bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2906: An effective anti-EpCAM antibody EpAb2-6 for the treatment of colon cancer and its underlying mechanisms
Authors: Wu Han-Chung , Chi Yi-Hsuan , Liao Mei-Ying
Published: May 2015

Epithelial cell adhesion molecule (EpCAM) is known to be overexpressed in epithelial cancers associated with enhanced malignant potential, particularly colorectal carcinoma (CRC) and head and neck squamous cell carcinoma (HNSCC). However, it is yet to be determined whether targeting EpCAM can directly inhibit the progression of malignance. Here, we have generated six novel monoclonal antibodies (mAbs) against EpCAM; these antibodies specifically bind to CRC and HNSCC cells, but not normal cells. One EpCAM mAb, EpAb2-6, was found to induce cancer cell apoptosis in vitro, inhibit tumor growth, and prolong the overall survival of mice with human colon or oral carcinoma xenografts. EpAb2-6 also increases the therapeutic efficacy of irinotecan, fluorouracil, and leucovorin (IFL) therapy in tumor-bearing mice. Subsequent analysis suggested that EpAb2-6 can potentially inhibit the growth and proliferation of tumorspheres. Investigation of the mechanism by which EpAb2-6 inhibits tumor growth revealed that EpAb2-6 inhibits tumor-necrosis-factor α converting enzyme (TACE/ADAM17) and γ-secretase activation and blocks EpCAM intracellular domain (EpICD) cleavage, thus preventing the nuclear localization of EpICD. Furthermore, EpAb2-6, which binds to positions in the EGF-II/TY domain of EpCAM, inhibits production of EpICD, thereby decreasing translocation of β-catenin into the nucleus and activation of p53 and p21 expression. Collectively, our results indicate that novel EpCAM mAbs can potentially be used for cancer-targeted therapy.

Citation Format: Han-Chung Wu, Yi-Hsuan Chi, Mei-Ying Liao. An effective anti-EpCAM antibody EpAb2-6 for the treatment of colon cancer and its underlying mechanisms. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2906. doi:10.1158/1538-7445.AM2014-2906
Abstract 2907: Everolimus enhances the cytotoxic action of the immunotoxin, HA22, directed to CD22 on B-cell malignancies
Authors: Antignani Antonella , Pasetto Matteo , Choo Chris , Angelus Evan , Mathews Lesley , Guha Rajarshi , Shinn Paul , Ferrer Marc , Thomas Craig J. , Pastan Ira , Fitzgerald David J.
Published: May 2015

Recombinant immunotoxins, antibody-toxin fusion proteins, are therapeutic proteins designed as targeted anti-cancer agents. Clinical trials of immunotoxins directed to surface CD22 or mesothelin are ongoing with a 50% complete response rate in patients with refractory Hairy Cell Leukemia as well as dramatic responses in some patients with advanced mesothelioma. To identify agents that could improve clinical efficacy further, a screen of 500 approved or near-approved compounds (MIPE-4) was conducted, revealing everolimus as a potential enhancer of Pseudomonas exotoxin-based recombinant immunotoxins. When the immunotoxin HA22 (moxetumomab pasudotox), which targets CD22, was added to various B-cell lines, everolimus enhanced cytotoxicity by 2-10-fold. Kinetic analyses indicated that combinations of HA22 and everolimus (0.25 uM) accelerated the cessation of protein synthesis in Nalm-6 and CA46 cells compared to either agent alone. Likewise, in CellTiterGlo assays of cell viability, combinations of HA22 and everolimus resulted in a greater loss of ATP from cells. Western blot analysis of key survival proteins indicated an early loss of Mcl-1 in combination treatments. Additional experiments are ongoing to extend these observations with the goal of uncovering insights in the mechanism of synergy seen with these two compounds.

Citation Format: Antonella Antignani, Matteo Pasetto, Chris Choo, Evan Angelus, Lesley Mathews, Rajarshi Guha, Paul Shinn, Marc Ferrer, Craig J. Thomas, Ira Pastan, David J. Fitzgerald. Everolimus enhances the cytotoxic action of the immunotoxin, HA22, directed to CD22 on B-cell malignancies. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2907. doi:10.1158/1538-7445.AM2014-2907
Abstract 2916: Childhood BMI is associated with risk of adult pancreatic cancer
Authors: Nogueira Leticia M. , Stolzenberg-Solomon Rachael , Gamborg Michael , Sørensen Thorkild , Baker Jennifer
Published: May 2015

The incidence and death rate for pancreatic cancer, the fourth leading cause of cancer death in the U.S., have been increasing. Excess weight through adulthood is one of the few modifiable risk factors for pancreatic cancer. Childhood obesity rates have increased more rapidly that adulthood obesity in the U.S, but its effects on pancreatic cancer risk are not clear.

Hence, the objective of this study is to test the hypothesis that childhood weight is associated with pancreatic cancer risk later in life.

We evaluated the association of body mass index (BMI) measured at ages 7-13 with risk of pancreatic cancer later in life in 288,423 children from the Copenhagen School Health Records Register who were born between 1930-1981. These data were linked with the Danish Cancer Registry to identify incident pancreatic cancer cases from 1977-2011. Follow-up ended on the date of pancreatic cancer diagnosis, death, emigration, 70th birthday, or December 31st 2010, whichever came first.

Hazard Ratios (HR) and 95% confidence intervals (CI) were calculated for age specific BMI z-scores using Cox proportional hazards stratified on birth cohort and gender. Restricted cubic splines analysis was used to test for linearity. There were 1163 pancreatic cancer cases during the 6,918,420 person-years of follow up. Childhood BMI was associated with pancreatic cancer risk in males and females. At age 10, for example, HR per one unit increase in BMI z-scores was 1.17 (CI: 1.08, 1.26). The observed association did not deviate from linearity.

Our results support the hypothesis that higher childhood body weight is associated with increased pancreatic cancer risk later in life.

Citation Format: Leticia M. Nogueira, Rachael Stolzenberg-Solomon, Michael Gamborg, Thorkild Sørensen, Jennifer Baker. Childhood BMI is associated with risk of adult pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2916. doi:10.1158/1538-7445.AM2014-2916
Abstract 2922: Prediagnostic smoking, alcohol intake, physical activity and survival after colorectal cancer diagnosis in Western European populations
Authors: Fedirko Veronika , Riboli Elio , Norat Teresa , Tjønneland Anne , Jenab Mazda
Published: May 2015

Background: Cigarette smoking, high intake of alcohol, and lack of physical activity are associated with increased colorectal cancer (CRC) risk, but little research has been conducted to investigate the association between these modifiable lifestyle factors and survival after CRC diagnosis.

Methods: Therefore, we prospectively assessed the association between smoking status and intensity, baseline alcohol intake, lifetime pattern of alcohol intake, physical activity and all-cause and CRC-specific death among 3,916 participants diagnosed with colorectal cancer (colon = 2,454, rectum = 1,462) between 1992 and 2008 in the EPIC cohort. Multivariable Cox proportional hazards models were used to calculate hazard ratios (HRs) and corresponding 95% confidence intervals (CIs).

Results: There were 1,303 deaths, 1,039 (80%) due to CRC. Mean follow-up after CRC diagnosis was 4 years. In multivariable analysis, current vs. never smokers had an increased risk of all-cause death (HR = 1.22, 95% CI: 1.04-1.44). Greater intensity of smoking (26+ cigarettes per day) was associated with almost a doubling in the risk of all-cause death when compared with never smokers (HR = 1.75, 95% CI: 1.12-2.73). Alcohol consumption was not statistically significantly associated with survival (HR = 1.00, 95% CI: 0.98-1.02 per 6 g per day), however former heavy drinkers had an increased risk of all-cause death (HR = 2.68, 95% CI: 1.13-6.39). Total physical activity was not statistically significantly associated with CRC survival. Additional subgroup analyses were conducted by sex, age at diagnosis, cancer stage and grade, location of tumor, time between blood collection and cancer diagnosis, year of diagnosis, follow-up period, and body mass index. Results for CRC-specific death were similar to those reported for all-cause death.

Conclusions: Our preliminary results suggest that cigarette smoking and being a former heavy drinker before cancer diagnosis are associated with worse survival among Western European patients with CRC, but further research is needed to confirm these results.

Citation Format: Veronika Fedirko, Elio Riboli, Teresa Norat, Anne Tjønneland, Mazda Jenab, on behalf of the EPIC CRC Working Group. Prediagnostic smoking, alcohol intake, physical activity and survival after colorectal cancer diagnosis in Western European populations. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2922. doi:10.1158/1538-7445.AM2014-2922
Abstract 2930: Global chromatin profiling reveals NSD2 mutation in pediatric ALL
Authors: Chan Ho Man , Jaffe Jacob D. , Wang Yan , Zhang Jinghui , Huether Robert , Kryukov Gregory V. , Bhang Hyo-eun C. , Taylor Jordan E. , Hu Min , Englund Nathan P. , Yan Feng , Wang Zhaofu , McDonald E Robert , Wei Lei , Ma Jing , Easton John , Yu Zhengtian , deBeaumount Rosalie , Gibaja Veronica , Venkatesan Kavitha , Schlegel Robert , Sellers William R. , Keen Nicholas , Liu Jun , Caponigro Giordano , Barretina Jordi , Cooke Vesselina G. , Mullighan Charles , Carr Steven A. , Downing James R. , Garraway Levi A. , Stegmeier Frank
Published: May 2015

Epigenetic dysregulation is an emerging hallmark of cancers. We developed a high-information-content mass spectrometry approach to profile global histone modifications in human cancers. When applied to 115 lines of the Cancer Cell Line Encyclopedia1, this approach identified distinct molecular chromatin signatures. One signature was characterized by increased H3K36 dimethylation, exhibited by several lines harboring NSD2 translocations. A novel NSD2 p.E1099K variant was identified in non-translocated acute lymphoblastic leukemia (ALL) lines sharing this signature. Ectopic expression of the variant induced a chromatin signature characteristic of NSD2 hyperactivation and promoted transformation. NSD2 knockdown selectively inhibited the proliferation of NSD2-mutant lines and impaired the in vivo growth of an NSD2-mutant ALL xenograft. Sequencing analysis of >1000 pediatric cancer genomes identified the NSD2 p.E1099K mutation in 14% of t(12;21)[ETV6-RUNX1]-containing ALLs. These findings identify NSD2 as a potential therapeutic target for pediatric ALL and provide a general framework for the functional annotation of cancer epigenomes.

1.

Barretina,J. et al. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483, 603-607 (2012).

Citation Format: Ho Man Chan, Jacob D. Jaffe, Yan Wang, Jinghui Zhang, Robert Huether, Gregory V. Kryukov, Hyo-eun C. Bhang, Jordan E. Taylor, Min Hu, Nathan P. Englund, Feng Yan, Zhaofu Wang, E Robert McDonald III, Lei Wei, Jing Ma, John Easton, Zhengtian Yu, Rosalie deBeaumount, Veronica Gibaja, Kavitha Venkatesan, Robert Schlegel, William R. Sellers, Nicholas Keen, Jun Liu, Giordano Caponigro, Jordi Barretina, Vesselina G. Cooke, Charles Mullighan, Steven A. Carr, James R. Downing, Levi A. Garraway, Frank Stegmeier. Global chromatin profiling reveals NSD2 mutation in pediatric ALL. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2930. doi:10.1158/1538-7445.AM2014-2930
Abstract 2933: Assessment of ABT-263 activity across a comprehensive cancer cell line collection leads to a novel, potent combination therapy for small cell lung cancer
Authors: Faber Anthony C. , Costa Carlotta , Dastur Anahita , Benes Cyril , Engelman Jeffrey
Published: May 2015

BH3 mimetics like ABT-263 induce apoptosis in subsets of cancer. However, they have shown limited clinical efficacy as single agents, and rational combination strategies remain underexplored. Toward the development of a novel combined therapy, we examined efficacy of ABT-263 across 331 cancer cell lines. Cell lines with high expression levels of the pro-apoptotic gene BIM were more sensitive to ABT-263 activity. This was particularly evident in small cell lung cancer (SCLC), which tends to have higher BIM levels and is among the most sensitive to ABT-263. However, MCL-1 expression mitigated the impact of ABT-263-induced apoptosis by sequestering BIM released from BCL-2 and BCL-XL. Since MCL-1 expression is dependent on TORC1 activity in some cancers, we attempted to overcome the apoptotic block in SCLC by combining ABT-263 with the TORC1/2 inhibitor AZD8055. AZD8055 treatment alone downregulated MCL-1 expression; combination AZD8055 and ABT-263 induced marked apoptosis and growth arrest in vitro, as well as tumor regressions in multiple SCLC xenograft models. Strikingly, low-dose AZD8055 was sufficient to promote tumor regressions in the combination strategy. Therefore, addition of a TORC1/2 inhibitor offers a therapeutic strategy to markedly improve single-agent ABT-263 activity in SCLC.

Citation Format: Anthony C. Faber, Carlotta Costa, Anahita Dastur, Cyril Benes, Jeffrey Engelman. Assessment of ABT-263 activity across a comprehensive cancer cell line collection leads to a novel, potent combination therapy for small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2933. doi:10.1158/1538-7445.AM2014-2933
Abstract 2937: Localization of sipuleucel-T- expanded T cell clones into tumors of treated prostate cancer patients
Authors: Fong Lawrence , DeVries Todd , Cham Jason , Zhang Li , Frohlich Mark , Trager James , Sheikh Nadeem
Published: May 2015

Sipuleucel-T is an FDA-approved autologous cellular therapy demonstrated to prolong overall survival in subjects with asymptomatic or minimally symptomatic metastatic castrate resistant prostate cancer (mCRPC). Sipuleucel-T is generated by culturing patients' own peripheral blood mononuclear cells (PBMC) with a fusion protein of prostatic acid phosphatase and granulocyte macrophage colony stimulating factor. While antigen presenting cells take up this antigen and can prime antigen specific T cells in vivo, the majority of the cells in the sipuleucel-T product are in fact T cells. Moreover, antigen-specific T cells in the product become activated during production of sipuleucel-T, particularly for the 2nd and 3rd infusions.

To examine whether T cells contained in sipuleucel-T might be homing to the site of the tumor, we used a next-generation sequencing-based method to assess T cell repertoire diversity in the blood, sipuleucel-T product, and resected tissues of patients participating in a neoadjuvant study of sipuleucel-T treatment (P07-1; NCT00715104). In this trial, patients with localized prostate cancer received sipuleucel-T prior to undergoing radical prostatectomy (RP). DNA extracted from the different samples was amplified using a multiplex PCR assay to amplify the CDR3 region of the T cell receptor spanning the variable region formed by the junction of the V, D and J segments and their associated non-templated insertions. Sequence reads were used to quantitate absolute TCR frequencies using standardized clonotype determination algorithms. We found that a subset of T cell clones was enriched in the generation of sipuleucel-T. We also found that T cell clones were expanded in patient blood after each successive infusion. When we examined the T cell clonotypes in the prostate tissues resected after sipuleucel-T treatment, we found that there were T cell clones in the prostate in common with the sipuleucel-T product. Interestingly a number of the clones present in the pre-treatment blood were reduced or lost in the blood by the second and third infusion but were present in the post-treatment prostate tissue, consistent with trafficking of the T cell clones from the peripheral circulation. These results indicate that ex vivo production of sipuleucel-T induces the expansion of certain T cell clones that are ultimately present in the prostates of patients treated prior to RP. Sipuleucel-T may therefore also function as an adoptive T cell therapy.

Citation Format: Lawrence Fong, Todd DeVries, Jason Cham, Li Zhang, Mark Frohlich, James Trager, Nadeem Sheikh. Localization of sipuleucel-T- expanded T cell clones into tumors of treated prostate cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2937. doi:10.1158/1538-7445.AM2014-2937
Abstract 2968: FTY720 inhibits mutant Kras-induced lung cancer via disrupting Stat3-S1PR1 vicious cycle and downregulating tumor PD-L1 expression
Authors: Yeh Hsuan-Heng , Hsu Tsung-I , Hung Jan-Jong , Su Wen-Pin , Su Wu-Chou
Published: May 2015

Aberrant activation of signal transducer and activator of transcription 3 (Stat3) occurs in many cancers and plays a critical role in tumor progression. The system that Stat3-induced Sphingosine-1-phosphate receptor 1 (S1PR1) expression and the S1P-S1PR1 pathway reciprocally regulate Stat3 activity was considered a major positive feedback loop for persistent Stat3 activation in cancer cells and the cells of tumor microenvironment. Tumor cells expressing the ligand for the receptor programmed death-1 (PD-1), PD-L1, have been shown to increase apoptosis of antigen-specific human T-cell to evade the immune system. In this study, we determined the influence of disrupting Stat3-S1PR1 vicious cycle on tumor formation and PD-L1 expression in lung cancer. We have established mutant Kras-induced lung cancer mice model and found that Stat3 phosphorylation was elevated in tumor tissues of lung cancer. Treatment of lung cancer cells with AG490 (JAK2 inhibitor) inhibited Stat3 activation and AG490 administration to the mice decreased the numbers of lung cancer nodules, indicating that blockage of Stat3 activation could inhibit mutant Kras-induced lung cancer in mice. Similar effects were also detected by using Stat3 inhibitor, S3I-201. We found that lung cancer cells expressed more pStat3 showed higher PD-L1 expression levels than that of lower pStat3 expressing cells. Inhibition of Stat3 activation by AG490 decreased PD-L1 expression in vitro and in vivo, indicating that PD-L1 expression could be regulated by Stat3. Moreover, FTY720 (S1P antagonist) could inhibit S1PR1 expression, down-regulate Stat3 activity, pJAK1 expression and inhibit IL-6 secretion in lung cancer cell lines and causes inhibition of the mutant Kras-induced lung cancer in mice. Furthermore, FTY720 administration dose-dependently suppressed tumor PD-L1 expression in mice lung cancer tissues. Taken together, our study indicated that FTY720 could suppress mutant Kras induced-lung cancer formation via inhibiting persistent Stat3 activation and down-regulating tumor PD-L1 expression.

Citation Format: Hsuan-Heng Yeh, Tsung-I Hsu, Jan-Jong Hung, Wen-Pin Su, Wu-Chou Su. FTY720 inhibits mutant Kras-induced lung cancer via disrupting Stat3-S1PR1 vicious cycle and downregulating tumor PD-L1 expression. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2968. doi:10.1158/1538-7445.AM2014-2968
Abstract 2980: Maximizing the efficacy of anti-angiogenesis cancer therapy: A multi-targeting strategy by tyrosine kinase inhibitors
Authors: Nakazawa Youya , Kawano Satoshi , Matsui Junji , Funahashi Yasuhiro , Tohyama Osamu , Muto Hiroki , Nakagawa Takayuki , Matsushima Tomohiro
Published: May 2015

Although vascular endothelial growth factor (VEGF) inhibitors provide significant clinical benefit, they often require dose reductions or even withdrawals due to their severe toxicities. Furthermore, almost all cancers show intrinsic and/or evasive resistance to VEGF inhibitors by multiple mechanisms. Serum angiopoietin-2 (Ang2) level has been proposed as a potential biomarker of VEGF inhibitor response in several cancers. Response to lenvatinib (E7080; VEGFR1-3 inhibitor) is also reported to correlate with low Ang2 level in differentiated thyroid cancer and endometrial cancer. From these clinical observations, Ang2 and its receptor Tie2 has been focused as promising targets. Here, we demonstrated mechanisms of resistance induced by Ang2 and a novel strategy to circumvent the resistance by combination of multi-tyrosine kinase inhibitors (TKIs), lenvatinib and golvatinib (E7050; c-Met, Tie2, EphB4 inhibitor). Tie2 defines a highly pro-angiogenic macrophage subset, Tie2-expressing macrophage (TEM). Ang-Tie2 and EphB4-EphrinB2 signaling play critical roles in pericyte-mediated vessel stabilization. Ectopic expression of Ang2 in thyroid cancer conferred resistance to lenvatinib and enhanced pericyte-associated endothelial network development and TEM infiltration. In vitro analyses suggested that golvatinib/lenvatinib combination inhibited pericyte-mediated vessel stabilization and TEM differentiation. In thyroid and endometrial cancer models, golvatinib/lenvatinib combination inhibited pericyte network development and TEM infiltration, resulting in severe perfusion disorder and massive apoptosis. Body weight loss was tolerable in mice, and no macroscopic change was observed. These results suggest that modulation of tumor microenvironment by strategic and well-tolerated combination of multi-targeting TKIs sensitizes cancer to VEGF inhibitors, which warrants further clinical investigation to determine the clinical benefit of anti-angiogenesis cancer therapy.

Citation Format: Youya Nakazawa, Satoshi Kawano, Junji Matsui, Yasuhiro Funahashi, Osamu Tohyama, Hiroki Muto, Takayuki Nakagawa, Tomohiro Matsushima. Maximizing the efficacy of anti-angiogenesis cancer therapy: A multi-targeting strategy by tyrosine kinase inhibitors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2980. doi:10.1158/1538-7445.AM2014-2980
Abstract 2983: Bisphosphonates: New strategies for targeting angiogenesis in ovarian cancer
Authors: Dalton Heather J. , Reusser Nicole M. , Zien Alexander , Jackson David , Previs Rebecca , Rupaimoole Rajesha , Zand Behrouz , Lopez-Berestein Gabriel , Coleman Robert L. , Sood Anil K.
Published: May 2015

Objectives:

Bisphosphonates, known to have beneficial effects on bone and soft tissue metastases in breast cancer, are potent inhibitors of macrophages, but effects on the microenvironment are not fully understood. We investigated the biological effect of clodronate on macrophage and endothelial cell-driven angiogenesis in ovarian cancer.

Methods:

Using the FDA Adverse Event Reporting System (FAERS), we examined the effects of bisphosphonate use on overall cancer mortality. We examined the in vitro (endothelial cell migration, capillary formation and cytokine secretion) and in vivo (orthotopic mouse models) effects of clodronate on angiogenesis, macrophage infiltration, and tumor growth.

Results:

Using (FAERS) data, out of ∼17,000 patients with a cancer diagnosis co-medicated with a bisphosphonate, overall reported death rate was 36% lower (17.6% vs 27.7%, p<0.0001) than those not receiving bisphosphonates, independent of anti-tumor therapy. In vitro, treatment with clodronate resulted in decreased endothelial cell secretion of IL-6 (p < 0.001), IL-8 (p < 0.001), and a 2.5-fold reduction in cell migration compared to controls (p < 0.001), with only partial recovery after VEGF stimulation. Clodronate reduced capillary formation by 1.5-fold compared to controls. In the ID8-VEGF model, clodronate-treated mice had a 5-fold decrease in tumor weight (0.05g vs. 0.3g, p < 0.001) compared to controls. In the SKOV3ip1 model, clodronate treatment resulted in a 3-fold decrease in tumor weight (0.35g vs. 0.96g, p = 0.003) and tumor nodules (5.2 vs. 16, p=0.005) compared to controls. Immunohistochemical staining of clondronate-treated tumor sections showed a 2-fold and 1.5-fold (ID8-VEGF and SKOV3ip1, respectively) reduction in macrophage density (p=0.001 and p < 0.001, respectively) compared to controls. Clodronate-treated tumor sections showed a 3-fold decrease in capillary density (p < 0.001) compared to controls.

Conclusions:

Bisphosphonates modulate tumor angiogenesis through effects on macrophages and endothelial cells and are associated with overall decreased mortality in cancer patients, independent of chemotherapeutic agents. Bisphosphonates represent an unexplored, but attractive clinical strategy in ovarian cancer, with potential for synergistic combination with other anti-angiogenic agents.

Citation Format: Heather J. Dalton, Nicole M. Reusser, Alexander Zien, David Jackson, Rebecca Previs, Rajesha Rupaimoole, Behrouz Zand, Gabriel Lopez-Berestein, Robert L. Coleman, Anil K. Sood. Bisphosphonates: New strategies for targeting angiogenesis in ovarian cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2983. doi:10.1158/1538-7445.AM2014-2983
Abstract 2985: In vivo evaluation of regorafenib and liposomal-doxorubicin in a panel of low passage uterine START-PDX tumor models
Authors: Meade Justin , Wick Michael J. , Vaught Teresa , Gamez Lizette , Chavez Roger , Tolcher Anthony , Rasco Drew , Patnaik Amita , White Allan J. , Papadopoulos Kyriakos
Published: May 2015

Background: Uterine and ovary cancers account for a majority of gynecologic cancer-related deaths in Western countries. While initial cure rates are high, recurrent disease is often chemoresistant with few treatment options. Regorafenib is a recently approved multi-tyrosine kinase targeting VEGFR2 and Tie2. As Tie2 has been implicated in the pathogenesis of uterine cancers, we evaluated regorafenib as a potential therapy in our panel of uterine PDX models and compared activity to the approved therapy liposomal-doxorubicin.

Methods: PDX uterine models were established in immune-deficient mice from primary or metastatic patient tissue and once established were confirmed by histologic comparative analysis with patient tumor tissue, linked with patient treatment and outcome data and characterized for activating mutations and known amplifications. Drug sensitivity studies were performed in each model evaluating daily oral regorafenib or weekly intravenous liposomal-doxorubicin versus control; group sizes were n=2-3 for control and n=1 for treatment arms. Study endpoints included a set day following treatment initiation with tumor growth inhibition or delay and regression reported at study completion.

Results: For this project we evaluated twenty-five uterine models. Statistically significant (p<0.05) regorafenib activity was reported in a subset of evaluated endometrial models; however, activity was not limited to a particular tumor subtype or mutation profile. Liposomal-doxorubicin also demonstrated differential but significant tumor growth inhibition towards models in the evaluated panel.

Conclusion: We have screened preclinical activity of regorafenib and liposomal-doxorubicin towards a panel of uterine START-PDX models and identified sensitive and resistant models. Additional genomic and characterization studies are underway to better understand activity of regorafenib with the goal of identifying a sensitivity profile for patient stratification.

Citation Format: Justin Meade, Michael J. Wick, Teresa Vaught, Lizette Gamez, Roger Chavez, Anthony Tolcher, Drew Rasco, Amita Patnaik, Allan J. White, Kyriakos Papadopoulos. In vivo evaluation of regorafenib and liposomal-doxorubicin in a panel of low passage uterine START-PDX tumor models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2985. doi:10.1158/1538-7445.AM2014-2985
Abstract 2990: Notch1 monoclonal antibody inhibits tumor growth and modulates angiogenesis
Authors: Okamura Heidi , Proia Theresa , Bell Alisa , Liu Qing , Siddiquee Zakir , Lin Jie , Gyuris Jeno
Published: May 2015

AVEO Oncology has developed a series of inducible mouse models of cancer, which through the preservation of critical tumor/stromal interactions, facilitate identification of cell-surface and secreted proteins that represent viable targets for therapeutic antibodies and other biologics. Functional genetic screens performed in vivo in these models identified the Notch pathway as a critical regulator of tumor maintenance. This finding was consistent with emerging evidence that activation of Notch signaling via receptor point mutation, receptor amplification, and elevated receptor and ligand expression, plays a key role in various human cancers. Moreover, the Notch pathway controls diverse aspects of tumorigenesis and tumor maintenance, regulating tumor autonomous processes and interactions with the microenvironment, including angiogenesis. To further understand the role of the Notch pathway in tumor maintenance, and to assess the therapeutic potential of targeting the Notch pathway in cancer, we have generated monoclonal antibodies that inhibit the Notch1 receptor. Characterization of monoclonal antibodies targeting Notch1 through cell-based and biochemical studies demonstrated that the Notch1 monoclonal antibody bound with high affinity and high specificity to the Notch1 receptor, prevented ligand-mediated activation of the target receptor, and specifically repressed Notch-dependent signaling with high potency. Significantly, specific inhibition by the Notch1 monoclonal antibody did not result in the dose-limiting gut toxicity observed with Notch1 mAb resulted in tumor inhibition that was accompanied by increased vascular density, inhibition of Notch target gene expression, and increased expression of angiogenesis-related genes, consistent with expected changes resulting from Notch1/Dll4 pathway inhibition. In addition, several tumor models treated in combination with Notch1 mAb and an investigational VEGF pathway inhibitor, tivozanib, demonstrated increased tumor inhibition compared to either agent alone. Interestingly, combination treatment resulted in decreased vascular density and downregulated expression of angiogenesis genes, similar to treatment with VEGF inhibitor alone. Collectively, our work suggests that combination therapy of Notch1 antibodies with VEGF inhibitors should be evaluated as a potential viable therapeutic option for VEGF-resistant tumors.

Citation Format: Heidi Okamura, Theresa Proia, Alisa Bell, Qing Liu, Zakir Siddiquee, Jie Lin, Jeno Gyuris. Notch1 monoclonal antibody inhibits tumor growth and modulates angiogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2990. doi:10.1158/1538-7445.AM2014-2990
Abstract 2991: Triple combination of bevacizumab, anti-DLL4 (delta like ligand 4) and trebananib gives enhanced therapeutic effects in three xenograft tumor models
Authors: Mattson Bethany , Moriguchi Jodi , Jun H. Toni , Coxon Angela , Cordover Dave , Kaufman Steve , Oliner Jon , Starnes Charlie
Published: May 2015

Bevacizumab is a monoclonal antibody targeting VEGF that is clinically approved for the treatment of multiple indications in oncology, including colorectal, lung and renal cancer as well as glioblastoma. Clinical activity has also been demonstrated in breast and ovarian cancer as well, however, the time periods of benefit for all of these indications is in months (1-5). Multiple pathways are known to participate in the process of angiogenesis, and the limited effects of bevacizumab may represent the maximum activity of targeting just one of them. The purpose of the present study was to examine the effect of simultaneously targeting multiple angiogenic axes including angiopoietins 1 and 2, Dll4 and VEGF. Trebananib is a recombinant peptide-Fc fusion protein that selectively blocks the Tie2 interactions of both Ang1 and Ang2. Trebananib, as well as a recombinant antibody to Dll4 were produced at Amgen, Inc., Thousand Oaks, CA. Bevacizumab was purchased from a local pharmacy. These reagents were employed in the combination treatment of three different preclinical murine tumor models (U87, glioblastoma; MiaPaca and BxPC3, pancreatic carcinoma), one of which (BxPC3) was specifically selected for its relative resistance to VEGF/KDR inhibition. Harlan Athymic nu/nu female mice at 6-7 weeks of age were injected subcutaneously on the right flank with 5 x 106 tumor cells in the presence of Matrigel and treated when the tumors were approximately 300 mm3. The results, as supported by histomorphometric analysis (percent blood vessel area, viable tumor burden), demonstrate that under conditions of optimal biological dosing, the addition of trebananib to a two-way combination of bevacizumab and anti-Dll4 resulted in a statistically significant enhancement of therapeutic effect for the three-way combination, an observation made consistently over 3 different tumor models and 3 different dose levels of bevacizumab and anti-Dll4 (p = or < 0.0024; RMANOVA). Furthermore, the therapeutic effect in all three models approximated prolonged stasis, the ultimate theoretical goal of antiangiogenesis in the treatment of cancer.

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Citation Format: Bethany Mattson, Jodi Moriguchi, H. Toni Jun, Angela Coxon, Dave Cordover, Steve Kaufman, Jon Oliner, Charlie Starnes. Triple combination of bevacizumab, anti-DLL4 (delta like ligand 4) and trebananib gives enhanced therapeutic effects in three xenograft tumor models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2991. doi:10.1158/1538-7445.AM2014-2991
Abstract 2997: Vessel co-option in colorectal cancer liver metastases mediates resistance to VEGF-targeted therapy
Authors: Frentzas Sophia , Thompson Victoria L. , Vermeulen Peter B. , Foo Shane , Brown Gina , Cunningham David , Reynolds Andrew R.
Published: May 2015

The purpose of this study is to: (1) examine whether vessel co-option is associated with a lack of response to anti-angiogenic therapy in advanced colorectal cancer patients, (2) identify whether inhibition of pathways associated with vessel co-option sensitizes to anti-angiogenic therapy in vivo.

Sprouting angiogenesis is the process whereby new blood vessels are induced to sprout from existing vessels. The growth of metastatic tumors is considered to require sprouting angiogenesis. This hypothesis has driven the development and clinical application of vascular endothelial growth factor (VEGF) targeted agents, such as bevacizumab. However, responses to such agents in cancer patients, including metastatic colorectal cancer (CRC), are variable. It is not currently clear why this is the case and we have no way of selecting patients suitable for such therapies.

Importantly, it is now evident that tumors can also utilize a number of putative ‘VEGF-independent’ angiogenesis mechanisms, the existence of which may limit the efficacy of VEGF-targeted therapy. One such mechanism is ‘vessel co-option,’ whereby tumors engulf existing local blood vessels as they invade into surrounding host tissue. Published studies have reported that this mechanism occurs in ∼28-30% of CRC liver metastases.

Here we present data from a retrospective study of patients with CRC liver metatases that were treated with bevacizumab. We demonstrate that, in this patient cohort, the presence of vessel co-option in liver metastases was strongly associated with lack of response to bevacizumab. We also show that a similar result is observed in a mouse model of colorectal cancer liver metastasis. Interestingly, when we disable actin nucleation activity in colorectal cancer cells, vessel co-option is compromised and sensitivity to VEGF-targeted therapy is restored in vivo. These data demonstrate a potential role for vessel co-option as a negative predictive biomarker for anti-angiogenic therapy and also have consequences for the development of novel therapies for targeting tumor angiogenesis.

Citation Format: Sophia Frentzas, Victoria L. Thompson, Peter B. Vermeulen, Shane Foo, Gina Brown, David Cunningham, Andrew R. Reynolds. Vessel co-option in colorectal cancer liver metastases mediates resistance to VEGF-targeted therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2997. doi:10.1158/1538-7445.AM2014-2997
Abstract 300: Expression profiling of miRNA in asbestos-related lung adenocarcinoma
Authors: Mahmut Yasen , Ninomiya Hironori , Motoi Noriko , Itoh Satoru , Fujiwara Mutsunori , Ishikawa Yuichi
Published: May 2015

Lung cancer has the highest mortality rate of all of the cancers in the world and asbestos-related lung cancer is one of the leading occupational cancers. The identification of asbestos-related molecular changes has long been a topic of increasing research interest. Microarray technology is a powerful high-throughput tool capable of monitoring the expression of thousands of small noncoding RNAs at once within tens of samples processed in parallel in a single experiment. The aim of this study was to construct a genome-wide expression profile of miRNA of asbestos-related lung cancers, and was to identify novel asbestos-related molecular biomarkers. The miRNA profiling was performed on 31 patients (13 patients with asbestos-exposed and 18 patients with non- asbestos-exposed) undergoing lung resection for primary adenocarcinoma, both fresh and FFPE materials of whom were available. Data analyses on miRNA expression revealed four novel asbestos-related miRNAs (three over-expressed miR-210, miR-200a, miR-429, and one under-expressed miR-140-3p). Also, 16 target genes, expression of which were inversely correlated, were identified. Among them we obtained a set of miRNAs that were involved in hypoxia-regulated and oxidative stress responders, seemingly related to asbestos burden. For example, miR-210 is involved in a wide-range of processes, including cell proliferation, migration, apoptosis, differentiation, DNA repair and cell metabolism, and more recently in the regulation of antitumor immune responses, suggesting roles of this miRNA as a central master regulator of the hypoxic response. According to gene ontology analyses of the target genes were mostly transcription factors, cyclins and cyclin-dependent kinases. For validation, we compared the expression levels of miRNA between fresh and FFPE materials, resulting in good correlations (R=0.76-0.90). The miRNAs/genes presented here may be interesting targets for further investigation and could eventually have potential diagnostic implications.

Citation Format: Yasen Mahmut, Hironori Ninomiya, Noriko Motoi, Satoru Itoh, Mutsunori Fujiwara, Yuichi Ishikawa. Expression profiling of miRNA in asbestos-related lung adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 300. doi:10.1158/1538-7445.AM2014-300
Abstract 3006: Serum insulin growth factor/insulin growth factor receptor1 (IGF-I, IGF-I-1R) and VEGF-A as prognostic factors and surrogate biomarkers in triple negative breast cancer patients (TNBC)
Authors: Zekri Abdel-Rahman N. , Bahnassy Abeer
Published: May 2015

Background: We evaluated the prognostic and predictive values of serum IGF-I, IGF-I-1R, VEGF and TGF-β1 in triple-negative and non-triple-negative breast cancer (TNBC&NTNBC) patients through correlation with standard prognostic factors and survival.

Methods: serum expression of VEGF-A, TGF-β1, IGF-I and IGF-I-1R proteins was measured in of 43 TNBC, 53 NTNBC and 20 normal control (NC) by quantitative enzyme linked immune-sorbant assay (ELISA). Results were correlated to patients’ characteristics, disease free and overall survival (DFS&OS).

Results: The cutoff points that discriminate between TNBC and NTNBC were: 10.09 ng/ml, 412.54 pg/ml and 106.96 ng/ml for IGF-I-1R, VEGF-A, and IGF-I. VEGF, IGF-I and IGF-I-1R differed significantly between groups (p <0.001) whereas TGF-β1 differed significantly between NC and the other groups (p<0.001). TNBC was associated with positive lymph nodes (p<0.001), relapse and metastasis (p<0.001). Response rate (RR) was significantly lower in TNBC patients (p<0.001) with high serum VEGF-A, IGF-I-I/IGF-I-IR (p= 0.004, 0.001/0.003). Reduced DFS associated with high tumor grade, high VEGF-A (p<0.001), high IGF-I/ IGF-I-IR (P<0.001/ p<0.001). Reduced OS associated with relapse (p<0.001), high VEGF-A (p=0.001) and high IGF-I/ IGF-I-IR (p= 0.012/p= 0.02). in multivariate analysis positive nodes (P< 0.001), high VEGF-A (P = 0.02) and IGF-I/ IGF-I-1R (P= 0.03, P= 0.01) were independent factor for disease recurrence.

Conclusions: TNBC represents an aggressive subset of breast tumors. IGF-I, IGF-I-IR, VEGF-A could be used as surrogate biomarkers for TNBC, as prognostic marker since their expression level correlates with reduced OS and high incidence of relapse. They also represent promising candidates for targeted therapy in TNBC patients.

Citation Format: Abdel-Rahman N. Zekri, Abeer Bahnassy. Serum insulin growth factor/insulin growth factor receptor1 (IGF-I, IGF-I-1R) and VEGF-A as prognostic factors and surrogate biomarkers in triple negative breast cancer patients (TNBC). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3006. doi:10.1158/1538-7445.AM2014-3006
Abstract 3024: Cancer stem-like cells in ovarian clear cell carcinoma are enriched in ALDH-high population associated with the accelerated scavenging system of reactive oxygen species
Authors: Mizuno Tomoko , Suzuki Noriko , Mori Minako , Makino Hiroshi , Furui Tatsuro , Ito Naoki , Yamamoto Akio , Morishige Kennichiro
Published: May 2015

In ovarian cancer cases, disease recurrence after chemotherapy is frequently observed, suggesting ovarian cancer stem-like cells (CSCs) are involved. Ovarian clear cell carcinoma is especially chemo-resistant among epithelial ovarian cancer subtypes. We first, examined the CSC markers (CD133,CD44, ALDH1) using the ovarian clear cell carcinoma tissues (n=81). Immunohistochemical staining revealed that high ALDH1 expression levels are related to advanced stage in clear cell carcinoma cases. Other makers are not related to clinical staging and prognosis. Clear cellcarcinoma cells (KOC-7C) are separated to ALDH-high and ALDH-low population by ALDEFLUOR assay and fluorescence-activated cell sorting. ALDH-high cells showed more quiescent character (G0/G1 phase) than ALDH-low cells. Recent studies have suggested that CSCs might have a high antioxidant capacity to keep cellular reactive oxygen species (ROS) at a low level that allows them to survive adverse conditions. We examined the cellular ROS level using Cell-ROX assay. ALDH-high cells contained the lower levels of ROS than ALDH-low cells. Also, ALDH-high cells showed an increased expression of in nuclear factor-E2-related factor 2 (Nrf2), a key transcriptional factor of the antioxidant system, associated with the increased antioxidant enzymes (SOD2, HO-1). Thus, ALDH-positive CSCs might have increased Nrf2-induced antioxidant scavengers, which lower ROS level relevant to chemo-resistance in ovarian clear cell carcinoma.

Citation Format: Tomoko Mizuno, Noriko Suzuki, Minako Mori, Hiroshi Makino, Tatsuro Furui, Naoki Ito, Akio Yamamoto, Kennichiro Morishige. Cancer stem-like cells in ovarian clear cell carcinoma are enriched in ALDH-high population associated with the accelerated scavenging system of reactive oxygen species. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3024. doi:10.1158/1538-7445.AM2014-3024
Abstract 3039: The MELK/FOXM1 axis is a master regulator of proneural to mesenchymal transition (PMT) in glioma stem cells by controlling EZH2 transcriptional activity
Authors: Joshi Kaushal , Kim Sunghak , Lee Jeongwu , Waclaw Ronald , Salcini Lisa , Nakano Ichiro
Published: May 2015

In the Central Nervous System, stem cells are present in the normal brain and primary brain tumor, glioblasotoma multiforme (GBM). Both normal and oncogenic stem cells play central roles in organogenesis and tumorigenesis, respectively. Previously we identified that the mitotic kinase MELK binds and phosphorylates the oncogenic transcription factor FOXM1 to regulate the growth of both brain-derived and GBM-derived stem cells (GSCs). Here we demonstrate that the Enhancer of Zeste Homolog 2 (EZH2), the lysine methyltransferase of Polycomb repressive complex 2, is the major transcriptional target of the MELK-FOXM1 complex in GSCs. PCR-based analysis of FOXM1 immunoprecipitated chromatin determined the binding of FOXM1 to the EZH2 promoter region to drive its transcriptional activity. This signaling is mediated through the protein complex formation of FOXM1 and MELK in a MELK kinase-dependent manner. The MELK-FOXM1 driven EZH2 signals are required to retain the immature stem cell trait and mesenchymal identity in GBM, as well as GSCs' radiation resistant phenotype in a MELK kinase-dependent manner. Clinically, GBM tumors co-express MELK and EZH2 with significant elevation in post-irradiation recurrent tumors. This post-irradiation elevation of MELK and EZH2 was observed specifically in GSCs. Irradiation induces transformation of proneural (PN) GSCs into a Mesenchymal (Mes)-like GSC phenotype (PMT) that is highly resistant to radiation treatment, and inhibition of EZH2 by shRNA reverses the radiation-resistant phenotype of Mes GSCs. Elimination of MELK or FOXM1 by shRNA reduced EZH2 expression resulting in attenuated stemness and radiation resistance in vitro and in vivo. These phenotypes were completely restored by exogenous expression of EZH2. To our knowledge, this study provides the first evidence of the role of MELK-FOXM1-EZH2 signaling axis for maintenance of stem cell trait and PMT regulation in GSCs. Identification of the role of transcriptional regulator EZH2 in PMT by the mitotic kinase MELK and transcription factor FOXM1 complex sheds lights on a novel therapeutic target for aggressive cancers including GBM.

Citation Format: Kaushal Joshi, Sunghak Kim, Jeongwu Lee, Ronald Waclaw, Lisa Salcini, Ichiro Nakano. The MELK/FOXM1 axis is a master regulator of proneural to mesenchymal transition (PMT) in glioma stem cells by controlling EZH2 transcriptional activity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3039. doi:10.1158/1538-7445.AM2014-3039