bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P4-04-04: Immunohistochemical analysis of Cancer Testis antigens and Topoisomerase 2-alpha expression in triple negative breast carcinomas: a retrospective study
Authors: Juretic A , Mrklic I , Spagnoli GC , Pogorelic Z , Tomic S 
Published: August 2014

Abstract
Aim. The aim of this study is to assess the expression of cancer testis (C/T) antigens in the TNBC group, expression of topoisomerase 2-alpha (TOPO2A), basal-like (BL) immunophenotype defined by basal markers (CK5/6, CK14, and EGFR), BL morphology and conventional clinicopathologic factors as well as to demonstrate their prognostic relevance with respect to this group of tumours.
Methods. The study includes 83 patients who underwent surgery between January 2003 and December 2009. Slides were stained immunohistochemically with CK5/6, CK14, EGFR, Ki-67, TOPO2A, MAGE-A1, MAGE-A10, NY-ESO and multi-MAGE. Survival-time and multivariate survival analyses were performed for the purpose of identifying prognostic markers for tumours with more aggressive behaviour.
Results. Of the 83 triple negative tumours, 55 (66.3%) have BL immunophenotype and 40 (48.2%) BL morphology. MAGE-A1 show the most frequent expression (69.0%), followed by multi-MAGE (58.0%), NY-ESO (27.1%) and MAGE-A10 (13, 2%). MAGE-A10 expression is significantly correlated with tumour size (p = 0.026). The expressions of MAGE-A1, MAGE-A10 and multi-MAGE are significantly correlated with clinical stage (p = 0.024, p = 0.041, p = 0.031, respectively). A significant correlation has been found between the expressions of MAGE-A10 and NY-ESO and the expression of TOPO2A (p = 0.005, p = 0.013). There is no significant correlation between the expression of C/T antigens and disease-free survival (DFS) or overall survival (OS).
Conclusions. High expression levels of MAGE and NY-ESO observed in TNBC are of potential clinical relevance, especially in the adjuvant setting of treatment, and may be of therapeutic value as a vaccine-based treatment in the TN group of breast tumours for which therapeutic options are limited.
Key words: triple negative breast cancer, basal-like breast cancer, cancer-testis antigen, topoisomerase 2-alpha, prognosis, survival analysis.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-04.
Abstract P4-04-05: Listeria monocytogenes-based bivalent Lm-LLO immunotherapy for the treatment of HER2/neu positive and triple negative breast cancer and its impact on immunosuppression
Authors: Wallecha A , Ramos K , Malinina I , Singh R 
Published: August 2014

Abstract
Overexpression of tumor associated antigens (TAA) such as HER2/neu and high molecular weight melanoma associated antigen (HMW-MAA) are associated with aggressive high-grade tumors leading to disease progression and reduced survival. HMW-MAA has been reported as a TAA in triple negative breast tumors and is also expressed at high levels by activated pericytes and pericytes in tumor angiogenic vasculature that are associated with neovascularization in vivo. Previously published reports have shown that bioengineered Listeria monocytogenes (Lm) secreting either a chimeric HER2/neu antigen (cHER2), or HMW-MAA fragment, fused to a truncated highly immunogenic fragment of the adjuvant protein listeriolysin O (LLO) were found to impact the growth of both transplantable and transgenic mouse breast tumor models. Therefore, we hypothesized that an Lm-LLO immunotherapy capable of delivering two different antigens would likely have a synergistic effect of decreasing tumor growth by targeting two independent mechanisms that support tumor growth; 1) tumor angiogenesis, and 2) tumor cell surface marker, thus improving the therapeutic efficacy of the agent. Here we report the construction of a bivalent recombinant Lm-LLO immunotherapy that expresses and secretes both the cHER2 and HMW-MAA antigens concomitantly as fusion proteins. Initial characterization of the bivalent immunotherapy indicates that both the antigens cHER2 and HMW-MAA are stably expressed and secreted after two in vivo mouse passages. Preliminary studies support the hypothesis that bivalent immunotherapy causes efficient reduction of tumor growth in both HER2 expressing and triple negative breast tumors. Further, mechanistic studies demonstrate that treatment with Lm-LLO immunotherapy resulted in a decreased proportion of regulatory T cells (Tregs) as well as Myeloid Derived Suppressor Cells (MDSC) particularly in the tumor microenvironment and not in spleen, tumor draining lymph nodes, or peripheral blood. Interestingly, both residual Tregs and MDSCs isolated from the tumor microenvironment were found to have a decreased ability to suppress the division of T cells after Lm-LLO immunotherapy, a finding which warrants further investigation. We will present data on the efficacy of bivalent Lm-LLO immunotherapy and the mechanisms that are likely responsible for the observed tumor regression. Currently Lm-LLO immunotherapies are being evaluated in Phase 2 clinical trials for HPV-associated dysplasia and malignancies such as CIN2/3, cervix cancer and head & neck cancer.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-05.
Abstract P4-04-07: Tartrate-resistant Acid Phosphatase (TRAcP)-Expressed Tumor-Associated Macrophages Promote Breast Cancer Progression
Authors: Dai M-S , Wu C-C , Chang P-Y , Ho C-L , Hsieh Y-F , Lo K-Y , Kao W-Y , Chao T-Y , Yu J-C 
Published: August 2014

Abstract
Purpose: Tumor-associated macrophages (TAMs) can promote or dampen breast cancer progression, depending on phenotypic changes within the cancer microenvironment. Two contrasting activation states were described in the literature: the M1 anti-tumoral and M2 pro-tumoral subtype of TAMs. In clinical observation, we analyzed the TRAcP expression in breast cancer TAM and correlated to patient's chemotherapy responses and survival. In animal models, we aim to investigate the tumor-promoting or inhibiting properties among tartrate-resistant acid phosphatase (TRAcP) expressed TAMs.
Methods: TRAcP expression, T lymphocytes, and other macrophage relevant markers were determined by immunohistochemical staining in the human breast cancer tissue from the patients before neoadjuvant chemotherapy. The expression of TRAcP within the TAMs were calculated and correlated to the patient's chemotherapy responsiveness and outcome. The polarized M1/M2 macrophages and breast cancer hematogenous metastasis were created in the animal model. Activated macrophages from TRAcP+/+ or TRAcP−/− mice were injected with JC (Balb/c breast cancer) cells in the metastatic model.
Results: The presence of Foxp3+ regulatory T cell (Treg) correlates to the tumor proliferation (Ki-67 and Her2 expression) and higher Foxp3+ Treg prevalence and tumor Ki-67 expression inferred poorer chemotherapy response. TRAcP staining within the CD68+ macrophages were observed with better chemotherapy response. In animal model, M2 polarized macrophages have higher TRAcP expression, promote breast cancer metastasis, and attract Treg infiltrating in tumor, consistent with the clinical observation.
Conclusion: TRAcP-expressing TAMs, representing M2-polarized macrophage, demonstrat local tumor-promoting and immune-suppressive effects, and chemotherapy resistance. TRAcP expression within TAM can be a potential prognostic marker for breast cancer.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-07.
Abstract P4-04-08: A Potential non-viral vector to transfect dendritic cell and thereby MHC-Class I antigen presentation might be a potential use in carcinoma
Authors: Shaheen S , Akita H , Souichirou I , Miura N , Harashima H 
Published: August 2014

Abstract
To date almost no remarkable potential non-viral vector was developed to transfect promisingly primary dendritic cell like Bone Marrow Derived Dendritic Cell (BMDC) so that it might generate antigen presentation while epitopic-plasmid was delivered. Here we have introduced a potential non-viral vector named Stearyl-KALA MEND (100–130 nm, zeta potential 35–40 mV), which expressed to date the highest transgene expression while firefly luciferase was introduced (sub-cloned), and thereby promising antigen presentation in vitro while OVA plasmid containing MHC Class-I restricted SIINFEKL epitope was introduced and co-cultured with B3Z T-cell hybridoma. On the basis of these results, we used this vector to transfect BMDC and harvested the DC to chase the corresponding EG7-OVA induced tumor ex vivo. Later we further immunized mice directly with STR-KALA MEND containing OVA plasmid and challenged against the tumor (EG7-OVA induced). In this in vivo study we found also a significant antitumor activity. To evaluate the promptness of our vector we further sub-cloned Mart-1 gene, in our sub-cloned plasmid vector and immunized the mice as before. Thereafter inserting B16F10 melanoma cells to the immunized mice we found also a significant antitumor activity after 24 days of inoculation. Thus the vector, STR-KALA MEND might be a future potential use as DNA vaccine in anti-tumor methodology.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-08.
Abstract P4-05-01: Oncolytic Herpes Simplex Virus Vector G47Δ Effectively Targets Breast Cancer Stem Cells
Authors: Liu R , Zeng W , Hu P , Wu J , Li J , Wang J , Lei L 
Published: August 2014

Abstract
Breast tumors can be classified according to their gene-expression profiles into different molecular subtypes with distinct biological and clinical features. Moreover, different subtypes show distinct responses to different therapeutic regimens, thereby resulting in markedly different outcomes. Recently, accumulating evidence has suggested that breast cancer originates from cancer stem cells (CSCs), which comprise a small percentage of the overall tumor but are highly tumorigenic and pluripotent with unlimited proliferation potential. Furthermore, cancer stem cells are highly resistant to conventional treatment, which potentially explains certain difficulties in treating cancer with current therapy options. The use of oncolytic viruses is a relatively new strategy in cancer therapy, oncolytic herpes simplex virus (oHSV) has been shown to be a safe and effective therapeutic approach for a variety of different cancers.
In this study, different types of breast cancer cell lines were cultured in anchorage-independent conditions, the cells typically form mammospheres within 10 days of anchorage-independent culture, which showed properties of CSCs. The mammospheres are capable of being passaged indefinitely and single dissociated mammosphere cells were able to regenerate spheres. Moreover, compared to the parental cell lines, mammoshpere cells were enriched for CD44+CD24− cells and highly expression of the alternative breast cancer stem cell marker ALDH-1.
The third generation oHSV vector G47Δ effectively killed different subtypes of breast cancer cells in vitro, with more than 98% of the tumor cells killed by day 5, even at multiplicities of infection(MOI) 0.01. Moreover, G47Δ equally targeted non-cancer stem cells (NCSCs) and CSCs which showed resistance to paclitaxel. We also demonstrated that G47Δ effectively replicated and spread among CSCs. Moreover, G47Δ impaired the self-renewal ability of CSCs, as the viable cells unable to form secondary tumor spheres. We also showed that G47Δ was able to induce the regression of tumor xenografts in BALB/c nude mice and demonstrated the ability of G47Δ to synergize with paclitaxel for killing both NCSCs and CSCs. This is the first report that oHSV effectively targets different breast cancer NCSCs and CSCs, thereby suggesting that oHSV may be an effective treatment modality for patients with breast cancer.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-05-01.
Abstract P4-06-06: RNAi screen of the breast cancer genome identifies KIF14 and TLN1 as genes that modulate chemosensitivity in breast cancer
Authors: Singel SM , Cornelius C , Batten K , Fasciani G , Wright WE , Lum L , Shay JW 
Published: August 2014

Abstract
We conducted a RNA-interference screen for genes whose loss-of-function enhanced doxorubicin and docetaxel chemosensitivity in a “triple-negative,” estrogen receptor negative, progesterone receptor negative, and Her2 negative (ER− PR− Her2−) breast cancer cell line, MDA-MB-231. From ranking chemosensitivity of 334 short-hairpin RNA (shRNA) MDA-MB-231 cell lines (targeting 134 genes with known somatic mutations in breast cancer), we focused on two genes, kinesin family member 14 (KIF14) and talin (TLN1) that not only enhanced chemosensitivity but also have oncogenic annotations. KIF14 has robust expression in breast cancer cells compared to normal mammary cells. TLN1 expression is important for migration of breast cancer cells. In TLN1-deficient cells, CK19 and ZO-1 are upregulated while CK14 and snail are downregulated, suggesting that TLN1 is important for the maintenance of epithelial-mesenchymal transition in MDA-MB-231 cells. KIF14 and TLN1 loss-of-function also enhanced chemosensitivity in 3 other triple negative breast cancer cell lines (HCC1937, HCC38, and Hs578T) but not in normal human mammary epithelial cells (HMECs). Examining protein-protein-interaction networks, we identified RIP2 as a target whose inhibition via SB203580 or PP2 can further enhance chemosensitization in KIF14-deficient cells. Knock-down of a number of other known protein-protein interaction partners of KIF14 and TLN1, including FAK, CIT, ARRB2, PSTPiP1, PRC1, SVIL, ITGA2B, ITGB3, VCL and PXN, do not significantly alter doxorubicin or docetaxel chemosensitivity in MDA-MB-231 cells. Mammary fat pad xenografts of KIF14- and TLN1- deficient MDA-MB-231 cells into NOD/SCID mice demonstrated significantly less tumor mass compared to control MDA-MB-231 cells after chemotherapy. Expressions of KIF14 and TLN1 from breast cancer expression arrays improve prognostic predictions compared to clinicopathological features alone. In summary, screenings for therapeutic targets using chemotherapy and genes with known somatic mutations in breast cancer not only provide a rational targeted screen, but also present possible up-front novel treatment combinations for patients with triple negative breast cancer.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-06.
Abstract P4-06-11: Expression of genes spanning a breast cancer susceptibility locus on 6q25.1 is modulated by epigenetic modification
Authors: Dunbier AK , White J , Van Huffel S 
Published: August 2014

Abstract
Background: Genome wide association studies have identified single nucleotide polymorphisms around C6ORF97, an open reading frame (ORF) immediately upstream of the gene encoding ERα (ESR1) on 6q25.1, to be associated with increased risk of breast cancer1,2. C6ORF97 and two neighbouring genes, RMND1 and C6ORF211, are also co-expressed with ESR1 in oestrogen receptor positive (ER+ve) breast cancers and C6ORF97 expression is inversely associated with proliferation and recurrence free survival in a tamoxifen-treated patient cohort3. Epigenetic modification represents one potential mechanism through which expression of co-localised genes could be modulated.
Aim: To investigate the role of epigenetic modification in the co-expression of ESR1, RMND1, C6ORF211 and C6ORF97.
Methods: Methylation status and expression of genes in the ESR1/C6ORFs locus was assessed in postmenopausal breast cancers and ER+ve cell lines. Two ER+ve cell lines, MCF7 and T47D, were cultured in oestrogen-depleted media and samples were taken fortnightly for 6 months. DNA and RNA from ER+ve tumours was also analysed in addition to control ER-ve cell lines. Methylation analysis of CpG Islands within the ESR1/C6ORFs locus was carried out using methylation sensitive sequencing analysis. Expression levels of ESR1, RMND1, C6ORF211 and C6ORF97 were determined using quantitative real-time PCR.
Results: Variation in methylation patterns was observed at a number of key CpG islands in the ESR1/C6ORFs region. ER+ve cell lines and highly ER+ve tumours were hypomethylated. Hypomethylation was associated with increased expression of ESR1, RMND1, C6ORF211 and C6ORF97. Temporal changes in methylation and expression of these genes were also observed over the period of oestrogen deprivation in both MCF7 and T47D.
Conclusions: Methylation at the ESR1/C6ORFs locus is associated with expression levels of ESR1, RMND1, C6ORF211 and C6ORF97. This suggests that methylation is involved in modulating the expression of these four genes and that it could partially explain the previously observed co-expression of the genes. Our finding that hypomethylation in this region occurs in tumours with high ERα expression suggests that it could be a mechanism through which tumours increase ERα levels to maximise growth stimulation from oestradiol. Our observation that RMND1, C6ORF211 and C6ORF97 also increase in expression suggests that the proteins encoded by these genes may contribute to the phenotype exhibited by ER+ve breast tumours.
References:
1. Turnbull C et al. Nature Genetics 2010;42:504–7. 2. Zheng W et al. Nature Genetics 2009;41:324–8. 3. Dunbier A et al. PLoS Genetics 2011;7:e1001382.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-11.
Abstract P4-06-16: TGF-β2, A Novel Target of CD44-Promoted Breast Cancer Invasion
Authors: Gupta I , Madani S , Abdraboh M , Al Riyami H , Muzumdar S , AbdElmageed Z , Shanmuganathan S , Bhat S , Ramkumar A , Hakkim L , Ouhtit A 
Published: August 2014

Abstract
Background: Hyaluronan (HA) mediates communication between cancer cells and the environment via interactions with the cell surface receptor CD44. We have previously shown that CD44-HA interaction in BC cells promotes adhesion to bone marrow endothelial cells. This suggests that HA/CD44 signalling may be correlated with breast cancer (BC) metastasis. The long-term objective of this study is to increase our understanding of the mechanisms by which CD44-HA interaction promotes BC metastasis, and further identify and validate CD44-downstream transcriptional targets for anti-metastatic therapy.
Materials & Methods: Pursuant to this goal, we have developed a tetracycline (tet)-regulated expression of CD44 gene in the BC cell line MCF-7 (B5 clone) and identified TGF-β2 (Transforming Growth Factor beta-2; 3 fold induction) as a potential CD44s-downstream transcriptional target by microarray analysis. To further validate this finding, the same RNA samples, used for microarray analysis and their corresponding protein lysates collected from the BC cell line MCF-7-B5, were examined for CD44 expression in the presence of HA for 18, 24, and 48 hr post-tet withdrawal. Expression of TGF-β2 was examined using RT-PCR and western-Blot analyses.
Results: Our results showed that TGF-β2 mRNA levels were significantly elevated following the removal of tet at 18, 24, and 48 h post-HA stimulation compared to the parental cells. Furthermore, the TGF-β2 precursor protein increased in a time-dependent pattern upon HA-stimulation and in the absence of tet. More interestingly, inhibition of CD44 gene by RNAi method decreased TGF-β2 expression upon HA-stimulation and in the absence of tet.
Conclusion: Our data strongly support the hypothesis that TGF-β2 is a potential target of HA/CD44- downstream-signaling mediating BC cell invasion. Ongoing investigation aims to elucidate the signal transduction pathways coupling CD44 to the regulation of TGF-β2 expression, and further validate CD44/TGF-β2 axes in breast tumor invasion and metastasis.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-16.
Abstract P4-06-17: Antitumor activity of the novel mithramycin analog EC8042 in triple negative breast cancer
Authors: Pandiella A , Montero JC , Cuenca D , Re-Louhau F , Nuñez LE , Moris F , Ocana A 
Published: August 2014

Abstract
Background: Triple negative breast cancer (TNBC) represents around 15% of all breast tumors. Therapeutic strategies in TNBC are mainly based on classical chemotherapeutic agents, with limited success in the metastatic setting. Here we have explored the action of a novel mithramycin analog named EC-8042 in TNBC.
Methods: we used a panel of cell lines representative of triple negative tumors including BT549, HBL100, HCC-1037, HS578T, MDA-MB213, HCC-1187, HCC-3153 and HCC-70. Proliferation studies with EC-8042 were measured by MTT assays. Propidium iodide (PI) staining followed by FACS was used for cell cycle analyses and AnnexinV-fluorescein for analysis of apoptosis. Proteins involved in cell cycle control and apoptosis were evaluated by western blot. Mitochondrial membrane potential was assessed by measuring the fluorescence of tetramethylrhodamineethylester (TMRE) dye.
Results: The IC50 values obtained were in the nM range, indicating that the drug had a potent effect on these cell lines. Mechanistically, the drug caused accumulation of cells in the S phase of the cell cycle at doses of 100 and 500 nM in MDA-MB231. Biochemically, EC-8042 caused an increase in cyclin E, cyclin A, cyclin D3, cyclin B, and p27, and decreased cyclin D1 and Rb phosphorylation at serine 780. These data are consistent with an action of EC-8042 on the cell cycle. Moreover, the increase in wee1 and pY15-CDK1 are in line with an effect of the drug blocking cell cycle progression through the S phase. Annexin V staining indicated that treatment with 500 nM of EC8042 provoked apoptosis, which was also revealed by PARP processing. The treatment reduced the amounts of MCL1, BCLXL, and XIAP, but did not affect BCL2 levels.
Conclusion: our results indicate that EC-8042 has a strong antitumoral action on TNBC cells, which is caused by increased apoptosis and a decrease in cell cycle progression, and opens the possibility for further development of this drug in TNBC.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-17.