bluuebunny/crossref_metadata_2025 huggingface dataset:
Telomere dysfunction in prostatic carcinogenesis
Authors: Joshua A. M. , Vukovic B. , Braude I. , Evans A. , Srigley J. , Squire J. A.
Published: March 2020

10021

Background: Telomeres are composed of tandemly repeated DNA sequences (TTAGGG) and specific binding proteins located at the ends of eukaryotic chromosomes. They stabilize chromosomal ends; telomere shortening is an important mechanism of genomic instability and can lead to end-to-end chromosomal fusion, rearrangements and cell death. Here we evaluate the hypothesis that telomere shortening contributes to the development of prostate cancer (CaP). Methods: We used telomeric, centromeric and chromosome specific peptide-nucleic acid probes with z-stacked quantitative fluorescence in-situ hybridisation analysis to initially analyse 15 radical prostatectomy specimens and then subsequently sextant core biopsies from 80 men obtained in 1998–2001 containing high-grade prostate intraepithelial neoplasia (HPIN) only. The biopsy cohort outcome is blinded to prevent experimental bias and has a minimum follow-up of 2 years with 41 men diagnosed subsequently with CaP and 39 men without CaP on rebiopsy. Regions of interest were identifying with an overlying haematoxylin and eosin slide. Results: We found a significant decrease in telomere length in both HPIN and CaP in comparison to normal prostatic epithelium accompanied by elevated rates of aneusomy. Telomere erosion in HPIN was more common in regions of the prostate-containing CaP. We now have analyzed ∼4000 cells of matching HPIN and surrounding stroma. Preliminary analysis demonstrated that the median telomere length in HPIN is approximately 27% of the surrounding stroma with upper and lower quartiles being 16% and 38% respectively. Logistic regression analysis is in progress to determine whether the length of the shortest telomeres or the average telomeric length in a sample predicts for subsequent diagnosis of CaP. Secondary analyses are examining the effect of telomere length on the interval to the diagnosis of CaP, the effect of age on telomere length and the eventual Gleason score. Conclusions: Analysis of telomere length holds great promise for developing improved prognostic markers in prostatic carcinogenesis. This is a first of its kind study in the field.

No significant financial relationships to disclose.
The gene coding for secreted phosphoprotein1/osteopontin is the most overexpressed gene in cholangiocarcinoma—detected by oligonucleotide arrays and LightCycler analysis
Authors: Hass H. G. , Jobst J. , Nehls O. , Frilling A. , Hartmann J. T. , Kaiser S.
Published: March 2020

10036

Background: Cholangiocarcinomas (CCC) are the second most common primary hepatic malignancy with a still poor prognosis and arise from biliary epithelia or cholangiocytes. Until now, less is known about the molecular pathways leeding to CCC. Methods: Oligonucleotide arrays were used to analyze gene expression profiles of 8 intrahepatic CCCs. After isolation of tRNA and transcription into cDNA, biotin-labelled cRNA probes were hybridized to GeneArrays (Affymetrix U 133A) containing probes of more than 22.000 genes/ESTs. For two-dimensional cluster analysis we used special software programs (Genexplore, GeneSpring). Dysregulated genes were determined by presence in more than 70% and a 2-fold change in relation to the corresponding non-malignant liver tissue. Lightcycler analysis were performed to validate the expression datas of dysregulated genes. Results: A total of 694 dysregulated genes (330 up-/364 down-regulated, compared with corresponding non-malignant tissue) were detected. As the gene with the highest and most consistent upregulation we were able to identify osteopontin (OPN) with an average 5-fold overexpression in all CCC tissues. OPN is an acidic phosphoprotein that is secreted by osteoblasts, macrophages and many other cell types and binds to a variety of cell surface receptors (integrins/CD44). OPN is multifunctional, with activities in cell migration, regulation of bone metabolism, immune cell function and control of tumor cell phenotype. Elevated OPN levels were seen in different tumors but until now no data exist about the expression in CCCs. As one possible interaction in human carcinogenesis, OPN has recently been shown to be a novel substrate for some MMPs, which play an importand role in tumor invasion and metastasis. Conclusions: This is the first report about an overexpression of OPN in CCC and our data indicate an important role in cholangiocarcinogenesis. Further studies are needed to illucidate the moleculargenetic mechanisms of OPN interactions in CCC.

No significant financial relationships to disclose.
Transcription factor NFkB a potential molecular marker for predicting and improving treatment efficacy in esophageal cancer
Authors: Izzo J. G. , Wu T. , Malhotra U. , Ensor J. , Luthra R. , Chao C. K. , Swisher S. G. , Liao Z. , Aggarwal B. B. , Hittelman W. N. , Ajani J. A.
Published: March 2020

10065

Background: Esophageal/gastroesophageal junction adenocarcinoma (E/GEJAC) remains one of the most aggressive malignancies. Chemoradiotherapy (CTXRT) followed by surgery has been used for localized E/GEJAC. Patients (pts) achieving pathologic complete response (pathCR) have an improved survival, but ≈70% of pts exhibit at surgery resistant residual, highly aggressive tumors despite CTXRT. There is a need to identify this high-risk population and target molecular pathways associated with cancer resistance. We have shown that nuclear NFκB was associated with poor clinical outcome of E/GEJAC pts undergoing 5FU, Docetaxel and Cisplatinum therapy. To validate our findings, we examined the impact of nuclear NFκB on clinical outcome of pts undergoing diverse CTXRT regimens. Methods: Pre- treatment tumor biopsies and post-treatment resected residual tumors were analyzed from pts receiving neo-adjuvant CT or CTXRT. NFκB protein expression was assessed by immunochemistry and correlated to pathCR and clinical outcome. Tumors were considered NFκB positive (pos) when ≥5% of cells expressed nuclear NFκB. Results: 80 pts, clinically staged II, III and IVA, were studied. All pts received antifolates, and 80%, 65% & 31% received taxanes, topo-1 inhibitors and/or platinum analogues, respectively. Radiation therapy was 50.4 Gy at 1.8 Gy once a day to all pts. 75/80 pts had available pre-treatment biopsies, all 58 pts with <pathCR had available residual tumors. Pre-treatment NFκB was predictive for lack of response to CTXRT [NFκB pos: 2/22 pathCR vs 27/53 <pathCR; P=.006]. In multivariate analysis, including clinical stage, tumor histology, pathCR and lymph nodes metastasis, pre-treatment NFκB was an independent prognostic factor of progression-free (P=.0029, HR=2.90, 95%CI:1.44–5.86) and overall (P=.0073,HR=2.70, 95%CI:1.30–5.60) survivals. NFκB was associated with recurrent disease [pre-treatment NFκB pos 14/29 (48%) vs NFκB negative11/46 (24%), P=.04; pre- or post- NFκB pos 22/47 (47%) vs NFκB neg 4/33 (12%), P=.003]. Conclusions: our data suggest that NFκB defines cancer biology and patterns to therapy response irrespective of the type of chemoradiation used. NFκB may serve as potential molecular target to improve treatment efficacy.

No significant financial relationships to disclose.
A preoperative model evaluating the tissue effects of chemopreventive agents in low-volume prostate cancer
Authors: Kim J. , Wang X. , Babaian R. , Pisters L. , Pettaway C. , Wood C. , Sabichi A. , McDonnell T. , Logothetis C. , Troncoso P.
Published: March 2020

1007

Background: To identify and assess biomarkers modulated by prostate cancer chemopreventive agents, we created a model that, by exploiting the time between histologic diagnosis of prostate cancer and definitive therapy (prostatectomy), expedites tissue cross comparison and confronts the multifocality and multizonal heterogeneity of prostate cancer. Methods: Between February 2001 and April 2002, 48 presurgical patients were enrolled in a single-institution, randomized, double-blind trial of vitamin E (VE) and L-selenomethionine (SeMET) based on the framework of the ongoing Selenium and Vitamin E Cancer Prevention Trial. Patients were treated with 400 IU VE, 200 μg SeMET, a combination of VE and SeMET, or placebo for 3 to 6 weeks before prostatectomy. All patients also received a multivitamin and vitamin C 250 mg each day. Ex vivo simulated sextant biopsies on the radical prostatectomy specimen (RPS) were performed, and 36 of 39 evaluable patients had RPS sections that were suitable for pathologic evaluation. Blood components were collected at baseline and prior to prostatectomy. We derived an apoptotic index (AI) morphologically and a proliferation index (PI) by using immunohistochemistry and counting Ki-67-positive nuclei. Cells were counted and identified by cell type (normal and cancerous) and by zone (peripheral [PZ], transition [TZ], and central [CZ]). Results: In normal epithelial cells when the Wilcoxon rank sum test was used to compare AI and PI changes between groups, the AI was higher in the TZ than in the CZ, a difference independent of treatment effects (p = .01). Between the largest PZ and TZ tumor foci, a statistically significant difference in PI was observed (p = .006). Using a multiple linear regression model fitted for AI from the largest RPS PZ tumor focus, we found a statistically significant difference between the SeMET group and the placebo group (p = .02). Conclusions: Besides introducing new findings, the clinicopathologic model proved itself a platform for biomarker discovery by leveraging molecular technologies to full advantage, including serum protein profiling (Kim et al., CEBP 2005) and gene expression profiling of different cell types. Supported by NIH grant CA88761.

No significant financial relationships to disclose.
Preclinical rationale for therapeutic targeting of mTOR by CC-I779 and rapamycin in Hodgkin lymphoma
Authors: Georgakis G. V. , Yazbeck V. Y. , Li Y. , Younes A.
Published: March 2020

10070

Background: Although conventional chemotherapy has been the golden standard for the treatment of Hodgkin Lymphoma (HL), significant long-term toxicities necessitate the need for development of targeted therapy that is potentially less toxic to normal cells. We and others have recently reported that the PI3K/AKT pathway is aberrantly activated in primary and cultured HL cells, and its inhibition induces apoptosis and cell cycle arrest. Because only mTOR inhibitors are currently being evaluated in clinical trials, we examined the biologic and molecular activity of two mTOR inhibitors CCI-779 and rapamycin in HL cell lines, and determined the molecular mechanisms of their activity. Methods: Three HL-derived cell lines were used (HD-LM2, L-428, and KM-H2). Cell viability was determined by the MTS assay. Cell cycle arrest was determined by propidium iodide staining and FACS analysis. Autophagy was determined by staining of intracellular organelles with Acridine Orange and FACS analysis. Molecular changes were determined by western blot. Results: The antiproliferative effect of CCI-779 and rapamycin in all HL cell lines was evident as early as 24 hour and was time- and dose-dependent. CCI-779 induced cell cycle arrest in all cell lines at the G0/G1 phase. CCI-779 and rapamycin were equally effective in HL cell lines. Cell cycle arrest was not followed by apoptosis but rather by autophagy, as evident by the increase of acidic autophagic intracellular organelles. On the molecular level, mTOR inhibition by CCI-779 dephosphorylated the downstream ribosomal S6 in all HL cell lines, but had no significant effect on AKT or phospho AKT. Cell cycle arrest was attributed to the upregulation of p27, but cyclin D1 remained unchanged. CCI-779 also enhanced the antiproliferative effect of the PI3K inhibitor LY294002, but did not change the effect of doxorubicin chemotherapy. Conclusions: Inhibition of mTOR by CCI-779 or rapamycin in HL cell lines has antiproliferative effect, by inducing cell cycle arrest followed by autophagy. CCI-779 enhances the activity of PI3K inhibitor LY294002. These data suggest that CCI-779 may have a therapeutic value in patients with HL.

No significant financial relationships to disclose.
Targeting bladder tumor cells in vivo and in the urine by a peptide identified using phage display
Authors: Kwon T. , Lee B. , Yoo E. , Chung S. , Park Y. , Kim B.
Published: March 2020

10071

Background: Bladder cancer is one of the most common tumors of genitourinary tract. Selective delivery of drugs to tumor tissues is important for effective tumor therapy. Here we identified a peptide targeting bladder tumor cells using phage display. Methods: Phage peptide library containing CX7C (C-cysteine and X-any amino acid residue) was based on T7 415–1b phage vector (Novagen). Tumor xenografts were prepared by subcutaneously injecting BALB/c Nu/nu female nude mice with HT-1376 bladder tumor cells. For a carcinogen-induced tumor model, Fischer 344 female rats were supplied ad libitum with tap water containing 0.05% N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) for 8 weeks. Results: A phage library containing CX7C random peptides was screened for selective binding to cells from human bladder tumor xenografts. Selected phage clones were individually evaluated for binding to cultured bladder tumor cells and for binding to cells from human tumor tissues of six patients. The peptide displayed by the most promising clone was synthesized and designated as Bld-1. Fluorescein-conjugated Bld-1 peptide showed selective binding to frozen sections of human bladder tumor tissues of three patients. In vivo tumor targeting was examined in a carcinogen-induced rat tumor model (n=20). When the fluorescent peptide was introduced into the bladder lumen, it selectively bound to tumor epithelium. Next, when the fluorescent peptide was intravenously injected into the tail vein, it homed to the bladder tumor, but was not detectable in normal bladder and control organs such as lung. Moreover, the fluorescent peptide bound to cells from urinary specimens of tumor patients (n=10), whereas little binding was observed in cells from healthy individuals (n=3). Conclusions: The Bld-1 peptide may be useful for targeting bladder tumor cells in vivo and in the urine.

No significant financial relationships to disclose.
Validation of a five reagent immunohistochemistry assay for prognostication of estrogen-receptor expressing breast cancer
Authors: Ross D. T. , Frost A. R. , Beck R. , Seitz R. S. , Ring B. Z.
Published: March 2020

10081

Background: We have previously reported the translation of gene expression data into a five reagent polyclonal antibody immunohistochemistry assay targeting p53, SLC7A5, NDRG1, HTF9C, and CEACAM5 whose staining results are combined using a Cox proportional hazard model for prognostication of estrogen receptor expressing breast cancer. Monoclonal antibodies targeting these genes have now been obtained and or produced. Methods: Paraffin blocks from 229 patients seen at the University of Alabama at Birmingham Clinical Cancer Center were assembled into tissue arrays and clinical follow-up data was collected by chart review. Immunohistochemical stains were performed and cases scored as positive or negative with the five monoclonal antibody assay. In order to further test the assays predictive value for assessing outcome, the prospectively defined Cox proportional hazard model and high, moderate, and low risk cutoffs were applied to the staining results on the estrogen receptor positive (ER+) patients for whom a assessment of death due to disease (DOD) and staining results for all five biomarkers were available. Results: The Kaplan-Meier estimates of outcome confirmed that the Cox model distinguished ER+ patients with poor outcomes. The five antisera algorithm identified high risk patients with a five year estimate of DOD of 41% compared to 20% for moderate and 9% for good (p=0.006) (79 patients total). In ER+ node negative patients the model identified high risk patients with a five year estimate of DOD of 50% compared to 18% for moderate and 5% for good (p=0.01) (40 patients total). The prognosticator was independent of age, pathological stage, and tumor size. Conclusions: A panel of five antibodies can significantly improve upon traditional prognosticators in predicting outcome for estrogen receptor positive breast cancer patients. This represents the third independent validation study performed on cohorts from different institutions. Retrospective studies on clinical trial cohorts and prospective studies should be performed to further establish the utility of this test in identifying early stage breast cancer patients at high risk for recurrence.

[Table: see text]
Hierarchical clustering analysis of tissue microarray immunostaining data identifies prognostically significant subsets of stage IIIA N2 non-small cell lung cancer patients. An Alpe-Adria Thoracic Oncology Multidisciplinary group study (ATOM 014)
Authors: Grossi F. , Bordo D. , Spizzo R. , Consiglieri C. , Aita M. , Morelli A. , Di Terlizzi S. , Ceschia T. , Beltrami C. A. , Belvedere O.
Published: March 2020

10085

Background: Based on gene expression profiling, prognostically relevant cluster groups of patients have been recently identified for breast cancer, lung cancer, and lymphoma. The development of tissue microarray (TMA) technology provides methodology for the concomitant analysis of multiple proteins on large numbers of tumor samples. Methods: Here, we report the hierarchical clustering analysis of TMA immunostaining data from 87 patients with stage IIIA pN2 non-small cell lung cancer (NSCLC), treated with surgery alone between 1985 and 1997. A panel of 10 markers was tested by immunohistochemistry: EGFR, ErbB-2, c-kit, COX-2, survivin, bcl-2, cyclin D1, cyclin B1, MMP-2 and MMP-9. Univariate, multivariate analyses and unsupervised hierarchical clustering analysis by using these 10 markers were performed. Results: Bcl-2 (p<0.0001) and cyclin D1 (p=0.0036) are more expressed in squamous cell carcinoma (SCC), while MMP-2 (p=0.0115), MMP-9 (p=0.0075) and survivin (p=0.02) show increased expression levels in other histological subtypes. In univariate analysis, only squamous cell histology, bcl-2 and cyclin D1 expression were favorable prognostic factors (p=0.0149, p=0.0013, p<0.0001, respectively), while MMP-2 expression showed a significant correlation with poor outcome (p=0.013). In multivariate analysis, cyclin D1 and MMP-2 were the only positive and negative prognostic factors, respectively (p<0.0001, p=0.06). Unsupervised hierarchical clustering analysis produced 5 distinct cluster groups and the deduced dendrogram identified 2 prognostically significant subsets of patients, with a median survival of 51 (groups 1–2) and 10 (groups 3–4-5) months, respectively (p<0.0001). Notably, groups 1–2 were mostly composed of SCC (80%). Conclusions: This study suggests that hierarchical clustering of TMA immunostaining data by using a limited set of markers might provide a useful tool for the identification of radically resected NSCLC patients at high risk of recurrence, likely to benefit from more aggressive treatment.

No significant financial relationships to disclose.
Elevated plasma endoglin (CD105) predicts decreased response and survival in metastatic breast cancer patients
Authors: Vo M. N. , Leitzel K. , Ali S. M. , Demers L. , Wilson M. , Evans D. , Lipton A.
Published: March 2020

10091

Background: Endoglin (CD105) is a proliferation-associated surface protein expressed by human vascular endothelial cells. It is a co-receptor for transforming growth factor (TGF) -β1 and TGF-β3, and plays a major role in angiogenesis. Upregulation of endoglin has been reported in endothelial cells of breast and colorectal cancers, and elevated levels of serum endoglin have been associated with tumor metastasis in breast cancer patients. Materials and Methods: Pretreatment EDTA plasma from 224 metastatic breast cancer patients enrolled in a phase III 2nd-line hormone therapy trial and 50 healthy post-menopausal female controls were assayed for soluble endoglin using a double-antibody sandwich ELISA from R&D Systems (Minneapolis, MN). Results: In the post-menopausal female control group (n=50), the mean ± SD for plasma endoglin levels were 5.00 ± 1.84 ng/ml, with a range of 2.38–9.83 ng/ml. The upper limit of normal was defined as the mean + 2 SD (8.7 ng/ml) for plasma endoglin. In the breast cancer patient group, the mean ± SD for plasma endoglin was 6.40 ± 2.23 ng/ml, with a range of 3.00–19.79 ng/ml. Elevated plasma endoglin levels were detected in 26 of 224 patients (11.6%). Patients with elevated vs. normal plasma endoglin levels had a reduced clinical benefit rate (CR +PR+Stable)(15% vs. 42%)(p=0.01) to hormone therapy. Time to progression (TTP) was shorter for patients with elevated vs. normal plasma endoglin levels, but did not reach statistical significance (p=0.2). Patients with elevated plasma endoglin had decreased overall survival (median 645 days vs. 947 days)(p=0.005). Another plasma angiogenic marker, soluble Tie-2 receptor, did not significantly correlate with clinical outcome. A weak correlation existed between plasma endoglin and Tie-2 (r =.54, p<0.00001). There was no correlation between plasma endoglin levels and the presence of bone metastasis vs. other sites of metastases. Conclusions: Elevated pretreatment plasma endoglin levels predicted for decreased clinical benefit rate to 2nd-line hormone therapy and a shorter overall survival in metastatic breast cancer patients. Circulating endoglin levels deserve further study for monitoring anti-angiogenic therapy.

[Table: see text]
Clinical significance of HuR expression, an mRNA-binding protein in non-small cell lung cancer (NSCLC)
Authors: Tanaka F. , Ogawa E. , Wada H. , Shanker M. , Garcia-Soto A. E. , Branch C. D. , Wistuba I. , Roth J. A. , Ramesh R.
Published: March 2020

10098

Background: HuR is a nucleo-cytoplasmic shuttling protein that specifically binds to mRNA that has AU rich (ARE) sites at the 3’end and transports the RNA to the cytoplasm for protein translation. In addition to mRNA transportation, HuR plays a role in mRNA stabilization and protein translation. Preclinical studies have shown mRNA’s of several growth factors, cell-cycle regulators, and transcription-regulating proteins have ARE’s at the 3’end and bound by HuR. However, there has been reported no clinical data on HuR expression in NSCLC. Thus, in the present study, we assessed clinical significance of HuR expression in NSCLC. Patients and Methods: A total of 236 patients with completely resected p-stage I-IIIA, NSCLC, were reviewed, and HuR expression was evaluated immunohistochemically. Results (Table): HuR expression was seen in the nucleus and cytoplasm of tumor cells. Cytoplasmic HuR expression was positively correlated with tumor progression (p-stage, P<0.01), especially nodal metastasis, microvessel density (MVD), and COX-2 expression. Enhanced nuclear HuR expression was also correlated with tumor progression and COX-2 expression, but not with MVD. Positive cytoplasmic HuR expression was a significant factor to predict a poor prognosis in all patients (5-year survival rates: 85% for HuR-negative and 43% for HuR-positive patients; P<0.01) and in any p-stage/histology subset patients. Nuclear HuR expression status was also a significant prognostic factor in all patients, but not in all subsets. A multivariate analysis confirmed that cytoplasmic HuR status was an independent prognostic factor (hazard ratio [95% CI], 4.261 [2.109–8.609]; P<0.001). Conclusions: Cytoplasmic HuR expression was correlated with tumor progression, and was a significant and independent prognostic factor in correlation with enhanced COX-2 expression and increased tumor angiogenesis.

[Table: see text]

No significant financial relationships to disclose.
Regulation of IKKe expression by androgen receptor and NF-kB transcriptional factor in prostate cancer
Authors: Péant B. , Diallo J. , Lessard L. , Mes-Masson A. , Saad F.
Published: March 2020

10106

Background: In unstimulated cells, NF-kB transcription factor is sequestered in the cytoplasm as an inactive p65/p50 dimer through interaction with a member of the inhibitor of kB protein family (IkBa). Prominent constitutive activation of NF-kB was observed in prostate cancer (PCa) cell lines lacking androgen receptor (AR) expression (PC3 and DU145) whereas only very low levels of NF-kB activity were seen in androgen-dependent cell lines (LNCaP and CWR22Rv1). As IkB kinase-e (IKKe) has recently been shown to be controlled by NF-kB, we hypothesize that IKKe may be involved in PCa progression based on its interaction with the NF-kB protein, and that these interactions are influenced by AR signaling. Methods: LNCaP cells were used to study IKKe expression with or without stimulation by the analog of androgen R1881 and by the tumor necrosis factor (TNF)-a. IKKe protein and RNA expression were characterized by immunoblot assay and quantitative PCR, respectively. IKKe expression was then correlated with p65 nuclear localisation. NF-kB activity was inhibited using an IkBa dominant negative construction. Inhibition of AR synthesis was performed using a siRNA against AR. Results: IKKe gene expression was stimulated by TNF-a treatment in LNCaP cells and inhibited by transfection of a dominant negative form of IkBa which prevented the nuclear translocation of p65. We also observed constitutive IKKe expression in hormone-refractory cells. Furthermore, we showed that TNF-a-induced IKKe expression is inhibited by R1881 in hormone-responsive PCa cells and this inhibition was correlated with the modulation of IkBa expression by R1881. Finally, we observed that the expression of IKKe is constitutively induced after blocking AR expression in LNCaP cells. Conclusions:. Our results show that IKKe expression is regulated by NF-kB in PCa cell lines. Moreover, IKKe appears to be down-regulated by ligand-dependent AR signaling through the control of IkBa expression. Further studies will be needed in order to determine the implications of this phenomenon with regard to NF-kB regulation, androgen resistance and effect on PCa progression.

No significant financial relationships to disclose.
Lymphatic mapping of ductal carcinoma in situ diagnosed by stereotactic core needle biopsy
Authors: Meijnen P. , Van Rijk M. C. , Oldenburg H. S. , Loo C. E. , Valdés Olmos R. A. , Nieweg O. E. , Peterse J. L. , Van de Vijver M. J. , Rutgers E. J.
Published: March 2020

10507

Background: About 20% of ductal carcinoma in situ (DCIS) lesions diagnosed by stereotactic core needle biopsy (SCNB) are shown to be invasive on postoperative pathology examination. Sentinel lymph node biopsy (SLNB) is an accurate method of evaluating axillary lymph nodes in patients with invasive breast cancer. The aim of this study was to review our experience with lymphatic mapping in patients with a SCNB diagnosis of pure DCIS, to evaluate the DCIS underestimation rate and consequently the risk of lymph node metastases. Methods: Files from 160 patients diagnosed with pure DCIS by SCNB between July 1999 and March 2005 were retrieved from our database. Patients with DCIS were selected for SLNB if there was concern for presence of an invasive component on the basis of size, palpability or imaging. Results: The median age of the study group was 55 years (range 29–85 years) and median DCIS size on mammography was 25 mm (range 4–96 mm). Thirty-six (23%) out of the 160 women underwent a SLNB. Macrometastases in the sentinel node were detected in seven (19%) patients and one (3%) patient was found to have a micrometastasis. Twenty (56%) of these 36 patients had invasive lesions on final pathology. Of the 124 women who did not receive SLNB, 29 (24%) turned out to have invasive lesions on postoperative evaluation. In total, 49/160 (31%) patients with pure DCIS diagnosed by SCNB had invasive breast cancer (range pT1mic-pT2) on final pathology. Finally, 88 patients underwent lymphatic staging by SLNB, basal node sampling or complete axillary lymph node dissection. Nodal involvement was present in 14 (16%) out of these 88 patients with initially diagnosed DCIS: 36% in patients with invasive breast cancer and 2% with pure DCIS on final pathology. Conclusion: Postoperative pathology examination of the specimen demonstrates DCIS underestimation in nearly one third of SCNB diagnosed DCIS patients. SLNB is of benefit for these patients as in 19% of the patients who undergo a SLNB macrometastatic disease in the sentinel node can be found.

No significant financial relationships to disclose.
hTert, MMP1, Her2 expressions by preneoplastic lesions and breast cancer risk
Authors: Hernandez J. , Mathieu M. , Taranchon E. , Spielmann M. , Delaloge S. , Soria J. , André F.
Published: March 2020

10510

Background: Although it is well established that preneoplastic lesions are associated with an increased risk of breast cancer, there is no available tool to identify which patients will develop breast cancer. Telomerase, matrix metalloprotease 1 (MMP1) and Her2 have all been involved in the early steps of carcinogenesis. In the present study, we have looked at whether the expression of hTert, MMP1 and her2 in preneoplastic lesions were associated with higher risk of breast cancer. Methods: hTert, MMP1 and Her2 expressions by preneoplastic lesions were determined by immunohistochemistry in 34 patients who have subsequently developed a breast cancer (cases), and in 32 patients who did not present breast cancer in the follow-up (control). Patients were matched for age, length of follow-up and type of preneoplastic lesion. The expression of the three biomarkers was compared in the two groups. The initially planned sample size of the study was 90 matched patients, but only 66 samples could be proceed for technical reasons. Results: Median age was 47 and 49 years old in patients with and without further cancer respectively. In the group of patients who subsequently developed breast cancer (cases), preneoplastic lesions consisted in lobular hyperplasia or lobular in situ carcinoma in 17 cases, ductal atypical hyperplasia in 12 cases and mixed lesions in 5 cases. In the control group, preneoplastic lesions consisted in lobular hyperplasia or lobular in situ carcinoma in 18 cases, ductal atypical hyperplasia in 12 cases, mixed lesions in 2 cases. The median interval between the diagnosis of preneoplastic lesion and the occurrence of breast cancer was 72 months (17–291). hTert was expressed in 8 (27%) and 2 (7%) assessable lesions in cases and controls respectively (p = 0.04). MMP1 was expressed in 21 (65%) and 22 (73%) assessable lesions in cases and control respectively (p = 0.49). Her2 was expressed in 6 preneoplastic lesions both in cases and controls (20%). Conclusions: This study suggests that hTert expression by preneoplastic lesions could be associated with an increased risk breast cancer.

No significant financial relationships to disclose.
Biological effects of pegfilgrastim after myelosuppressive chemotherapy in breast cancer
Authors: Danova M. , Bencardino K. , Grasso D. , Manzoni M. , Collovà E. , Ferrari S. , Travaglino E. , Rovati B. , Porta C. , Invernizzi R.
Published: March 2020

10521

Background: The dose and schedule of chemotherapy (CT), that play a crucial role in the outcome of pts with chemosensitive tumors, were made feasible with the primary use of colony stimulating factors. Few data are available on the in vivo biological effects of the “long-lasting” cytokine pegfilgrastim. We have focused on the cytokinetic effects on the CD34+/38+ peripheral blood (PB) progenitor cell subset and on its influence on neutrophils functional parameters. Methods: We studied PB samples from 14 breast cancer pts (median age 47 yrs; 35–61 yrs), treated with Docetaxel (80 mg/sqm, d 1) + Epirubicin (75 mg/sqm, d 1) + pegfilgrastim (6 mg s.c. on d +1). The % of CD34+/38+ circulating progenitor cells (CPCs) (sorted with immunomagnetic procedure) undergoing G0/G1, S and G2/M phases of the cell cycle or showing apoptotic features were evaluated using flow cytometry. Annexin V was quantitated at a single cell level and correlated with cell cycle phases. On PB buffy coat smears, alkaline phosphatase activity by cytochemistry, actin polymerization using FITC-labelled phalloidin and apoptosis by TUNEL technique, were evaluated on neutrophils. Results: Seven days following CT + pegfilgrastim the CD34+/38+ absolute numbers were: 46 (27–74) and 41 (25–66) on day 14+ from CT.On day 7+ the % of CD34+/38+ CPCs in S-phase was 13.0 ± 9 while 3.7% ± 6 of this cell subset showed apoptotic features. One week later, these values were 8.4% ± 7 and 7.7% ± 5, respectively. We also observed: stability of the absolute neutrophil count for all the duration of treatment; a significant increase of the leucocyte alkaline phosphatase; abnormalities of actin assembly in neutrophils, indicative of changes in cytoskeleton organization, and a significant reduction of neutrophil apoptosis. Conclusions: 1) pegfilgrastim exerts stimulatory effects on cell cycle status of PB CD34+/38+ CPCs, protecting them from apoptosis; this is evident 7 days after its administration and tends to decrease one week later 2) pegfilgrastim improves the neutrophil function by inhibiting their accelerated apoptosis and prolonging survival. These data could be useful when dose-dense CT is planned with pegfilgrastim support.

No significant financial relationships to disclose.
Phase I/II trial in patients with metastatic breast cancer (MBC) previously treated with a taxane and an anthracycline: Final safety data
Authors: Vahdat L. T. , Klimovsky J. , Bunnell C. A.
Published: March 2020

10528

Background: Ixabepilone, a semi-synthetic analog of the natural product epothilone B, has shown anti-tumor activity in a broad range of tumor types including breast cancer. Preclinical studies suggest synergy between ixabepilone and capecitabine in breast cancer models and has fuelled interest in the clinical setting. Methods: This open-label Phase I/II study was conducted to determine the recommended Phase II (and III) doses of ixabepilone and capecitabine using a 3-hour infusion of ixabepilone given on Day 1 (Schedule A) or a 1-hour infusion of ixabepilone given for 3 days (Schedule B) in combination with capecitabine given orally on Days 1–14 every 21 days in patients (pts) with MBC previously treated with a taxane and an anthracycline. Eligible pts were aged ≥18 years with pathologic diagnosis of breast cancer and evidence of MBC, who had received prior treatment with a taxane and an anthracycline in the adjuvant or metastatic setting. Pts were excluded if they had received >3 prior chemotherapy regimens in the metastatic setting or had ≥Grade 2 neuropathy. Toxicity was continuously monitored using NCI CTC v2. Results from the 62 pts treated with 40 mg/m2 as a 3 hr-infusion ixabepilone and 2000 mg/m2 capecitabine are shown here. Results: 56 of the 62 pts (90%) were aged <65 years. 80% of pts had visceral disease; 44% were ER/PR/HER-2 negative. 44% of pts had received ≥ two prior chemotherapies in the metastatic setting. Grade 3 neuropathy (sensory and motor) was reported in 12 pts and 1 pt (19%, 2%), respectively. Hand-foot syndrome, reported in 39 pts (63%), was primarily Grade 2 (26%) or 3 (34%). Hematologic toxicities included Grade 4 leukopenia (7 pts, 12%) and Grade 4 neutropenia (16 pts, 26%). Dose reductions were used to manage sensory neuropathy and hematologic abnormalities. Conclusions: The recommended Phase II (and III) doses are 40 mg/m2 ixabepilone (3-hour infusion on Day 1 every 21 days) and 2000 mg/m2 capecitabine (divided BID on Days 1–14 every 21 days). This ixabepilone/capecitabine combination demonstrated a manageable safety profile and promising clinical anti-tumor activity in pts with MBC previously treated with a taxane and an anthracycline.

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Breast sentinel lymphadenectomy—Is immediate intra-operative assessment warranted?
Authors: Tafe L. J. , Tsongalis G. J. , Kaufman P. A. , Moore C. P. , Wells W. A.
Published: March 2020

10533

Background: Sentinel lymphadenectomy is the standard of care in breast cancer evaluation. Since conventional axillary dissection is only considered for patients (pts) with sentinel lymph node positive (SLN+) disease, there is increasing interest in immediate intra-operative assessment of SLN by frozen section or RT- PCR. Methods: Institutional SLN cases (2003–2004) were retrospectively reviewed. Each SLN (submitted in toto for processing with 21 tissue levels/block, H&E staining, and cytokeratin (CK) immunostudies) was evaluated for tumor deposit size, distribution and extranodal extension. Results: 80% (n = 222) of pts had SLN biopsy: 50% at primary excision, 21% at mastectomy, 9% at re-excision and 20% as a separate procedure. 1–4 SLN were sampled in 41%, 25%, 19%, and 14% respectively with no node identified in 1 case. Of all, 31% (n = 68) had >/=1 (+) node and 72% of those (n = 50) had a completion axillary dissection. 50% of pts required separate surgical procedure. Only 21% (n = 14) of pts with SLN+ had subsequent (+) axillary nodes on completion dissection. Nodal deposits ranged in size from </= 0.02 cm (13%) to >2.0 cm (1%). The majority (26%) were in the micrometastasis range of >0.02<0.2 cm, where CK-immunostaining was useful to confirm detection. 19% were >/=0.2< 0.5 cm, 25% >/= 0.5<1.0 cm, and 15% >/= 1.0<2.0 cm. 3% SLN showed CK(+)-only cells without H&E confirmation. Tumor sites were parenchymal (1%), subcapsular (31%), or both (68%). Only 7% of nodes with subcapsular metastases were associated with subsequent positive axillary nodes. In 91% (n = 62) of SLN+ cases, metastases were identified in the first 3 tissue levels. Extranodal extension was seen in 22%. Conclusions: Overall, 31% were SLN+; 21% of those were node + in subsequent axillary sampling. These findings do not support the labor and expense of immediate intraoperative SLN assessment. 3-level tissue processing with simultaneous RT-PCR could be performed on all SLN. Subsequent tissue leveling and immunostaining could potentially only be performed on PCR+ cases to provide metastasis size and extranodal extension. PCR probes can also facilitate detection of novel prognostic markers. Additionally, identification of only subcapsular deposits may warrant foregoing subsequent axillary sampling in some clinical scenarios.

No significant financial relationships to disclose.
Phase II study of docetaxel plus epirubicin versus docetaxel plus cisplatin as first-line chemotherapy for treatment of advanced breast cancer (CHN TAX-618)
Authors: Wang Y. J. , Fu Q. , Wu Q. , Su F. X. , Zhou L. Z. , Ye Z. B. , Yang J. Q.
Published: March 2020

10535

Background: The combination of docetaxel plus epirubicin (TE) have demonstrated a significant activity in advance breast cancer (ABC) as first-line chemotherapy. In the meantime, some emerging literatures suggest that docetaxel-cisplatin (TP) combination has a powerful synergistic interaction with less cardiotoxicity. The main objectives of this multicenter study are to evaluate the efficacy and safety of TE versus TP. Methods: From August 2003 to February 2005, 80 patients with at least one measurable lesion were randomized with ratio of 2:1 to receive TE or TP as as first-line chemotherapy. Dosages were as follows: docetaxel 75 mg/m2 plus epirubicin 60 mg/m2 or docetaxel 75 mg/m2 plus cisplatin 75 mg/m2 intravenously once every 3 weeks for a maximum of 6 cycles. Baseline characteristics were well balanced. Results: A total of 55 patients were assigned to TE (median age 48 yearas) and 25 patients were treated with TP (median age 50 yearas). The same of median 4 cycles treatment were administered for two arms. A complete clinical response was observed in 3.6% and 4.0% of patients treated with TE and TP, respectively (P = 1.00). Overall (complete and partial) clinical response rates for TE and TP were 48% and 56%, respectively (P =0.469). Disease control rates (CR + PR + SD) for TE and TP were 83.6% and 80%, respectively (P = 0.755). The median TTP were TE: 10.8months,TP: 11.2 months days (P = 0.247). Grade 3/4 toxicities included: nausea/vomiting (16.3% TE; 12.0% TP); diarrhea (1.8% TE; 0% TP); alopecia (32.7% TE; 12.0% TP); anemia (1.8% TE; 4.0%TP); neutropenia (65.5% TE; 4.8% TP); febrile neutropenia (3.6% TE; 0% TP). Conclusions: Both TE and TP were active and tolerant regimens for ABC. While there may have been a trend toward lower toxicities in favor of the docetaxel/cisplatin combination, although no significant difference was observed during our study.

No significant financial relationships to disclose.
High malignancy yield in core needle biopsy of BIRADS 3 breast lesions with calcification
Authors: Larke A. , O’Doherty A. , Quinn C.
Published: March 2020

10568

Background: Breast cancer screening programmes reduce mortality but result in increased numbers of breast biopsies. Stereotactic and ultrasound-guided core needle biopsy yield similar results to each other and to open biopsy. Study: Analysis of prospectively-acquired data relating to 174,338 screening mammograms performed in BreastCheck between Jan 2001 to Oct 2004. Relative use of ultrasound and stereotactic guidance for core needle biopsy is ascertained, and malignancy yields analysed. Results: In this time period, 174,338 screening mammograms discovered a total of 2782 lesions requiring further assessment. Of 2760 complete records, 1066 were BiRads R3–5 (39%) and 1033 of these (97%) had CNB. Guidance was provided exclusively by ultrasound in 737 (71%), and by stereotaxis in 270 (26%) of cases. There were no differences in malignancy yield of ultrasound versus sterotactically-guided CNB of R4 and R5 lesions (approximately 50% and 90% respectively). The malignancy yield in R3 lesions was significantly higher when performed with stereotactic guidance (12%) than when performed with ultrasound-guidance (5%) and both were significantly higher than R3 biopsy malignancy yields reported in the literature. Conclusion: Almost all BIRADS 3, 4 and 5 lesions are biopsied. The relatively high malignancy yield in R3 biopsies, particularly those with calcification, supports current practice.

No significant financial relationships to disclose.
Brachial plexopathy in breast cancer
Authors: Terstriep S. , Amrami K. , Spinner R. , Moynihan T.
Published: March 2020

10601

Background: Brachial plexopathy is a well-recognized complication of breast cancer, most attributed to late effects of radiation. However, direct involvement of brachial plexus by recurrent breast cancer occurs. Misdiagnoses may lead to inappropriate attribution of the plexopathy to radiation, and therefore inappropriate therapy. Methods: We report fourteen cases of brachial plexopathy secondary to breast cancer diagnosed at the Mayo Clinic-Rochester from 2003 to 2005. Results: Thirteen of fourteen primary tumors and 12/14 recurrences were ER/PR positive. Only 1/14 patient had Her 2 neu overexpression. The median time from the original breast cancer diagnosis to first brachial plexus symptom was 14 years. The median time from the development of symptoms to diagnosis was 8 months with the range being 1 month to 8 years. MRI revealed a distinct mass in 3/13 patients, plexus thickening in 5/13, and was normal in 5/13. Only 5/13 original MRI interpretations suggested tumor. In four patients that the 1.5 T MRI was either interpreted as normal or unlikely cancer the 3 Tesla MRI revealed abnormal uptake suggestive of malignancy. PET scan suggested malignancy in 3/6 patients. Two of these cases had other metastatic disease. In the other three cases that PET scans were done the uptake was mild, suggesting inflammation rather than malignancy. Reinterpretations of the PET scan and MRI in combination by an experienced musculoskeletal radiologist was highly suggestive of malignancy in all cases. Definitive diagnosis was eventually obtained by biopsy of the brachial plexus (7/14) or another site of metastasis in (4/14). Conclusions: The often slowly progressive nature of hormonally driven breast cancer can mimic radiation induced brachial plexopathy. In the cases reviewed, PET was not reliable in differentiating inflammation from tumor. MRI findings are nonspecific and higher resolution 3T MRI may be needed to detect an abnormality. Interpretation of MRI and PET together is useful. Patients with a history of breast cancer, and brachial plexus symptoms should be carefully evaluated with consideration for recurrent disease as a cause. Biopsy of the brachial plexus can be done safely and has an important role in diagnosis.

No significant financial relationships to disclose.
Micrometastasis in sentinel (SLN) and non-sentinel lymph nodes of breast cancer: An update including clinico-pathologic impact and survival
Authors: Masci G. , Di Tommaso L. , Del Prato I. , Orefice S. E. , Gullo G. , Rubino A. , Eboli M. W. , Morenghi E. , Roncalli M. , Santoro A.
Published: March 2020

10602

Aims: To assess the rate of positive axillary clearance when the SLN biopsy contains micro-metastatic disease and to evaluate the survival impact of local lymph nodes with micro-metastases. Materials and Methods: From 1997 to 2005 2,750 consecutive patients (pts) underwent breast surgery with SLN biopsy or axillary dissection. Of these, 87 pts had lymph node micro-metastases assessed with haematoxylin and eosin or cytokeratine staining and defined according to the current TNM classification. Lymph node micro-metastases were found after axillary dissection in 27 pts and after SLN biopsy in 60 cases; 55 (63.2%) were micro-metastases 0.2–1 mm, 22 (25.3%) measured 1–2 mm, 7 were isolated tumor cells and in 3 cases size was not available; micro-metastases were located within lymph node sinuses in 43 and in nodal parenchyma in 37 cases. Age: median 53 years (range 34–78), surgery: 77 (88.5%) conservative and 10 radical mastectomy; histotype: 79 (90.8%) ductal, 4 lobular, 2 tubular and 2 other histotypes. T stage: pT1a/pT1b in 16 (18.4%), pT1c in 54 (62.1%) and pT2 in 17 (19.5%) pts; grading 3 in 24 pts (27.6%); 76 pts (87.3%) had positive Er/Pgr; vascular invasion: present in 30 pts (34.5%); HER-2/neu overexpression in 16 (18.4%) pts; 64 pts (73.6%) received adjuvant chemotherapy (49 AC, 11 CMF ev, and 4 FEC); 69 pts (79.3%) had radiotherapy and hormonal therapy. Results: Among 60 pts with SLN micro-metastases, 8 (13%) had further axillary involvement; in 5/8 cases size of micro-metastases was >1 mm, in 7/8 cases micro-metastases were located in the nodal parenchyma. These 8 pts were not considered for survival analysis. Of the remaining 79 patients with micro-metastases, survival data were as follow: at median follow-up time of 32 months (range 2–158) 69 pts were alive without any evidence of disease, 4 pts died of disease and 6 pts were alive with disease. Three years DFS was 87.75% and OS 95.57%. Conclusions: Further axillary involvement was found in 13% of pts with micrometastases in the SLN biopsy supporting complete nodal dissection in all pts.Our preliminary results show that even minimal nodal involvement could correlate to worse prognosis and may require adjuvant treatment.

No significant financial relationships to disclose.