bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2706: Crizotinib, a selective c-Met inhibitor, prevents metastasis to the liver and lungs in a primary uveal melanoma mouse model.
Authors: Surriga Oliver , Vinagolu Rajasekhar , Ambrosini Grazia , Schwartz Gary
Published: August 2014

Uveal melanoma is the most common primary intraocular malignant tumor in adults. About half of patients with uveal melanoma will develop metastatic disease to the liver and the lung. These tumors are characterized by mutations in G-proteins (GNAQ and GNA11) and also the over-expression of c-Met. Crizotinib is a selective inhibitor for c-Met as well as anaplastic lymphoma kinase (ALK). We found that crizotinib inhibited cell proliferation of uveal melanoma cell lines in a dose dependent manner with an IC50 in the range of 0.75 μM to 2 μM. Phospho-c-Met was inhibited by crizotinib starting at 10 nM with inhibition of downstream signaling including p-ERK1/2, p-AKT and p-STAT. In our xenograft model there was only a modest inhibition of tumor growth after treatment with doses of ≥ 50 mg/kg crizotinib. As crizotinib also prevented cell migration regardless of G-protein status, we established a primary metastatic uveal mouse model in which we created a GFP-luciferase expressing uveal melanoma cell line that developed liver and lung metastases 6 weeks after primary implantation into the eye. At a dose of 50 mg/kg daily, starting 4 weeks after primary implantation, there was a dramatic reduction in the liver and lungs metastases when compared to untreated controls. These results indicate that the inhibition of c-Met alone may be sufficient to prevent metastasis of uveal melanoma and suggest there may be a potential for using crizotinib as an adjuvant therapy in uveal melanoma.

Citation Format: Oliver Surriga, Rajasekhar Vinagolu, Grazia Ambrosini, Gary Schwartz. Crizotinib, a selective c-Met inhibitor, prevents metastasis to the liver and lungs in a primary uveal melanoma mouse model. [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 2706. doi:10.1158/1538-7445.AM2013-2706
Abstract 2708: The TWEAK-FN14 pathway promotes prostate cancer bone metastasis through the NFκB pathway.
Authors: Yin Juan Juan , Liu Yen-Nien , Barrett Ben , Fang Lei , Lake Ross , Casey Orla , Tillman Heather , Ward Yvona , Kelly Kathleen
Published: August 2014

Advanced prostate cancer is highly associated with castration resistance and the development of bone metastases. Understanding the mechanisms contributing to prostate cancer bone metastasis is needed in order to develop novel therapeutic agents. NFκB signaling has been implicated in prostate cancer progression. The Tweak-Fn14 axis, which activates both canonical and non-canonical NFκB pathways, has been shown to be up-regulated in many cancer types. However, the role of the Tweak-Fn14 signaling in prostate cancer progression has not been investigated. In this study, we show that oncogenic Ras transformation of the AR negative, DU145 cell line led to the acquisition of bone metastatic capability and associated increased expression of TWEAK and FN14. Knocking down FN14 using shRNAs, as well as blocking the NFκB pathway using an IKBα super repressor, in DU145/Rasb1 cells, significantly inhibited bone metastasis and improved survival. This inhibitory effect of FN14shRNA was fully rescued by activating the NFκB canonical pathway, but not the NFκB non-canonical pathway. FN14 expression is also high in the bone metastatic, AR negative prostate cancer PC3 cell line. Similarly, knocking-down FN14 in PC3 cells inhibited bone metastatic capacity following intracardiac inoculation in a xenograft model. TWEAK and FN14 are relatively low or undetectable in AR positive prostate cancer cell lines including Lncap and 22RV1. AR binding was detected in both TWEAK and FN14 promoters as determined by chromatin immunoprecipitation analysis in Lncap. Reporter assays demonstrated that AR binding inhibited FN14 and TWEAK transcription. The above experimental models were further supported by analyses of publically available expression data sets for clinical prostate cancer samples that showed FN14 expression was inversely correlated with AR gene expression signatures. In addition, individual FN14 RNA mean expression was higher in clinical metastasis of castrate (n=11) as compared to non-castrate (n=8) patients. We propose that down-regulation of AR activity under castrate conditions may lead to increased FN14 expression and NFκB activation, which provides a survival benefit for prostate cancer cells associated with disease progression. Therefore, targeting the TWEAK-FN14 pathway in prostate cancer patients provides potential, new preventive and therapeutic approaches.

Citation Format: Juan Juan Yin, Yen-Nien Liu, Ben Barrett, Lei Fang, Ross Lake, Orla Casey, Heather Tillman, Yvona Ward, Kathleen Kelly. The TWEAK-FN14 pathway promotes prostate cancer bone metastasis through the NFκB pathway. [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 2708. doi:10.1158/1538-7445.AM2013-2708
Abstract 2712: Identification of tumor initiating genes RPL39 and MLF2 that mediate lung metastasis through nitric oxide signaling and mesenchymal to epithelial transition.
Authors: Dave Bhuvanesh , Granados Sergio , Mai Junhua , Choi Dong Soon , Gao Ding Cheng , Mitra Sucharita , Shen Haifa , Muthuswamy Senthil , Mittal Vivek , Ferrari Mauro , Chang Jenny
Published: August 2014

Triple negative breast cancer (TNBC) is the most aggressive and lethal form of cancer characterized by lack of estrogen, progesterone and Her2 receptors. It is prevalent in women of African American descent, often present in younger and premenopausal women. It shows high risk of recurrence and frequently metastasizes to lungs and brain, resulting in poor overall prognosis. There is no targeted therapy for TNBC as it does not respond to hormonal therapy and is intrinsically resistance to conventional chemotherapy. It is therefore imperative to understand the mechanism of survival of these cancers and unravel its biological pathways and modes of progression. We have previously described breast cancer stem cells (BCSC) to be intrinsically resistant to treatment which is further confirmed by recent publications by other groups that describe direct functional evidence for the same. Using genomic assays, we traced a BCSC gene signature comprising of 477 genes derived from patient biopsies. On selective shRNA knockdown of these genes we identified RPL39 and MLF2 as the top two candidates that affect BCSC self-renewal. Selective siRNA knockdown of RPL39 and MLF2 in human cancer xenografts, showed reduced tumor volume and lung metastases with a concomitant decrease in BCSC markers. Thus, targeting BCSCs in combination with chemotherapy should eliminate the heterogeneous populations within a tumor. Additionally, next generation RNA-seq confirmed mutations in RPL39 and MLF2 in 50% of lung metastases from breast cancer patients. In vitro and in vivo siRNA knockdown of RPL39 and MLF2 showed decrease in nitric oxide synthase, suggesting that these genes are driven by nitric oxide signaling. In conclusion this study reveals novel tumor initiating genes, RPL39 and MLF2 that target the breast cancer stem cells and also show impact on lung metastasis. Our findings enhance the understanding of treatment resistant breast cancer stem cells, the mutations that cause metastases and also lay foundation for developing new therapies for such cancers with poor prognosis.

Citation Format: Bhuvanesh Dave, Sergio Granados, Junhua Mai, Dong Soon Choi, Ding Cheng Gao, Sucharita Mitra, Haifa Shen, Senthil Muthuswamy, Vivek Mittal, Mauro Ferrari, Jenny Chang. Identification of tumor initiating genes RPL39 and MLF2 that mediate lung metastasis through nitric oxide signaling and mesenchymal to epithelial transition. [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 2712. doi:10.1158/1538-7445.AM2013-2712
Abstract 2727: Sulfiredoxin is required for AOM/DSS induced mouse colon carcinogenesis and invasiveness of human colon cancer cells.
Authors: Wei Qiou , Jiang Hong , Baker Alyson , Dodge Lisa K. , Gerard Matthieu , Young Matthew R. , Toledano Michel B. , Colburn Nancy H.
Published: August 2014

Sulfiredoxin (Srx) is the enzyme that reduces the overoxidized inactive form of peroxiredoxins (Prxs). To study the function of Srx in carcinogenesis in vivo, we tested whether loss of Srx protects mice from cancer development. Srx null mice were generated and colon carcinogenesis was induced by an azoxymethane (AOM) and dextran sulfate sodium (DSS) protocol. Compared to either wildtype or heterozygotes, Srx-/- mice had significantly reduced rates in both tumor multiplicity and volume. Mechanistic studies reveal that loss of Srx did not alter tumor cell proliferation; however, increased apoptosis and decreased inflammatory cell infiltration were obvious in tumors from Srx null mice compared to those from wildtype control. In addition to the AOM/DSS model, examination of Srx expression in human reveals a tissue-specific expression pattern. Srx expression was also demonstrated in tumors from colorectal cancer patients and the levels of expression were associated with patients’ clinic stage. These data provide the first in vivo evidence that loss of Srx renders mice resistance to AOM/DSS-induced colon carcinogenesis, suggesting that Srx has a critical oncogenic role in cancer development, and Srx may be used as a marker for human colon cancer pathogenicity.

Citation Format: Qiou Wei, Hong Jiang, Alyson Baker, Lisa K. Dodge, Matthieu Gerard, Matthew R. Young, Michel B. Toledano, Nancy H. Colburn. Sulfiredoxin is required for AOM/DSS induced mouse colon carcinogenesis and invasiveness of human colon 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 2727. doi:10.1158/1538-7445.AM2013-2727

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2752: Novel inhibitor of HSP90- PU-H71 exhibits significant activity in Ewing sarcoma pre-clinical models.
Authors: Ambati Srikanth R. , Caldas Lopes Eloisi , Mony Ullas , Kosugi Kohji , Moreira Andre , Meyers Paul , Chiosis Gabriela , Moore Malcolm
Published: August 2014

Ewing sarcoma is characterized by multiple deregulated pathways that mediate cell survival and proliferation. Heat shock protein-90 (HSP90) is a critical component of the multi-chaperone complexes that regulate the disposition and activity of a large number of proteins responsible for the assembly and regulation of cell-signaling systems. We tested the efficacy of PU-H71, a novel purine scaffold HSP90 inhibitor developed at Memorial Sloan-Kettering Cancer Center in several Ewing sarcoma cell lines and primary cultures established from patient samples. We investigated its effect on cell proliferation and cell viability on multiple Ewing cell lines, benign epithelial, endothelial and mesenchymal stromal cells and its effect on colony forming assay using CD34 hematopoietic stem cells . We performed cell cycle analysis, clonogenic assay, immunoblot analysis and in vivo experiments in NOD scid gamma (NSG) mice using A673 cell line transduced with GFP luciferase for bioimaging. We also established a metastatic model by calculating the mean fluorescence intensity (MFI) in various organs. Our results show that there is a significant therapeutic window in its activity against Ewing cell lines and benign cells. PU-H71 treatment resulted in arrest in G2 phase and diminished S phase in Ewing cell lines. We also noted that many of the deregulated pathway proteins including IGF1R, AKT, ERK, MYC, hTERT and EWS-FLI1 are stabilized by HSP90 and exposure to PU-H71 resulted in depletion of the above proteins. We also noted an increase in cleaved PARP and proapoptotic proteins and a decrease in antiapoptotic proteins on immunoblot analysis. Based on earlier PK and MTD studies, we treated mice at a dose of 75mg/kg by intraperitoneal injection, three times a week for 4 weeks. Our results indicated that Ewing's sarcoma tumor growth was significantly inhibited and metastatic burden was significantly decreased in the mice injected with PU-H71 compared to the mice injected with vehicle, based on MFI, weight and size of the tumors. Immunohistochemical staining showed that CD31 expression was significantly reduced in PU-H71 treated mice tumors. Our results suggest that PU-H71 has the ability to affect multiple critical oncogenic pathways simultaneously resulting in decreased tumor growth and metastases and has a strong rationale for clinical evaluation in Ewing sarcoma.

Citation Format: Srikanth R. Ambati, Eloisi Caldas Lopes, Ullas Mony, Kohji Kosugi, Andre Moreira, Paul Meyers, Gabriela Chiosis, Malcolm Moore. Novel inhibitor of HSP90- PU-H71 exhibits significant activity in Ewing sarcoma pre-clinical models. [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 2752. doi:10.1158/1538-7445.AM2013-2752
Abstract 2760: Identification of ET-743 analogs with improved selectivity and potency for EWS-FLI1 and Ewing sarcoma cells.
Authors: Harlow Matt , Maloney Nichole , Segars Laura , Woldemichael Girma , Helman Lee J. , Grohar Patrick J.
Published: August 2014

BACKGROUND: Ewing sarcoma is a pediatric malignancy characterized by the fusion of the EWS and FLI1 genes, which creates a constitutively activated transcription factor, EWS/FLI1. The transcriptional program of EWS/FLI1 drives malignant transformation as well as high-risk features such as increased angiogenesis, resistance to apoptosis, and increased metastatic potential. It is well known that perturbing the transcriptional activity of EWS/FLI1 leads to cell death in vitro and should be considered as a possible site of therapeutic intervention in Ewing sarcoma patients. To that end, we identified ET-743 (trabectedin, Yondelis), a natural product, which inhibits EWS/FLI1 activity in vitro. Using ET-743 as a lead compound, we have characterized a number of structural analogs of ET-743 to identify more potent and/or more selective inhibitors of EWS/FLI1 activity.

METHODS: We evaluated the potency of the ET-743 structural analogs using MTS viability assays using Ewing sarcoma, alveolar rhabdomyosarcoma, and osteosarcoma cell lines. The specific inhibition of EWS/FLI1 downstream transcriptional activity was measured using a luciferase-based approach and western blotting. We also evaluated each analog's ability to suppress a gene signature of EWS/FLI1, which included more than 20 genes known to be deregulated by EWS/FLI1 using a novel microfluidic based assay and the Fluidigm platform.

RESULTS: Several analogs proved to be more potent than ET-743 and selective for transcriptional suppression of EWS/FLI1. All of the analogs induced varying degrees of DNA damage and cell death, which were detected by biochemical and metabolic assays. We also show that the analogs sensitize ES cells to treatment with camptothecins to a variable degree based on analog relative to the parent compound.

CONCLUSIONS: In comparison to ET-743, we have shown that its structural analogs confer better pharmacological properties such as increased potency and specificity towards Ewing sarcoma cell lines. We are currently working to further characterize the mechanism of action for these analogs and evaluate the compounds in an in vivo model. The ultimate goal would be to translate these compounds to the clinic and improve patient outcomes with Ewing sarcoma.

Citation Format: Matt Harlow, Nichole Maloney, Laura Segars, Girma Woldemichael, Lee J. Helman, Patrick J. Grohar. Identification of ET-743 analogs with improved selectivity and potency for EWS-FLI1 and Ewing sarcoma 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 2760. doi:10.1158/1538-7445.AM2013-2760
Abstract 2775: Establishment of patient primary gastric cancer xenograft models for test of anticancer agents.
Authors: Zhang Ning , You Shengyi , Li Wenwei , Zhou Wen , Liu Yong , Zhou Rui , He Fang , Liu Changnian
Published: August 2014

Background. Human xenograft tumor models established by inoculating human tumor cell lines into immunodifficient mice have been routinely used for preclinical test of anticancer agents. But tumor cell lines have a relatively low transplantability, which resulted in a limited number of tumor models available for selection of right tumor modes for test of targeted therapeutics. Recently, we have developed a large number of patient primary gastric cancer xenograft models by transplanting human fresh gastric tumor fragments into nude mice, which have been employed for preclinical test of clinically used drugs and novel anticancer agents for their chemosensitivity screening. Mehtods. The fresh gastric tumor samples were collected from local hospitals. The tumor fragments of 2-3 mm were subcutaneously implanted in the flanks of nude mice by trocar needle. Sixteen tumor fragments were grafted into four mice from one patient tumor tissue (passage 0). The clinically used drugs included 5-FU, gemcitabine, and doxorubicin. The histology and gene sequence of the established primary gastric tumor models were analyzed and compared with patients’ original tumors. Results. A total of 110 patient gastric cancer samples were implanted into nude mice; and 30 primary tumor models have been established with a tumor taking rates of 27% for the first passage. The tumor taking rates were higher in the later passages ranged from approximately 70-95%. The therapeutic efficacy of the test anticancer drugs was consistent with their clinical findings. The patients’ primary gastric cancer xenografts from all 5 passages retained a similarity in architecture, histopathological morphology, and genomic mutation status to their patients’ original tumors. Conclusions. The patient primary tumor model system can provide a larger number of models within same tumor histological type for selection of right models for preclinical testing novel agents based on their anticancer mechanism. The primary tumor models retain a similarity in histology and genomic mutation status to their patients’ original tumors. They may predict more relevant clinical response rate and higher correlation with clinical findings than use of traditional xenograft models established from long-term cultured cancer cell lines. Especially, they have advantages for test of target-oriented therapeutics in new drugs discovery and development programs.

Citation Format: Ning Zhang, Shengyi You, Wenwei Li, Wen Zhou, Yong Liu, Rui Zhou, Fang He, Changnian Liu. Establishment of patient primary gastric cancer xenograft models for test of anticancer agents. [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 2775. doi:10.1158/1538-7445.AM2013-2775
Abstract 2792: Characterization of spontaneous in vivo cachexia models in Champions TumorGraft™ models.
Authors: Anderson Nathan , Khor Tin Oo , Feldhaus Andrew , Olson Katie , Latham John , Sidransky David , Bruckheimer Elizabeth M.
Published: August 2014

Cachexia, defined as a loss of adipose tissue and skeletal muscle mass not reversible by nutritional support, is present in the majority of patients with advanced cancer progression. This reduces the patient's quality of life and impacts their ability to function normally and undergo treatments. Reversing cachexia has been problematic due to a lack of quality pre-clinical models. Traditional xenograft models using cell lines are limited due to their inability to accurately replicate the whole body response to cancer. Champions Oncology overcomes this challenge through the use of its innovative Champions TumorGraft™ platform, where primary human tumors are implanted into immunocompromised mice in a manner that preserves the biological properties of the original human tumor. Champions has identified a panel of three TumorGraft models which spontaneously induce cachexia when implanted subcutaneously into nude mice: 1) CTG-0282, a pancreatic cancer model, 2) CTG-0765, a NSCLC model, and 3) CTG-0804, a renal cell carcinoma model. All three models have shown a greater than 90% cachexia rate, as demonstrated by lost body weight, over multiple passages. In addition, loss of body weight correlates with an increase in tumor volume size. Mice that have little or no tumor growth show no loss of body weight. Past studies have indicated a function for pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6) and interferon gamma (IFNγ), in the cachexia process. Champions is currently investigating the roles of these possible tumor-mediated factors in the aforementioned models. Principle among these factors is IL-6, which has been shown to be required in the regulation of cachexia in tumor-bearing mice and is correlated with weight loss and survival in patients. Preliminary results from our studies suggest a correlation between IL-6 levels and spontaneous cachexia in Champions TumorGraft models. In summary, we demonstrate that the Champions TumorGraft platform preserves the biological properties of the original human tumor, including spontaneous cachexia behavior in these models, and is therefore ideal for oncology drug development programs focused on the inhibition of cachexia.

Citation Format: Nathan Anderson, Tin Oo Khor, Andrew Feldhaus, Katie Olson, John Latham, David Sidransky, Elizabeth M. Bruckheimer. Characterization of spontaneous in vivo cachexia models in Champions TumorGraft™ models. [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 2792. doi:10.1158/1538-7445.AM2013-2792
Abstract 2793: Development of an androgen-dependent prostate Champions TumorGraft™ cancer model.
Authors: Khor Tin Oo , Paz Keren , Tiruchinapalli Dhanrajan , Sidransky David , Bruckheimer Elizabeth M.
Published: August 2014

Prostate cancer (PCa)is the second most frequently diagnosed cancer and the sixth leading cause of cancer death in males, with the highest incidence rates recorded in developed countries such as those in North America (Jemal et al., 2011). While the five year-survival rate for PCa patients with localized disease is 100%, it is only 30.6% once the cancer metastasizes (Vishnu et al., 2010). One of the biggest challenges for finding a better treatment for metastatic PCa is the lack of predictable and accurate preclinical models that closely recapitulate different stages of the pathogenesis of human PCa. Champions Oncology focuses on the development of Champions TumorGraft™ models derived from the direct implantation of patient tumors into immunocompromised mice. Compared to traditional cell line-based xenograft models, the patient-derived TumorGraft models maintain stable gene-expression patterns and mutational status and correlate to clinical predictability. To overcome the challenge for PCa, Champions Oncology has utilized its TumorGraft technology to establish and characterize a new androgen-dependent prostate cancer TumorGraft model, CTG-0488. This involved the implantation of a primary human prostate tumor in immunocompromised mice in a manner that preserves the biological properties of the original human tumor. In this preliminary work, the characteristics of Champion TumorGraft™ model CTG-0488 are showcased, including tumor histology, mutational status, and chemosensitivity profiles to several standard of care compounds. Overall the development of an androgen-dependent prostate cancer Champions TumorGraft™ model will significantly aid in the development of oncology compounds focused on androgen-dependent prostate cancer.

Citation Format: Tin Oo Khor, Keren Paz, Dhanrajan Tiruchinapalli, David Sidransky, Elizabeth M. Bruckheimer. Development of an androgen-dependent prostate Champions TumorGraft™ cancer model. [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 2793. doi:10.1158/1538-7445.AM2013-2793
Abstract 2804: Anti-tumor and anti-metastatic efficacy of TCM on prostate cancer in orthotopic model of metastatic cancer.
Authors: Zhang Lei , Wu Chengyu , Zhao Ming , Zhang Yong , Hoffman Robert M.
Published: August 2014

We report here on the efficacy of TCM formula TCM-formula LQ on patient-like orthotopic nude-mouse models of metastatic human cancer. The anti-tumor and anti-metastatic activity of LQ was determined on orthotopic models of human prostate and colon cancer. In the present study, human cancer cell lines PC-3 prostate and HCT-116 colon, labeled with green fluorescent protein (GFP), were used. The cells were initially implanted subcutaneously. Subcutaneous tumors were harvested to obtain tissue fragments which were implanted orthotopically in nude mice. The herbal mixture LQ (gavage, 600 mg/kg/day) significantly inhibited prostate cancer tumor growth as measured by imaging, tumor size and weight in subcutaneous, and orthotopic models. LQ was efficacious against primary and metastatic cancer without weight loss and cachexia. Fifty percent of mice in control group got the ascites, but only 20% had ascites in the LQ group. Survival of tumor-bearing mice was also prolonged by LQ treatment. The results indicate that LQ can have non-toxic efficacy against metastatic prostate cancer in clinically relevant orthotopic mouse models.

Citation Format: Lei Zhang, Chengyu Wu, Ming Zhao, Yong Zhang, Robert M. Hoffman. Anti-tumor and anti-metastatic efficacy of TCM on prostate cancer in orthotopic model of metastatic cancer. [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 2804. doi:10.1158/1538-7445.AM2013-2804
Abstract 2823: Molecular response of the axillary lymph node microenvironment to metastatic colonization.
Authors: Ellsworth Rachel E. , Valente Allyson L. , Kane Jennifer L. , Ellsworth Darrell L. , Shriver Craig D.
Published: August 2014

Breast stroma plays an active role in tumorigenesis, undergoing both phenotypic and molecular changes to facilitate and promote tumor development and growth. The metastatic microenvironment also plays a role in successful colonization; however, the genetic changes in these secondary microenvironments are not well described.To improve understanding of the molecular changes associated with metastatic colonization of a foreign tissue, gene expression data was generated from lymph node tissues from women with at least one lymph node with macrometastases and one lymph node with no detectable metastases. Lymph node tissue was microdissected and hybridized to U133A 2.0 gene expression arrays. Differential expression was detected using Partek® Genomics Suite™6.5 with P<0.001, >2-fold change defining significance. Thirty-four genes were differentially expressed, 23 genes, including EPCAM, KRT19 and MUC1, were expressed at significantly higher levels in colonized lymph nodes and 11 genes, such as CXCL2 and HPGDS, were expressed at significantly higher levels in non-metastatic lymph nodes. Thus, lymph node tissue differs in gene expression between those harboring metastatic tumors and those without metastasis. Genes expressed at higher levels in colonized lymph nodes are involved in tumorigenesis, suggesting that like the breast stromal microenvironment, the metastatic microenvironment undergoes cross-talk with the tumor cells, while genes involved in immune function are down-regulated in colonized lymph nodes, suggesting that suppression of proper immune response may be required for successful metastatic colonization.

Citation Format: Rachel E. Ellsworth, Allyson L. Valente, Jennifer L. Kane, Darrell L. Ellsworth, Craig D. Shriver. Molecular response of the axillary lymph node microenvironment to metastatic colonization. [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 2823. doi:10.1158/1538-7445.AM2013-2823
Abstract 2826: Expression of HBEGF enhances breast carcinoma cell motility, invasion and metastasis.
Authors: Zhou Zhen Ni , Sharma Ved P. , Beaty Brian T. , Kenny Paraic , Wiley H. Stephen , Condeelis John S. , Segall Jeffrey E.
Published: August 2014

Despite advances in screening and prevention, breast cancer is still associated with a high mortality rate that predominantly results from the development of metastatic disease. One of the key steps of the metastatic cascade is invasion. Previous work in our lab has discovered a paracrine loop between tumor cells and macrophages in which tumor cells are capable of stimulating macrophages through the secretion of CSF-1 and the tumor associated macrophages in turn aid in tumor cell invasion by secreting EGF. The goal of our project is to explore how expression of EGFR ligands, specifically HBEGF, by tumor cells affects the paracrine loop, invasion and metastasis. Previous studies have shown that HBEGF is often found to be coexpressed with the EGFR and that HBEGF expression in tumors is correlated with more biologically aggressive disease and poorer patient prognosis. An initial Oncomine cancer gene expression analysis showed that HBEGF expression was correlated with more rapid recurrence of disease in breast cancer upon removal of the primary breast tumor. Based on the published data, we decided to focus our study on this particular ligand. Using MTLn3 rat mammary adenocarcinoma cells and human MDA-MB 231 breast adenocarcimona cells that were transduced to overexpress HBEGF, we have shown that HBEGF expression enhanced tumor cell invasion by increasing tumor cell motility, invadopodium formation and matrix degradation. We have also discovered that expression of HBEGF enhanced metastasis by making tumor cell invasion independent of the paracrine loop. Our studies suggest that for tumors that express HBEGF, therapies that target tumor associated macrophages using CSF-1R inhibitors may not be effective in inhibiting tumor cell invasion. In addition, we have found that HBEGF expression causes an increase in the recruitment of tumor associated macrophages and tumor angiogenesis along with increased expression of VEGF, which suggests that anti-VEGF treatment could be particularly useful for treating patients whose tumors overexpress HBEGF.

Citation Format: Zhen Ni Zhou, Ved P. Sharma, Brian T. Beaty, Paraic Kenny, H. Stephen Wiley, John S. Condeelis, Jeffrey E. Segall. Expression of HBEGF enhances breast carcinoma cell motility, invasion and metastasis. [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 2826. doi:10.1158/1538-7445.AM2013-2826
Abstract 2834: Cancer testis antigens-specific CD4+ T-cell immunity augmented by CTL-epitopes vaccination in cancer patients.
Authors: Tomita Yusuke , Yuno Akira , Yoshitake Yoshihiro , Tsukamoto Hirotake , Irie Atsushi , Senju Satoru , Yoshida Koji , Tsunoda Takuya , Nakamura Yusuke , Shinohara Masanori , Nishimura Yasuharu
Published: August 2014

We identified novel cancer testis antigens (CTAs) including CDCA1, LY6K (URLC10) and KIF20A that are overexpressed in lung cancer, head and neck cancer (HNC), and other various malignancies, but not expressed in many adult normal tissues except for testis and fetal organs by using genome-wide cDNA microarray analyses. We identified CTAs-derived peptides that can activate tumor-reactive and HLA-A2 or -A24-restricted CTLs in both in vitro preclinical and in vivo clinical studies of peptide vaccine. It is well known that IL-2 and IFN-γ produced by Th1 cells have pivotal roles in efficient induction of memory CTLs and efficient infiltration of CTLs into the tumor. We identified these CTAs-derived promiscuous 20∼26-mer long peptides (LPs) that can induce both Th1 cells restricted by several frequent HLA class II and tumor-reactive CTLs restricted by HLA-A2 or -A24. The LPs were naturally processed from CTAs and presented by dendritic cells. In addition, these LPs induced propagation of CTA-specific CTLs by cross presentation in both human in vitro and HLA-class I transgenic mice in vivo. Furthermore, these LPs and Th1-cell clone enhanced induction of CTA-specific CTLs in vitro. Significant frequencies of CTA-specific Th1 cells were detected after short-term in vitro stimulation with LPs in HNC patients vaccinated with CTA-derived CTL-epitope peptides and cancer patients before vaccination but not in healthy donors; responses were augmented by repeated vaccinations. These results show for the first time the presence of novel CTAs-specific Th1 cell responses in cancer patients and suggest the potential utility of these LPs for propagation of CTAs-specific Th1 cells and CTLs.

Citation Format: Yusuke Tomita, Akira Yuno, Yoshihiro Yoshitake, Hirotake Tsukamoto, Atsushi Irie, Satoru Senju, Koji Yoshida, Takuya Tsunoda, Yusuke Nakamura, Masanori Shinohara, Yasuharu Nishimura. Cancer testis antigens-specific CD4+ T-cell immunity augmented by CTL-epitopes vaccination in cancer patients. [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 2834. doi:10.1158/1538-7445.AM2013-2834
Abstract 2853: Monoclonal antibodies derived in neuN mice following successful T cell therapy of spontaneous breast tumors are specific for tumor progenitor cells.
Authors: Wang Li-Xin , Patel Mona , Berk Michael , Plautz Gregory E.
Published: August 2014

There is considerable interest in developing approaches to specifically target tumor progenitor cells. Our previous results demonstrated that adoptive transfer of CD4+ and CD8+ T cells primed with a breast cancer progenitor vaccine inhibited the progression of spontaneous primary tumors in neuN mice. Interestingly, post-treatment serum but not pre-treatment serum was reactive with surface molecules on the vaccine cells. This indicates that the transferred T cells resulted in uptake of tumor antigens and stimulation of host B cells. To characterize the antigenic targets, we generated hybridomas from spleen cells derived from treated mice. Three candidate clones which were strongly reactive with a neu-negative variant tumor line were subjected to two rounds of limiting dilution cloning. The mAb prepared from hybridoma #1 (IgG1) and hybridoma #2 (IgG2b) were each reactive against two independent tumor lines derived from spontaneous breast tumors in neuN mice (C2D1 and MMC5) when these cells were maintained in serum-free culture medium as tumorspheres. There was no reactivity against the same tumor cell lines when cultured as adherent monolayers in serum-containing medium nor was there reactivity against 3T3 adherent or tumorsphere cultures. Our previous analysis of tumorsphere cultures of C2D1 and MMC5 indicated that they express a panel of markers associated with tumor progenitor cells and have markedly enhanced per-cell tumorigenicity. Two of these antibodies have been purified by protein G or Protein A columns and retain binding to C2D1-tumorsphere cells, but not to C2D1 adherent cells. Importantly, these two antibodies can also bind to cultured 4T1-tumorsphere cells, an additional breast cancer cell line arising initially in BALB/c mice. These studies will allow us to identity corresponding antigens and determine expression on a broader array of spontaneous breast carcinomas in neuN mice. The epitopes recognized by these mAbs, which are expressed on progenitor cells from multiple tumor lines, might have importance as an immunotherapy target.

Citation Format: Li-Xin Wang, Mona Patel, Michael Berk, Gregory E. Plautz. Monoclonal antibodies derived in neuN mice following successful T cell therapy of spontaneous breast tumors are specific for tumor progenitor 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 2853. doi:10.1158/1538-7445.AM2013-2853
Abstract 2878: Targeting the CXCL5-CXCR2-CREB signaling axis suppresses growth, survival, and invasion of lung adenocarcinoma.
Authors: Park Hee Sun , Lee Jong Woo , Saintigny Pierre , Kurie Jonathan M. , Herbst Roy S. , Koo Jaseok Peter
Published: August 2014

The role of chemokines and chemokine receptors has been evolving in cancer. Besides their functions in the immune system, they also play a critical role in tumor initiation, promotion and progression. Since the tumor microenvironment play a critical role in cancer development and progression, the immune components of the tumor microenvironment have gained attention in the recent decades for its critical role in tumorigenesis and tumor control. Recent studies have clearly demonstrated the importance of CXCR2 in the development of lung cancer. Our recent studies showed that CXCR2 expression in tumor cells was associated with poor prognosis in non-small cell lung cancer (NSCLC) patients (n=262). Analysis of gene expression profiles from the National Cancer Institute (NCI) Director's Challenge Cohort of in lung adenocarcinoma patients (n=442) revealed that overexpression of CXCR2 ligands and CXCR2 genes was clustered together in approximately 25% of lung adenocarcinoma patients. Expression of CXCR2 and its associated ligand gene cluster in tumor cells was associated with poor prognosis, smoking status, and poor histologic differentiation. Furthermore, CXCL5 was involved as an potent angiogenic factor in NSCLC. The objective of the current study was to establish the role of the CXCL5/CXCR2 axis in tumor growth and invasion and to determine its related molecular mechanisms. In the study, CXCL5 activated the CREB transcription factor via the pERK1/2-p90Rsk-pCREB signaling pathway. In addition, inhibitors of CXCR2 and downstream signal transduction molecules blocked the activation of pERK1/2-pRsk-pCREB signaling molecules. Blockade of the CXCR2 receptor, CXCR2-dependent signal molecules, or CREB resulted in suppression of NSCLC cell growth, survival and invasion. These results suggest that the CXCL5-CXCR2-CREB signaling axis may be a source of potential therapeutic targets in lung adenocarcinoma.

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Citation Format: Hee Sun Park, Jong Woo Lee, Pierre Saintigny, Jonathan M. Kurie, Roy S. Herbst, Jaseok Peter Koo. Targeting the CXCL5-CXCR2-CREB signaling axis suppresses growth, survival, and invasion of lung adenocarcinoma. [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 2878. doi:10.1158/1538-7445.AM2013-2878
Abstract 2881: E-FABP protects host from tumor development.
Authors: Zhang Yuwen , Rao Enyu , Li Bing
Published: August 2014

Epidermal fatty acid-binding protein (E-FABP) is constitutively expressed in many immune cells, including dendritic cells, macrophages and T lymphocytes. However, the role of E-FABP in host immunity against tumor and cancer development is unknown. We hypothesize that E-FABP plays an essential role in host immune responses to ward off tumor challenges. Here we utilize a murine breast cancer model and E-FABP knockout mice to study the function of E-FABP in host immune responses to tumor challenges. It is found that implanted breast cancer cells develops much faster into tumor in E-FABP knockout mice than in wild type ones. It is also discovered that, when compared to wild type mice, F4/80+CD11c+ macrophages in tumor stroma from E-FABP knockout mice show a defective protective immune response to breast cancer cells. Moreover, tumors in E-FABP knockout mice contained much lower numbers of NK cells, CD8+ and CD4+ T cells than those in WT mice. All these results suggest E-FABP protects host from tumor development, implicating a significant role of E-FABP in host immunity against tumor formation.

Citation Format: Yuwen Zhang, Enyu Rao, Bing Li. E-FABP protects host from tumor development. [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 2881. doi:10.1158/1538-7445.AM2013-2881
Abstract 2886: Multidimensional single-cell analysis of BCR signaling reveals proximal activation defect as a hallmark of Chronic Lymphocytic Leukemia B cells.
Authors: Palomba M. Lia , Piersanti Kelly , Ziegler Carly G. K. , Cotari Jesse , Balderas Robert , Malek Sami N. , Seymour Erlene , Zelenetz Andrew D. , van den Brink Marcel R. M. , Altan-Bonnet Gregoire
Published: August 2014

Chronic Lymphocytic Leukemia (CLL) is defined by a perturbed B-cell receptor-mediated signaling machinery. To assess the aberrant behavior of BCR signaling in CLL B cells, we developed an experimental methodology combining immunophenotyping, multiplexed phosphospecific flow cytometry, and multifactorial statistical modeling. Utilizing patterns of signaling network covariance, we modeled the complex nature of BCR signaling using Partial Least Squares Regression (PLSR) to gain new insights into the strongest predictors of discrimination between CLL and normal B cells. We identified a dynamic and variable imbalance between proximal (pSYK, pBTK) and distal (pPLCγ2, pBLNK, ppERK) phosphoresponses. Furthermore, PLSR identified the relationship between upstream tyrosine kinase SYK and its target, PLCγ2, as maximally predictive and sufficient to distinguish CLL from healthy samples, pointing to this juncture in the signaling pathway as key to the pathogenic properties of CLL B cells. Specific BCR pathway signaling signatures that correlate with the disease and its degree of aggressiveness were identified. Single-cell multidimensional analysis of BCR signaling permitted focused study of the variability and heterogeneity of signaling behavior from patient-to-patient, and from cell-to-cell. Heterogeneity in the PLSR response variable within the B cell population is both a characteristic mark of healthy samples and predictive of disease aggressiveness.

Citation Format: M. Lia Palomba, Kelly Piersanti, Carly G. K. Ziegler, Jesse Cotari, Robert Balderas, Sami N. Malek, Erlene Seymour, Andrew D. Zelenetz, Marcel R. M. van den Brink, Gregoire Altan-Bonnet. Multidimensional single-cell analysis of BCR signaling reveals proximal activation defect as a hallmark of Chronic Lymphocytic Leukemia B 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 2886. doi:10.1158/1538-7445.AM2013-2886

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2894: Integrated bioinformatic analysis to elucidate molecular mechanisms associated with reversal of osteosarcoma malignancy induced by CD99.
Authors: Sciandra Marika , Grilli Andrea , Marino Maria Teresa , Manara Maria Cristina , Picci Piero , Scotlandi Katia
Published: August 2014

Introduction: CD99 is a transmembrane glycoprotein which is physiologically involved in several biological processes, such as cell migration, adhesion, and differentiation. High levels of CD99 have been reported in mesenchymal stem cells and osteoblasts. In contrast, the molecule has been shown to be absent in osteosarcoma (OS), in which it acts as a tumour suppressor. The molecular mechanisms responsible for its functions are however still poorly defined. In this study we used bioinformatic studies to integrate gene expression and miRNA data to identify crucial mediators.

Methods: Forced expression of CD99 was stably induced in Saos-2 OS cell line. Microarray analyses of gene or miRNA expression were performed and integrated by using different bioinformatic approaches. TargetScan, Miranda, Diana-microT and PicTar prediction databases were used to identify miRNA targets.

Results: CD99-overexpressing cells displayed a massive down-regulation of gene expression. Only 19 probes were found to be significantly up-regulated in CD99 transfectants compared to parental cells. In contrast, 231 probes were found to be down-regulated, thus suggesting gene modulation by epigenetic mechanisms. We thus examined miRNA profile of these cells. A significant up-regulation of miR-1225-3p, miR-1238, miR-1305, miR-191*, miR-34a, and miR-425* was observed in CD99 overexpressing cells. Enrichment analyses pointed out an involvement of TGFβ signalling. Interestingly, SMAD-2 and -4, which emerged as targets of miR-34a, were among the main down-modulated genes. miR34 mimics confirmed the association between miR34a and SMAD-4. Functional studies are on-going to establish associations with osteosarcoma malignancy.

Conclusions: Besides inhibiting osteosarcoma malignancy, CD99 expression in osteosarcoma cells determines massive gene expression repression, likely due to changes in miRNA profile. Up-regulation of miR-34a together with down-regulation of TGFβ signalling mediators seem to be critical players.

Studies are granted by AIRC 10452 and Ministry Health RF-2009 to KS.

Citation Format: Marika Sciandra, Andrea Grilli, Maria Teresa Marino, Maria Cristina Manara, Piero Picci, Katia Scotlandi. Integrated bioinformatic analysis to elucidate molecular mechanisms associated with reversal of osteosarcoma malignancy induced by CD99. [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 2894. doi:10.1158/1538-7445.AM2013-2894