. , Koelbl H. , Gehrmann M.
Published: March 2020

21102

Background: MKI67 and MMP1 mRNA, which might reflect the biological processes of tumor proliferation and matrix degradation, respectively, have been proposed as prognostic marker in invasive carcinoma of the breast. We compared the expression of both genes in invasive lobular versus invasive ductal carcinoma, which represent the most common histological breast cancer subtypes. Methods: MKI67 and MMP1 mRNA expression was determined by microarray analysis of fresh frozen tumor tissue of 118 patients with invasive ductal and 39 patients with invasive lobular carcinoma. We compared the distribution of these markers between both histological subtypes as well as within each subtype between tumors from patients with or without subsequent distant metastasis or local recurrence by two tailed Mann-Whitney test. Results: MKI67 as marker of proliferation and MMP1 as marker of matrix degradation were significantly elevated in invasive ducal versus lobular carcinomas (p = 0.023 and p < 0.0001, respectively). Furthermore, elevation of both markers were significantly associated with the occurrence of metastasis in ductal (p = 0.023 and p = 0.012) but not in lobular carcinomas (p = 0.986 and p = 0.366). In addition, no significant association was found between these markers and development of local recurrence in patients with ductal (p = 0.304 and p = 0.899) or lobular (p = 0.891 and p= 0.153) carcinomas . Conclusions: Our data suggest that differences exist between ductal and lobular carcinoma of the breast regarding both the expression and the prognostic impact of MKI67 and MMP1. Furthermore, the prognostic relevance of proliferation and matrix degradation was more important for the development of distant metastasis than for the development of local recurrence in ductal carcinoma emphasizing differences in the underlying biology of local recurrence and distant metastasis.

No significant financial relationships to disclose.
Residual cancer burden (RCB) is practical and may be more informative than pathological response following neoadjuvant chemotherapy (NC) for locally advanced breast cancer (LABC)
Authors: Sivasubramaniam D. , Komrokji R. , Dhaliwal S. , Sundarajan V. , Nahleh Z.
Published: March 2020

21103

Background: Complete pathological response (pCR) has been considered a reliable endpoint to assess the benefit of NC. However, different pathological responses ranging from near complete response to resistance would likely indicate different prognostic groups. Method: We studied patients with locally advanced breast cancer (LABC) who received NC between 2001–2006 at the University of Cincinnati. Pathological response to therapy was evaluated. In addition, RCB was quantified according to MD Anderson RCB Calculator index that combines pathologic measurements of primary tumor (size and cellularity) and nodal metastases (number and size). We examined the correlation between pCR, RCB, event-free survival (EFS) and over all survival (OS) by Cox regression analyses. Result: Pathological slides of 32 patients were analyzed. Median age 52, 38% white and 62% African American. Stage IIB 12% , Stage IIIA 19%, Stage IIIB 53% and Stage IIIC 16% . 72% invasive ducal, 6% invasive lobular and 22% inflammatory cancer. Forty seven percent of tumors were ER +/or PR+ , 53% ER-/PR-, 28% HER-2 /neu + ( IHC 3+ or FISH HER2 gene to chromosome 17 ration > 2.2). Tumor response was as follows: 22% (n=7) achieved pCR , RCB scores ranged between 0- 4.87. By univariate Cox regression analysis, RCB correlated with EFS {Hazard ratio (HR) 1.57 (95% CI 1.04–2.38), p-value 0.018}, and with OS {HR 1.74 (95% CI 0.91 -3.32), p value-0.09}. However, pCR did not seem to correlate with EFS {HR 0 .24 (95%CI 0.03 -1.86–2.38), p-value .172} or OS {HR 0.03 (95% CI 0–89),p value-0.40}. By multivariate Cox regression analysis, RCB was noted to be an independent predictive variable for EFS {HR 1.59 (95% CI 1.04–2.43), p value-0.033} while pCR was not {HR 0.90 (95% CI 0.52–1.57), p value-0.7. Conclusion: RCB was easily quantifiable and appears to be a better predictor of outcome following neoadjuvant chemotherapy in LABC compared to pCR. Higher RCB scores were associated with higher EFS and lower rate of OS. Prospective trials are needed to further evaluate the role of RCB as an endpoint following NC.

No significant financial relationships to disclose.
Outcomes after breast cancer in an ethnically diverse cohort of high-risk patients: Differences in survival based on BRCA1/BRCA2 mutation status
Authors: Nanda R. , Huo D. , Cook M. , Chen L. , Hope K. , Cummings S. , Olopade O. I.
Published: March 2020

21116

Background: Most studies of hereditary breast cancer report that BRCA1 associated tumors are characterized by high grade and hormone receptor negativity, while those associated with BRCA2 are more similar to sporadic cases. Several groups have demonstrated that BRCA1 mutations, but not BRCA2 mutations, are associated with reduced survival in women with breast cancer. These studies have primarily included women of European and Ashkenazi Jewish heritage. No study to date has assessed outcomes in an ethnically diverse cohort of high-risk individuals. Methods: High-risk individuals were identified through the University of Chicago Cancer Risk Clinic (UCCRC). Clinicopathologic data was collected on all patients with breast cancer who had undergone genetic testing and consented to inclusion in research protocols. Recurrence-free survival (RFS) and overall survival (OS) for BRCA1, BRCA2 and non-carriers was compared using the log-rank test. Hazard ratios (HR) and 95% Confidence Intervals (95% CI) were calculated from Cox proportional hazard models. Results: 280 patients from 258 families were studied. 67 were BRCA1 mutation positive, 36 were BRCA2 positive and 177 were non-carriers. 65% of patients were non-Hispanic non-Jewish whites, 15% African American, 15% Ashkanazi Jewish, 3% Hispanic and 2% Asian. At a median follow up of 5 years, 40 patients had relapsed and 22 had died. HRs (95% CI) for RFS of the BRCA1 and BRCA2 carriers relative to non-carriers were 1.6 (0.77–3.33) and 1.3 (0.55–3.09) respectively, when adjusted for year at diagnosis. The adjusted HRs (95% CI) for OS for the BRCA1 and BRCA2 carriers relative to non-carriers were 1.82 (0.48–3.02) and 0.67 (0.15–3.04), respectively. Conclusions: In this study of an ethnically diverse cohort of high-risk individuals, BRCA1 mutation carriers had a poorer outcome as compared to those with BRCA2 mutations or those without identifiable mutations in either gene, although this difference was not statistically significant. This observation is consistent with previous studies of women of predominantly Ashkenzi Jewish and European ancestry. This study was funded by the Falk Medical Research Trust, the Breast Cancer Research Foundation and the Entertainment Industry Fund.

No significant financial relationships to disclose.