bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1144: Detection of a novel ALK fusion variant in lung adenocarcinoma using a comprehensive genomic analysis
Authors: Shien Kazuhiko , Ruder Dennis , Ileana Ecaterina E. , Papadimitrakopoulou Vassiliki A. , Frampton Garrett M. , Behrens Carmen , Kalhor Neda , Lee J. Jack , Tang Ximing , Herbst Roy S. , Wistuba Ignacio I. , Izzo Julie G.
Published: July 2016

Background: Anaplastic lymphoma kinase (ALK) is a tyrosine kinase that is constitutively activated in a subset of non-small cell lung cancer (NSCLC), mainly as a result of chromosomal translocation. In ALK-rearranged NSCLC, the growth of tumors is strongly addicted to ALK activity, and suppression of ALK is a potent therapeutic strategy. To date, 6 known fusion partners of ALK onco-kinase have been reported in NSCLC, although the sensitivity to ALK inhibitor according to fusion variant except for EML4-ALK is still unclear. In this study, we reviewed clinical trial samples analyzed by comprehensive genomic profiling approach to detect unknown actionable ALK rearrangements.

Materials and methods: We reviewed gene alteration and expression profile of clinical samples in preliminary molecular studies from the ongoing BATTLE-2, phase II clinical trial of targeted drugs in chemo-refractory NSCLC.

Results: In 187 NSCLC samples, 3 samples harbored ALK fusion gene: 2 EML4-ALK and 1 novel ALK fusion variant PRKAR1A-ALK in a lung adenocarcinoma of male former smoker patient. The ALK gene expression levels of ALK-rearranged samples were significantly higher compared to other samples. Interestingly, the patient harboring PRKAR1A-ALK was treated with ALK inhibitor crizotinib and showed long time stable disease.

Conclusions: Our findings suggest that PRKAR1A-ALK fusion could be a novel oncogenic driver in lung adenocarcinoma and an indicator of ALK inhibitor response. (NHI-NCI CA155196; 2P50CA070907-16A1)

Citation Format: Kazuhiko Shien, Dennis Ruder, Ecaterina E. Ileana, Vassiliki A. Papadimitrakopoulou, Garrett M. Frampton, Carmen Behrens, Neda Kalhor, J. Jack Lee, Ximing Tang, Roy S. Herbst, Ignacio I. Wistuba, Julie G. Izzo. Detection of a novel ALK fusion variant in lung adenocarcinoma using a comprehensive genomic analysis. [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 1144.
Abstract 1152: MUC1: A metabolic regulator in triple-negative breast cancer
Authors: Goode Gennifer D. , Chaika Nina , Purohit Vinee , Gunda Venugopal , Singh Pankaj
Published: July 2016

Breast cancer, the second leading cause of cancer deaths in women, is the most common cancer among North American women, accounting for nearly 1 in 3 cancer cases diagnosed in the U.S. women. Triple negative breast cancer (TNBC) subtype accounts for approximately 15-25% of all breast cancer cases and has an increased incidence of metastasis, high recurrence within 1-3 years and a high mortality rate. Therefore, identifying factors that facilitate tumor growth and metastases have the potential to serve as novel molecular targets for breast cancer therapy. Mucin1 (MUC1), a glycoprotein associated with chemoresistance, is aberrantly overexpressed in TNBC and facilitates growth and metastasis of TNBC cells. This occurrence can be partially attributed to MUC1 interaction with hypoxia-inducible factor alpha (HIF1α), a key regulator of glycolysis. As metastasis is the leading cause of cancer related deaths, this process relies on the ability of tumor cells to adapt to the changing nutrient levels in the surrounding stroma. In the present study we examined how signaling through MUC1 facilitates metabolic phenotype in TNBC; thus facilitating tumor growth and metastasis. Our results indicate that MUC1 expression facilitates glucose and glutamine uptake in a panel of TNBC cell lines. Furthermore, metabolomic analysis through liquid chromatography-coupled tandem mass spectrometry approach revealed that MUC1 expression modulates metabolite levels in glycolysis, pentose phosphate pathway and TCA cycle in TNBC cells. Thus, our results support the notion that MUC1 serves as a metabolic regulator in TNBC, facilitating metabolic reprogramming that influences growth of TNBC.

Citation Format: Gennifer D. Goode, Nina Chaika, Vinee Purohit, Venugopal Gunda, Pankaj Singh. MUC1: A metabolic regulator in triple-negative 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 1152.
Abstract 1156: Downregulation of glycine decarboxylase contributes to paclitaxel resistance in ovarian cancer cells
Authors: Kim Jin Young , Shin Sojin , HA Eunyoung , Cha Soondo , Cho Chi-Heum
Published: July 2016

Glycine decarboxylase (GLDC) is a metabolic oncogene that links glycine metabolism with tumorigenesis. In humans, GLDC is part of a multienzyme complex that couples the decarboxylation of glycine to the biosynthesis of serine. The role of GLDC in relation to chemoresistance is unclear. We designed this study to determine the role of GLDC in chemoresistance in ovarian cancer cells. First, we found GLDC is downregulated in paclitaxel resistant ovarian cancer cells (OVCAR3/PTX). Prompted by decreased glucose transport in OVCAR3/PTX cells, we transplanted OVCAR3 and OVCAR3/PTX onto the back of the NOD-scid IL2Rgammanull mice and found that the OVCAR3/PTX tumor grew less rapidly than paclitaxel sensitive ovarian cancer cells (OVCAR3). We also found that GLDC is downregulated in OVCAR3/PTX tumor. In vitro study revealed that the inhibitory effect of siGLDC on proliferation was significantly greater in OVCAR3 than in OVCAR3/PTX. We also found that knockdown of GLDC in OVCAR3 cells rendered ovarian cancer cells PTX-resistant. Finally, we determined GLDC expressions in patients with chemosensitive and chemoresistant serous type ovarian cancer and found that GLDC expression was markedly downregulated in chemoresistant cancer patients. These results suggest GLDC is associated with ovarian cancer chemoresistance.

Citation Format: Jin Young Kim, Sojin Shin, Eunyoung HA, Soondo Cha, Chi-Heum Cho. Downregulation of glycine decarboxylase contributes to paclitaxel resistance in ovarian 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 1156.
Abstract 1170: EphA3 maintains radioresistance in head and neck cancers
Authors: Han Myung Woul , Kim Song Hee , Chang Hyo Won , Nam Hae Yun , Lee Myungjin , Kim Gui Chul , Lee Yoon sun , Kim Mi Ra
Published: July 2016

Radiotherapy is a well-established therapeutic modality used in treatment of many cancers. However, radioresistance remains a serious obstacle to successful treatment. Radioresistance can cause local recurrence and distant metastases in some patients after treatment by radiation. Thus, special emphasis has been given to the discovery of effective radiosensitizers. EphA3 receptors functions contribute to tumor development, modulating cell-cell adhesion, invasion, neo-angiogenesis, tumor growth and metastasis. However, the role of EphA3 in radioresistance remains to be elucidated. Here, we established the stable radioresistant head and neck cancer cell line (AMC HN3R cell line) and identified that EphA3 was overexpressed predominantly in the AMC HN3R cancer cell line through DNA microarray, real time PCR and Western blotting. Additionally, we identified that EphA3 was overexpressed in recurred laryngeal cancer specimen after radiation therapy. And we found that EphA3 mediated the tumor invasiveness and migration and EMT (epithelial mesenchymal transition) related protein expression in AMC HN3R cancer cell line. To investigate the role of EphA3 in modulating the radiosensitivity, we observed the change of survival fraction after transfection EphA3 siRNA. And we found that inhibition of EphA3 enhances radiosensitivity in AMC HN 3R cell line. In conclusion, these results suggest that EphA3 is overexpressed in radioresistant head and neck cancer and can play a crucial role in development radioresistance in head and neck cancers through regulation of EMT pathway.

Citation Format: Myung Woul Han, Song Hee Kim, Hyo Won Chang, Hae Yun Nam, Myungjin Lee, Gui Chul Kim, Yoon sun Lee, Mi Ra Kim. EphA3 maintains radioresistance in head and neck cancers. [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 1170.
Abstract 1172: Expression of notch1, numb, and musashi1 in non-small cell lung cancer
Authors: Kikuchi Hajime , Sakakibara-Konishi Jun , Ikezawa Yasuyuki , Takashina Taichi , Mizugaki Hidenori , Kikuchi Eiki , Kikuchi Junko , Shinagawa Naofumi , Oizumi Satoshi , Hida Yasuhiro , Kaga Kichizo , Kinoshita Ichiro , Dosaka-Akita Hirotoshi , Nishimura Masaharu
Published: July 2016

Background: Deregulation of Notch pathway is associated with carcinogenesis, and Notch signaling is identified as tumor activator in non-small cell lung cancer(NSCLC). The function of Musashi1 has been found to activate Notch signaling through the translational repression of Numb, which represses an intracellular Notch signaling. In NSCLC, the association between Numb or Musashi1 expression and clinicopathological factors or prognosis has remained unclear. In this study, we evaluated the expression of Notch1, Numb, and Musashi1 in NSCLC. Methods: A total of 135 surgically resected NSCLCs were immunohistochemically assessed for Notch1, Numb, and Musashi1 expression. Only cytoplasm and nuclear staining was considered in the evaluation of activated Notch1 expression. The immunoscores were determined, and then its correlation with either the clinicopathological variables or the survival outcomes was analyzed. Results: Immunohistochemical reactivity for Numb, and Musashi1 was detected in the cytoplasm and nuclear of the tumor cells. Notch1 expression was associated with several clinicopathological factors, including pathological histology (p = 0.002), and differentiation of tumor (p = 0.024). Numb expression was also associated with several clinicopathological factors, including sex (p = 0.037), smoking habit (p = 0.033), pathological histology (p = 0.002), and differentiation of tumor (p = 0.015). No correlations were noted between Musashi1 expression and any of the variables. Analysis of cellular biological expression demonstrated a link between low Numb expression and high Notch expression. Multivariate Cox regression analysis showed that positive Numb expression was an independent favorable prognostic factor. Conclusions: We demonstrate that Numb expression was associated with better prognosis in NSCLC. Numb might have the function as tumor suppressor in NSCLC.

Citation Format: Hajime Kikuchi, Jun Sakakibara-Konishi, Yasuyuki Ikezawa, Taichi Takashina, Hidenori Mizugaki, Eiki Kikuchi, Junko Kikuchi, Naofumi Shinagawa, Satoshi Oizumi, Yasuhiro Hida, Kichizo Kaga, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Masaharu Nishimura. Expression of notch1, numb, and musashi1 in non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1172.
Abstract 1176: The role of KIF3A in the suppression of canonical Wnt signaling through the KIF3A and β-arrestin complex, independent of the ciliary mechanism, in non-small cell lung cancer (NSCLC)
Authors: Kim Minsuh , Suh Yong-Ah , Oh Ju-hee , Lee Bo Ra , Kim Joon , Jang Se Jin
Published: July 2016

Aberrant Wnt/β-catenin signaling is implicated in the progression of several human cancers, including non-small-cell lung cancer (NSCLC). However, mutations in Wnt/β-catenin pathway com-ponents are uncommon in NSCLC, and epigenetic mechanisms controlling the Wnt/β-catenin path-way remain unclear. Here, we show that KIF3A, a member of the kinesin-2 motor family, plays a key role in suppressing Wnt/β-catenin signaling in NSCLC cells. Knockdown of KIF3A increases both β-catenin levels and transcriptional activity, with a concomitant promotion of malignant phenotypes, such as enhanced proliferation and migration, and upregulation of stemness markers. KIF3A binds to β-arrestin, and KIF3A depletion allows β-arrestin to form a complex with DVL2 and AXIN, result-ing in β-catenin stabilization. Although primary cilia, of which the biogenesis requires KIF3A, are thought to restrain the Wnt response, pharmacological inhibition of ciliogenesis does not enhance β-catenin activity in NSCLC cells. A correlation between KIF3A loss and worse NSCLC prognosis as well as upregulation of β-catenin and Cyclin D1 further suggests that KIF3A is a suppressor of Wnt/β-catenin signaling and tumorigenesis in NSCLC.

Citation Format: Minsuh Kim, Yong-Ah Suh, Ju-hee Oh, Bo Ra Lee, Joon Kim, Se Jin Jang. The role of KIF3A in the suppression of canonical Wnt signaling through the KIF3A and β-arrestin complex, independent of the ciliary mechanism, in non-small cell lung cancer (NSCLC). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1176.
Abstract 1193: Treatment of breast cancer xenografts with paclitaxel enriches for cancer stem cells that can be targeted by a ROR1-specific antibody
Authors: Zhang Suping , Liu Grace , Lai Sam H. , Zhang Han , Ghia Emanuela M. , Cui Bing , Wu Rongrong , Widhopf George , Yu Jian , Schwab Richard , Messer Karen , Parker Barbara , Kipps Thomas J.
Published: July 2016

Although initially responsive to hormone therapy and chemotherapy, many patients with metastatic breast cancer develop resistant and potentially fatal disease. One of the mechanisms of resistance is the emergence of post-treatment survival of a small number of breast cancer stem cells (CSC), which are relatively resistant to chemotherapy, have self-renewal capacity, and can repopulate the tumor and spread to distal sites. New therapies are required to eliminate breast CSC that could be used alone or in combination with standard therapy to mitigate the risk of developing relapsed disease.

In this study, treatment of immune-deficient mice bearing breast-cancer patient-derived xenografts (PDX) with paclitaxel reduced tumor volumes, but enhanced the proportion of tumor cells that expressed ROR1, an RTK-like orphan receptor that is ordinarily expressed during embryogenesis. We found that breast cancer tumors with high levels of ROR1 expression were enriched for stem-cell gene-expression signatures. Moreover, cells isolated from breast cancer PDX that were positive for CSC markers such as aldehyde dehydrogenase 1 (ALDH1) were enriched for ROR1 expression, and ROR1+-cells isolated from breast cancer PDX had a greater capacity to form spheroids in vitro or re-engraft immune-deficient mice, than did ROR1-negative (ROR1Neg) cells isolated from the same tumor population. Treatment of breast cancer cells with a humanized anti-ROR1 mAb, UC-961, suppressed expression of the polycomb-ring-finger oncogene Bmi-1, and genes encoding proteins implicated in the epithelial-mesenchymal transition. UC-961 also impaired the capacity of breast-cancer cells to form spheroids or to re-engraft immune-deficient mice. Finally, combined therapy with UC-961 and paclitaxel appeared more effective in reducing re-engraftment potential than either agent alone (Table1). Collectively, this study implies that patients with advanced breast cancer may benefit from anti-CSC therapy, either alone or in combination with conventional anti-cancer drugs.

Citation Format: Suping Zhang, Grace Liu, Sam H. Lai, Han Zhang, Emanuela M. Ghia, Bing Cui, Rongrong Wu, George Widhopf, Jian Yu, Richard Schwab, Karen Messer, Barbara Parker, Thomas J. Kipps. Treatment of breast cancer xenografts with paclitaxel enriches for cancer stem cells that can be targeted by a ROR1-specific antibody. [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 1193.
Abstract 1212: Effect of metformin and insulin-/IGF1-receptor inhibitor combination drug therapy in triple negative breast cancer
Authors: Xue Lei , Camacho Laura , Kothapalli Sushma , Jiralerspong Sao
Published: July 2016

Background: Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer with poor survival outcomes. Chemotherapy is the only approved treatment, as there are no targeted therapies. Metformin is an anti-diabetic drug for Type 2 Diabetes. It has been shown to have anti-tumor effects by lowering serum levels of the mitogen insulin, and by pleiotropic effects on cancer cell signaling pathways. BMS-754807 is a potent and reversible inhibitor of the insulin receptor (IR)/insulin-like growth factor 1 receptor (IGF-1R) family kinases that has been shown to have activity in a subset of TNBC. The aim of this study is to evaluate the hypothesis that the combination of metformin and BMS-754807 is more effective than single drug treatment in TNBC.

Experimental design and methods: We treated TNBC cell lines with metformin and BMS-754807 alone and in combination and tested cell viability using MTS assays. Then we used CompuSyn software to analyze the dose-effect of drug combinations for additivity, synergism, or antagonism. We also examined the mechanism by performing Western blots of candidate pathways.

Results: 5 out of the 9 cell lines tested (56%) showed synergism of metformin and BMS-754807 in the drug combination assays. Western blots showed significant inhibition of IR/IGF-1R and PI3K/Akt pathways with the drug combination.

Conclusion: We conclude that the combination of metformin and BMS-754807 is more effective than single drug in a significant proportion of triple negative breast cancer cell lines. This combination may represent effective targeted therapy for a subset of TNBC. Further studies to evaluate this possibility are underway.

Citation Format: Lei Xue, Laura Camacho, Sushma Kothapalli, Sao Jiralerspong. Effect of metformin and insulin-/IGF1-receptor inhibitor combination drug therapy in triple negative 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 1212.
Abstract 1213: Thymidine kinase 1 is on the cell surface of prostate cancer cells
Authors: Weagel Evita G. , Chu Roger P. , Meng Wei , Brog Rachel A. , Townsend Michelle H. , Robison Richard A. , O’Neill. Kim L.
Published: July 2016

Thymidine Kinase 1 (TK1) is a salvage pathway enzyme that assists in DNA repair and is characteristically expressed in the cytosol, but it also presents at an abnormally high level in the serum of cancer patients. We developed a unique mouse-anti-human TK1 antibody (A72), and used it in conjunction with Flow Cytometry, Scanning Electron Microscopy (SEM), and Confocal Microscopy, to explore the presence of TK1 on the cell surface of two prostate cancer cell lines, namely PC3 and DU145. Flow Cytometry was performed using a standard protocol. Briefly, cells were stained with A72 conjugated to FITC. Anti-NF-κB-FITC and a non-specific IgG-FITC were used as negative controls. Anti-Na+/K+-ATPase-FITC was used as positive control. Cells were then analyzed using a Flow Cytometer. SEM was carried out by first incubating cells with anti-Na+/K+-ATPase and anti-TK1 conjugated to biotin. After probing with streptavidin-gold solution, cells were imaged to examine the presence of gold particles on cell surface. Our data demonstrated TK1-positive populations after flow cytometry analysis, and the presence of gold particles on the membrane of prostate cancer cells. The presence of TK1 on plasma membrane of PC3 and DU145 was further confirmed by Confocal Microscopy. Cells were first stained with A72-FITC and a red plasma membrane stain, then consecutively imaged with a Confocal Microscope. Results showed that A72-FITC associate with the cells in the same region that the membrane had been stained red, concluding that the A72 binds to the cell membrane. These data strongly suggest that TK1 is located on the surface of prostate cancer cells, and indicate that TK1 may be used as a new immunotherapy target for prostate cancer treatment.

Citation Format: Evita G. Weagel, Roger P. Chu, Wei Meng, Rachel A. Brog, Michelle H. Townsend, Richard A. Robison, Kim L. O’Neill. Thymidine kinase 1 is on the cell surface of prostate 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 1213.
Abstract 1214: Development of anti-5T4 antibody-drug conjugates, ZV05-ADCs for targeted cancer therapy in different type of cancers
Authors: Li Zhaohui , Xie Zhangming , Zhang Qian , Fang Jie , Wu Liqin , Fang Hedi , Zhang Hong , Zhu Tong , Chen Gang , Miao David , Cao Sheldon
Published: July 2016

5T4 is an N-glycosylated transmembrane 72 kDa glycoprotein expressed in a number of carcinomas, and has been explored as a target for cancer therapy in different type of cancers, including NSCLC, RCC, colorectal, ovarian, pancreatic and gastric cancers. The expression of 5T4 in normal tissue is very limited, but widespread in malignant tumors throughout their development, making it an attractive target for antibody-drug conjugate (ADC). In this aspect, a 5T4-MMAF ADC from Pfizer is under clinical investigation in phase I/II trials.

We have developed novel anti-5T4 ADCs, using our proprietary conjugation technologies. A highly potent anti-5T4 antibody ZV05 was conjugated to a variety of internally developed linkers and toxins and purified to populations containing defined DAR (Drug-Antibody Ratio) numbers. The ADCs were studied both in vitro and in vivo for their activity against a variety of tumors. The results showed that the lead ZV05 ADCs were potent towards a bunch of cancer cell lines that express 5T4, and also dramatically inhibited tumor growth in several models in vivo. In the xenograft studies, a single dose of the lead ZV05 ADCs completely wiped out the tumors and kept the tumor volume down for the whole period of studies. ZV05 ADCs are currently under further evaluation to select a candidate for clinical treatment of 5T4-positive cancers.

Citation Format: Zhaohui Li, Zhangming Xie, Qian Zhang, Jie Fang, Liqin Wu, Hedi Fang, Hong Zhang, Tong Zhu, Gang Chen, David Miao, Sheldon Cao. Development of anti-5T4 antibody-drug conjugates, ZV05-ADCs for targeted cancer therapy in different type of cancers. [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 1214.
Abstract 1216: Assays for the selection and functional characterization of antibody-drug conjugates at the National Research Council of Canada
Authors: Jaramillo Maria L. , Meury Luc , Jolicoeur Normand , Banville Myriam , Tao Limei , O’Connor McCourt. Maureen
Published: July 2016

One of the most promising and fastest growing classes of cancer therapeutics builds on the molecular targeting abilities of antibodies by combining them with drugs to generate highly specific antibody-drug conjugates (ADCs). However, the development of ADCs requires time-consuming selection of the antibody for every target and cancer type. Screening technologies based on the use of conjugated secondary antibodies provide a fast and efficient surrogate assay from which to identify which antibodies are best internalized and suitable for immunoconjugate development into ADCs.

As part of its integrated antibody development initiative, NRC has isolated and characterized anti- mouse Fc and anti-human Fc monoclonal antibodies to serve as very selective detective reagents for various IgG isoforms. We have shown that these secondary antibodies are species specific, selective and of high affinity. Furthermore, they exhibit high specific potency and low background toxicity after conjugating them to drugs (DM1, MMAE) or immunotoxins(saporin) in cell viability studies. By combining this methodology with our proprietary mRNA and DNA expression database for the selection of appropriate cell lines, we plan on screening thousands of NRC antibodies generated against variety of cancer associated cell surface targets for ADC development.

In addition to secondary conjugate cytotoxicity assays, we have developed a suite of assays based on cellular accumulation, endosomal routing and activation (intracellular drug release) of antibody drug conjugates. These assays can all contribute to our mechanistic understanding of ADCs under development. Combined with our strength in biologic production and characterization, this expertise promotes the integration and advancement of NRC's capabilities and strengths in the area of Antibody-Drug Conjugates (ADCs), and can be used to establish strategic collaborations with other Canadian or international partners to develop external or internal NRC antibodies into novel ADC biologics.

Citation Format: Maria L. Jaramillo, Luc Meury, Normand Jolicoeur, Myriam Banville, Limei Tao, Maureen O’Connor McCourt. Assays for the selection and functional characterization of antibody-drug conjugates at the National Research Council of Canada. [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 1216.
Abstract 1220: A novel PTK7-targeted antibody-drug conjugate eliminates tumor-initiating cells and induces sustained tumor regressions
Authors: Damelin Marc Isaac , Bankovich Alex , Bernstein Jeff , Lucas Justin , Chen Liang , Williams Sam , Park Albert , Aguilar Jorge , Ernstoff Elana , Charati Manoj , Dushin Russell , Jackson-Fisher Amy , Aujay Monette , Lee Christina , Ramoth Hanna , Milton Milly , Hampl Johannes , Lazetic Sasha , Pulito Virginia , Armellino Douglas , Rosfjord Edward , Guffroy Magali , Falahatpisheh Hadi , King Lindsay , Barletta Frank , Stull Robert , Pysz Marybeth , Escarpe Paul , Liu David , Foord Orit , Gibson Brenda , Powell Eric , O’Donnell Christopher , Xin Xiaohua , Gerber Hans Peter , Sapra Puja , Dylla Scott
Published: July 2016

Disease relapse after treatment is common in triple-negative breast cancer (TNBC), ovarian cancer and non-small cell lung cancer (NSCLC). Therapies that target tumor-initiating cells (TICs) should improve patient survival by eliminating the cells that can drive tumor regrowth and metastasis. Here we identify Protein Tyrosine Kinase 7 (PTK7), a highly conserved but catalytically inactive receptor tyrosine kinase, as an antigen that is enriched on TICs in low-passage patient-derived xenografts (PDX) of TNBC, NSCLC and other tumor types. An anti-PTK7 antibody-drug conjugate (ADC) was generated from a humanized anti-PTK7 monoclonal antibody, a cleavable valine-citrulline-based linker and the Aur0101 auristatin microtubule inhibitor. The anti-PTK7 ADC induced sustained regressions of TNBC, NSCLC and ovarian cancer PDX, with improved activity over standard-of-care chemotherapy, and reduced the frequency of TICs as determined by serial transplantation experiments. Moreover, the ADC may have additional mechanisms of action, including an anti-angiogenic effect, that promote anti-tumor immune responses. Together these preclinical results indicate the potential of the anti-PTK7 ADC to improve the long-term survival of cancer patients. The ADC is currently being tested in a Phase 1 clinical trial, from which interim results will be presented.

Citation Format: Marc Isaac Damelin, Alex Bankovich, Jeff Bernstein, Justin Lucas, Liang Chen, Sam Williams, Albert Park, Jorge Aguilar, Elana Ernstoff, Manoj Charati, Russell Dushin, Amy Jackson-Fisher, Monette Aujay, Christina Lee, Hanna Ramoth, Milly Milton, Johannes Hampl, Sasha Lazetic, Virginia Pulito, Douglas Armellino, Edward Rosfjord, Magali Guffroy, Hadi Falahatpisheh, Lindsay King, Frank Barletta, Robert Stull, Marybeth Pysz, Paul Escarpe, David Liu, Orit Foord, Brenda Gibson, Eric Powell, Christopher O’Donnell, Xiaohua Xin, Hans Peter Gerber, Puja Sapra, Scott Dylla. A novel PTK7-targeted antibody-drug conjugate eliminates tumor-initiating cells and induces sustained tumor regressions. [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 1220.
Abstract 123: Multiplatform molecular profiling of invasive lobular breast cancer
Authors: Nunes Raquel A. , Arguello David , Gatalica Zoran , Reddy Sandeep , Swain Sandra M.
Published: July 2016

Introduction: Invasive lobular breast cancer (ILC) is the second most common subtype of invasive breast cancer accounting for 10% of breast cancer diagnosis. ILC has particular histological and clinical characteristics and a distinct response to therapy. Characterizing the molecular alterations in ILC may lead to an improved understanding of its biology and provide new therapeutic options. The purpose of this study is to describe the molecular profile of ILC and compare it to the one of invasive ductal cancer (IDC).

Methods: Three-hundred and thirty nine pure ILC specimens profiled from January 2012 - November 2015 were evaluated (Caris Life Sciences, Phoenix, AZ). Multiplatform profiling consisted of gene sequencing (next generation sequencing [NGS]), gene amplification (CISH or FISH), and protein expression (immunohistochemistry [IHC]). Molecular characteristics of estrogen receptor (ER) positive and human epidermal growth receptor factor 2 (HER2) negative pure ILC (n = 236) and IDC (n = 286) were compared.

Results: By IHC, ER expression was present in 87.7% (277/316), progesterone receptor in 59.6% (198/313), HER2 in 3.5% (11/313), androgen receptor in 87% (262/301), PD-L1 in 8.1% (12/148) and PTEN in 63.3% (198/313). Amplifications were detected in MYC (7.7%, 2/26), EGFR (8.3%, 2/24), ERBB2 (4.5%, 13/290) and TOP2A (1.3%, 3/236). Mutations were detected in AKT1 (4.7%, 9/191), ATM (3.7%, 7/190), BRCA1 (4.2%, 4/96), BRCA2 (9.5%, 9/95), ERBB2 (7.5%, 14/186), PIK3CA (54.5%, 103/189), PTEN (7.9%, 15/189), and TP53 (13.4%, 25/186). A comparison of ER-positive/HER2-negative invasive lobular and ductal carcinomas revealed significant differences in AR expression (89.7% vs 79.6%, p = 0.0022), and mutations in CDH1 (10.1% vs 0.0%, p = 0.0001), ERBB2 (8.2% vs 2.1%, p = 0.0079), and TP53 (10.3% vs 31.8%, 0.0001).

Conclusions: Multiplatform testing of this large series of ILC reveals recurrent alterations and a distinct molecular profile when compared to IDC. These support the definition of ILC as biologically distinct entity. High AR expression and high rates of dysregulation along the PIK3CA/AKT/mTOR pathway are consistent with recent reports in the literature.

Citation Format: Raquel A. Nunes, David Arguello, Zoran Gatalica, Sandeep Reddy, Sandra M. Swain. Multiplatform molecular profiling of invasive lobular 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 123.
Abstract 1236: ONC201 sensitivity profiling indicates pronounced sensitivity in lymphoid, prostate, colon and brain tumors
Authors: Tarapore Rohinton , Garnett Mathew , McDermott Ultan , Benes Cyril , Allen Joshua
Published: July 2016

ONC201 is a first-in-class small molecule inducer of the integrated stress response that is currently in phase II clinical trials in select advanced cancers and exhibits a benign safety profile and promising early clinical results. The efficacy of this novel agent has been demonstrated in a variety of preclinical advanced cancer models in multiple indications with an exceptional safety profile that has translated well to the clinic. To better characterize the differential sensitivity of tumor cells to ONC201 we performed in vitro efficacy assays in the Genomic of Drug Sensitivity in Cancer collection of cell lines (>1,000 cell lines), in addition to profiling in NCI60 panel. Sensitivity profiling was assessed by cell viability assays using dose responses curves at concentrations up to 20uM and at 72 hours post-treatment. Ranking the sensitivity dataset by tumor type revealed that lymphoma, colon, prostate and brain cancers were highly responsive to ONC201. The strongest predictor of sensitivity to ONC201 was tumor type, with lymphoma being the strong predictor. Within lymphomas, diffuse histiocytic lymphoma was the most sensitive subtype. Mutations that are frequent in lymphoma were not associated with ONC201 sensitivity.

Recent mechanistic studies have implicated the ER stress response in the early stage mechanism of ONC201 that triggers its downstream antitumor effects. The mutation-agnostic efficacy that is pronounced in lymphomas, prostate, colon, and brain cancers is in accordance with the recent findings that ONC201 induces the integrated stress response through a novel target to trigger is downstream late apoptotic effects. Cancers of the prostate and brain (glioblastoma multiforme) are among some of the solid tumors that are susceptible to induction of apoptosis via the integrated stress response, as they have relatively high basal activation of this pathway due to ER stress. Confirmatory studies revealed that brain cancer cell lines and ex vivo samples indeed possess pronounced sensitivity with nanomolar GI50s, unlike most other tumor types, which is particularly encouraging given the systemic concentrations observed in the first-in-man study. Together, these studies suggest specific advanced cancer indications, such as non-Hodgkin's lymphoma, multiple myeloma, and glioblastoma as promising lead indications for this novel agent that are being evaluated in phase II clinical trials.

Citation Format: Rohinton Tarapore, Mathew Garnett, Ultan McDermott, Cyril Benes, Joshua Allen. ONC201 sensitivity profiling indicates pronounced sensitivity in lymphoid, prostate, colon and brain 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 1236.
Abstract 1241: Podophyllotoxin acetate (PA) as an anti-cancer drug candidate regulates various malignancy traits of non-small cell lung cancer
Authors: Cho Jeong Hyun , Hong Wan Gi , Hwang Sang-Gu , Um Hong-Duck , Park Jong Kuk
Published: July 2016

Here, we reported that podophyllotoxin acetate (PA) showed various aniti-cancer effects: induction of several cell death pathways, inhibition of γ-ionizing radiation (IR)-induced cancer invasion, chemo-sensitizer effect with topoisomerase (TOP) inhibitors against non-small cell lung cancer (NSCLC) cell lines as follows. (1) Stimulation of cell death via various mechanisms. First, PA dose-dependently induced cell cycle arrest at G2/M phase, as shown by accumulation of the mitosis-related proteins, p21, survivin and aurora B. This arrest was due to the PA-induced inhibition of microtubule polymerization that related to DNA damage (reflected by accumulation of γ-H2AX) resulting induction of intrinsic/extrinsic damage apoptotic pathways. PA also activated the pro-apoptotic ER stress pathway, as evidenced by increased expression levels of BiP, CHOP, IRE1-a, phospho-PERK, and phospho-JNK. Next, PA induced autophagy that reflected the expression of beclin-1, Atg3, Atg5 and Atg7, and the cleavage of LC3. (2) Inhibition of IR-induced cancer invasion. IR increased the invasion/migration of A549 cells, and this effect was decreased by PA treatment that the expressions/activities of matrix metalloprotase (MMP) -2, MMP-9, and vimentin, suggesting that PA could block the IR-induced epithelial-mesenchymal transition (EMT). In this study, we also identified IR-induced new signaling pathway, EGFR - p38/ERK - CREB-1/STAT3 - EMT pathway, that was inhibited by PA. (3) Chemo-sensitizer effect conjugated with TOP inhibitors - etoposide (Eto) and camptothecin (Cpt). Combination Index (CI) values of the combinations showed synergistic effects between PA and topoisomerase inhibitors. Combination with PA and Eto/Cpt promotes disruption of dynamics of actin filaments, which subsequently enhanced apoptotic cell death that accompanied to increased phosphorylation of p38. We also observed these combinations also inhibited activation of CREB-1, synergistically. Therefore, this study indicated that PA functions as a chemosensitizer by enhancing apoptosis through activation of p38/caspases apoptotic axis and suppression of CREB-1.Taken together; these results suggest PA is a new anti-cancer drug candidate that could control several malignancy traits of cancer including uncontrolled cell cycle and IR-induced invasion.

Citation Format: Jeong Hyun Cho, Wan Gi Hong, Sang-Gu Hwang, Hong-Duck Um, Jong Kuk Park. Podophyllotoxin acetate (PA) as an anti-cancer drug candidate regulates various malignancy traits of non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1241.
Abstract 1245: RRx-001 is effective in temozolomide-sensitive and resistant GBM
Authors: Steri Veronica , Oronsky Bryan , Scicinski Jan , Bergers Gabriele
Published: July 2016

Glioblastoma Multiforme (GBM) is one of the most aggressive malignancies in humans, being highly invasive and drug-resistant, limiting the effectiveness of surgery, chemotherapy and radiotherapy. Concurrent Temozolomide (TMZ) and radiation is the standard of care for newly diagnosed GBM patients, however, patients respond poorly and prognosis remains dismal. TMZ response is limited to patients with hypermethylation of the methylguanine methyltransferase (MGMT) promoter. Despite recent advances in therapeutic strategies, treating GBM is still an unsolved clinical challenge requiring the investigation of novel approaches with better responses over approved therapies. RRx-001 is a novel ROS-mediated systemically non-toxic chemo-sensitizing epigenetic agent with vascular normalizing properties under investigation in Phase II clinical trials in brain metastases and GBM.

We assessed whether RRx-001 is an effective approach for treating GBM by testing its effects alone and in combination with TMZ both in vitro as well as in GBM mouse models. RRx-001 induced cytotoxicity in human TMZ-sensitive as well as resistant patient-derived GBM cell lines. In addition, its effect in TMZ-sensitive GBM cells was greater than the one induced by TMZ alone.

We then asked whether the in vitro effects of RRx-001 translated to therapeutic efficacy in vivo. To this end, we injected mice intracranially with a TMZ-resistant murine mesenchymal GBM cells, and treated tumor-bearing mice with RRx-001 alone and in combination with TMZ. RRx-001 treatment alone modestly prolonged survival compared to control cohorts, whereas the combination of RRx-001 and TMZ substantially prolonged survival compared to controls and either treatment alone. Moreover, histological evaluation of GBM tumors revealed that combination of RRx-001 and TMZ induced vessel hyperdilation and apoptosis compared to RRx-001 alone. Thus, in vivo RRx-001 combined to TMZ may also indirectly regulate tumor growth by modifying the tumor stroma.

These data suggest that TMZ may still affect the microenvironment in TMZ-resistant tumors and that RRx-001 enhances these effects by both directly and indirectly restricting GBM growth, making it a promising therapeutic agent alone and in combination with TMZ.

Citation Format: Veronica Steri, Bryan Oronsky, Jan Scicinski, Gabriele Bergers. RRx-001 is effective in temozolomide-sensitive and resistant GBM. [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 1245.
Abstract 1250: Antitumor activities of CD161, a structurally novel and orally bioavailable BET inhibitor, in leukemia and triple negative breast cancer cells in vitro and in vivo
Authors: Bai Longchuan , Zhao Yujun , Liu Liu , Yang Chao-Yie , McEachern Donna , Stuckey Jeanne , Meagher Jennifer , Wen Bo , Sun Duxin , Wang Shaomeng
Published: July 2016

Bromodomain and extraterminal (BET) proteins, such as bromodomain-containing protein 2 (BRD2), BRD3 and BRD4, bind lysine acetylated proteins and regulate gene transcription. Several BET inhibitors are currently under clinical development for hematological malignancies and solid tumors. We have recently developed a novel class of BET inhibitor, CD161, which binds the bromodomain 1 (BD1) and BD2 domains of BRD2, BRD3, BRD4 and BRDT with Ki less than 10 nM. Co-crystal structure of CD161 and BRD4 BD2 at 2.5 Å resolution confirmed that the docked CD161 occupies the acetyl-lysine binding pocket of BD2. CD161 has >1000-fold selectivity over other bromodomain containing proteins including CREBBP, ATAD2B, TRIM24 and ASH1L. In human leukemia cell lines MV4-11 and MOLM-13, CD161 strongly inhibits cell growth and induces apoptosis, with IC50s <100 nM. Furthermore, CD161 exerts potent growth-inhibitory activities in a panel of breast cancer cell lines representative of triple-negative breast cancer (TNBC), with IC50s in the sub micromolar range. At 20-50 mg/kg daily oral dosing for 2 weeks, CD161 significantly inhibited the growth of MV4-11 and MDA-MB-231 tumors (TGI: 55-80%) in mouse xenograft models and had no significant effect on body weights. Pharmacokinetics studies show that CD161 has a T1/2 of 2-3 hours and oral bioavailability (F%) of 93% in male Sprague−Dawley rats. Transcriptional profiling revealed that CD161 causes broad transcriptional changes in TNBC cell lines. Our preliminary results suggest the therapeutic potential of CD161 in human leukemia and TNBC.

Citation Format: Longchuan Bai, Yujun Zhao, Liu Liu, Chao-Yie Yang, Donna McEachern, Jeanne Stuckey, Jennifer Meagher, Bo Wen, Duxin Sun, Shaomeng Wang. Antitumor activities of CD161, a structurally novel and orally bioavailable BET inhibitor, in leukemia and triple negative breast cancer cells in vitro and in vivo. [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 1250.
Abstract 1251: Significant growth suppressive effects of TOPK and MELK inhibitors in kidney cancer
Authors: Kato Taigo , Inoue Hiroyuki , Imoto Seiya , Tamada Yoshinori , Miyamoto Takashi , Matsuo Yo , Nakamura Yusuke , Park Jae-Hyun
Published: July 2016

Kidney cancer is the seventh most common cancer and the tenth most common cause of cancer death for men, and it is the tenth most common cause of cancer for women. The 5-year disease-specific survival has improved from about 50% in 1975-1977 to 65% in 2000-2005, although that of patients at an advanced stage still remains poor with less than 10%; 30% of patients who underwent surgery of localized kidney cancer develop distant metastasis and have limited therapeutic options such as tyrosine kinase inhibitor (TKI) and mammalian target of rapamycin (mTOR) inhibitors. In addition, the clinical effects of these drugs are limited and patients often discontinue these drugs due to severe side effects including hand-foot syndrome, liver dysfunction and interstitial pneumonia. Therefore, development of more effective therapy for kidney cancer is eagerly expected. T-lymphokine-activated killer cell-originated protein kinase (TOPK) and maternal embryonic leucine zipper kinase (MELK) are promising molecular targets that play critical roles in cancer cell proliferation and maintenance of their stemness. In this study, we investigated growth promotive effect of TOPK and MELK in kidney cancer and found a complex feedback mechanism of these two molecules. Blockade of either of these two kinases effectively caused downregulation of forkhead box protein M1 (FOXM1) activity which is known as an oncogenic transcriptional factor in various types of cancer cell. On the other hand, knockdown and overexpression of FOXM1 caused downregulation and upregulation of both TOPK and MELK mRNA levels, respectively, suggesting that FOXM1 is a critical transcriptional factor for these two molecular targets. Small molecular compound inhibitors against TOPK (OTS514) and MELK (OTS167) effectively suppressed the kidney cancer cell growth and the combination of these two compounds additively induced the very strong growth suppressive effect on kidney cancer cells. Collectively, our results suggest that both TOPK and MELK are promising molecular targets for kidney cancer treatment and that dual blockade of OTS514 and OTS167 should produce synergistic anti-tumor effects with low risk of side effects.

Citation Format: Taigo Kato, Hiroyuki Inoue, Seiya Imoto, Yoshinori Tamada, Takashi Miyamoto, Yo Matsuo, Yusuke Nakamura, Jae-Hyun Park. Significant growth suppressive effects of TOPK and MELK inhibitors in kidney 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 1251.