bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 836: Effect of voluntary running on metastasis in a mouse model of breast cancer
Authors: Mijwel Sara , Rundqvist Helene , Lindholm Carolin , Strell Carina , Roswall Pernilla , Pietras Kristian , Johnson Randall S. , Östman Arne
Published: July 2016

Introduction: Emerging evidence states that regular physical exercise provides a risk reduction for breast cancer by approximately 25%. In addition, recent studies suggest that physical activity may also have an effect on recurrence and mortality. Mouse models are suitable tools to study anti-neoplastic mechanisms.

Aim: to evaluate the effect of voluntary running on tumor initiation, growth and metastasis in the Polyoma Middle T (PyMT) model of breast cancer.

Methods: PyMT mice were housed with access either to wirelessly recording running wheels or locked control wheels. Running distances and tumor volumes were recorded. At 12 weeks, mice were sacrificed. Mammary glands were histologically staged and pulmonary metastases were quantified. In a follow up study, pre-trained mice were injected intravenously with tumor cells derived from the PyMT model and after an additional 10 weeks of voluntary running, pulmonary metastases were quantified.

Results: PyMT mice ran significantly more than wildtype mice (6.4 vs 3.4 km/day). No significant effects of voluntary running on tumor- initiation, volume or stage were found. However, a trend for reduced metastasis was observed. After intravenous injections of tumor cells, runners had a significantly lower pulmonary metastasis frequency than non-runners.

Conclusion: In this aggressive breast cancer model, an average of 6 km/day of voluntary running did not induce any effect on tumor formation or growth. However, the findings suggest that physical activity has impact on the metastatic process.

Citation Format: Sara Mijwel, Helene Rundqvist, Carolin Lindholm, Carina Strell, Pernilla Roswall, Kristian Pietras, Randall S. Johnson, Arne Östman. Effect of voluntary running on metastasis in a mouse model of breast 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 836.
Abstract 86: Functional effects of MutL homolog 1 promoter polymorphism in prostate cancer
Authors: Fukuhara Shinichiro , Mitsui Yozo , Hashimoto Yutaka , Kato Taku , Wong Ryan K. , Shahryari Varahram , Yamamura Soichiro , Majid Shahana , Saini Sharanjot , Deng Guoren , Dahiya Rajvir , Tanaka Yuichiro
Published: July 2016

The mutL homolog (MLH1) gene protects the integrity of the genome by correcting erroneous insertions, deletions and mismatches that occur during DNA replication. Proper MLH1 expression is thus essential for the health of the cell and deficiency can result in mutation and genomic instability, which ultimately may lead to cancer. Polymorphisms in the promoter region can lead to altered gene expression and we tested the hypothesis that MLH1 promoter variants can affect levels as well as be a risk for prostatic cancer. Two polymorphic sites were analyzed, rs1800734 (A to G) and rs4647203 (G to A). A luciferase reporter construct containing the MLH1 promoter region was utilized and site-directed mutagenesis was performed to create rs1800734 and rs4647203 variants. Activity was tested using DU145, PC3 and LNCaP prostatic cells and interestingly, the rs1800734 G variant resulted in significantly decreased luciferase levels in all three cell lines compared to A (P<0.01). No difference however, was seen for the rs4647203 variant. To determine whether rs1800734 can affect clinical expression levels, benign prostatic hyperplasia specimens were evaluated for MLH1 expression by immunohistochemistry. A significantly lower H score was observed for the GG genotype compared to AA (P<0.01). To further explore the clinical relevance of rs1800734, a case-control study was performed using DNA extracted from normal healthy, BPH, and prostate cancer patients from a Japanese population by sequence-specific PCR and sequencing. Results of experiments showed a trend for the variant G allele to be associated with prostate cancer when compared to normal controls (P<0.1). However, no differences were observed between BPH and controls as well as between stages or grades of prostate cancer. The MLH1 promoter polymorphism is thus capable of altering expression and associated with prostate cancer, and these results are important in understanding its role in this disease.

Citation Format: Shinichiro Fukuhara, Yozo Mitsui, Yutaka Hashimoto, Taku Kato, Ryan K. Wong, Varahram Shahryari, Soichiro Yamamura, Shahana Majid, Sharanjot Saini, Guoren Deng, Rajvir Dahiya, Yuichiro Tanaka. Functional effects of MutL homolog 1 promoter polymorphism in prostate 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 86.
Abstract 877: Evolution of resistance to EGFR inhibition from drug tolerant cancer cells
Authors: Hata Aaron N. , Drier Yotam , Gomez-Caraballo Maria , Siddiqui Faria , Sequist Lecia , Bernstein Bradley , Engelman Jeffrey
Published: July 2016

Acquired drug resistance to targeted cancer therapies remains a significant clinical problem. Although mechanisms of acquired resistance of EGFR mutant non-small cell lung cancers to EGFR inhibitors have been identified, little is known about how resistant clones evolve during drug therapy. We demonstrate that acquired resistance caused by the T790M gatekeeper mutation can occur either by selection of pre-existing T790M clones or via evolution of T790M-negative drug tolerant cells that develop the mutation during drug treatment. Additionally, the path to resistance impacts the biology of the resistant clone, as those that evolved from drug tolerant cells have a diminished apoptotic response to third generation EGFR inhibitors that target T790M EGFR. To characterize drug tolerant cells in patients undergoing EGFR inhibitor therapy, we performed single-cell RNA-Seq on EGFR mutant NSCLC samples prior to and after initiation of EGFR inhibitor. These results offer novel insights into the evolution of acquired resistance to EGFR inhibitors and point to new therapeutic opportunities to prevent or overcome resistance in the clinic.

Citation Format: Aaron N. Hata, Yotam Drier, Maria Gomez-Caraballo, Faria Siddiqui, Lecia Sequist, Bradley Bernstein, Jeffrey Engelman. Evolution of resistance to EGFR inhibition from drug tolerant cancer cells. [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 877.
Abstract 879: Single cell analysis resolves combinatory targeted therapy for arresting the BRAFi-induced cellular dedifferentiation of metastatic melanomas
Authors: Su Yapeng , Wei Wei , Xue Min , Robert Lidia , Tsoi Jennifer , Graeber Thomas , Ribas Antoni , Heath James
Published: July 2016

Targeted therapy that inhibits the BRAF (V600E) protein has shown impressive initial responses in metastatic melanoma patients. However, acquired drug resistance, derived from tumor heterogeneity, always limits its efficacy and compromises its treatment outcomes. Melanocyte to neural crest transition (MNT) has been observed to play crucial role in early stage adaptive drug tolerance, and the mathematic modeling indicates the transition is contributed from both stochastic phenotypic transition and non-genetic selection. Whole transcriptomic analysis enriched important signaling proteins that are involved in the resistance development process. Single cell proteomic analysis of those proteins was performed on patient-derived melanoma models to further study the early stage drug tolerance. This resolves how protein signaling network coordination rewires in the process, which cannot be resolved from proteomic analysis of bulk tumor tissue. Signaling network coordination analysis indicates that co-inhibiting the hubs of the network (MEK, NFKB-p65) could keep cells in the drug-susceptible stages. The combination therapy that co-inhibits mutant BRAF, MEK and NFKB-p65 together successfully arrested the MNT transition and induced a sustained tumor growth suppression in multiple patient-derived melanoma cell lines. This study provides us a brand-new way of using single cell proteomic analysis to infer effective combinations by resolving protein signaling network coordination that is inaccessible through traditional bulk-level proteomic measurement.

Citation Format: Yapeng Su, Wei Wei, Min Xue, Lidia Robert, Jennifer Tsoi, Thomas Graeber, Antoni Ribas, James Heath. Single cell analysis resolves combinatory targeted therapy for arresting the BRAFi-induced cellular dedifferentiation of metastatic melanomas. [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 879.
Abstract 886: Notch signaling activates B-cell specific enhancers to drive oncogene targets in B-cell lymphoma
Authors: Ryan Russell J. , Petrovic Jelena , Rausch Dylan M. , Lee Winston , Kluk Michael , Donohue Laura , Gillespie Shawn , Aster Jon C. , Pear Warren S. , Bernstein Bradley E.
Published: July 2016

Recurrent gain-of-function mutations in genes encoding Notch receptors are associated with poor clinical outcome in chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL), but genome-wide functional targets of Notch signaling in B cells have not been characterized.

We identified genome-wide binding sites for Notch transcription factor (TF) complexes, using antibodies specific for intracellular NOTCH1 and its binding partner RBPJ, in CLL and MCL lymph node biopsies, and lymphoma cell lines. We employed a gamma-secretase inhibitor washout strategy to rapidly modulate mutant NOTCH1 protein activation in complementary lymphoma cell line models of ligand-dependent and ligand-independent Notch signaling, and performed RNA-sequencing and genome-wide enhancer acetylation profiling in both the “notch on” and “notch off” states. These data identify a common set of functional Notch target genes in B-cell lymphoma that includes both canonical Notch targets as well as B-cell-specific target genes. The latter are frequently associated with adjacent B-cell-specific enhancers that show direct Notch TF binding and Notch-signaling-dependent histone acetylation. Notch-activated target genes include MYC and other TFs implicated in B-cell lymphoma, and are significantly enriched for mediators of targetable oncogenic lymphoma signaling pathways, (B-cell receptor, toll-like receptor, JAK-STAT, MAP kinase, and G-protein signaling pathways, all FDR q-value < 1e-3 in KEGG and / or Reactome analyses). Many B-cell Notch target genes show significantly increased expression in lymph nodes from patients with NOTCH1 mutant vs. wild-type CLL. Notch target genes are also up-regulated in lymph node- versus peripheral blood-derived CLL cells, consistent with immunohistochemical evidence for microenvironment-dependent Notch signaling activation in the CLL lymph node. Combined small-molecule inhibition of Notch and B-cell receptor signaling showed a synergistic anti-proliferative effect in Notch-dependent lymphoma cells.

Our data reveal the functional regulome of Notch signaling in small B-cell lymphoma, and may have implications for the role of Notch signaling in normal B-cell development. These findings suggest novel strategies for rational combination therapy in CLL and MCL.

Citation Format: Russell J. Ryan, Jelena Petrovic, Dylan M. Rausch, Winston Lee, Michael Kluk, Laura Donohue, Shawn Gillespie, Jon C. Aster, Warren S. Pear, Bradley E. Bernstein. Notch signaling activates B-cell specific enhancers to drive oncogene targets in B-cell lymphoma. [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 886.
Abstract 891: Dual defects of MDM2/MDMX-p53 pathways cause global metabolic disruption and enhance tumorigenesis
Authors: Lu Hua , zhang Yi-wei , Liao Peng , Liao Wenjuan , Chen Jiaxiang , Zeng Shelya X.
Published: July 2016

Although recent discoveries have linked the tumor suppressing functions of p53 to its regulation on cellular metabolism, the exact signaling pathways underlying how p53 is activated in response to various metabolism stresses, and how p53 coordinates the systemic metabolism homeostasis and further blocks tumorigenesis remain largely unclear. Previous studies have shown that the mice containing a single knock-in at cysteine residue 305 of MDM2 (MDM2C305F) is defective in ribosomal stress (RS) induced p53 activation and display severe nutrient shortage-induced hepatosteatosis. And very recently we found that in response to metabolic stress (MeS), AMPK directly phosphorylates serine 342 of MDMX, leading to p53 activation, which is defective in MDMX3SA mice harboring a mutant MDMX with S341A, S367A and S402A mutations. In the current study, we want to determine if the dual defects in the MDM2/MDMX-p53 pathways might thoroughly impair p53's ability to regulate metabolic homeostasis and thus favor metabolism-related tumorigenesis, by generating a double knock-in (DKI) MDMX3SA/MDM2C305F mouse line. Our results indicate that DKI mice on normal chow diet exhibit mild, but significant, metabolism abnormalities, including increased body weight, higher fasting-glucose level and impaired glucose tolerance, which is exacerbated by high-fat-diet feeding. Consistently, MEFs isolated from DKI mice possess higher capacity of differentiation into adipocytes, and less beige fat is induced in the subcutaneous white adipose tissue of DKI mice after chronic cold exposure. Furthermore, we observe that the survival rate of DKI mice is significantly lower compared to either control or single knock-in groups, which may be due to the higher morbidity of cancers in DKI mice, including hepatoma, lymphoma and lung carcinoma. Because of these observations, we further specifically investigate if dual defects of these p53 activation pathways will accelerate hepatocellular carcinogenesis or not. We indeed find that treatment of carcinogen DEN causes more and bigger tumor nodules in the liver of DKI mice. These results suggest that both RS-MDM2-p53 and MeS-MDMX-p53 pathways play important roles in regulation of global metabolism, and dual defects of these pathways benefit metabolism-related tumor formation and growth. Further study is required to establish the direct effect of the p53 pathway defects-mediated metabolic disruption on tumorigenesis, and determine the underlying mechanisms.

Citation Format: Hua Lu, Yi-wei zhang, Peng Liao, Wenjuan Liao, Jiaxiang Chen, Shelya X. Zeng. Dual defects of MDM2/MDMX-p53 pathways cause global metabolic disruption and enhance 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 891.
Abstract 899: Cancer protection associated with dietary methyl donor deficiency is characterized by persistent changes to epithelial proliferation and metabolism
Authors: Hanley Matthew P. , Rosenberg Daniel W.
Published: July 2016

The role of folate one-carbon metabolism in colorectal cancer development is incompletely understood, and nutritional intervention studies have produced conflicting results. We previously demonstrated that a diet deficient in the methyl donors folic acid, methionine, choline and vitamin B12, and supplemented with homocysteine, reduces intestinal tumor incidence by greater than 95% in ApcMin/+ mice. Here we extend these findings to a second mouse tumor model, ApcÄ14, and further show that the cancer protection afforded by even short-term dietary methyl donor deficiency (MDD) is long-lasting. Eleven weeks of MDD followed by methyl donor repletion was sufficient to maintain tumor suppression for at least 7 additional weeks (22.2 ± 3.5 vs 70.2 ± 4.6 intestinal tumors; p < 0.001). Sustained tumor protection was associated with altered intestinal crypt homeostasis. MDD increased the proportion of intestinal epithelial cells undergoing apoptosis in normal crypts and in tumors (4.9- and 3.2-fold, respectively), while reducing cell proliferation (Ki-67) and mitosis (PHH3). In addition, metabolomic profiling and metabolite set enrichment analysis (MSEA) revealed that tumor protection is associated with persistent alterations to metabolic pathways related to cellular proliferation, including glutamine metabolism, biogenic amine synthesis, nucleotide salvage and glutathione synthesis. Taken together, these results indicate that even a temporary dietary methyl donor restriction in cancer-prone mice can induce persistent changes to the intestinal epithelium that provide long-lasting protection in adult mice.

Citation Format: Matthew P. Hanley, Daniel W. Rosenberg. Cancer protection associated with dietary methyl donor deficiency is characterized by persistent changes to epithelial proliferation and metabolism. [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 899.
Abstract 903: IFN-ã induced PD-L1 on tumor and host cells co-operatively prevents tumor immune elimination after cancer immunoediting
Authors: Noguchi Takuro , Ward Jeffrey P. , Gubin Matthew M. , Arthur Cora , Schreiber Robert D.
Published: July 2016

Tumor cells can escape immune destruction via engaging the PD-1/PD-L1 inhibitory pathway. However since PD-L1 is induced on many different cell types by IFN-ã, the importance of PD-L1 on tumor versus host cells in mediating immune escape remains unclear. In this study, we assessed the role of PD-L1 expression on two closely-related but distinct clones derived from the same d42m1 methylcholanthrene (MCA)-induced sarcoma line. Edited d42m1-T3 sarcoma cells grow progressively in wild type (WT) mice but are rejected in mice treated with anti-PD-L1. Upregulation of PD-L1 was observed on both immune and tumor cells in vivo. To assess the role of PD-L1 specifically on tumor cells, we generated d42m1-T3 cells lacking PD-L1 (T3ÄPDL1) using CRISPR-Cas9. T3ÄPDL1 cells were spontaneously rejected in naïve syngeneic WT mice but formed progressively growing tumors in Rag2-/- mice. To explore the role of host-cell expressed PD-L1, we challenged WT mice with 3-fold higher numbers of T3ÄPDL1 cells and found that the tumor cells formed progressively growing tumors in WT mice. However, when tumor bearing mice injected with the high dose of T3ÄPDL1 cells were treated with blocking PD-L1 mAb, they all rejected their tumors. Thus, PD-L1 expression on both tumor and host cells plays a critical role in preventing immune elimination of edited MCA sarcoma cells. Alternatively highly immunogenic, unedited d42m1-T9 sarcoma cells were spontaneously rejected in WT mice due to the presence of the strong rejection antigen, mutant Spectrin-â2. Enforced expression of PD-L1 on unedited d42m1-T9 cells to levels similar to those induced under physiologic in vivo conditions did not alter the regressor phenotype of these cells. In contrast, enforced overexpression of PD-L1 on d42m-T9 cells to levels that were 20 fold higher than physiologic led to formation of progressively growing tumors in WT mice. Thus, physiologic levels of PD-L1 expression on highly immunogenic unedited tumor cells are not sufficient to counteract immune effector mechanisms driven by “strong” tumor antigens. This result shows that adaptive immune resistance occurs only on tumor cells that have undergone immunoediting.

Citation Format: Takuro Noguchi, Jeffrey P. Ward, Matthew M. Gubin, Cora Arthur, Robert D. Schreiber. IFN-ã induced PD-L1 on tumor and host cells co-operatively prevents tumor immune elimination after cancer immunoediting. [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 903.
Abstract 926: Pentraxin 3 is responsive to the activation of CCAAT/enhancer binding delta and promotes cancer progression in the tumor microenvironment
Authors: Chi Jhih-Ying , Wang Ju-Ming
Published: July 2016

This lack of understanding especially impacts the design of anticancer treatments. It is a common observation in the clinic that recurrent cancers, and particularly anticancer drug-resistant cancers, are more difficult to treat because they spread faster than the original tumor. CCAAT/enhancer binding protein delta (CEBPD) was activated in M2 macrophages and myofibroblasts/cancer-associated fibroblasts (CAFs) and contributed to metastasis, invasion and recurrence of cancer cells in response to PGE2 and the anticancer drugs. We further found that pentraxin 3 (PTX3) was activated in response to CEBPD induction and involved in migration and invasion of breast cancer cells, and the same phenomenon was also observed in drug-resistant cancers. Interestingly, we found that a recombinant PTX3 protein containing the C-terminal region (pPTX3) showed an opposite tumorigenic effect to the mammalian PTX3. The results suggest that PTX3 could be a promising target for preventing the metastasis/invasion of cancers and treating drug-resistant cancers.

Citation Format: Jhih-Ying Chi, Ju-Ming Wang. Pentraxin 3 is responsive to the activation of CCAAT/enhancer binding delta and promotes cancer progression in the tumor microenvironment. [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 926.
Abstract 938: Activation of ATR/CHK1 checkpoint is critical for mediating p53-dependent autophagy and pro-inflammatory VEGF production in human bronchial epithelial cells under fine particulate matters (PM2.5) exposure
Authors: Song Lun
Published: July 2016

The epidemiological and clinical studies have increasingly shown that fine particulate matters (PM2.5) exposure is associated with exacerbation of airway inflammation, which might results in the occurrence of lung cancer. Thus, understanding how PM2.5 triggers inflammatory responses in the respiratory system is a crucial issue for the study of PM2.5 toxicity. In the current study, we found that exposure of human bronchial epithelial cells (Beas-2B) to the winter PM2.5 collected in Wuhan, the south city of China, induced an significant up-regulation of VEGF production, a typical signaling event functionally in controlling chronic airway inflammation and vascular remodeling. Further investigations showed that autophagy was induced upon PM2.5 exposure and then mediated VEGF up-regulation by activating Src/STAT3 pathway in the Beas-2B cells. When exploring the upstream signaling events responsible for autophagy induction, we disclosed the requirement of p53 activation and its downstream target DRAM expression for the induction of autophagy, which results thus have extended the role of p53/DRAM-dependent autophagy beyond cell fate determination under genotoxic stresses and to the control of pro-inflammatory cytokine production. Moreover, PM2.5 exposure strongly induced activation of ATR/CHK1 axis, which subsequently triggered p53-dependent autophagy and VEGF production in Beas-2B cells. Therefore, these findings suggest a novel link between processes regulating genome integrity and the airway inflammation via autophagy induction in bronchial epithelial cells under PM2.5 exposure.

Citation Format: Lun Song. Activation of ATR/CHK1 checkpoint is critical for mediating p53-dependent autophagy and pro-inflammatory VEGF production in human bronchial epithelial cells under fine particulate matters (PM2.5) exposure. [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 938.
Abstract 944: The effects of GnRH-(1-5) on endometrial cancer cell lines
Authors: Larco Darwin O. , Cho-Clark Madelaine J. , Sorini Maya , Lee Cameron , Wu. T. John
Published: July 2016

The Gonadotropin-Releasing Hormone (GnRH) is a central regulator of reproductive function and behavior. In the periphery, GnRH is secreted by gynecologic tissues to exert local effects. Furthermore, GnRH is processed in the extracellular environment by the metalloendopeptidase EP24.15 to generate the bioactive metabolite, GnRH-(1-5). We have previously demonstrated that GnRH-(1-5) transactivates the epidermal growth factor receptor (EGFR) pathway to promote cell migration by binding the orphan receptor GPR101 in the Ishikawa and ECC-1 endometrial cancer cell lines. In this study, we sought to determine whether the effects of GnRH-(1-5) were dependent on the stage of the endometrial cancer. Cell lines ACI-181, ACI-52, and ACI-80 derived from endometrial cancers representative of stages 1, 2, and 3 respectively were tested in their response to GnRH-(1-5) treatment. Furthermore, the expression of GPR101 was confirmed by western blot analysis. Treatment with 100nM GnRH-(1-5) did not have significant changes in phospho-EGFR (pEGFR) levels in all ACI cell lines. However, subsequent measurement of ERK phosphorylation demonstrated that ACI-181 cells treated with GnRH-(1-5) had significantly (p < 0.05) increased levels in the pERK2 subunit but not pERK1, which parallels our initial findings in the Ishikawa cell line. No changes in pERK1/2 levels were determined in the other cell lines. To measure the effect of GnRH-(1-5) on cell migration, we implemented a wound closure assay. Interestingly, ACI-181 and ACI-52 cells had increased migration in response to GnRH-(1-5) treatment. To determine whether another pro-migratory pathway is regulated by GnRH-(1-5) in these cells, we measured TGF-β bioactivity using a cell-based assay from conditioned media of cells treated with GnRH-(1-5). We found that ACI-181 and ACI-52 cells had significantly (p<0.05) increased TGF-β bioactivity in response to GnRH-(1-5), indicating that these cells produced higher levels of bioactive TGF-β. It is likely that TGF-β and/or other growth factors may promote pro-survival and/or pro-migratory signaling cascades via an autocrine mechanism. These results further suggest that GnRH-(1-5) may exert differential effects on endometrial cancer cells that are not dependent on cancer progression.

Funding Sources: This research was supported by grants from the John P. Murtha Cancer Center at Walter Reed National Military Medical Center through the Uniformed Services University and the National Science Foundation (IOS-1052288).

Citation Format: Darwin O. Larco, Madelaine J. Cho-Clark, Maya Sorini, Cameron Lee, T. John Wu. The effects of GnRH-(1-5) on endometrial cancer cell lines. [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 944.
Abstract 956: Anticancer effects of silibinin-induced small nucleolar RNA 11B on bladder cancer cells
Authors: Yamamura Soichiro , Mitsui Yozo , Majid Shahana , Nip Hannah , Bucay Nathan , Saini Sharanjot , Deng Guoren , Shahryary Varahram , Dahiya Rajvir , Tanaka Yuichiro
Published: July 2016

Silibinin is the major active constituent of silymarin, an extract of milk thistle seeds. Silibinin has been shown to have significant anti-cancer effects in a variety of malignancies. However, the molecular mechanisms of silibinin action in bladder cancer have not been reported. In the present study, we found that silibinin (10 μM) significantly suppressed proliferation and invasion of T24 and UM-UC-3 bladder cancer cells. We analyzed various pathways as possible mechanisms of silibinin action using RNA-Seq technology in bladder cancer cell lines. Analysis of RNA-seq data identified silibinin targets that down-regulate the actin cytoskeleton pathway. Analysis of RNA-seq data also identified the mitogen-activated protein (MAP) kinase signaling pathway as a silibinin-down-regulated pathway. These pathways were also found to crosstalk through Ras and phosphatidylinositide 3-kinase (PI3K). These data show that silibinin anti-cancer effects are through down-regulation of actin cytoskeleton and MAP kinase pathways in bladder cancer. We also found that silibinin induced small nucleolar RNA 11B (SNORD11B) and that SNORD11B suppressed proliferation and invasion of bladder cancer cells. We will discuss the pathway of the anti-cancer effects of SNORD11B.

Citation Format: Soichiro Yamamura, Yozo Mitsui, Shahana Majid, Hannah Nip, Nathan Bucay, Sharanjot Saini, Guoren Deng, Varahram Shahryary, Rajvir Dahiya, Yuichiro Tanaka. Anticancer effects of silibinin-induced small nucleolar RNA 11B on bladder cancer cells. [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 956.
Abstract 965: Exosome-derived microRNAs contributes to prostate cancer chemoresistance
Authors: Li JIng , Xu Xiaozhen , Guan Hao , Mizokami Atsushi , Keller Evan T. , Yang Xin , Liu Xia , Tan Jiyong , Hu Longyuan , Lu Yi , Zhang Jian
Published: July 2016

Accumulating evidence has demonstrated that certain miRNAs play important roles in cancer cell chemoresistance. However, the pleotropic functions of exosome-derived miRNAs on developing chemoresistance remain unknown. In this study, we aimed to build potential networks of miRNAs, which derived from the exosome of chemoresistant prostate cancer (PCa) cells, with their known target genes using miRNA expression profiling and bioinformatic tools. Global miRNA expression profiles were measured by microarray. Twelve miRNAs were initially selected and validated by qRT-PCR. Known targets of deregulated miRNAs were utilized using DIANA-TarBase database v6.0. The incorporation of deregulated miRNAs and target genes into KEGG pathways were utilized using DIANA-mirPath software. To construct potential miRNA regulatory networks, the overlapping parts of two selected KEGG pathways were visualized by Cytoscape software. We identified 29 deregulated miRNAs, including 19 up and 10 down, in exosome samples derived from two kinds of paclitaxel resistance PCa cells (PC3-TXR and DU145-TXR) compared with their parental cells (PC3 and DU145). The enrichment results of deregulated miRNAs and known target genes showed that a few pathways were correlated with several critical cell signaling pathways. We found that hub hsa-16-5p, hsa-miR-3915, hsa-miR-488-3p, hsa-miR-5004-5p, hsa-miR-23c, hsa-miR-3673, hsa-miR-3654, and hsa-miR-32-5p were potential targets to hub androgen receptor (AR) and phosphatase and tensin homolog (PTEN). Hub TCF4 target genes were mainly regulated by hub hsa-miR3176, hsa-miR-141-3p, hsa-miR-606, hsa-miR381, and hsa-miR-429 miRNAs. These results may provide a linkage between PCa chemoresistance and exosome regulatory networks and thus lead us to propose that PTEN and TCF4 genes may be the important genes which regulated by exosome miRNAs in chemoresistance cancer cells. Supported by NSFC Key Project 81130046; NSFC Project 81171993 and NSFC81272415; Guangxi Key Projects 2013GXNSFEA053004 and 2012GXNSFCB053004; Guangxi Ministry of Education grants 201202ZD022 and 201201ZD004.

Citation Format: JIng Li, Xiaozhen Xu, Hao Guan, Atsushi Mizokami, Evan T. Keller, Xin Yang, Xia Liu, Jiyong Tan, Longyuan Hu, Yi Lu, Jian Zhang. Exosome-derived microRNAs contributes to prostate cancer chemoresistance. [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 965.
Abstract 982: Long non-coding RNA PICSAR promotes growth of cutaneous squamous cell carcinoma by regulating ERK1/2 activity
Authors: Piipponen Minna , Nissinen Liisa , Farshchian Mehdi , Riihilä Pilvi , Kivisaari Atte , Kallajoki Markku , Peltonen Juha , Peltonen Sirkku , Kähäri. Veli-Matti
Published: July 2016

Keratinocyte-derived cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer, and its incidence is increasing globally. Long non-coding RNAs (lncRNAs) are involved in various biological processes, and their role in cancer progression is emerging. The aim of this study was to find and characterize new biomarkers for evaluating the risk of progression and metastasis of cSCC and identify novel therapeutic targets for recurrent and metastatic cSCC. RNA-sequencing of cSCC cells (n = 8) and normal human epidermal keratinocytes (NHEKs, n = 4) revealed overexpression of a previously uncharacterized long intergenic non-coding RNA (lincRNA), LINC00162 in cSCC cells. Analysis of tissue sections by RNA in situ hybridization showed that LINC00162 is specifically expressed by tumor cells in cSCCs but not by keratinocytes in normal skin in vivo. The expression of LINC00162 in cSCC cells was upregulated by inhibition of the p38α and p38δ MAPKs using BIRB796 or p38α and p38δ specific siRNAs. Knockdown of LINC00162 inhibited proliferation and migration of cSCC cells, and suppressed the growth of human cSCC xenografts in vivo. Furthermore, knockdown of LINC00162 inhibited ERK1/2 activity and upregulated expression of dual specificity phosphatase DUSP6 in cSCC cells. Based on these observations, LINC00162 was named PICSAR (P38 Inhibited Cutaneous Squamous cell carcinoma Associated lincRNA). Our results provide mechanistic evidence for the role of PICSAR in promoting cSCC progression via activation of ERK1/2 signaling pathway by downregulating DUSP6 expression. These results also identify PICSAR as a novel biomarker and putative therapeutic target in cSCC.

Citation Format: Minna Piipponen, Liisa Nissinen, Mehdi Farshchian, Pilvi Riihilä, Atte Kivisaari, Markku Kallajoki, Juha Peltonen, Sirkku Peltonen, Veli-Matti Kähäri. Long non-coding RNA PICSAR promotes growth of cutaneous squamous cell carcinoma by regulating ERK1/2 activity. [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 982.
Abstract 986: Long noncoding RNA CCAT2 cooperates with miR-145 and miR-21 to regulate colon cancer stem cells
Authors: Yu Yingjie , Nangia-Makker Pratima , Farhana Lulu , Majumdar Adhip P. N.
Published: July 2016

Colon cancer-associated transcript-2 (CCAT2), a long noncoding RNA (lncRNA), has been reported to be over-expressed in colorectal cancer to promote tumor growth and metastasis and also reduces sensitivity to chemotherapy, a property related to cancer stem cells (CSCs). MicroRNAs, small endogenous noncoding RNAs, negatively control the expression of target genes by cleaving mRNA or through translation repression. We have recently reported cooperation of miR-21 and miR-145 to regulate colon CSCs proliferation and differentiation. Down-regulation of miR-21 enhances CSC enriched chemo-resistant colon cancer cells’ susceptibility to chemo- therapeutic regimens. Although both CCAT2 and miR-21 increase WNT signaling activity and reduce chemosensitivity to 5-FU, a critical evidence on how CCAT2 regulates stemness leading to chemo-resistance is lacking. The primary goal of this investigation was to examine whether CCAT2 associates with miR-145 and miR-21 and whether this association plays a role in regulating stemness of colon cancer cells. We have observed that CCAT2 transcript level is dependent on p53 expression and activity, as evidenced by a marked increase in CCAT2 expression in p53 wild type harboring HCT-116 colon cancer cells compared with p53 null HCT-116 and p53 mutant HT-29 cells. We further revealed that knockdown of CCAT2 increased the expression of miR-145 and negatively regulates miR-21 in HCT-116 cells and impairs proliferation and differentiation of these cells. On the other hand, upregulation of CCAT2 induces proliferation of HCT-116 cells and also increases the expression of CSC markers CD44 and CD166. We have also observed that miR-145 and miR-21 cooperation plays a role in regulating colon cancer stem cells by down-regulating Sox2 and increased activity of Wnt/β-catenin pathway. Our current results suggest that lncRNA CCAT2 cooperates with miR-145 and miR-21 to regulate colon cancer stem cells, which provides a potential therapeutic target for chemoresistance in colorectal cancer.

Citation Format: Yingjie Yu, Pratima Nangia-Makker, Lulu Farhana, Adhip P. N. Majumdar. Long noncoding RNA CCAT2 cooperates with miR-145 and miR-21 to regulate colon cancer stem cells. [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 986.
Abstract 998: LncRNA WDFY3-AS2 contributes to the EMT and metastasis in breast cancer
Authors: Richards Edward , Challa Sridevi , Li Yajuan , Permuth-Wey Jennifer , Bui Marilyn , Coppola Domenico , Sellers Thomas , Cheng Jin
Published: July 2016

The role of TGFβ in promoting mammary cell EMT and breast cancer metastasis was well established. However, the involvement of long non-coding RNA (lncRNA) in this process remains elusive. Here we demonstrated that mouse lncRNA BB179049 and its human ortholog WDFY3-AS2 were significantly increased upon TGFβ-induced EMT. WDFY3-AS2 was frequently elevated in invasive, metastatic and triple-negative breast cancer. Depletion of WDFY3-AS2 abrogated TGFβ-induced EMT and breast cancer metastasis. WDFY3-AS2 positively regulates STAT3 and WDFY3 through interaction with hnRNP-R. These data indicate that WDFY3-AS2 could play a pivotal role in TGFβ-induced EMT and metastasis and serves as a critical therapeutic target in aggressive breast cancer.

Citation Format: Edward Richards, Sridevi Challa, Yajuan Li, Jennifer Permuth-Wey, Marilyn Bui, Domenico Coppola, Thomas Sellers, Jin Cheng. LncRNA WDFY3-AS2 contributes to the EMT and metastasis in breast 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 998.
Abstract CT029: The effect of tazemetostat on CYP3A-mediated metabolism of midazolam in patients with solid tumors
Authors: Smith Sherri , Suttle Benjamin , Waters Nigel L. , Ribrag Vincent , Italiano Antoine , Michot Jean-Marie , Postel-Vinay Sophie , Toulmonde Maud , Cousin Sophie , Roche Maria , Pimentel Patricia , Ho Peter
Published: July 2016

Background: Tazemetostat is a selective oral small molecule inhibitor of enhancer of zeste homolog 2 (EZH2). Results from in vitro studies using human liver microsomes, recombinant CYP isoforms, and hepatocytes demonstrated that tazemetostat is an inhibitor, inducer, and substrate of CYP3A4. Previous results indicated that systemic exposure to tazemetostat decreased after multiple dose administration in patients with cancer, consistent with net induction of CYP3A-mediated metabolism. Therefore, a drug-drug interaction sub-study was performed as part of the ongoing study E7438-G000-101 (NCT01897571).

Purpose: To investigate the effect of tazemetostat on CYP3A-mediated metabolism in patients with solid tumors using midazolam as a sensitive CYP3A probe substrate.

Methods: This was a single-sequence, open-label, crossover study. Patients with solid tumors (n = 13) received a single oral dose of 2 mg midazolam on Day -1 and on Day 15. Tazemetostat 800 mg twice daily (BID) was administered continuously starting on Day 1. Serial blood samples for the analysis of plasma midazolam and metabolites were collected over 24 h on Day -1 (midazolam alone) and Day 15 (midazolam plus tazemetostat).

Results: A summary of midazolam pharmacokinetic parameters after administration alone and with tazemetostat 800 mg BID in 12 evaluable patients is presented below: ParameterDay -1aDay 15aGLSMRb (90% CI)AUC(0-∞) ng*h/mL51.9 (87.2)31.1 (48.1)0.60 (0.46, 0.78)Cmax ng/mL16.5 (94.3)13.0 (48.3)0.78 (0.57, 1.08)t1/2 h5.67 (34.3)4.25 (32.4)NCaData presented as geometric mean (%CV)bGLSMR = geometric least squares mean ratioNC = not calculated

Plasma midazolam AUC(0-?) and Cmax decreased approximately 40% and 22%, respectively, after administration with tazemetostat 800 mg BID relative to administration of midazolam alone. Geometric mean t1/2 for midazolam decreased approximately 25%, after administration of midazolam with tazemetostat 800 mg BID relative to administration of midazolam alone.

Conclusions: Administration of tazemetostat 800 mg BID resulted in net induction of CYP3A-mediated metabolism in patients with cancer. Tazemetostat 800 mg BID resulted in a less than 50% decrease in midazolam AUC and therefore is a weak inducer of CYP3A-mediated metabolism.

Citation Format: Sherri Smith, Benjamin Suttle, Nigel L. Waters, Vincent Ribrag, Antoine Italiano, Jean-Marie Michot, Sophie Postel-Vinay, Maud Toulmonde, Sophie Cousin, Maria Roche, Patricia Pimentel, Peter Ho. The effect of tazemetostat on CYP3A-mediated metabolism of midazolam in patients with solid tumors. [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 CT029.
Abstract CT118: Efficient basket trial designs
Authors: Cunanan Kristen M. , Iasonos Alexia , Shen Ronglai , Hyman David , Begg Colin , Gonen Mithat
Published: July 2016

Background: Basket trials are used to study targeted treatments across a range of tumor sites. In these designs all patients have the same target altered and receive the same regimen but they are placed in baskets defined by the site of primary tumor. This design is particularly appealing since the prevalence of the target mutation is typically low in any given tumor site, and the drug sponsor is naturally interested in as broad an indication for use as possible. Given the multiple baskets, this design requires the investigation of whether the activity of the regimen applies uniformly to all baskets, i.e., homogeneity of treatment effect, and if not, which baskets show sufficient response. Many basket trial designs to date have resembled parallel phase II studies, using Simon's two-stage design separately for each basket. The inclusion of multiple baskets increases the overall false positive rate in the trial, an aspect that is commonly not taken into account in current designs. More importantly, the use of independent, parallel phase II trials fails to take advantage of the opportunity to combine baskets in the event that the interim results suggest common responsiveness across baskets, either common evidence of efficacy or common evidence of lack of efficacy.

Methods: We develop a novel, adaptive study design that makes use of an interim assessment of the heterogeneity of treatment effect across baskets to determine the most appropriate statistical analysis, while controlling the overall false positive rate in the trial. Our proposed design allows for aggregation of baskets in stage 2 when evidence points to a common effect of the drug, while retaining the option to maintain selected individual baskets in stage 2, if the interim evidence supports this strategy. Similar to conventional designs, our proposed design allows for early termination of baskets for lack of responsiveness. Using simulations we demonstrate the potential reduction in the number of patients required compared with the traditional strategy based on parallel Simon two-stage designs, while maintaining similar levels of Type I and II errors.

Citation Format: Kristen M. Cunanan, Alexia Iasonos, Ronglai Shen, David Hyman, Colin Begg, Mithat Gonen. Efficient basket trial designs. [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 CT118.
Abstract CT124: A phase II study of bevacizumab and irinotecan as second-line therapy for patients with metastatic colorectal cancer previously treated with fluoropyrimidine, oxaliplatin, and bevacizumab
Authors: Nakajima Go , Kuramochi Hidekazu , Ando Masayuki , Itabashi Michio , Kawakami Kazuyuki , Hamano Mie , Hirai Eiichi , Iino Takayuki , Yokomizo Hajime , Okuyama Ryuji , Araida Tatsuo , Yoshimatsu Kazuhiko , Kameoka Shingo , Hayashi Kazuhiko
Published: July 2016

Background: FOLFIRI plus bevacizumab (BV) is widely used as second-line chemotherapy for patients with metastatic colorectal cancer (mCRC) previously treated with FOLFOX/XELOX plus bevacizumab. However, there is insufficient data characterizing the effectiveness of administration of fluoropyrimidine beyond first disease progression. FOLFIRI requires using a CV catheter. Oral fluoropyrimidine may cause severe diarrhea when administered with irinotecan (CPT-11). CPT-11+BV may overcome those problems.

Methods: Patients with mCRC previously treated with at least four courses of fluoropyrimidine, oxaliplatin, and bevacizumab were designated to receive 150 mg/m2 of CPT-11 and 10 mg/kg of BV, every 2 weeks as second-line therapy. The primary endpoint was progression-free survival (PFS), and secondary endpoints included response rate (RR), overall survival (OS), and adverse events.

Results: Thirty patients from six institutes were enrolled from March 2011 to January 2014. Twenty-five (83.3%) patients were refractory to first-line therapy and eight patients (16.6%) were unable to tolerate first-line therapy. The median PFS was 5.7 months (95% CI; 4.2-7.3 months) and the RR was 6.7% (range, 0.8-22.1%. Grades 3-4 adverse events included leucopenia (36.7%), neutropenia (50%), thrombocytopenia (26.7%), anemia (30%), diarrhea (3.3%), anorexia (6.7%), and hypertension (3.3%). Relative dose intensities were 94.5% and 96.3% for CPT-11 and BV, respectively. Median OS was 11.8 months (6.3 months-not reached).

Conclusions: Administration of CPT-11 plus BV to patients with mCRC achieved comparable efficacy with relatively lower toxicities compared with the results of previous studies using FOLFIRI plus BV as second-line therapy. The dose intensity of CPT-11 was judged satisfactory. Clinical trial information: UMIN000005228

Citation Format: Go Nakajima, Hidekazu Kuramochi, Masayuki Ando, Michio Itabashi, Kazuyuki Kawakami, Mie Hamano, Eiichi Hirai, Takayuki Iino, Hajime Yokomizo, Ryuji Okuyama, Tatsuo Araida, Kazuhiko Yoshimatsu, Shingo Kameoka, Kazuhiko Hayashi. A phase II study of bevacizumab and irinotecan as second-line therapy for patients with metastatic colorectal cancer previously treated with fluoropyrimidine, oxaliplatin, and bevacizumab. [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 CT124.