bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 700: Regulation of Fn14 expression by EGFRvIII-STAT signaling enhances glioblastoma cell invasion and survival
Authors: Roos Alison , Mayo Zachary , Sonnemann Heather , Pineda Michael , Lambert Gil , Dhruv Harshil D. , Winkles Jeffrey A. , Berens Michael E. , Tran Nhan L.
Published: July 2016

Glioblastoma Multiforme (GBM) is the most common malignant brain tumor in adults. Most GBM patients succumb to the disease less than one-year post diagnosis due to the highly invasive nature of the tumor, which prevents complete surgical resection and gives rise to tumor recurrence. The invasive phenotype also confers radio-and chemoresistant properties to the tumor cells; therefore, there is a need to develop new therapeutics that target drivers of GBM invasion. Amplification of EGFR is observed in over 50% of GBM tumors, of which half concurrently overexpress the variant EGFRvIII, and expression of both receptors confers a worse prognosis. EGFR and EGFRvIII cooperate to promote tumor progression and invasion, in part, through activation of the JAK/STAT-signaling pathway. Here we report that GBM cells expressing EGFRvIII show increased expression of a previously established mediator of glioma cell invasion and survival, fibroblast growth factor-inducible 14 (Fn14), at the mRNA and protein level. Treatment with STAT3, STAT5, JAK, or Src inhibitors decreased Fn14 mRNA and protein expression. Finally, knockdown of Fn14 levels in the EGFRvIII-expressing glioma cells decreased both cell survival after temozolomide (TMZ) treatment and cell invasion, which suggests that Fn14, in part, mediates the oncogenic phenotypes conferred by EGFRvIII signaling. Since EGFR inhibitors display limited therapeutic efficacy in GBM patients, we hypothesize that Fn14-targeted therapies could potentially limit invasiveness and chemoresistance in EGFRvIII-dependent GBM tumors.

Citation Format: Alison Roos, Zachary Mayo, Heather Sonnemann, Michael Pineda, Gil Lambert, Harshil D. Dhruv, Jeffrey A. Winkles, Michael E. Berens, Nhan L. Tran. Regulation of Fn14 expression by EGFRvIII-STAT signaling enhances glioblastoma cell invasion and survival. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 700.
Abstract 704: Hyperglycemia stimulates glycolysis and enhances migratory/invasive potential in tongue squamous cell carcinoma
Authors: Wang Anxun , Wang Wei , Zhao Luodan , He Qianting , Sun Jingjing , Liu Zhonghua , Chen Zujian , Zhou Xiaofeng
Published: July 2016

Objective: The aim of this study was to investigate the role of glycolysis (HK2/PKM2) in the development of tongue squamous cell carcinoma (TSCC) and elucidate how hyperglycaemia stimulate HK2 and PKM2 and enhance migration/invasion potential in TSCC.

Methods: A total of 501 patients which diagnosed as TSCC between 1998 and 2004 were enrolled in the retrospective study. The expression of HK2/PKM2 was examined by IHC and the relationship between TSCC and diabetes mellitus (DM) was investigated. We then investigated the role of HK2/PKM2 in the migration and invasion of TSCC and relevant pathways (the SOD2-H2O2 pathway and miR-138 pathway). Finally, we investigated whether hyperglycaemia enhance migratory/invasive potential of TSCC through stimulating glycolysis.

Results: The prevalent rate of TSCC patients with diabetes was 13.97%, which was significantly higher than the incidence rate of DM (11.6%) in China. Cox analysis revealed that the HR for patients with DM vs without DM was 1.32. IHC results confirmed that the up-regulation of PKM2/HK2 expression was correlated with tumor stage, clinical stages, diabetes, lymph node metastasis and associated with reduced overall survival. PKM2/HK2/AKT1 overexpression increased cell migration and invasion, SOD2 activities and intracellular H2O2. The siRNA-based PKM2/HK2 knockdown inhibited cell migration and invasion, reduced activities of SOD2 and intracellular H2O2, and also inhibited tumor growth and lung metastasis in vivo. Luciferase assays revealed that PKM2/AKT1 was directly targeted by miR-138 and miR-138 can reduce the expression of the PKM2/AKT1 gene. High glucose enhanced the migration/invasion ability, miR-138/PKM2/HK2 expression and increased SOD2 activities and intracellular H2O2.

Conclusions: Our results confirm that DM may be a risk factor in the development of TSCC. PKM2/HK2 plays an important role in the progression of TSCC, and may serve as a biomarker or therapeutic target for patients with TSCC. High glucose stimulates glycolysis (PKM2/HK2) and enhances migration/invasion potential in TSCC.

Citation Format: Anxun Wang, Wei Wang, Luodan Zhao, Qianting He, Jingjing Sun, Zhonghua Liu, Zujian Chen, Xiaofeng Zhou. Hyperglycemia stimulates glycolysis and enhances migratory/invasive potential in tongue squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 704.
Abstract 713: Determining the role of tumor-infiltrating B cells in NSCLC
Authors: Centuori Sara M. , Kim Samuel , Gomes Cecil , Putnam Charles , Mount David , Garland Linda , Larsen Brandon , Martinez Jesse D.
Published: July 2016

Recent studies in human non-small cell lung cancer (NSCLC) have shown that upregulation of B cell-associated genes strongly correlate with early stage patient survival. After analyzing a gene expression database of lung tumors we showed that CD79A, a pan-B cell marker, had the strongest predictive value, suggesting that B cells are playing a crucial role in NSCLC immunity. In order to evaluate this we first examined where this genetic signature was originating from. We observed that patients with high levels of B cell-related genes also showed high numbers of CD79A+ B cells intimately associated with the tumor, but not being expressed by tumor cells themselves. A closer look at tumor-infiltrating B cells (TIL-B cells) by IHC and flow cytometry of fresh patient tumor samples has confirmed their presence and allowed us to begin elucidating their phenotype. Interestingly, we have found that not all early stage patients have detectable levels of TIL-B cells, but in those that do, TIL-B cells represent a marked proportion of the lymphocytic infiltration. Further evaluation of T cell populations in the same samples indicate that T cell numbers remain relatively consistent but that the numbers of B-cells, especially CD79A+ B-cells, does vary from patient to patient. These data indicate that CD79A+ TIL-B cells are contributing to the generation of an efficient immune response in some but not all patients, and are greatly influencing disease survivability. This information may allow us to stratify patients into low and high risk groups based on the presence or absence of TIL-B cells, respectively.

Citation Format: Sara M. Centuori, Samuel Kim, Cecil Gomes, Charles Putnam, David Mount, Linda Garland, Brandon Larsen, Jesse D. Martinez. Determining the role of tumor-infiltrating B cells in NSCLC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 713.
Abstract 715: Adipose tissues derived exosomal microRNAs and their variants in ovarian cancer progression
Authors: Au Yeung Chi Lam , Tsuruga Tetsushi , Co Ngai Na , Yeung Tsz-Lun , Leung Cecilia S. , Wong Kwong K. , Mok Samuel C.
Published: July 2016

Most ovarian cancers are diagnosed at an advanced stage when the tumor is widely metastatic. The 5-year survival drops to 50% for the cancer cases that spread beyond the pelvis to the omentum. However, the mechanisms underlying the effect of omental adipose tissue on ovarian cancer progression are poorly understood. Recent studies showed that exosomes also contain non-coding RNAs such as microRNAs (miRNAs). Thus, we hypothesize that the transfer of microRNAs and their variants from ovarian cancer-associated omental adipose tissues to ovarian cancer cells via exosomes may contribute to the nearby microenvironment for ovarian cancer metastasis and cancer progression.

Ion Torrent next generation sequencing was performed on miRNAs isolated and enriched from exosomes and cell lysates of ovarian cancer cell lines (OVCA), the epithelial component of microdissected omental ovarian cancer tissues (CT), normal omental adipose tissues (OMN) and ovarian cancer-associated omental adipose tissues (OMT). By integrating the miRNA expression profiles, 65 miRNAs were expressed at significant higher levels in OMT-derived exosomes compared with those in OMN-derived exosomes and OVCA-derived exosomes. A set of miRNAs (miR-32a, miR-221 and miR320a), which had been implicated in controlling cell growth and chemoresistance, was identified. Also, the Ion Torrent results were validated and exosomal transfer of OMT-derived miRNAs was confirmed in vitro. The exosomal communication between adipose tissues and ovarian cancer cells in the omental tumor microenvironment is verified. The transferable miRNAs and their variants may remain functional in the recipient ovarian cancer cells and confer more aggressive phenotypes in these cells.

Citation Format: Chi Lam Au Yeung, Tetsushi Tsuruga, Ngai Na Co, Tsz-Lun Yeung, Cecilia S. Leung, Kwong K. Wong, Samuel C. Mok. Adipose tissues derived exosomal microRNAs and their variants in ovarian cancer progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 715.
Abstract 718: Macrophage PI3Kgamma signaling promotes cancer immune suppression
Authors: Kaneda Megan M. , Varner Judith A.
Published: July 2016

Innate immune cells can switch from a pro-inflammatory state that defends against pathogens and to an anti-inflammatory state that repairs damaged tissue. Pro-inflammatory myeloid cells activate cytotoxic T cells to eliminate pathogens, but anti-inflammatory myeloid cells inhibit T cell mediated immunity, stimulate angiogenesis, and stimulate fibrosis, all of which promote tumor progression. The predominant isoform of PI(3)Kinase in myeloid cells, PI3Kgamma, controls the switch between immune stimulation and immune suppression during inflammation and cancer. Myeloid cell PI3Kgamma and its effectors Akt1, mTor, and S6Kinase stimulate C/EBPbeta-dependent expression of immunosuppressive factors, including Arginase and TGFbeta, but suppress NFkappaB-dependent expression of pro-inflammatory cytokines such as IL12 and IFNgamma, thereby inducing tumor immune suppression and consequent tumor growth and metastasis.

In contrast, inhibition of PI3Kgamma activity by genetic or pharmacological means suppresses C/EPBbeta-mediated transcription of anti-inflammatory factors and stimulates NFkappaB-mediated transcription of the pro-inflammatory cytokines IL12 and IFNgamma, thereby restoring CD8+ T cell-dependent cytotoxicity that inhibits tumor growth and metastasis. Therefore, inhibitory targeting of PI3Kgamma indirectly restores cytotoxic T cell immune responses that elicit tumor suppression without unwanted side effects. Our studies highlight the strong therapeutic potential of targeting this kinase to control inflammation and cancer.

This work was supported by grants to JAV from the NIH (5R01CA126820), the Landon Foundation-AACR (12-60-27-VARN), the Lustgarten Foundation for Pancreatic Research and the Whitworth Foundation for Cancer Research.

Citation Format: Megan M. Kaneda, Judith A. Varner. Macrophage PI3Kgamma signaling promotes cancer immune suppression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 718.
Abstract 722: MYB promotes pancreatic tumor associated-desmoplasia by up-regulating Sonic hedgehog and adrenomedullin
Authors: Bhardwaj Arun , Srivastava Sanjeev K. , Tyagi Nikhil , Arora Sumit , Singh Seema , Carter James E. , Singh Ajay P.
Published: July 2016

Pancreatic cancer (PC) stands as the fourth leading cause of cancer-related death in the United States. Unfortunately, even with the emergence of tumor-targeted therapies, the 5-year survival rate of PC continues to remain very dismal (∼ 7.0%). The presence of dense desmoplastic region at the tumor site is a prominent pathological characteristic of PC, and tumor-associated desmoplasia is suggested to limit the therapeutic efficacy by acting as a physical barrier for drug delivery. In addition, it plays pivotal roles in promoting the survival and aggressive phenotypes of tumor cells. Therefore, it is of utmost importance that we identify novel gene targets and associated molecular mechanisms accountable for the development of desmoplasia in pancreatic tumors. Our recent study provided first evidence for a role of MYB, an oncogenic transcription factor, in pancreatic tumor growth and metastasis. Upon histological examination of orthotopic tumor xenografts derived from MYB-overexpressing and -silenced pancreatic tumor cells, we noticed a positive association of MYB expression with the extent of desmoplastic reaction. This finding was further confirmed by staining the tumor xenograft sections with Collagen-I, fibronectin and á-SMA, the well-characterized markers of desmoplasia. In a co-culture experiment, we observed that MYB-overexpressing pancreatic tumor cells promoted greater proliferation of pancreatic stellate cells (PSCs) as compared to the MYB-silenced cells further supporting a role of MYB in desmoplasia. From the mechanistic standpoint, we identified MYB to be novel direct regulator of Sonic hedgehog (SHH) and Adrenomedullin (ADM) expression in PC cells as confirmed by chromatin immunoprecipitation (ChIP) analysis. Treatments of pancreatic tumor (high or low MYB) and stellate cells co-culture with specific inhibitors and/or recombinant SHH and ADM demonstrated that both SHH and ADM cooperatively mediated the effect of MYB on PC and stellate cells via autocrine and/or paracrine mechanisms. Together, our studies suggest that MYB promotes malignant phenotype of PC not only by directly impacting the tumor cells, but also by altering the tumor microenvironment.

Citation Format: Arun Bhardwaj, Sanjeev K. Srivastava, Nikhil Tyagi, Sumit Arora, Seema Singh, James E. Carter, Ajay P. Singh. MYB promotes pancreatic tumor associated-desmoplasia by up-regulating Sonic hedgehog and adrenomedullin. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 722.
Abstract 730: The roles of FGF-2-NCAM/FGFR1 interplay in esophageal squamous cell carcinomas and its microenvironment including tumor-associated macrophages
Authors: TAKASE NOBUHISA , Koma Yuichiro , Arai Noriaki , Kodaira Himiko , Hosono Masayoshi , Ichihara Yumi , Nishio Mari , Shigeoka Manabu , Yokozaki Hiroshi
Published: July 2016

Tumor-associated macrophages (TAMs) have important roles in the angiogenesis, tumor immunosuppression and tumor infiltration of various cancers. Previously, we reported that a large number of infiltrated tumor promoting CD204+ TAMs within esophageal squamous cell carcinomas (ESCCs) contributed to the significant association of clinicopathological malignancy and poor disease free survival. However, the relevant malignant progression mechanisms have not been studied explicitly yet. To elucidate roles of TAMs in ESCCs, we established an in vitro macrophage model using human peripheral blood acquired by healthy volunteer donors. Peripheral blood monocytes were treated with 25 ng/mL recombinant human M-CSF for 6 days to induce macrophage-like differentiation (macrophage-like cells), then exposed to the conditioned media of TE series ESCC cell lines (TECM) for 2 days to induce TAM-like cells. From the cDNA microarray data between macrophages-like cells treated with and without TECM, we interested in upregulated genes. Among highly expressed genes in TAM-like cells, we focused on neural cell adhesion molecule (NCAM) which was a possibility to act on the regulation between tumor and its surrounding stroma. Gene knockdown of NCAM by siRNA in TAM-like cells revealed significant suppression of cell survival and migration via decrease of PI3K/Akt signaling and downregulation of fibroblast growth factor receptor 1 (FGFR1) expression. Interestingly, fibroblast growth factor-2 (FGF-2) derived from ESCCs promotes FGF2/FGFR1 autocrine loop, and activates FGFR1 signaling in TAM-like cells. Furthermore, we analyzed 70 ESCC tissue samples by immunohistochemistry whether the expression levels of FGF-2 were associated with clinicopathological background factors of ESCC patients. Relationships of the expression levels of FGF-2 in human ESCC stromal cells including macrophages had a correlation with the depth of invasion, blood vessel invasion, TNM classification UICC cancer staging and high number of infiltrating CD163 positive and CD204 positive macrophages as a marker for the M2 macrophage phenotype. These findings may indicate novel roles of FGF-2-NCAM/FGFR1 interplay in ESCCs and its microenvironment including TAMs.

Citation Format: NOBUHISA TAKASE, Yuichiro Koma, Noriaki Arai, Himiko Kodaira, Masayoshi Hosono, Yumi Ichihara, Mari Nishio, Manabu Shigeoka, Hiroshi Yokozaki. The roles of FGF-2-NCAM/FGFR1 interplay in esophageal squamous cell carcinomas and its microenvironment including tumor-associated macrophages. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 730.
Abstract 734: Role of TGF-â in myeloid regulation of tumor-induced bone disease
Authors: Buenrostro Denise , Merkel Alyssa , Sterling Julie
Published: July 2016

Bone is one of the most common metastatic sites for breast cancer, where nearly 70% of patients that die from disease have reported skeletal metastases upon autopsy. Specifically in the bone microenvironment, TGF-â has a complex role in the regulation of tumor growth and is critical for the initiation and continuation of bone destruction, an important parameter of tumor-induced disease. Previous publications have demonstrated that TGF-â can affect myeloid cells by increasing their expansion in the tumor microenvironment and modulating host immune surveillance, thus supporting a tumor promoting environment. Therefore, we hypothesize that inhibition of TGF-â will decrease tumor-induced bone disease by reducing myeloid expansion and tumor growth in bone. To address this we injected mice with the TGF-â inhibitory antibody, 1D11, one week prior to tumor inoculation and continuously treated until sacrifice where myeloid expansion and parameters of tumor-induced bone disease were measured. Two mouse models were used to assess the importance of the immune system, an immunodeficient model inoculated with human breast cancer cells (MDA-MB-231) and a syngeneic model using murine breast cancer cells (4T1). Flow cytometry analysis revealed that TGF-â inhibition decreased a triple positive myeloid population (CD11b+F4/80+Gr-1+) in mice sacrificed at day 28 by five percent. This triple positive myeloid cell population has been associated with supporting tumor progression. Surprisingly our flow cytometry analysis demonstrated that myeloid-derived suppressor cells (MDSCs: CD11b+GR-1+) remained unchanged when TGF-â was inhibited. Micro-CT analysis demonstrated that 1D11 treated mice had a 2-fold increase in bone volume and connective density compared to control mice. X-ray analysis demonstrated that control mice had greater than 2-fold osteolytic bone destruction compared to mice treated with the TGF-â inhibitory antibody. Histomorphometric analyses suggests that TGF-â inhibition reduces tumor burden, increases osteoclast cell numbers and modulates macrophage cell populations. Together these results suggest that inhibiting TGF-â prior to tumor inoculation and subsequently after may prevent expansion of a tumor-promoting myeloid population and promote a bone-phenotype that does not support tumor growth in bone. Future studies will focus on investigating specific myeloid factors and assessing whether TGF-â inhibition decreases expression of these pro-tumorigenic factors.

Citation Format: Denise Buenrostro, Alyssa Merkel, Julie Sterling. Role of TGF-â in myeloid regulation of tumor-induced bone disease. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 734.
Abstract 741: Insulin-like growth factor binding protein-3 (IGFBP-3) enhances obesity-related breast tumorigenesis
Authors: Scully Tiffany , Scott Carolyn D. , de Silva Hasanthi C. , Firth Sue M. , Twigg Stephen M. , Pintar John E. , Baxter Robert C.
Published: July 2016

Introduction and Aim: Epidemiological studies have shown an association between obesity and poor breast cancer prognosis. We have reported that IGFBP-3 influences both breast cancer growth and adipocyte maturation. This study investigated the role of endogenous IGFBP-3 on the development of obesity and subsequently on breast tumor growth. Methods: Wild-type (WT) C57BL/6 or IGFBP-3 knock-out (BP3KO) mice were fed either a high-fat diet (HFD) or control chow diet for 15 weeks, then injected with the syngeneic mouse breast cancer cell line E0771 in the 4th left mammary gland. When the largest tumor reached 1000 mm3, tissues including the mammary fat pad were excised, formalin-fixed and paraffin-embedded for immunohistochemical analysis. Results and Conclusion: Compared to WT mice, BP3KO mice showed both significantly reduced weight gain and mammary fat pad mass (contralateral to tumor) in response to HFD (p < 0.0001, 1-way ANOVA), despite similar food intake between WT and BP3KO mice on HFD. E0771 tumor weight and volume were overall increased by HFD independent of IGFBP-3 status (p = 0.051), and decreased in BP3KO mice independently of diet (p < 0.001, 2-way ANOVA). Despite differences in tumor volumes and weights, tumors from WT and BP3KO mice did not show differences in either the number of mitotically active (Ki67+) or apoptotic (cleaved caspase-3+) cells. Tumors from BP3KO mice however, showed greater infiltration of CD3+ T-cells compared to WT mice (p = 0.0299, 2-way ANOVA). WT and BP3KO mice showed similar intra-tumoral mean vessel densities (CD31+ area) but differed in terms of the relationship between vessel density and final tumor volume. Increased vessel density was associated with decreased tumor volume in BP3KO mice (r = -0.62, p = 0.03, Spearman's correlation test) in contrast to the increased vessel density associated with increased tumor volume in wild-type mice (r = 0.63, p = 0.02, Spearman's correlation test). These data suggest that endogenous (circulating and/or stromal) IGFBP-3 (i) has a potentially stimulatory role in the expansion of adipose tissue, (ii) can enhance mammary tumor growth in immune-competent mice, potentially by influencing T-cell infiltration into the tumor and (iii) may influence the relationship between tumor vascularity and growth.

Citation Format: Tiffany Scully, Carolyn D. Scott, Hasanthi C. de Silva, Sue M. Firth, Stephen M. Twigg, John E. Pintar, Robert C. Baxter. Insulin-like growth factor binding protein-3 (IGFBP-3) enhances obesity-related breast tumorigenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 741.
Abstract 742: A novel multidrug metronomic chemotherapy significantly delays tumor growth in mice
Authors: Tagliamonte Maria , Petrizzo Annacarmen , Tornesello Maria Lina , Luciano Antonio , Rea Domenica , Barbieri Antonio , Arra Claudio , Napolitano Maria , Ciliberto Gennaro , Buonaguro Franco M. , Buonaguro Luigi
Published: July 2016

The tumor immunosuppressive microenvironment represents a major obstacle to an effective tumor-specific cellular immune response.

We have previously shown that a novel multi-drug chemotherapy administered in a metronomic fashion was able to increase immune response to peptides. The chemotherapy consisted of a cocktail including taxanes (paclitaxel and docetaxel) and alkylating (cyclophosphamide) agents. The newly designed strategy was shown to be safe, well tolerated and significantly efficacious (Tagliamonte et al., CII 2015).

In the present study, the effect on the immunosuppressive tumor microenvironment by the same multi-drug cocktail was evaluated in a mouse model upon sub-cutaneous ectopic implantation of B16 melanoma cells.

Treated animals showed a remarkable delay in tumor growth and prolonged survival as compared to control group. Such an effect was directly correlated with CD4+ T cell reduction and CD8+ T cell increase. A significant reduction in the percentage of both CD25+FoxP3+ and CD25+CD127low regulatory T cell population was found in the spleens as well as in the tumor lesions. An intrinsic CD8+ T cell response specific to B16 naturally expressed Trp2 TAA was observed. The same metronomic chemotherapy combined to a vaccine based on mutated antigens, was subsequently shown to increase immune response to vaccine resulting in a significant delay of B16 tumor growth as well as animal survival.

The novel multi-drug daily metronomic chemotherapy evaluated in the present study was very effective in counterbalancing the immunosuppressive tumor microenvironment and enhancing vaccine efficacy. Consequently, the intrinsic as well as vaccine-induced anti-tumor T cell immunity could exert its function containing tumor growth.

Overall, the described metronomic chemotherapy may represent a promising adjuvant approach to enhance anti-tumor cellular immunity and amplify the biological effects of therapeutic cancer vaccines.

Citation Format: Maria Tagliamonte, Annacarmen Petrizzo, Maria Lina Tornesello, Antonio Luciano, Domenica Rea, Antonio Barbieri, Claudio Arra, Maria Napolitano, Gennaro Ciliberto, Franco M. Buonaguro, Luigi Buonaguro. A novel multidrug metronomic chemotherapy significantly delays tumor growth in mice. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 742.
Abstract 75: CellMiner, a systems pharmacological web-application for the NCI-60 cancerous cell-lines: Updates, data integration, and translationally relevant results
Authors: Reinhold William C. , Sunshine Margot , Varma Sudir , Doroshow James , Pommier Yves
Published: July 2016

Understanding the influences of molecular alterations on pharmacological responses in the omic sense is at the fore of the effort to make oncology more effective and specific. At present, however, this remains a field in its infancy. The NCI-60 cancerous cell lines provide a premier set of databases for systems molecular pharmacological studies. This is due to the availability of both the robust, high-quality activity profiles generated by the Developmental Therapeutics Program (https://dtp.cancer.gov), and the panoply of molecular and phenotypic information available. The CellMiner web-application (http://discover.nci.nih.gov/cellminer) provides the user access to both of these. Activity data is available for 20,861 compounds, including 159 Food and Drug Administration (FDA)-approved, 85 clinical trial, and 443 known mechanism-of-action drugs. The molecular data includes transcript expression for 25,772 genes, genetic variants for 16,568 genes, transcript expression for 360 microRNAs, protein levels for 94 genes, DNA copy number (from aCGH) for 23,413 genes, and soon DNA methylation levels for 17,559 genes. “Cell line signatures” are provided for each of these data types, facilitating their comparison. It also provides “Pattern comparisons” for any input pattern of interest to 87,419 activity, molecular and phenotypic patterns (For CellMiner 1.7).

Here we present our CellMiner updates, examples of data integration, and translationally relevant results.

Citation Format: William C. Reinhold, Margot Sunshine, Sudir Varma, James Doroshow, Yves Pommier. CellMiner, a systems pharmacological web-application for the NCI-60 cancerous cell-lines: Updates, data integration, and translationally relevant results. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 75.
Abstract 759: Sunitinib has opposite roles to regulate the myeloid-derived suppressor cells in tumors and peripheral blood
Authors: Chen Fang-Hsin , Fu Sheng-Yung , Wang Chun-Chieh , Chiang Chi-Shiun , Hong Ji-Hong
Published: July 2016

Recent studies indicate that sunitinib, a tyrosine kinase inhibitor, could decease accumulation of myeloid-derived suppressor cells (MDSC) in tumors and also in peripheral blood. This study aims to investigate the behavior and dynamic profile of myeloid-derived suppressor cells in tumor tissues and peripheral blood of sunitinib-treated mice bearing prostate adenocarcinoma TRAMP-C1 tumors. To this end, 3×106 TRAMP-C1 tumor cells were inoculated at shank muscle of C57BL6/J mouse and sunitinib was administered by intraperitoneal injection of a daily dose of 20mg/kg into mice bearing 4 mm diameter tumor. Comparing to tumors in untreated group, sunitinib administration slightly delayed tumor growth delay for 2 days, but significantly decreased micro-vascular density and induced chronic hypoxia in tumors, resulting in the specific accumulation of CD11b+Gr-1+ MDSCs at the tumor necrotic region within CA-IX positive chronic hypoxic area. Flow cytometry was used to analyze the change of CD11b+ myeloid cells within tumor tissues and peripheral blood. Results showed that sunitinib treatment decreased the percentage of CD11b+Ly6G-LyC- tumor-associated macrophages (TAMs), but increased the percentage of CD11b+Ly6G-Ly6C+ monocytic MDSCs (M-MDSCs) while had no effect on CD11b+Ly6G+Ly6C+ neutrophilic MDSCs (N-MDSCs) within tumor tissues. In contrast, both the percentage of N-MDSCs and M-MDSC were expanded in peripheral blood of sunitinib-treated mice. Multiplex immunoassay showed significant increase of CCL2, CCL3, CCL5, CXCL5, IL-17a, GM-CSF, G-CSF and VEGF-A in tumor tissues, but only CXCL5, IL-á, IL-6 and G-CSF in the peripheral blood of sunitinib-treated mice were affected. In conclusion, this study shows that sunitinib has anti-vascular effect and could disrupt the recruitment of CD11b+Ly6G-Ly6C- TAMs into tumors. We also propose that sunitinib might exert different effects on tumor microenvironment and peripheral blood.

Citation Format: Fang-Hsin Chen, Sheng-Yung Fu, Chun-Chieh Wang, Chi-Shiun Chiang, Ji-Hong Hong. Sunitinib has opposite roles to regulate the myeloid-derived suppressor cells in tumors and peripheral blood. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 759.
Abstract 761: Specific inhibition of prostaglandin E2 production in medulloblastoma
Authors: Bergqvist Filip , Ljungblad Linda , Wickström Malin , Larsson Karin , Kock Anna , Korotkova Marina , Johnsen John Inge , Kogner Per , Jakobsson Per-Johan
Published: July 2016

Introduction: Prostaglandin E2 (PGE2) is a key lipid mediator of inflammation and carcinogenesis. Apart from promoting direct growth of cancer cells, PGE2 has been shown to be crucial in maintaining the immunosuppressive tumor microenvironment. Drugs targeting cyclooxygenase (COX)-1 and -2, enzymes upstream in the production of PGE2 and other prostanoids, have successfully reduced tumor growth in many cancer models but these drugs are all associated with adverse long-term effects. The aim of this study is to demonstrate an anti-tumor effect in medulloblastoma by selective inhibition of microsomal prostaglandin E1 (mPGES-1), the enzyme downstream of COX-1/2 specifically required for the synthesis of PGE2.

Experimental procedures: Human medulloblastoma cell lines (DAOY and D283) were treated with a selective small-molecule inhibitor against mPGES-1 or COX-2 in the presence or absence of pro-inflammatory interleukin (IL)-1â. Prostaglandins in supernatants were extracted and quantified with LC-MS/MS. Drug efficacy was determined and cell toxicity was measured with cell viability assay. Mice with human medulloblastoma xenograft will function as proof-of-concept for the anti-tumor activities associated with inhibition of PGE2 production. Key inflammatory enzymes were quantified with Western blot.

Results: Medulloblastoma cells express high levels of COX-2, mPGES-1, and produce PGE2. The increased production of PGE2 by IL-1â was completely blocked when cells were co-treated with inhibitor for mPGES-1 or COX-2. Both inhibitors showed cell toxicity. We did not observe any shunting towards other prostanoids.

Conclusion: We investigate the effect of selective inhibition of mPGES-1 or COX-2 on medulloblastoma growth. We aim to further characterize the mPGES-1 inhibitor and compare to selective COX-2 inhibitor in preclinical medulloblastoma models. Specific inhibition of PGE2 production, rather than general inhibition of prostanoid production, would potentially prove useful as a complement to current medulloblastoma treatments.

Citation Format: Filip Bergqvist, Linda Ljungblad, Malin Wickström, Karin Larsson, Anna Kock, Marina Korotkova, John Inge Johnsen, Per Kogner, Per-Johan Jakobsson. Specific inhibition of prostaglandin E2 production in medulloblastoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 761.
Abstract 77: Establishment of lung cancer patient derived xenograft model from circulating tumor cells: Identification of somatic mutations associated with metastatic potentials
Authors: Lee Chansu , Kim Youngwook , Bae Eunkyung , Cho Sunghoon , Youn Sohyun , Ahn Kwang-Sung , Kim Bong-Seog
Published: July 2016

Carcinoma metastasis is initiated by a subpopulation of circulating tumor cells (CTCs) found in the blood of patients. However, the existence and genetic alternations of metastasis-initiating cells (MICs) among CTCs has not been experimentally demonstrated. The establishment of xenograft model using CTCs should help us to understand the biologic behaviors and genetic alternations of MICs associated the promotion of metastasis. Here, we demonstrate that xenograft model can be established by CTCs obtained from lung cancer patients and somatic mutations analyzed from xenograft model in which MICs develop metastatic tumor. After injecting mononuclear cells of lung cancer patients via tail vein, mouse was sacrificed at four weeks. After primary cultured cells were isolated from mouse lung tissues, the MICs were selected by using FASC sorting (EpCam+) and then re-injected via tail vein. And tumor formations in mouse were examined at 8 weeks. For identifying somatic mutations, target sequencing with NGS was performed in tumor DNA and cell free DNA (CF DNA) of lung cancer patients bloods. Our results indicated that nonsynonymous IDH, EGFR, KIT, ABL1, PTEN, ATM, P53, and SMAD4 were found in both tumor tissues and CF DNA of lung cancer patients’ bloods. IDH and EGFR was alternative homo. Others newly found in CF DNA. For further examination of the biologic behaviors of tumor cells which harvested from xenograft model using CTCs, we cultured primary cells from lung tissues and analyzed the anti-cancer drug resistance. Our results showed that primary cultured cells (BH-007) were resistant against cisplatin and erlotinib. In assay of migration and invasion, BH-007 cells possessed high metastatic potential when compared with A549 and H23 cell lines. These data suggested that several somatic mutations affect the anti-cancer drug resistance in lung cancer patients. Our future researches will include development of patient-derived xenograft models for preclinical testing of novel therapeutics. And we develop the possible integration of CF DNA analysis and PDX model from CTCs in the design of co-clinical studies for testing in individual lung cancer patients.

Citation Format: Chansu Lee, Youngwook Kim, Eunkyung Bae, Sunghoon Cho, Sohyun Youn, Kwang-Sung Ahn, Bong-Seog Kim. Establishment of lung cancer patient derived xenograft model from circulating tumor cells: Identification of somatic mutations associated with metastatic potentials. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 77.
Abstract 773: Analysis of oncogene affected networks in tumorigenesis of lung cancer
Authors: Sathe Adwait , Chen Yong , Yoo Hyuntae , Zhang Michael Q.
Published: July 2016

Genetic abnormality is one of major causes of tumorigenesis by driving the progressive transformation of normal human cells into highly malignant derivatives. Tracing the downstream pathways through which and how the effects of the oncogenic-activating mutations are transmitted is an important and challenging problem in studying disease mechanisms, diagnoses and treatments. In this study, we designed a network-based computational method, called MAXMAP, to not only predict mutation downstream affected pathways, but also quantitatively measure the dynamics of the pathway genes.

We first constructed a heterogeneous network by integrating protein-protein interaction networks and signaling and regulatory networks from available up-to-date curated databases, as well as RNASeq expression data. The RNAseq dataset of these mutated cell lines has 18,096 genes whereas the Protein Protein Interaction and Signalling, regulation dataset has 14,864 genes. We then used the maximal information flow method to globally optimize the effects from the mutations to target downstream genes. The method was applied to analyze the biological network dynamics that is caused by single (TP53), double (TP53+KRAS) and triple (TP53+KRAS+STK11) mutations of HBEC30KT, an immortalized lung cell line, and revealed the core dynamical subnetworks of these mutations. Only the triple mutant is known to have tumor-like properties and as shown in earlier studies these sequential mutation accumulations lead to tumorigenesis in mouse xenograft models. The experimental validation of a small subnetwork was performed using transient knock down of the pathway genes and expression was verified using qPCR.

We showed the dynamical subnetworks through which a gene, SLC7A5 was affected by sequential single, double or triple mutations. We have experimentally validated the predicted small subnetwork for SLC7A5 which is an important transporter of neutral amino acids. It is associated with bladder and bile duct adeno carcinomas and overexpressed in lung cancers. We selected the subnetwork affected due to the single mutation (TP53) for validation, including THBS1, TNFRSF11B and MAPK1 and SLC7A5.

Hence, using the MAXMAP method we have successfully identified a dynamical subnetwork affected by changes in the known network of genes. These results suggest that this computational method may allow us to predict drug targets or biomarkers or to study interesting alternative paths and predict the most likely path leading to certain phenotypic outcomes.

Citation Format: Adwait Sathe, Yong Chen, Hyuntae Yoo, Michael Q. Zhang. Analysis of oncogene affected networks in tumorigenesis of lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 773.
Abstract 794: A genetic polymorphism of ALDH2 and breast cancer risk: an analysis of 6,624 cases and 5,750 controls from the Breast Cancer Association Consortium
Authors: Ugai Tomotaka , Milne Roger L. , Ito Hidemi , Matsuo Keitaro
Published: July 2016

Background.

Numerous epidemiological studies consistently indicate that consumption of alcoholic beverages is an independent risk factor for female breast cancer. Although the mechanism underlying this effect remains unknown, it has been hypothesized that ethanol is mutagenic via its first metabolite, acetaldehyde. An impact of acetaldehyde on the carcinogenesis of several types of cancer has been shown in experimental models and in epidemiological studies in East Asian countries where a functional aldehyde dehydrogenase 2 (ALDH2) polymorphism Glu504Lys (rs671) is prevalent. However, its association with breast cancer risk has not been fully elucidated. We conducted an association analysis of original data from Asian studies participating in the Breast Cancer Association Consortium (BCAC).

Patients and Methods.

Nine studies were used for this analysis, including 6,624 cases and 5,750 controls. The pooled odds ratio (OR) with 95% confidence interval (CI) for breast cancer risk associated with the ALDH2 polymorphism was estimated using logistic regression models. We estimated unadjusted ORs and adjusted ORs for potential confounders.

Results.

In the unadjusted (crude) analysis, no significant association between ALDH2 polymorphism and breast cancer was observed, with ORs relative to Glu/Glu, of 1.02 (95% CI = 0.95-1.11; P = 0.51) for Glu/Lys, 1.13 (0.96-1.32; 0.14) for Lys/Lys and 1.04 (0.97-1.12; 0.29) for carriers of any Lys allele. In addition, after adjustment for all potential confounding factors, this lack of significant association was consistently observed, with ORs relative to Glu/Glu, of 0.95 (95% CI = 0.86-1.04; P = 0.25) for Glu/Lys, 0.99 (0.82-1.20; 0.93) for Lys/Lys and 0.95 (0.87-1.04; 0.29) for carriers of any Lys allele. These findings were consistent across ER, PR and HER2 status.

Conclusions.

No significant association between ALDH2 polymorphism and breast cancer was observed in this study. Considering the established impact of ALDH2 504Lys on the carcinogenesis of several types of alcohol-induced cancer, our findings suggest that acetaldehyde is less influential to the breast carcinogenesis induced by alcohol, as is the case for upper-aerodigestive tract cancers.

Citation Format: Tomotaka Ugai, Roger L. Milne, Hidemi Ito, Keitaro Matsuo. A genetic polymorphism of ALDH2 and breast cancer risk: an analysis of 6,624 cases and 5,750 controls from the Breast Cancer Association Consortium. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 794.
Abstract 811: Mosaic chromosome Y loss is not associated with testicular germ cell tumor risk
Authors: Machiela Mitchell J. , Dagnall Casey L. , Pathak Anand , Loud Jennifer T. , Chanock Stephen J. , Greene Mark H. , McGlynn Katherine A. , Stewart Douglas R.
Published: July 2016

Background

Testicular germ cell tumors (TGCTs) affect approximately 6 in 100,000 men with peak incidence in men 15 to 40 years of age. Established risk factors include family history of disease, undescended testis, contralateral TGCT, impaired fertility, and 21 GWAS susceptibility loci. Recent evidence suggests a 1.6 Mb deletion in the AZFc region (“gr/gr” deletion) of chromosome Y previously associated with male infertility is also associated with two- and three-fold increased risks of sporadic and familial TGCT, respectively. Furthermore, population-based studies have suggested mosaic chromosome Y loss in blood-derived DNA may be associated with increased risk of various solid tumors. Our aim was to investigate if mosaic loss across the entire Y chromosome is a TGCT risk factor.

Methods

We obtained blood and buccal-derived DNA from two studies: the NCI Familial Testicular Cancer Study (FTC, Cases = 172, Controls = 163) and NCI's US Servicemen's Testicular Tumor Environmental and Endocrine Determinants Study (STEED, Cases = 506, Controls = 611). We designed a quantitative polymerase chain reaction (qPCR) with 15 probes spanning the Y chromosome to assess Y loss. Samples were run in triplicate across three batches and normalized to a standard curve. We quantified probe signals as a target to reference (T/R) ratio, and averaged ratios across the 15 probes to yield an overall Y chromosome signal.

Results

On average, men with TGCT had slightly lower mean T/R ratios compared with control men lacking TGCT (1.02 vs. 1.03). Multivariate logistic regression models adjusted for study batch effects detected no significant relationship between mean Y chromosome T/R ratio and TGCT risk (OR = 0.34, 95% CI = 0.10-1.17, P = 0.09). A similar null relationship was observed when using a threshold of 1.5 standard deviations below the mean T/R ratio to dichotomize between mosaic Y loss and no mosaic Y loss (OR = 0.59, 95% CI = 0.21-1.63, P = 0.31).

Conclusion

Our study suggests that mosaic chromosome Y loss, as measured by 15 probes spanning the Y chromosome, is not associated with either sporadic or familial TGCT risk. Further studies aimed at investigating the AZFc region of the Y chromosome are needed to better assess whether this locus is directly associated with TGCT risk or is indirectly associated by means of a shared relationship with impaired fertility.

Citation Format: Mitchell J. Machiela, Casey L. Dagnall, Anand Pathak, Jennifer T. Loud, Stephen J. Chanock, Mark H. Greene, Katherine A. McGlynn, Douglas R. Stewart. Mosaic chromosome Y loss is not associated with testicular germ cell tumor risk. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 811.
Abstract 832: Preventing the progression of estrogen receptor-negative/HER2-overexpressing early stage breast disease by lapatinib
Authors: Acharya Sunil , Xu Jia , Wang Xiao , Jain Shalini , Wang Hai , Zhang Qingling , Bower Joseph , Richards-Kortum Rebecca , Yu Dihua
Published: July 2016

Tamoxifen and aromatase inhibitors (AIs) have shown efficacy to prevent estrogen receptor positive (ER+) breast cancer; however, there is no effective prevention strategy for ER- breast cancer. HER2 overexpression (HER2+) can be detected in up to 40% of ER- breast cancers and it has been reported to promote the progression of early stage breast cancer. Here, we tested the feasibility of lapatinib to prevent ER-/HER2+ early stage breast disease. Our data showed that lapatinib reduced the proliferation and increased the apoptosis in an in vitro ER-/HER2+ early breast disease model. Furthermore, lapatinib treatment in the HER2-driven MMTV-neu mouse model from early stage of disease development (atypia, at 10 weeks of age) significantly delayed tumor initiation and progression as detected by mammary gland biopsy and fiber optic micro-endoscope (FOME) imaging. Mechanistically, we found that lapatinib inhibited GLUT4 upregulation and increased glucose up-taking in HER2-overexpressing cells. These results suggested that lapatinib is a promising agent for the prevention/intervention of ER- and HER2+ breast cancers. Our study brings better understanding of the mechanism underlying ER-/HER2+ early stage breast disease progression to cancer and our strategy of cancer prevention with low-dose kinase inhibitor could be rapidly translated into the clinic for other cancer types in the near future.

Citation Format: Sunil Acharya, Jia Xu, Xiao Wang, Shalini Jain, Hai Wang, Qingling Zhang, Joseph Bower, Rebecca Richards-Kortum, Dihua Yu. Preventing the progression of estrogen receptor-negative/HER2-overexpressing early stage breast disease by lapatinib. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 832.