bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract B23: Citral reduces ALDH1A3 activity in breast cancer: Potential applications in targeting breast cancer stem cells
Authors: Thomas Margaret L. , Coyle Krysta M. , Cruickshank Brianne , Giacomantonio Michael , Wallace Melissa , Giacomantonio Carman , Marcato Paola 
Published: May 2016

Abstract
Breast tumors contain a subpopulation of cells known as cancer stem cells (CSCs) which are highly tumorigenic, resistant to typical chemo- and radiotherapy, and express high levels of aldehyde dehydrogenase (ALDH) isoforms ALDH1A3 and ALDH1A1. These enzymes initiate retinoic acid (RA) signaling and potentially contribute to detoxification of chemotherapy. A compound that inhibits these enzymes could be used as an adjuvant therapy to target breast CSCs and/or re-sensitize this population to chemotherapy. In particular, there are no currently characterized inhibitors of the ALDH1A3 isoform, so twelve general ALDH inhibitors were tested for their effectiveness in targeting ALDH1A3 activity. Ability to directly inhibit ALDH activity was quantified with the Aldefluor assay on a patient tumor xenograft and on breast cancer cell lines MDA-MB-231, MDA-MB-468, and SKBR3. Inhibition of RA signaling was quantified by measuring RA-inducible gene expression after 24 hours treatment with ALDH inhibitor. To investigate the anti-cancer properties of these compounds, apoptosis was quantified with the annexin V assay after 24 hours treatment. To determine if increased ALDH1A3 expression is involved in chemoresistance, MDA-MB-231 cells overexpressing ALDH1A3 and MDA-MB-468 cells with ALDH1A3 shRNA knockdown were treated for 72 hours with doxorubicin, 4-hydroperoxycyclophosphamide, or paclitaxel. Increased ALDH1A3 expression did not affect the amount of chemotherapy-induced apoptosis or cell proliferation rates. Citral significantly reduced ALDH1A3-mediated expression of RA-inducible genes and significantly reduced the ALDH activity of the patient-derived xenograft as well as ALDH1A3-overexpression MDA-MB-231 cells. Citral induced apoptosis in an ALDH1A3-dependant manner in breast cancer cell lines, and its ability to reduce MDA-MB-231 in vivo growth was investigated by implanting MDA-MB-231 cells in female NOD/SCID mice and injecting with 0.1mg/kg citral. Tumor size was not affected by citral injections, however recent data suggests that micelle-encapsulated citral has the potential to reduce tumor bulk. Should citral prove to be an effective breast cancer and ALDH1A3 inhibitor in future tumor xenograft experiments, this compound could potentially be used as adjuvant therapy for breast cancer patients.
Citation Format: Margaret L. Thomas, Krysta M. Coyle, Brianne Cruickshank, Michael Giacomantonio, Melissa Wallace, Carman Giacomantonio, Paola Marcato. Citral reduces ALDH1A3 activity in breast cancer: Potential applications in targeting breast cancer stem cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B23.
Abstract B41: HGFL-Ron signaling enhances breast cancer stem cell populations
Authors: Ruiz-Torres Sasha J. , Waltz Susan E. 
Published: May 2016

Abstract
The Ron receptor tyrosine kinase is overexpressed in a high fraction of human breast cancers (BCs). Ron and its ligand, the Hepatocyte Growth Factor-Like protein (HGFL), play important roles during BC initiation, progression, and therapeutic resistance, with overexpression of this signaling cascade associated with increased BC metastasis and poor prognosis in humans. Reports demonstrate that a small subpopulation of cancer cells, called the breast cancer stem cells (BCSCs), are essential for the development and maintenance of BC, leading to tumor formation and recurrence. Despite the role of HGFL-Ron signaling as a key regulator of BC, nothing is known about this signaling pathway in BCSC populations. To investigate the function of HGFL-Ron signaling in BCSCs, the expression levels of Ron and HGFL were analyzed in BCSCs. mRNA analyses show increased levels of Ron and HGFL in BCSC-enriched Mammospheres (MSs) compared to parental BC epithelial cells. Next, the association between Ron signaling and the BCSC population was examined. Aldefluor assays of several Ron replete and depleted human and murine BC epithelial cell lines show that Ron overexpression increases the BCSC population. In vivo, flow cytometry analyses of breast tumors for BCSC markers show that Ron signaling is associated with increases in the BCSC population. MS formation assays using several Ron replete and depleted human and murine BC cell lines also demonstrate that cells expressing high levels of Ron have increased MS formation. Ex vivo studies of sorted BCSCs further support that Ron signaling enhances the MS formation ability of BCSCs. Interestingly, our data suggest that both genetic ablation and chemical inhibition of Ron signaling is sufficient to decrease BCSC MS formation ability. Mammosphere culture over several passages also suggest that Ron signaling BCSCs have higher self-renewal capabilities than Ron deficient BCSCs, highlighting the role of HGFL-Ron signaling in regulating BCSC function. Recent preliminary studies correlate Ron signaling in BCSCs with increased β-catenin and NF-κB pathways, suggesting these as the potential molecular mechanisms through which Ron signaling promotes BCSC function in BC. Overall, our data provide (i) the first evidence demonstrating both HGFL and Ron as critical factors regulating the BCSC population and (ii) that this regulation might occur in a β-catenin and NF-κB pathway dependent manner. These studies suggest both Ron and HGFL as potential therapeutic targets to eradicate BCSCs and improve the outcome of BC patients.
Citation Format: Sasha J. Ruiz-Torres, Susan E. Waltz. HGFL-Ron signaling enhances breast cancer stem cell populations. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B41.
Abstract B48: Intratumor ploidy heterogeneity as a chemoresistance mechanism in triple-negative breastcancer
Authors: Gyanchandani Rekha , Lee Adrian 
Published: May 2016

Abstract
Background: Triple negative breast cancers (TNBC) are aggressive, heterogeneous tumors, and currently lack targeted therapies. Chemotherapy is the mainstay treatment, but tumor recurrence is a major cause of mortality and morbidity. Chemotherapy may select for resistant subclones that have a survival advantage over the bulk population of cancer cells and cause tumor relapse. DNA aneuploidy (abnormal number of chromosomes) is a common genetic feature in breast cancer and has been frequently associated with poor prognosis. Although, aneuploid or polyploid cancer cells can contribute to intratumor ploidy heterogeneity, their functional role in tumorigenesis and chemoresistance is largely undefined. Traditionally, polyploid cells were thought to arise due to defects in cell division and were considered to be senescent. However, recent reports have shown that polyploid cells represent a viable and proliferating subpopulation within a tumor and may have a direct role in regulating chemoresistance.
Methods: In this study, we isolated and functionally characterized the diploid and polyploid sub-populations derived from TNBC cell lines. DNA ploidy was assessed in Hoechst 33342 stained cells using flow cytometry. Cells with greater than 4N DNA content were considered as polyploid. Next, we analyzed cell proliferation in HCC1937 cells using CFSE and DNA labeling. Live-cell imaging was performed on flow-sorted diploid and polyploid cells using IncuCyte Zoom to monitor cell proliferation in the absence and presence of mitotic inhibitor (docetaxel).
Results: DNA ploidy analysis identified presence of polyploid cells in HCC1395 and HCC1937 cell lines at a frequency of 6.7% and 5.8% respectively. Polyploid cells have higher forward-scatter than diploid cells suggesting that they are larger in size. In addition, nuclear staining with Hoechst 33342 showed that giant polyploid cells contain enlarged and/or multiple nuclei. Time-lapse microscopy in polyploid cells revealed mitotic structures suggestive of multipolar cell division. Carboxyﬂuorescein diacetate, succinimidyl ester (CFSE) labeling showed that polyploid cycle slower than the predominant diploid population. Further, the IncuCyte cell proliferation assay also confirmed that sorted polyploid cells divide slower than the diploid cells and displayed resistance to docetaxel.
Concusions: These findings indicate that there are functional differences between diploid and polyploid sub-populations. In our future studies, we will examine the isolated diploid and polyploid cells for differences in their tumorigenic potential and role in tumor recurrence.
Citation Format: Rekha Gyanchandani, Adrian Lee. Intratumor ploidy heterogeneity as a chemoresistance mechanism in triple-negative breastcancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B48.
Abstract B56: Prolactin promotes breast cancer to bone metastasis and breast cancer cell-mediated osteoclast differentiation
Authors: Forsyth Amanda , Sutherland Ashley , Cong Yingying , Grant Laurel , Juan Tzu-Hua , Lee Jae K. , Klimowicz Alexander , Petrillo Stephanie K. , Hu Jinghui , Chan Angela , Boutillon Florence , Goffin Vincent , Egan Cay , Tang Patricia A. , Cai Li , Morris Don , Magliocco Anthony , Shemanko Carrie S. 
Published: May 2016

Abstract
The hormones prolactin (PRL), estrogen and progesterone have long been studied for their role in the primary breast tumor but not yet in modulating the secondary tumor microenvironment of the bone. Metastasis to the bone is a debilitating aspect of many cancers, including breast cancer, where it is a preferred site of metastasis that results in bone loss. Breast cancer cells release osteolytic factors that induce the breakdown of bone, which releases growth factors and calcium that create a vicious cycle of metastatic tumor growth. Using quantitative immunohistochemistry (AQUA) (n=134), we determined that high PRL-receptor expression in the primary tumor was associated with a shorter time to bone metastasis (PRLR AQUA Max Hazard ratio=1.04, 95% Hazard Ratio confidence limits 1.00-1.07, p=0.03/multivariable Cox proportional hazards model), indicating their treatment failure may be related to the PRL-receptor. We also identified the PRL-receptor on rare samples of matched primary and bone metastases. In an analysis of advanced breast cancer patients, we also detected the PRL-receptor in circulating tumor cells of the blood. PRL treatment of breast cancer cells induced osteoclast differentiation and bone lysis via presumed secreted factors, and interestingly these effects were abrogated by a PRL-receptor-antagonist (delta1-9-G129R-hPRL). We identified sonic hedgehog as part of the molecular mechanism by which PRL and the PRL-receptor induce breast cancer cells to directly promote the differentiation of osteoclast cells capable of bone resorption. This molecular mechanism identifies key potential therapeutic targets to ameliorate the devastating effects of breast cancer to bone metastasis and potential predictive biomarkers.
Citation Format: Amanda Forsyth, Ashley Sutherland, Yingying Cong, Laurel Grant, Tzu-Hua Juan, Jae K. Lee, Alexander Klimowicz, Stephanie K. Petrillo, Jinghui Hu, Angela Chan, Florence Boutillon, Vincent Goffin, Cay Egan, Patricia A. Tang, Li Cai, Don Morris, Anthony Magliocco, Carrie S. Shemanko. Prolactin promotes breast cancer to bone metastasis and breast cancer cell-mediated osteoclast differentiation. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B56.
Abstract B59: Age and estrogen-dependent inflammation in breast adenocarcinoma and normal breast tissue
Authors: Quigley David A. , Tahiri Andliena , Lüders Torben , Riis Margit H. , Balmain Allan , Børresen-Dale Anne-Lise , Bukholm Ida R. , Kristensen Vessela 
Published: May 2016

Abstract
Chronic inflammation promotes breast tumor growth and invasion by accelerating angiogenesis and tissue remodeling in the tumor microenvironment. There is a complex relationship between inflammation and estrogen, which drives the growth of 70 percent of breast tumors. Low levels of estrogen exposure stimulate macrophages and other inflammatory cell populations, but very high levels are immune suppressive. Breast tumor incidence is increased by obesity and age, which interact to influence inflammatory cell populations in normal breast tissue. To characterize the impact of these factors on tumors and the tumor microenvironment, we measured gene expression profiles in 195 breast adenocarcinomas and 195 matched adjacent normal breast tissue samples. Age and Body Mass Index (BMI) were independently associated with inflammation in normal tissue but not tumors. We developed a novel statistic, the Rewiring Coefficient, to quantify the rewiring of gene co-expression networks at the level of individual genes. Estrogen Receptor (ER)-negative tumors had elevated macrophage expression compared to matched normal tissue, but ER-positive tumors showed an unexpected decrease in macrophage expression. We found an inverse relationship between the increase in tumor estrogen pathway expression compared to adjacent normal tissue and that tumor's macrophage score and validated this finding in 126 matched breast tumor-normal pairs from the previously published METABRIC cohort. Our data support an immune suppressive effect of high doses of estrogen signaling in breast tumor microenvironment, suggesting that this effect contributes to the greater presence of prognostic and therapeutically relevant immune cells in ER-negative tumors.
Citation Format: David A. Quigley, Andliena Tahiri, Torben Lüders, Margit H. Riis, Allan Balmain, Anne-Lise Børresen-Dale, Ida R. Bukholm, Vessela Kristensen. Age and estrogen-dependent inflammation in breast adenocarcinoma and normal breast tissue. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B59.
Abstract IA10: Sox10 regulates stem/progenitor and mesenchymal cell states in mammary epithelial cells
Authors: Dravis Christopher , Wahl Geoffrey M. 
Published: May 2016

Abstract
To discover mechanisms that mediate the initiation and progression of aggressive and stem-like breast cancers, we characterized signaling networks that are present in the mammary stem cells responsible for fetal and adult mammary development. These analyses identified a signaling axis between FGF signaling and the transcription factor, Sox10. Here we report that Sox10 is specifically expressed in mammary cells that exhibit the highest levels of stem/progenitor activity. This includes fetal and adult mammary cells in vivo and mammary organoids in vitro. Sox10 is functionally relevant, as its deletion reduces stem/progenitor competence, while its overexpression increases stem/progenitor activity. Intriguingly, we also discover that Sox10 overexpression elicits epithelial-to-mesenchymal transition (EMT) in mammary organoids in an FGF signaling-dependent manner. Further, modulation of Sox10 levels can induce sequential EMT, migratory, and clonogenic behaviors that strikingly resemble proposed mechanisms of metastasis. Consistent with these findings, we find that Sox10 is preferentially expressed in the most stem- and EMT-like triple negative breast cancer subtypes, and that Sox10+ tumor cells utilize FGF signaling for growth and invasive behaviors, which suggest that Sox10 may reprise these same functions during tumorigenesis. Collectively, these results demonstrate a signaling mechanism through which stem and mesenchymal-like states are acquired in mammary cells, and indicate possible therapeutic targets to counter these functions in breast cancers for which targeted therapies are currently unavailable.
Citation Format: Christopher Dravis, Geoffrey M. Wahl. Sox10 regulates stem/progenitor and mesenchymal cell states in mammary epithelial cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr IA10.
Abstract IA16: Clinical relevance of host immunity in breast cancer
Authors: Loi Sherene 
Published: May 2016

Abstract
The clinical relevance of the host immune system in breast cancer has long been unclear. Recently an appreciation of the biological heterogeneity of breast cancer has prompted investigation of the immune system's role in the different molecular breast cancer subtypes. A robust association between a more favourable prognosis and the presence of high levels of lymphocytic infiltration has been found in early-stage triple negative and HER2-positive breast cancer. These infiltrates seem to represent favorable anti-tumor immune responses, suggesting that activation of host immunity will be important for improving survival outcomes. The biological influences underlying the immune response are incompletely understood.My talk will review recent data supporting the clinical relevance of host anti-tumor immunity as represented by lymphocytic infiltration, the composition of the infiltrate in breast cancer and how this biomarker could be used in the clinical setting. Lymphocytic infiltration may be important as a biomarker in breast cancer prognosis: efforts to standardize the methodology for evaluating TILs in clinical breast cancer specimens in routine histopathological practise are ongoing. I will also discuss the rationale for enhancing immunity in breast cancer, including data on the efficacy of T-cell checkpoint inhibition..
References:
1. The evaluation of tumor-infiltrating lymphocytes (TILs) in breast cancer: recommendations by an International TILs Working Group 2014. Ann Oncol. 2015 Feb;26(2):259-71. doi: 10.1093/annonc/mdu450. Epub 2014 Sep 11.
2. Host antitumor immunity plays a role in the survival of patients with newly diagnosed triple-negative breast cancer. J Clin Oncol. 2014 Sep 20;32(27):2936-8.
3. Tumor-Infiltrating Lymphocytes and Associations With Pathological Complete Response and Event-Free Survival in HER2-Positive Early-Stage Breast Cancer Treated With Lapatinib and Trastuzumab: A Secondary Analysis of the NeoALTTO Trial. JAMA Oncol. 2015 Jul 1;1(4):448-54. doi: 10.1001/jamaoncol.2015.0830.
Citation Format: Sherene Loi. Clinical relevance of host immunity in breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr IA16.
Abstract A33: A role of SET/I2PP2A in HNSCC angiogenesis and tumor-promoting microenvironment
Authors: Sobral Lays , Santos Leandro F. , Curti Carlos , Leopoldino Andreia M. 
Published: May 2016

Abstract
This study addressed a potential role of SET/I2PP2A protein on angiogenesis and immune response in head and neck squamous cell carcinoma (HNSCC). A HNSCC cell line with stable SET knockdown (HN12shSET/HN12shControl), previously established in our laboratory, was used to perform (1) qPCR array analysis of genes related to angiogenesis (Qiagen), and (2) tube-formation assay using Human Umbilical Vein Endothelial cells (HUVEC) cultured with HN12shSET cells conditioned medium. Moreover, several chemokines and cytokines were analyzed by Multi-Analyte ELISA array (Qiagen), and the specific levels of TGF-B1 and RANTES were validated by a Single-Analyte ELISA array (Qiagen) in cells exposed or not to TNF-alpha; stimulus. The cellular distribution of NF-kB, p-NF-kB and p-STAT3 was assessed by immunofluorescence, cell fractionation and western blotting assays. Our results showed higher levels of genes that induce angiogenesis, in particular VEGFC, in HN12shSET cells; these cells presented, in addition, higher TGF-B1 and lower RANTES/CCL-5 secretion levels compared to controls, as well as dowregulation of CCL-2 expression. Moreover, the stimulation of HUVEC tube-formation by the HN12shSET cells conditioned medium was higher (1.7 fold) compared to control, and p-NF-kB and p-STAT3 were found predominantly in the nucleus of HN12shSET cells; this suggests a negative balance between SET and regulation of these transcription factors. Therefore, SET knockdown in HN12 cells stimulates angiogenesis through VEGFC and TGF-B1 secretion, which could contribute to create a tumor-promoting microenvironment. These results support a SET role in HNSCC development and progression
Citation Format: Lays Sobral, Leandro F. Santos, Carlos Curti, Andreia M. Leopoldino. A role of SET/I2PP2A in HNSCC angiogenesis and tumor-promoting microenvironment. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr A33.
Abstract B12: March E3 ubiquitin ligase is required for head formation by mediating Dishevelled degradation during Xenopus development
Authors: Lee Hyeyoon , Cheong Seong-Moon , Han Jin-Kwan 
Published: May 2016

Abstract
Wnt signaling controls numerous biological events including vertebrate development, adult tissue homeostasis and organogenesis. In addition, dysregulation of Wnt itself or its downstream signaling components causes human birth defects and cancer. Dishevelled (Dvl) is a crucial scaffolding protein that exerts activation of Wnt signaling pathway. Recently, several Dvl-interacting proteins have been suggested to mediate the stability of Dvl. However, the molecular mechanisms by which Dvl undergoes degradation remain to be elucidated. Here we identify March, a RING domain-containing ubiquitin ligase, targeting Dvl protein and show that it regulates canonical Wnt signaling in Xenopus. During Xenopus embryogenesis, March transcripts were ubiquitously expressed. We intriguingly found that March depletion by microinjection of morpholino oligonucleotide caused defects in the anterior head formation, including loss of the forebrain, cement gland and eye structures. We also found that these defective phenotypes were caused by the Wnt antagonistic function of March in the head organizer region. Knockdown of March suppressed expression of head organizer genes Gsc, Chd and Otx2 at the dorsal marginal region of the gastrula embryo. Wnt reporter assay, western blot analysis measuring phosphorylation of LRP6 and activation of β-catenin, RT-PCR and axis duplication assay in Xenopus revealed that March inhibits canonical Wnt signaling pathway. Furthermore, we identified that March protein associates with Dvl and mediates its poly-ubiquitination for lysosomal degradation in Xenopus embryos. Taken together, these results suggest that March retains cytosolic pools of Dvl protein and leads subsequent limitation of Wnt signaling for vertebrate head development.
Citation Format: Hyeyoon Lee, Seong-Moon Cheong, Jin-Kwan Han. March E3 ubiquitin ligase is required for head formation by mediating Dishevelled degradation during Xenopus development. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B12.