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  • 1
    In: Neuro-Oncology, Oxford University Press (OUP), Vol. 17, No. suppl 5 ( 2015-11), p. v34.5-v35
    Type of Medium: Online Resource
    ISSN: 1522-8517 , 1523-5866
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2015
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  • 2
    In: Molecular Cancer Research, American Association for Cancer Research (AACR), Vol. 14, No. 8 ( 2016-08-01), p. 707-719
    Abstract: In estrogen receptor (ER)–negative breast cancer, high tumor glucocorticoid receptor (GR) expression has been associated with a relatively poor outcome. In contrast, using a meta-analysis of several genomic datasets, here we find that tumor GR mRNA expression is associated with improved ER+ relapse-free survival (RFS; independently of progesterone receptor expression). To understand the mechanism by which GR expression is associated with a better ER+ breast cancer outcome, the global effect of GR-mediated transcriptional activation in ER+ breast cancer cells was studied. Analysis of GR chromatin immunoprecipitation followed by high-throughput sequencing in ER+/GR+ MCF-7 cells revealed that upon coactivation of GR and ER, GR chromatin association became enriched at proximal promoter regions. Furthermore, following ER activation, increased GR chromatin association was observed at ER, FOXO, and AP1 response elements. In addition, ER associated with GR response elements, suggesting that ER and GR interact in a complex. Coactivation of GR and ER resulted in increased expression (relative to ER activation alone) of transcripts that encode proteins promoting cellular differentiation (e.g., KDM4B, VDR) and inhibiting the Wnt signaling pathway (IGFBP4). Finally, expression of these individual prodifferentiation genes was associated with significantly improved RFS in ER+ breast cancer patients. Together, these data suggest that the coexpression and subsequent activity of tumor cell GR and ER contribute to the less aggressive natural history of early-stage breast cancer by coordinating the altered expression of genes favoring differentiation. Implications: The interaction between ER and GR activity highlights the importance of context-dependent nuclear receptor function in cancer. Mol Cancer Res; 14(8); 707–19. ©2016 AACR.
    Type of Medium: Online Resource
    ISSN: 1541-7786 , 1557-3125
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2016
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  • 3
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 72, No. 8_Supplement ( 2012-04-15), p. 1409-1409
    Abstract: Orthotopic mouse models of human disease are absolutely essential for validating therapeutic targets and assessing efficacy of emerging therapies. Early detection of in vivo tumor growth is a critical benchmark in humanized mouse models, for it reveals tumor take at a time of reasonable tumor burden. To develop an optimal mouse model of human metastatic breast cancer, we utilized TMD231 breast cancer cells (derived from MDA-MB-231) that have a high propensity to metastasize to the lung. TMD231 tumor take and improved kinetics of tumor growth and metastatic foci formation appeared more consistently and at a faster rate in NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) versus NOD/SCID mice. For in vivo detection of small tumor burden at study initiation and metastatic lesions over time, TMD231 breast cancer cells were transduced with lentiviral vectors that express either mCherry or E2-Crimson fluorescent proteins (FPs). Flow cytometric analysis indicated transduction efficiencies for the lentiviral vectors expressing mCherry and E2-Crimson in the TMD231 cells was & gt;99% and & gt;80%, respectively. TMD231 cells expressing either the mCherry or E2-Crimson FPs were implanted into NSG mice and were imaged weekly using the Optix MX3 (ART Technologies, Montreal, Canada) to assess to what degree fluorescently tagged cells could be longitudinally and non-invasively visualized at primary and secondary sites within the NSG mice. While photon emission from TMD231 tumors expressing mCherry could be detected, levels were low and did not correlate with tumor growth overtime. In contrast, high levels of photon emission were detected using TMD231-E2-Crimson cells. Non-palpable tumors expressing the E2-Crimson FP could be detected in NSG mice as early as 7 days post-implant into the mammary fat pad. Additionally, metastatic foci in the lung were detected via optical imaging as early as 2-3 weeks post-implant. Following imaging analysis, TMD231-E2-Crimson tumor size correlated to increased fluorescent intensity, tumor depth, and fluorescent concentration longitudinally. In vitro imaging analysis of TMD231-E2-Crimson cells confirmed the in vivo imaging results showing a linear relationship between cell number and fluorescent intensity. The discrepancy in the in vivo detection of mCherry versus E2-Crimson proteins is most likely due to the greater brightness of E2-Crimson, as well as the closer alignment of the excitation/emission spectra of E2-Crimson with the laser/filter set of the Optix MX3 imaging system. To confirm optical imaging results, H & E staining indicated the presence of numerous foci in the lungs post-mortem. The optimized orthotopic model described here is being used to investigate mechanisms of metastasis as well as novel therapeutic strategies in metastatic breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1409. doi:1538-7445.AM2012-1409
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2012
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  • 4
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    Online Resource
    American Association for Cancer Research (AACR) ; 2013
    In:  Cancer Research Vol. 73, No. 8_Supplement ( 2013-04-15), p. 4639-4639
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 73, No. 8_Supplement ( 2013-04-15), p. 4639-4639
    Abstract: Metastatic breast cancer is highly refractive to current treatment strategies and new multi-targeted treatments need to be elucidated. In metastatic disease, blocking key protein-protein interactions with the murine double minute 2 (MDM2) could be beneficial for developing new treatment modalities since this signaling pathway is a critical regulatory point in cancer progression. Inhibition of protein binding to the hydrophobic pocket of MDM2 by Nutlin-3a can result in increases in pro-apoptotic proteins such as p53, p73, and E2F1. Since the DNA damaging agent carboplatin is being studied in clinical trials of triple-negative breast cancers (TNBCs), our objective was to evaluate the effects of carboplatin and Nutlin-3a in combination in TNBC cancer in a mutant p53 background. We utilized TMD231 breast cancer cells derived from the MDA-MB-231 human TNBC cancer line. TMD231 cells are highly metastatic in vivo and readily metastasize from the primary tumor to the lung. Combination studies were performed using different ratios of carboplatin to Nutlin-3a to evaluate the range of carboplatin-mediated DNA damage required to obtain synergism with inhibition of MDM2 function. A fixed ratio of 1:1 carboplatin:Nutlin-3a was strongly synergistic with a combination index of & lt; 0.5. In cell proliferation assays there was increased sensitivity to the drugs when given together and a significant reduction in number of colonies in colony formation assays. In order to understand how DNA damage and inhibition of MDM2 signaling leads to enhanced cell death, dose-response and time-course studies were performed. Western analyses demonstrated increases in E2F1 when treated with carboplatin and Nutlin-3a alone, and a time-dependent increase in E2F1 following combination treatment at a fixed 1:1 ratio. Following DNA damage, E2F1 has been shown to promote the transcription of pro-apoptotic genes and repression of survival genes once E2F1 is released from interaction with Rb. Here, E2F1 levels correlated with Rb phosphorylation at serine 780, a phosphorylation site which inhibits binding of E2F1 to Rb. The E3 ubiquitin ligase Itch, which ubiquitinates p73, increased following treatment with carboplatin alone or in combination with Nutlin-3a in a dose- and time-dependent manner but appeared not to regulate p73 since p73 levels remained unchanged following combination treatment. Studies are ongoing to further understand how modulation of DNA damage by blockade of MDM2 results in a robust synergistic response and how upregulation of E2F1 affects downstream targets. The impact of the combination of DNA damaging carboplatin and MDM2 blockade by Nutlin-3a on DNA repair kinetics is currently being evaluated in an orthotopic TMD231 model. These studies will lead to a better understanding of how to potentiate DNA damage and may lead to new clinical therapies in the future. Citation Format: Eva Tonsing-Carter, Harlan E. Shannon, Barbara J. Bailey, Lindsey D. Mayo, Karen E. Pollok. Blockade of MDM2-mediated signaling in context of DNA damage increases E2F1 expression and enhances cell death in triple-negative breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4639. doi:10.1158/1538-7445.AM2013-4639
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2013
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  • 5
    In: Clinical Cancer Research, American Association for Cancer Research (AACR), Vol. 24, No. 14 ( 2018-07-15), p. 3433-3446
    Abstract: Purpose: Although high glucocorticoid receptor (GR) expression in early-stage estrogen receptor (ER)-negative breast cancer is associated with shortened relapse-free survival (RFS), how associated GR transcriptional activity contributes to aggressive breast cancer behavior is not well understood. Using potent GR antagonists and primary tumor gene expression data, we sought to identify a tumor-relevant gene signature based on GR activity that would be more predictive than GR expression alone. Experimental Design: Global gene expression and GR ChIP-sequencing were performed to identify GR-regulated genes inhibited by two chemically distinct GR antagonists, mifepristone and CORT108297. Differentially expressed genes from MDA-MB-231 cells were cross-evaluated with significantly expressed genes in GR-high versus GR-low ER-negative primary breast cancers. The resulting subset of GR-targeted genes was analyzed in two independent ER-negative breast cancer cohorts to derive and then validate the GR activity signature (GRsig). Results: Gene expression pathway analysis of glucocorticoid-regulated genes (inhibited by GR antagonism) revealed cell survival and invasion functions. GR ChIP-seq analysis demonstrated that GR antagonists decreased GR chromatin association for a subset of genes. A GRsig that comprised n = 74 GR activation-associated genes (also reversed by GR antagonists) was derived from an adjuvant chemotherapy-treated Discovery cohort and found to predict probability of relapse in a separate Validation cohort (HR = 1.9; P = 0.012). Conclusions: The GRsig discovered herein identifies high-risk ER-negative/GR-positive breast cancers most likely to relapse despite administration of adjuvant chemotherapy. Because GR antagonism can reverse expression of these genes, we propose that addition of a GR antagonist to chemotherapy may improve outcome for these high-risk patients. Clin Cancer Res; 24(14); 3433–46. ©2018 AACR.
    Type of Medium: Online Resource
    ISSN: 1078-0432 , 1557-3265
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2018
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  • 6
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 79, No. 13_Supplement ( 2019-07-01), p. 4341-4341
    Abstract: The mission of the National Cancer Institute’s (NCI) Office of Cancer Genomics (OCG) is to advance the molecular understanding of cancers in order to improve clinical outcomes through precision medicine. Although vast amounts of genomic data are available for many types of cancers, identifying genetic alterations in rare and pediatric cancers is still a challenge. Efficient bioinformatics tools to analyze, manage, store, and access data are also necessary for the research community. To develop effective and targeted treatments, clinically accurate genotypic and phenotypic research models are much needed. OCG’s programs focus on addressing these challenges through multidisciplinary, collaborative research efforts. The four initiatives of OCG support research on structural, functional, and translational genomics, as well as development of next-generation cancer models. The Cancer Genome Characterization Initiative (CGCI) and the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) programs use transcriptomic, genomic, and epigenomic approaches to examine genetic alterations between various tumors and matched normal tissues. CGCI projects focus on HIV-associated and rare cancers such as Burkitt lymphoma, while TARGET focuses on high-risk childhood cancers. The goals of these programs are to attain insights into key mutations that drive tumors and genetic abnormalities specific to cancer subtypes and to develop effective and less toxic therapies for patients. CGCI and TARGET data are available to the research community through data matrices on the OCG website. OCG also recently launched the Pediatric Genomic Data Inventory (PGDI) as a new resource for investigators to access molecular characterization data.The Cancer Target Discovery and Development (CTD2) Network advances cancer research by bridging the knowledge gap between cancer genomics and precision oncology. The Network aims to understand the cancer metastasis, tumor heterogeneity, and drug resistance to develop optimal combinations of small-molecules or immunotherapy with small molecules. As a community resource program, the CTD2 Network develops and provides access to data, tools, methods, and reagents through the Data Portal and the Dashboard. The Human Cancer Models Initiative (HCMI) is an international consortium that is generating novel human tumor-derived culture models from a wide variety of cancer types including rare and understudied cancers. The models, together with related clinical and genomic data, will be available as a resource to the world-wide research community. OCG’s policies on data usage, as well as guides to accessing data, are explained on the OCG website (https://ocg.cancer.gov/). Researchers, potential collaborators, and interested members of the public are encouraged to visit the OCG webpages or contact OCG at ocg@mail.nih.gov. Citation Format: Cindy W. Kyi, Pamela C. Birriel, Tanja M. Davidsen, Martin L. Ferguson, Patee Gesuwan, Nicholas B. Griner, Yiwen He, Subhashini Jagu, Eva Tonsing-Carter, Daniela S. Gerhard. NCI Office of Cancer Genomics: Promoting multidisciplinary research to translate findings into the clinic and advance precision oncology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4341.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2019
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  • 7
    In: Breast Cancer Research, Springer Science and Business Media LLC, Vol. 21, No. 1 ( 2019-12)
    Type of Medium: Online Resource
    ISSN: 1465-542X
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2019
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  • 8
    In: Molecular Cancer Therapeutics, American Association for Cancer Research (AACR), Vol. 14, No. 12 ( 2015-12-01), p. 2850-2863
    Abstract: Triple-negative breast cancers (TNBC) are typically resistant to treatment, and strategies that build upon frontline therapy are needed. Targeting the murine double minute 2 (Mdm2) protein is an attractive approach, as Mdm2 levels are elevated in many therapy-refractive breast cancers. The Mdm2 protein–protein interaction inhibitor Nutlin-3a blocks the binding of Mdm2 to key signaling molecules such as p53 and p73α and can result in activation of cell death signaling pathways. In the present study, the therapeutic potential of carboplatin and Nutlin-3a to treat TNBC was investigated, as carboplatin is under evaluation in clinical trials for TNBC. In mutant p53 TMD231 TNBC cells, carboplatin and Nutlin-3a led to increased Mdm2 and was strongly synergistic in promoting cell death in vitro. Furthermore, sensitivity of TNBC cells to combination treatment was dependent on p73α. Following combination treatment, γH2AX increased and Mdm2 localized to a larger degree to chromatin compared with single-agent treatment, consistent with previous observations that Mdm2 binds to the Mre11/Rad50/Nbs1 complex associated with DNA and inhibits the DNA damage response. In vivo efficacy studies were conducted in the TMD231 orthotopic mammary fat pad model in NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. Using an intermittent dosing schedule of combined carboplatin and Nutlin-3a, there was a significant reduction in primary tumor growth and lung metastases compared with vehicle and single-agent treatments. In addition, there was minimal toxicity to the bone marrow and normal tissues. These studies demonstrate that Mdm2 holds promise as a therapeutic target in combination with conventional therapy and may lead to new clinical therapies for TNBC. Mol Cancer Ther; 14(12); 2850–63. ©2015 AACR.
    Type of Medium: Online Resource
    ISSN: 1535-7163 , 1538-8514
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2015
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  • 9
    Online Resource
    Online Resource
    American Association for Cancer Research (AACR) ; 2014
    In:  Cancer Research Vol. 74, No. 19_Supplement ( 2014-10-01), p. 1680-1680
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 74, No. 19_Supplement ( 2014-10-01), p. 1680-1680
    Abstract: Metastatic breast cancer is highly refractive to current treatment strategies, and new multi-targeted treatments need to be elucidated. In metastatic disease, inhibiting key protein-protein interactions with the murine double minute 2 (MDM2) could be beneficial for developing new treatment modalities since this signaling pathway is a critical regulatory point in cancer progression. Inhibition of protein binding to the hydrophobic pocket of MDM2 by Nutlin-3a can activate pro-apoptotic proteins such p73 and E2F1 as well as decrease pro-angiogenic Hif-1α. Since the DNA damaging agent carboplatin is currently being studied in clinical trials of triple-negative breast cancers (TNBCs), our objective was to evaluate the effects of carboplatin and Nutlin-3a in combination in TNBC in a mutant p53 background. Using a TNBC cell line TMD231 derived from the MDA-MB-231 human breast cancer cell line, we performed combination studies using different ratios of carboplatin to Nutlin-3a in vitro to evaluate the range of carboplatin-mediated DNA damage required to obtain synergism with inhibition of MDM2 function. A fixed ratio of 1:1 carboplatin:Nutlin-3a was strongly synergistic with a combination index of & lt;0.5. In cell proliferation assays there was increased sensitivity to the drugs when given in combination (p & lt;0.05). TMD231 cells implanted into the mammary fat pad of NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice showed enhanced tumor growth, and metastasis was evident in the lungs. Dose-finding studies were performed to determine an optimal carboplatin dosing schema. NSG mice were randomized based on fluorescent imaging of E2-crimson expressing TMD231 cells allowing for a sensitive measurement of early tumor burden. Following Nutlin-3a and carboplatin combination treatment in vivo, there was a statistically significant reduction in tumor volume and lung metastases compared to vehicle and single drug treated mice (p & lt;0.001). Following Kaplan-Meier analysis, the combination treated mice had a significant increase in survival, (54.3±1.5 days) compared to the vehicle (39.3±0.6 days) and each single drug (Nutlin-3a: 39±1 and carboplatin: 47.5±1.8 days) (p & lt;0.001). While there was a decrease in bone-marrow cellularity, this did not lead to bone-marrow aplasia, and body weights recovered to normal levels within 7 days post-treatment. Pharmacodynamic studies are ongoing to further understand at the molecular level how the DNA damage response and repair is modulated by MDM2 resulting in a robust synergistic response. These studies will lead to a better understanding of how to potentiate DNA damage and may lead to new clinical therapies in the future for metastatic breast cancer. Citation Format: Eva Tonsing-Carter, Harlan E. Shannon, Barbara J. Bailey, Anthony L. Sinn, Kacie M. Peterman, Lindsey D. Mayo, Karen E. Pollok. Modulation of MDM2 in context of DNA damage enhances cell death in a metastatic breast-to-lung xenograft model. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1680. doi:10.1158/1538-7445.AM2014-1680
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2014
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  • 10
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 75, No. 15_Supplement ( 2015-08-01), p. 5196-5196
    Abstract: Breast cancer will affect 1 in 8 women in their lifetime. Approximately 20-30% of all breast cancer cases become metastatic. This study was designed to evaluate the effect of tumor met knockdown with individual and combination drug therapy. In this mouse model, with 3 per group, we used the following chemotherapeutic agents: a vehicle control, carboplatin (20mg/kg i.p.), Nutlin-3a (200mg/kg p.o.) and a combination group with the same doses. Nutlin-3 is a small-molecule inhibitor that binds to the p53-binding pocket of MDM2, and activates the p53 pathway. This has been shown to be effective in mouse models of cancer that have a wild-type p53, including prostate, leukemia and multiple myeloma. The lungs were removed and examined for metastatic lung lesions, fixed in 10% neutral buffered formalin (NBF), processed into a paraffin block, microtomed, and slides were stained with H & E. Ki67 immunostain was performed to clearly define the met lesions and the whole slide was imaged with an Aperio Digital Imaging System. The Image Scope positive pixel algorithm was used to quantitate the met lesions in the lungs. The data revealed upon drug treatment the number of mets in the lungs were unchanged in the carboplatin and nutlin-3a individual treatment groups compared to the vehicle group. The lungs in the combination group had the lowest number of tumors with both visual and quantitative image analysis compared to the vehicle control group. The Aperio automated positive pixel count for Ki67 immunostain was consistent with the H & E pathology read. The values decreased with combination treatment (control vs. combination 20mg/kg carboplatin, 200mg/kg nutlin 3a) and were 6% and 1.7% respectively. It was concluded that mice treated with carboplatin and Nutlin-3a combination therapy had fewer metastatic breast cancer mets in the lungs. The individual drug doses were low compared to previous studies and did not have an effect as was expected. This finding suggests that the combination of a traditional chemotherapy with a small protein inhibitor could inhibit the spread of metastatic breast cancer. Citation Format: Margaret Strack, Eva Tonsing-Carter, Tony Sinn, Tiaishia Spragin, Kacie Peterson, Barbara Bailey, Karen Pollok, George Sandusky. Histologic and whole slide quantitative image analysis of lung metastases in a mouse breast cancer model using carboplatin and Nutlin 3A alone and in combination. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5196. doi:10.1158/1538-7445.AM2015-5196
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2015
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