bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P4-16-06: Histone deacetylases inhibitor SAHA enhances anti-tumor effects of poly (ADP-ribose) polymerase (PARP) inhibitor olaparib in breast cancer cells
Authors: Min A , Im S-A , Kim S , Song S-H , Lee K-H , Han S-W , Oh D-Y , Kim T-Y , Bang Y-J 
Published: August 2014

Abstract
Background: The poly (ADP-ribose) polymerase (PARP) inhibitor, olaparib, has been found to have a therapeutic potential for treating cancers that have an impaired DNA repair ability. However, some cancer presents acquired resistance to PARP inhibitor or platinum. Histone deacetylases (HDACs) are important to enable functional homologous recombination (HR) by regulating the expression of HR-related genes and promoting the accurate assembly of HR-directed subnuclear foci. Thus, HDAC inhibitors have emerged recently as a class of therapeutic agents for the treatment of cancer by inhibiting DNA repair. For this mechanism, HDAC inhibition would enhance the anti-tumor effect of PARP inhibitor in cancer cells by blocking DNA repair pathway.
Materials and Methods: We determined whether SAHA, a HDAC inhibitor could enhance the growth inhibition of olaparib on breast cancer cell lines using MTT assay. We examined whether exposure to SAHA affects the expression level of genes involved in HR. The accumulation of DNA double strand breaks (DSBs) induced by combination treatment was accessed by the comet assay. Cell cycle analysis and molecular changes induced by combination of olaparib plus SAHA were also performed. These in vitro data were validated in the in vivo xenograft model as well.
Results: Triple-negative breast cancer cell lines showed heterogeneous response to dual inhibition of PARP and HDACs. SAHA enhanced olaparib-induced cell death of MDA-MB-157 and HCC1143 but not of HCC70 and MDA-MB-468. Combination of olaparib plus SAHA caused a greater decrease of pAKT, pERK, and pSTAT3 in MDA-MB-157 and HCC1143 cells compared with monotherapy either olaparib or SAHA. Furthermore, inhibition of PARP increased the accumulation of DNA DSBs induced by SAHA in these two cell lines, MDA-MB-157 and HCC1143. There was no change in proliferative pathway activation in HCC70 and MDA-MB-468 cells with combination of olaparib plus SAHA. Our findings showed that triple-negative breast cancer cells are differentially effective to combination of SAHA plus olaparib which increased levels of unrepaired DNA DSBs.
Conclusion: HDAC inhibitor SAHA enhances growth inhibitory activity of PARP inhibitor olaparib in several triple-negative breast cancer cells with increased accumulation of DNA DSBs. Our results provide a rationale for the future clinical trials of olaparib combined with SAHA in the treatment of cancers that have an impaired DNA repair ability.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-16-06.
Abstract P4-16-12: CARE: A pilot study of the effects of short-term fasting on tolerance to (neo) adjuvant chemotherapy in breast cancer patients
Authors: de Groot S , Vreeswijk MPG , Gravesteijn G , Boei JJWA , Jochems A , Houtsma D , Nortier HWR , van der Hoeven KJM , Pijl H , Kroep JR 
Published: August 2014

Abstract
Background:
Preclinical evidence shows that short-term fasting (STS) protects normal cells and makes cancer cells more vulnerable to chemotherapy. This pilot study examines the feasibility and the effects of STS on tolerance to chemotherapy in patients with breast cancer.
Patients and methods:
Eligible patients had histologically confirmed, HER2-negative, early stage breast cancer and adequate bone marrow, liver and renal function. Women receiving (neo) adjuvant TAC courses (docetaxel/adriamycin/cyclophosphamide; day 1, q 3 weeks with G-CSF support at day 2) were randomized to fast 24 hours before and 24 hours after start of chemotherapy or to eat according to the guideline for healthy nutrition. The primary endpoint of the study was to compare neutrophil count after therapy. Secondary endpoints were side effects of chemotherapy, other hematologic counts and chemotherapy-induced DNA damage in leukocytes.
Results:
A total of 13 patients were included of which 7 patients fasted for 48 hours around the chemotherapy infusion (arm A) and 6 patients had a normal diet according to healthy nutrition guidelines (arm B). The median age was 52 years versus 53 years, BMI was 25.5 kg/m2 versus 22.9 kg/m2 and stage III was 43% versus 17% of patients in arms A and B, respectively. Patients were generally motivated to fast and the fasting was well tolerated. Plasma glucose levels were significant lower in fasting patients compared to controls. However, other metabolic parameters showed no significant difference. Fasting did not result in significant differences in neutrophil count or side effects of chemotherapy. Hemoglobin levels and erythrocyte counts after therapy were significantly higher in patients who fasted. Leukocytes of the patients which were isolated at various time points during therapy will soon be analysed for chemotherapy-induced DNA damage and presented at San Antonio.
Conclusion:
This is the first study evaluating the feasibility of 48 hours STS and its impact on side effects of chemotherapy in a homogeneous group of cancer patients. STS was well tolerated and had a beneficial effect on hemoglobin level, but not on experienced side effects. DNA analysis will follow. Larger studies are required to produce more insight into the possible benefits of STS during chemotherapy.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-16-12.
Abstract P4-19-05: Breast cancer campaign tissue bank human cell culture programme: Generating new biomaterials for breast research
Authors: Gomm JJ , Smith SG , Allen MD , Morgan AS , Haywood LA , Sarper M , Dreger S , Ironside A , Cortes Lopez EJ , Del Rio Hernandez A , Speirs V , Jones LJ 
Published: August 2014

Abstract
Background:
In-vitro model systems provide an important tool in breast cancer research and, over recent years, there have been considerable advances in their construction. However, despite these advances, the majority of models still employ a narrow range of established cell lines, which frequently are not appropriate for the in-vivo cell type they are representing1.
Method:
As part of the Breast Cancer Campaign Tissue Bank, we have developed a cell culture programme which feeds into the main tissue collection and involves the systematic generation of materials from normal, high-risk, cancer-containing and malignant breast tissue in order to make available a breadth of material to the research community.
Following informed patient consent, fresh tissue is retrieved and, using tissue digestion and magnetic bead technology2, purified cell populations are isolated, characterized and frozen down, or further processed for DNA and RNA isolation.
Results:
We have isolated purified populations of luminal epithelial, myoepithelial and fibroblast cells from normal breast, risk-reduction (BRCA1 and BRCA2 mutated) breast and from morphologically normal breast tissue surrounding breast cancer. We have generated matched surround and tumour-associated fibroblasts, as well as matched tumour epithelial and tumour-associated fibroblast cell isolates.
These cells have been incorporated into 2- and 3-D culture models and we have demonstrated successful genetic manipulation, with siRNA gene knockdown and gene over-expression through retroviral transduction. Immortalisation of selected cell populations is currently underway.
Intact organoids and tissue explants have also been generated for in-vitro experimentation. Matched frozen and FFPE samples are available, together with full clinicopathological data. The research community can access these biomaterials via a web-based search portal, with full user support.
Conclusion:
Through this cell culture programme we aim to make available a wider and more appropriate range of materials for breast research, which should allow more clinically relevant model systems to be developed and add value to breast cancer biobanking.
References:
1. Thompson A., et al. Evaluation of the current knowledge limitations in breast cancer research: a gap analysis. Breast Cancer Research 2008; 10: R26.
2. Gomm J.J. et al. Isolation of pure populations of epithelial and myoepithelial cells from the normal human mammary gland using immunomagnetic separation with Dynabeads. Analytical Biochemistry 1995; 226: 91-99.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-19-05.
Abstract P5-01-05: Personalized peptide vaccines in combination with conventional chemo- or endocrine therapy for metastatic breast cancer: A single-arm phase II study
Authors: Toh U , Iwakuma N , Mishima M , Takenaka M , Takahashi R , Koura K , Fujii T , Nakagawa S , Ogo E , Tanaka M , Sasada T , Itoh K , Shirouzu K 
Published: August 2014

Abstract
Background: Immunotherapy using multiple personalized peptide vaccines (PPV) has been reported to be effective for boosting anticancer immunity in patients (pts) with advanced prostate cancer and glioblastoma. The PPVs binding HLA-A2, A24, A3 family and A26 molecules were selected from 31 candidate peptides derived from various tumor associated antigens. Currently, we performed a phase II trial to evaluate the benefit of the PPVs in combination with conventional chemo-or endocrine therapy for pretreated metastatic breast cancer (MBC). Methods: Seventy-one pretreated pts had histologically confirmed measurable MBC and their HLA A molecules matched each of HLA-A2, A24, A26 and A3 family. Pre-vaccination plasma was measured for their IgG levels reactive to each of 31 peptides followed by administration subcutaneously of the four peptides at maximum showing higher levels of IgG in order with schedule of every 1-2 week. The concurrent conventional chemotherapy or endocrine therapy was available for the combination of PPVs. The primary endpoint was anti-tumor immunologic effect and safety, and the secondary endpoints were clinical responses and survival. Levels of IgG reactive to each of the 31 peptides in the pre- and post-treatment plasma at every 6 times of vaccination were measured using LUMNEX system. The CTL responses were simultaneously evaluated using ELISPOT method. Clinical response was evaluated by RECIST criteria. Results: Between May 2009 and December 2012, a total of 71 pts (Luminal-Her2- cases: 39; Her2+cases: 18; Triple negative(TN) cases:14) were enrolled in this study. The median duration of follow-up was 20.7 months. Concurrent chemotherapies (capecitabine, gemcitabine, eribulin, vinorelbine etc.) and endocrine therapies (aromatase inhibitor, toremifen or fulvestrant) were administered to 36 and 24 pts, respectively. Trastuzumab, Laptinib or Bevacizumab was simultaneously used for 13 pts. After 6th vaccination, the IgG reaction and CTL response were identified in 53 of 71 (74.6%) pts and 33 of 63 (52.3%) pts. There were no significant differences between TN cases and the other cases of MBC with regard to immunological humoral and/or cellular responses. No vaccine-related severe adverse events were observed. Clinical response rate of assessable 61 pts was 14.8% (CR 3, PR 6, SD 24, PD 28). The median PFS and OS were 7.6 m and 20.7 m, respectively. In contrast, the PFS and OS of luminal type, Her2 type or TN type were 12, 4.6 or 8.3 m and 24, 15 or 12 m. Conclusions: The phase II PPVs study for MBC demonstrated the promising response and safety, and further studies are essential to identify the clinical benefits of this novel therapy. Clinical trial information: UMIN000001482.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-05.
Abstract P5-01-06: Remodeling the tumor immune microenvironment via intratumoral administration of immunomodulatory agents
Authors: Campbell MJ , Senman B , Esserman LJ 
Published: August 2014

Abstract
Background: Immune dysregulation has been implicated in the poor outcomes associated with high grade and/or triple negative (Tneg) breast cancers. This suggests that a multi-modal approach to stimulating the immune system could be an effective therapeutic strategy for Tneg breast cancer. However, many immunotherapeutic agents demonstrate significant toxicities when administered systemically at high doses. Therefore, we investigated the therapeutic efficacy of local, intratumoral administration of immunomodulators in a mouse model of Tneg breast cancer.
Methods: 4T1, an aggressive, Tneg mammary carcinoma, was implanted orthotopically into syngeneic BALB/c mice. Treatment was initiated when tumors were 0.5-1.0 cm in the largest dimension. We examined various combinations of Toll-like receptor agonists (CpG oligonucleotide [TLR9] and imiquimod [TLR7]) with immunomodulatory antibodies (anti-CD40, anti-CTLA-4, anti-OX40). These agents were administered intratumorally (i.t.), either in PBS or formulated in an in situ depot forming drug delivery system. Growth of the injected tumors was monitored over time. In some experiments, tumors and spleens were harvested 3-10 days post therapy and analyzed for the presence of CD4+ T cells, CD8+ T cells, immunoregulatory T cells (Treg), and myeloid derived suppressor cells (MDSC). To test whether local immunotherapy generates a systemic antitumor immune response, 4T1 tumors were implanted at 2 sites on opposite sides of the mice, with one (left) used as the site of in situ immunomodulation (i.t. injections) and the other (right) observed to assess the systemic antitumor immune response.
Results: Intratumoral administration of various combinations of immunomodulators resulted in complete regression of the injected tumor. These regressions were associated with a reduction in the number of MDSCs in the tumor as well as activation of both CD4+ and CD8+ T cells. Furthermore, local immunotherapy generated a systemic antitumor immune response that inhibited tumors growing at a distant site.
Conclusions: Our findings demonstrate that remodeling the tumor immune microenvironment via intratumoral administration of immunomodulatory agents elicits antitumor immunity that eradicates tumor cells locally as well as systemically at distant sites. The prospect of delivering immunotherapeutics only where it matters most, namely within the tumor microenvironment, is extremely attractive as this would likely be substantially less toxic and more specific than many of the current cancer treatments. Ongoing studies include identifying companion diagnostics that can indicate which patients would benefit most from such an immunotherapeutic approach.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-06.
Abstract P5-01-07: Select neutrophils inhibit breast cancer metastases in patients via chemokine-mediated mechanisms
Authors: Comen EA , Granot Z , Blum B , Coker C , Shah R , Seshan V , Norton L , Benezra R 
Published: August 2014

Abstract
Background: In both breast cancer patients and healthy women, we have previously demonstrated that select neutrophils found in breast cancer patients as opposed to healthy women are cytotoxic to breast cancer cell lines. (Granot Z et al. Cancer Cell. 2010) This work stemmed from our prior research in murine breast cancer models indicating that primary breast tumors can mobilize select neutrophils, termed Tumor Entrained Neutrophils (“TENS”); these entrained neutrophils have the unique capacity to inhibit metastatic seeding in the lung through cell-kill mechanisms. (Granot Z et al. Cancer Cell. 2010) In this study, we evaluated the relationship between select chemokines and neutrophil cytotoxicity in breast cancer patients versus healthy volunteers.
Methods: Neutrophils were purified from the blood of 75 randomly selected newly diagnosed pre-operative breast cancer patients without evidence of metastatic disease, and 47 healthy female volunteers with no history of cancer. Cytotoxicity was evaluated by incubating neutrophils with luciferase labeled MDA-MB-231 cells. Luciferase activity was measured as a reflection of% cytotoxicity. Serum was also isolated from breast cancer patients and healthy volunteers. Based on prior experiments, we used the Millipore® Milliplex Human Cytokine Plex Kit to evaluate Il-1Ra, MCP-1 and TNFa in our serum samples in 50/75 of the cancer patients and 25/47 controls. We used multiple linear regression to develop a model to predict cytotoxicity as a function of the chemokines.
Results: In comparison to healthy volunteers whose mean neutrophil cytotoxicity to MDA-MB-231 cells was 6.5%, pre-operative breast cancer patients demonstrated a mean neutrophil cytotoxicity of 12.7%, p&lt;0.0001. We then evaluated the serum chemokine levels of 50 of the breast cancer patients; 31/50 had high neutrophil cytotoxicity (&gt;10%), and 19/50 had low neutrophil cytotoxicity (&lt;10%). We compared the serum chemokine levels in the patients to the levels found in the 25 controls. Using a multiple linear regression model, we found that the levels of these three chemokines are associated with cytotoxicity (R2 = 0.126, p = 0.022). An ANOVA decomposition of the model suggested that Il.1RA was the most predictive (p = 0.018) followed by MCP.1 (p = 0.088) and TNF.alpha (p = 0.245).
Conclusion: Our work demonstrates the cytotoxic role of select neutrophils in the peripheral blood of breast cancer patients as contrasted with neutrophils from healthy women. We further demonstrate that select chemokines appear to be correlated with neutrophil cytotoxicity. We are currently evaluating the prognostic and therapeutic roles of cytotoxic neutrophils and their related chemokines in breast cancer patients.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-07.
Abstract P5-01-12: Prognostic implications of tumor-infiltrating lymphocytes (TIL) in association with PD-L1 expression, and serum cytokine levels in early breast cancer
Authors: Park IH , Kong S-Y , Ro JY , Kwon Y , Kang JH , Mo HJ , Jung S-Y , Lee S , Lee KS , Kang H-S , Lee E , Ro J 
Published: August 2014

Abstract
Background
Immune system has been known to influence the prognosis of breast cancer (BC). However, the relationship between immune modulating factor (PD-L1) and tumor infiltrating lymphocyte (TIL) profiles in breast cancer has yet to be revealed according to breast cancer subtypes. In addition, the effects of circulating cytokines on TILs have not been addressed.
Patients and methods
We investigated the relationship between the profiles of TILs and PD-L1 expression of the primary tumor tissue by immunohistochemistry with clinical outcomes in 253 patients who underwent surgery for early breast cancer at National Cancer Center from January 2001 to December 2005. Besides, the serum cytokines including IL-10, IL-18, IL-6, IFN-g, and TGF-β1 were measured at diagnosis. Clinical data including hormone receptors status, HER2 expression, disease free survival (DFS), and overall survival (OS) were collected.
Results
Median age of patients was 49 years (range, 32-74) and median follow-up was 8.5 years. One hundred eighty five (73.1%) patients had hormone receptor (HR) positive and 101 (39.9%) patients had node positive BC. CD8+ TILs were more abundant in low PD-L1 expressed tumor (P = 0.027), though there was no association between FOXP3+ TILs and PD-L1 expression (P = 0.585). A total number of TILs was higher in HR negative compared with HR positive BC (P = 0.061) and the expression of PD-L1 was more frequent in HR positive BC (P&lt;0.001). In HR negative BC, there was a trend of longer DFS in patients with higher CD8+ TILs and low PD-L1 expression (P = 0.097). However, such association was not detected in HR positive BC patients. Among serum cytokines we examined, the higher levels of IL-18 were significantly associated with shorter DFS in HR negative BC (P = 0.006). In HR negative BC, higher CD8+ TILs with low PD-L1 expression and lower IL-18 were significantly related with better clinical outcomes when adjusted with other clinical factors (DFS, P = 0.032; OS, P = 0.048).
Conclusions
Lower PD-L1 expression in breast tumor was associated with higher CD8+ lymphocyte infiltration. Especially in HR negative BC, increasing CD8+ TILs with lower PD-L1 expression and lower serum IL-18 level were good prognostic factors. Further validation will be needed to establish the role of immune profiles in BC patients.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-12.
Abstract P5-01-13: Poly(I:C), an innate adjuvant receptor ligand, can induce the antitumor effects on human breast cancer cells
Authors: Inao T , Harashima N , Yamamoto Y , Harada M , Iwase H 
Published: August 2014

Abstract
[Background] Innate adjuvant receptors are expressed in immune cells and some types of cancers. If antitumor therapies targeting adjuvant receptors in cancer cells are established, it could be therapeutically useful because antitumor effects and activation of the immune system can be induced simultaneously.
[Objective]Poly(I:C) is one of innate adjuvant receptors ligands and triggers signaling through not only endosome Toll-like receptor (TLR) 3 but also cytoplasmic melanoma differentiation-associated gene (MDA) 5. In this study, we investigated antitumor effects of poly(I:C) on human breast cancer cell lines, and then elucidated the mechanisms.
[Methods] Three breast cancer cell lines (MCF-7, MDA-MB-231, and BT-549) were used in this study. Poly(I:C) was transfected into these cancer cells, and cell viability and apoptosis were evaluated by WST8 assay and flow cytometry, respectively. The expression of adjuvant receptors and cell cycle-related molecules were examined by immunobotting. Autophagy was evaluated by immunoblotting and confocal imaging. In vivo antitumor effect was determined using a xenograft mouse model.
[Results] All three cell lines were positive for both TLR3 and MDA5. Poly(I:C) transfection drastically reduced their cell viabilities in a partially MDA5-dependent manner. Poly(I:C)-transfected cancer cells showed caspase-dependent apoptosis and growth arrest as a result of suppression of c-Myc and cyclinD1. Interestingly, poly(I:C) transfection was accompanied by autophagy, which is known to work cytoprotectively under starvation and stress, and knockdown of beclin-1, an essential molecule for autophagy, by RNA interference promoted poly(I:C) transfection-induced apoptosis. In vivo poly(I:C) transfection exerted the antitumor effect on xenografted MDA-MB-231 in nude mice.
[Conclusions] These results suggest that the innate adjuvant receptors are promising targets in human breast cancer and that autophagy plays a protective role in this treatment modality.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-13.