bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3200: Targeting PHF5A for the treatment of glioblastoma and other Myc-driven cancers
Authors: Mhyre Andrew J. , Turnbaugh Shanon , Morris Shelli M. , Xin Hu , Paddison Patrick J. , Ferrer Marc , Olson James M.
Published: July 2017

Glioblastoma multiforme (GBM) is one of the most aggressive and invasive types of brain cancer, but targeted treatment options remain elusive. The standard of care (surgery chemotherapy and radiation) falls far short of where it should be with two-year survival rates less than 10%. Using stem cell isolates from GBM patients, we found that perturbing PHF5A, a component of the spliceosome machinery, was lethal and caused hundreds of genes to be mis-spliced. These mis-splicing events included both exon skipping and intron inclusions. In contrast, similar levels of PHF5A suppression in normal control stem cells and astrocytes failed to induce cell death and mis-splicing, indicating that PHF5A plays a specific role in the cancer biology. Moreover, when normal astrocytes were transformed with the Myc oncogene, they became sensitive to PHF5A perturbation. Taken together, these results suggested that specifically inhibiting PHF5A would be an effective therapy for glioblastoma and other Myc-driven cancers. Specifically targeting PHF5A would also likely result in reduced side-effects seen with general spliceosome inhibitors. Unfortunately, there are currently no known inhibitors that target PHF5A.

In order to discovery novel PHF5A inhibitors, we created a mini-gene mis-splicing reporter assay that was sensitive to both general spliceosome inhibitors and PHF5A perturbation. In a 96-well assay format, the assay was robust with a 200-fold assay window and Z’ values over 0.8. Following miniaturization to a 1536-well format, we conducted a high throughput screening (HTS) campaign testing 450,000 small molecule compounds. The initial hits were retested and counter-screened yielding 381 confirmed actives and we are further interrogating these actives in secondary and tertiary assays. Future efforts will focus on developing an SAR of the lead and backup series and identifying potential liabilities that will be addressed, if necessary, in further lead optimization efforts. We are enthusiastic about the potential of developing a targeted PHF5A inhibitor as a novel and effective therapy for patients and their families fighting GBM and other Myc-driven cancers.

Citation Format: Andrew J. Mhyre, Shanon Turnbaugh, Shelli M. Morris, Hu Xin, Patrick J. Paddison, Marc Ferrer, James M. Olson. Targeting PHF5A for the treatment of glioblastoma and other Myc-driven cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3200. doi:10.1158/1538-7445.AM2017-3200
Abstract 3278: Particulate matter and traffic-related exposures in relation to breast cancer survival
Authors: DuPre Natalie , Poole Elizabeth M. , Holmes Michelle D. , Hart Jaime E. , James Peter , Beck Andrew , Kraft Peter , Laden Francine , Tamimi Rulla
Published: July 2017

Background: Outdoor air pollution is considered a human carcinogen by the International Agency for Research on Cancer, particularly for its influence on lung cancer incidence and prognosis. While air pollution has not been consistently associated with breast cancer incidence, only one study has examined the impacts of air pollution on survival from breast cancer and was unable to consider important predictors of breast cancer mortality. We assessed whether PM exposures (PM2.5, PM2.510, and PM10) were associated with breast cancer specific-death and all-cause mortality.

Methods: The Nurses’ Health Studies (NHS and NHSII) are ongoing nationwide cohorts with detailed longitudinal data on medical history, addresses, lifestyle factors, and causes of death. We included women with confirmed Stage I-III breast cancer and collected additional clinical information from medical records. PM was estimated using GIS-based spatio-temporal models linked to addresses between 1989-2007. We performed Cox proportional hazards models to assess associations of breast cancer specific-death and overall mortality with PM exposures starting at diagnosis and updated over follow-up through June 2014.

Results: There were 1,211 breast-cancer specific deaths among 8,936 women with Stage I-III breast cancer. Overall, increases in PM were not associated with breast cancer survival (PM2.5: HR=1.09 95% CI 0.87, 1.36; PM2.5-10: HR=1.03 95% CI 0.85, 1.24; PM10: HR=1.05, 95% CI 0.89, 1.24, each per 10 μg/m3). However, each 10 μg/m3 increase in PM2.5 was associated with breast cancer death among participants with Stage I disease after adjusting for clinical, demographics and pre-diagnostic lifestyle factors (HR=1.69 95%CI 1.16, 2.47) and after additionally adjusting for post-diagnostic lifestyle factors (HR=1.64 95% CI 1.11, 2.43). Furthermore, PM exposures were associated with a 9-12% increase in all-cause mortality.

Conclusion:

Overall, PM exposures were not associated with breast cancer survival; however, increases in PM2.5 did appear to adversely impact prognosis among Stage I breast cancer patients.

Acknowledgments:

This work was funded by the NIH T32 CA 09001 and by a grant from the Susan G. Komen for the Cure ® (IIR13264020). We thank the NHS and NHSII participants and coordinators.

Citation Format: Natalie DuPre, Elizabeth M. Poole, Michelle D. Holmes, Jaime E. Hart, Peter James, Andrew Beck, Peter Kraft, Francine Laden, Rulla Tamimi. Particulate matter and traffic-related exposures in relation to breast cancer survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3278. doi:10.1158/1538-7445.AM2017-3278
Abstract 3475: Inhibitory effects of ZFP36L1 and ZFP36L2 on the cell proliferation in human colorectal cancer cells
Authors: Suk Fat-Moon , Chen Ya-Ting , Liang Yu-Chih
Published: July 2017

ZFP36 family members include ZFP36 (also named TTP and TIS11), ZFP36L1 (also called BRF1, ERF1 or TIS11B) and ZFP36L2 (also called BRF2, ERF2 and TIS11D), which belong to the CCCH-type zinc finger protein with 2 tandem zinc finger region (TZF). These ZFP36 family proteins can function as RNA-binding protein through binding to AU-rich elements (AREs) in the 3’ untranslated region (3’ UTR) of mRNA, which promote the mRNA degradation or translation repression. Previously, ZFP36 has been found to inhibit cell proliferation through p53-dependent manner. In this study, we’d like to investigate whether ZFP36L1and ZFP36L2 have anti-proliferative activity as same as ZFP36 work. Tetracycline-inducible (Tet-On) system was used to induce the overexpression of ZFP36L1 or ZFP36L2 proteins in T-REx-293 cells by doxycycline (Dox) treatment. While ZFP36L1 or ZFP36L2 was overexpressed, we found that cell proliferation was dramatically inhibited, but didn’t cause cell death significantly. The importance of TZF was confirmed by using TZF mutants, and found that cell proliferation was deprived in HEK-293 cells with overexpression of ZFP36L1 mutant (C135/173R). Using western blot analysis, it was revealed that expression level of p53 protein was increased after the overexpression of ZFP36L1 or ZFP36L2 protein. Whereas, the levels of cell cycle-related proteins including cyclin B1, cyclin D1, cyclin A2 and p21 were decrease. Next, three human colorectal cancer cells HCT116 p53+/+, HCT116 p53-/- and SW620 (mutated p53) cells were used and ZFP36L1 or ZFP36L2 gene was transduced into these three cells by lentivirus. Overexpression of ZFP36L1 or ZFP36L2 also inhibited the cyclin D1 protein expression and cell proliferation in these three cells, however, increased the p53 and p21 protein expression in HCT116 p53+/+ cells, and decreased the c-Myc expression only in SW620 cells. On the other hand, knockdown of ZFP36L1 or ZFP36L2 increased cell proliferation, and mutation in TZF of ZFP36L1 (C135/173R) or ZFP36L2 (C174/212R) lost their anti-proliferative abilities in these three cells. Taken together, the results suggest that ZFP36L1 and ZFP36L2 play a negative role in cell proliferation of human colorectal cancer cells, and the underlying mechanisms might be mediated through downregulation of cyclin D1 and p53-independent pathway.

Citation Format: Fat-Moon Suk, Ya-Ting Chen, Yu-Chih Liang. Inhibitory effects of ZFP36L1 and ZFP36L2 on the cell proliferation in human colorectal cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3475. doi:10.1158/1538-7445.AM2017-3475
Abstract 3542: Coordination of glutamine and glucose metabolism in pancreatic cancer
Authors: Vernucci Enza , Gunda Venugopal , Shukla Surendra , Singh Pankaj K.
Published: July 2017

Pancreatic ductal adenocarcinoma is the fourth leading cause of cancer related deaths in the United States. Due to early metastasis by the time it is diagnosed it advances to advanced stages and becomes unresectable. Oncogene mediated metabolic reprogramming has been shown to promote the growth, maintenance and metastasis of tumors in pancreatic ductal adenocarcinoma. Pancreatic tumors route majority of glucose and glutamine for biomass generation and maintenance of redox potential. The aim of this study was to investigate the link between glutamine and glucose metabolism in pancreatic cancer cells. We primarily focused on the role of glutamine mediated regulation of glucose metabolism. We observed that glutamine deprivation reduces pancreatic cancer cell growth as suggested by the previous studies. We further investigated how pancreatic cancer cells adapt to glutamine deprived conditions. We observed that glucose uptake increases significantly upon glutamine deprivation. However, GLUT1 expression showed a contrasting decreased expression under conditions of glutamine deprivation. Using tandem mass spectrometry based metabolomic analysis we observed that glutamine deprivation does not alter glycolysis. However, the TCA cycle, amino acid metabolism and glutathione metabolism were significantly impacted in glutamine deprived cells. We noticed a significant increase in the levels of hydroxyglutarate and succinyl CoA levels in the TCA cycle. These changes were accompanied with an increase in GSH to GSSG ratio that reflects a possibility for alternative pathways involved in redox maintenance under glutamine deprived conditions. Hence, we conclude that glutamine metabolism significantly impacts the uptake and metabolism of glucose in pancreatic cancer cells.

Citation Format: Enza Vernucci, Venugopal Gunda, Surendra Shukla, Pankaj K. Singh. Coordination of glutamine and glucose metabolism in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3542. doi:10.1158/1538-7445.AM2017-3542
Abstract 358: LTB and LTBR mediates alternative Nf-kB activation through NIk and RELB/NF-kB2 to promote cell migration of HNSCC
Authors: Das Rita , Coupar Jamie , saleh Anthony , Clavijo Paul E. , Chen Zhong , VanWaes Carter
Published: July 2017

Nuclear factor-κB (NF-κB) includes a family of NF-κB/REL transcription factors that regulate key inflammatory and cancer genes, and which are aberrantly activated in head and neck squamous cell carcinomas (HNSCC). Lymphotoxin beta/Lymphotoxin beta receptor (LTβ/LTβR) can mediate activation of the “alternative” NF-κB pathway that includes NF-κB-inducing kinase (NIK), Inhibitor-κB kinase α (IKKα), and transcription factors RELB/NF-κB2. While LTβ is expressed in lymph nodes where HNSCC spread, the role of this pathway in pathogenesis of HNSCC is not well studied. Analysis of The Cancer Genome Atlas (TCGA) data reveals that LTβR is amplified or overexpressed in larynx or oral cavity HNSCC, while LTα/β, NIK, (MAP3K14) and RELB are overexpressed in oropharynx and tonsil cancers. Similarly, we observed overexpression of LTβR, NIK, and RELB proteins in subsets of HNSCC cell lines. In UM-SCC46, LTβ enhanced nuclear translocation of RELB/NF-κB2 (p52) subunit proteins, while LTβR siRNA knockdown decreased the expression of its target kinase NIK and RELB/NF-κB2 (p52). NIK knockdown also decreased RELB/NF-κB2 (p52) protein expression, whereas LTβ treatment stabilized NIK, and RELB/NF-κB2 (p52). Functional LTβ-mediated NF-κB activation was examined using a stably transfected NF-κB reporter cell line for β-lactamase/Fret activity. Knockdown of LTβR and NIK decreased NF-κB activity, whereas treatment with LTβ partially restored it. Treatment with a NIK inhibitor (1, 3[2H, 4H]-Iso-Quinoline Dione) reduced the protein expression of NIK, IKKα, and RELB/NF-κB2 (p52) in the cytoplasm and/or in the nucleus, and blocked LTβ induced translocation of RELB to nucleus. NIK and RELB knockdown by siRNA or using NIK inhibitor slowed HNSCC migration. Knockdown of NIK inhibited the LTβ-induced expression of MET, and SERPINE1 genes involved in migration, and knockdown of MET by siRNA inhibited the migration of HNSCC cell lines. Our findings show that LTβ/LTβ receptor promotes activation of the alternative NIK-IKKa-RELB/NF-κB2 pathway, to enhance MET-mediated cell migration in HNSCC. This may help explain why HNSCC exhibit migration and metastasis to local lymph nodes, where LTβ is expressed. The constitutive and LTβ induced activation of NIK and downstream NF-κB alternate pathway proteins, RELB and NF-κB2/p52 could be therapeutic targets in HNSCC.

(Supported by NIDCD intramural projects ZIA-DC-000016, 74)

Citation Format: Rita Das, Jamie Coupar, Anthony saleh, Paul E. Clavijo, Zhong Chen, Carter VanWaes. LTB and LTBR mediates alternative Nf-kB activation through NIk and RELB/NF-kB2 to promote cell migration of HNSCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 358. doi:10.1158/1538-7445.AM2017-358
Abstract 3630: Preclinical evaluation of a BiTE® antibody construct with extended half-life that targets the tumor differentiation marker mesothelin
Authors: Sternjak Alexander , Lee Fei , Wahl Joachim , Rock Dan , Thomas Oliver , Stienen Sabine , Lutterbuese Ralf , Hoffmann Patrick , Raum Tobias , Kufer Peter , Rattel Benno , Coxon Angela , Friedrich Matthias , Bailis Julie
Published: July 2017

Bispecific T cell engager (BiTE®) antibody constructs redirect T cells to induce lysis of tumor cells. The anti-CD19/CD3 BiTE® Blincyto® can deplete target cells in both the blood and tissue compartment in B cell malignancies, suggesting that the BiTE® mechanism of action will also be effective against solid tumors. The tumor differentiation antigen mesothelin (MSLN) is an attractive target for the BiTE® approach. MSLN is highly expressed in >80% of ovarian and pancreatic tumors and mesothelioma. Expression of MSLN in normal tissues appears restricted to mesothelial cell surfaces such as the pleural, pericardial, and peritoneal layer.

Here, we report the preclinical characterization of an anti-MSLN/CD3 BiTE® antibody construct with extended half-life. The BiTE® antibody construct binds MSLN and CD3 with low nM affinity and has low pM cytotoxic activity against MSLN-positive ovarian, pancreatic and lung cancer cell lines in vitro. Activity of the anti-MSLN/CD3 BiTE® antibody construct is observed at low effector to target cell ratios, and in cancer cells that express low levels of MSLN. This BiTE® antibody construct is also cytotoxic to cell lines that are resistant to chemotherapy. In mice, significant tumor growth inhibition of an established ovarian tumor xenograft model was achieved by IV dosing of the anti-MSLN/CD3 BiTE® antibody construct every 5 days. Pharmacokinetic evaluation of the anti-MSLN/CD3 BiTE® antibody construct in non-human primates demonstrated a half-life of 4-10 days. The potency and specificity of the anti-MSLN/CD3 BiTE® antibody construct, together with projected ability to dose once a week IV in humans, supports development of this BiTE® for treatment of MSLN-positive tumors.

Citation Format: Alexander Sternjak, Fei Lee, Joachim Wahl, Dan Rock, Oliver Thomas, Sabine Stienen, Ralf Lutterbuese, Patrick Hoffmann, Tobias Raum, Peter Kufer, Benno Rattel, Angela Coxon, Matthias Friedrich, Julie Bailis. Preclinical evaluation of a BiTE® antibody construct with extended half-life that targets the tumor differentiation marker mesothelin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3630. doi:10.1158/1538-7445.AM2017-3630
Abstract 3855: Excavation of therapeutic targets based on the molecular signature of patient-derived tumor xenografts in gastric cancer
Authors: Chen Zuhua , Gao Jing , Shen Lin
Published: July 2017

Objective: The optimal preclinical models in new targeted drug development are patient-derived tumor xenografts (PDXs) models with definite molecular signatures, which will be excavated and validated in this study. Methods: The genomic profiles of 50 PDXs from advanced gastric cancer (AGC) were analyzed by targeted next-generation sequencing. The Epstein-Barr virus (EBV) infection, microsatellite instability (MSI) status, and expressions of EGFR, HER3, c-Met, PTEN and c-Myc were determined by in situ hybridization and immunohistochemisty, respectively. Results: A total of 581 non-synonymous single nucleotide variations (SNV), 513 Indels, and 225 amplifications were identified in 50 PDXs. Pathway analysis revealed recurrent alterations in the MAPK signaling, ErbB signaling, VEGF signaling, mTOR signaling and cell cycle pathway. The ErbB family amplified in 8% (EGFR), 10% (ERBB2), 2% (ERBB3) and 2% (ERBB4) of the 50 PDXs respectively. Meanwhile, several activating mutations in EGFR (L858R and G719S), ERBB3 (P1212S) and ERBB4 (R1174Q) were also detected in our study. Moreover, we detected 7 PDXs harboring PIK3CA activating mutations (E545K, Q546K, H1047R) and 2 PDXs with PTEN loss. Whether PI3K/Akt/mTOR inhibitors could turn over the primary or acquired resistance of anti-HER2 therapies remains further researches. Genes in the cell cycle pathway altered in 70% of the 50 PDXs. The mutation rates of TP53, BRCA1 and BRCA2 were 28%, 8% and 14%. We also detected amplifications of CCNE1, CCND1 and MYC in 16%, 6% and 20% of the xenografts. In this study, 9 PDXs (18%) were EBV positive, with frequent alterations of ARID1A but rare mutations of TP53, and a robust expression of PD-L1. However, we didn’t observe any loss of MMR protein expressions. Conclusions: Several potential therapeutic targets were identified based on the molecular signature of PDXs in advanced gastric cancer, which will provide evidences for drug development.

Citation Format: Zuhua Chen, Jing Gao, Lin Shen. Excavation of therapeutic targets based on the molecular signature of patient-derived tumor xenografts in gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3855. doi:10.1158/1538-7445.AM2017-3855
Abstract 3970: Targeting the adenosine immunosuppressive pathway for cancer immunotherapy with small molecule agents
Authors: Fons Pierre , Esquerré Michael , Versluys Stéphanie , Mambrini Gigliola , Paillasse Michael , Bell Andy , Schreyer Adrian , Cox Richard , Bickerton Richard , Lisztwan Joanna , Whittaker Mark , Bono Françoise , Johnstone Craig , Hopkins Andrew
Published: July 2017

Standard Immune Checkpoint Therapies, such as anti-CTLA-4 or anti-PD-1/PD-L1 mAb, demonstrate long progression-free-survival for responding patients but with a low response rate. This can be due to additional immunosuppressive mechanisms developed by resistant tumors. The adenosine pathway is one of the main drivers of tumor progression by acting on two key features of the TME (Tumor Micro-Environment); immunity and angiogenesis. Expression of CD73 on tumor cells and on immunosuppressive cell subsets leads to the generation of adenosine from AMP. Adenosine inhibits the biological functions of T lymphocytes infiltrating the tumor by binding to the A2AR (receptor). Evotec and Exscientia are collaborating to accelerate small molecule drug discovery in the field of Immuno-Oncology by integrating a unique biophysical screening approach to the adenosine receptors and CD73 to drive automated medicinal chemistry design with a translational-focused screening cascade, to discover novel agents that target the adenosinergic pathway in immuno-oncology. We have firstly sought to generate novel A2AR antagonists and secondly to create A2AR/CD73 bi-specific inhibitory molecules with a dual pharmacological profile, from fragment screening and de novo design approaches. In vivo combination study demonstrate the advantages of targeting CD73 and A2AR. We have obtained active molecules illustrated by in vitro data on primary human T lymphocytes. These results, have paved the way to an optimized process to identify adenosinergic pathway inhibitors with properties optimized for immuno-oncology.

Citation Format: Pierre Fons, Michael Esquerré, Stéphanie Versluys, Gigliola Mambrini, Michael Paillasse, Andy Bell, Adrian Schreyer, Richard Cox, Richard Bickerton, Joanna Lisztwan, Mark Whittaker, Françoise Bono, Craig Johnstone, Andrew Hopkins. Targeting the adenosine immunosuppressive pathway for cancer immunotherapy with small molecule agents [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3970. doi:10.1158/1538-7445.AM2017-3970
Abstract 399: CPTAC phase II final report
Authors: Thiagarajan Mathangi
Published: July 2017

National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (CPTAC) is a comprehensive and coordinated effort to accelerate the understanding of the molecular basis of cancer through the application of robust, quantitative, proteomic technologies and workflows. The primary goal for this program is to systematically identify proteins that derive from alterations in cancer genomes and related biological processes, integrate the genomic and proteomic data from analysis of common cancer biospecimen and provide this data with accompanying assays and protocols to the community. The phase II effort managed by the Leidos Biomedical Research Inc. had a goal to procure and qualify a minimum of 100 cases from 3 cancer types (Breast, Ovarian and Colon). Tissues and analytes from the 100 cases were sent to proteomic and genomic characterization centers for analysis. The proteomic data were made available through the Data Coordinating Center (DCC) and the genomic data were made available through NCI’s Genomic Data Commons (GDC). The success from the phase II effort has set precedence for a much larger phase III effort that will focus on additional cancer types and cases. Preliminary analysis results confirm the need for the combination of the genomic and proteomic approaches to produce a more comprehensive inventory of the detectable proteins in a tumor and advance our understanding of cancer biology.

Citation Format: Mathangi Thiagarajan. CPTAC phase II final report [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 399. doi:10.1158/1538-7445.AM2017-399
Abstract 4: Inhibitory effects of aprotinin on survival and local invasion of human breast cancer cell lines
Authors: Soleyman-Jahi Saeed , Zendehdel Kazem , Sadeghi Fatemeh , Afshari Ziba , Barati Tahereh , Mohammadnezhad Samad , Ghasemi Sevil , Amanpour Saeed
Published: July 2017

Background: Aprotinin is a nonspecific serine protease inhibitor, which can inhibit kallikrein and many downstream extracellular matrix degrading enzymes. In this study, we assessed inhibitory effects of aprotinin on local invasion of human breast cancer cell lines.

Methods: We assessed effects of aprotinin on local invasion and survival of human breast cancer cell lines MDA-MB231 and SK-BR-3 in vitro. CHEMICON Cell Invasion Assay Kit was used to assess local invasion of tumoral cells. MTT assay was used to determine the anti-proliferative activity of aprotinin on different cancer cell lines. Mean optical densities of duplicate experiments were calculated for both assays. HDF-1 human fibroblast cell line was used as control normal cell. In addition, local invasion and survival of tumor cell lines treated with aprotinin were compared with untreated experiments.

Results: Cancer cell lines showed more invasion compared to HDF-1. Aprotinin significantly decreased the invasiveness of MDA-MB231 in concentration of 1 TIU/ml, 1.3 TIU/ml, and 1.7 TIU/ml in comparison with untreated group (ANOVA P <0.001). Treatment of SK-BR-3 with 1.3 TIU/ml aprotinin trended towards reduction of invasion optical density (P = 0.06). Treatment with different concentrations of aprotinin significantly decreased the surviving fraction and significantly inhibited the growth of all cell lines tested in this study (ANOVA P <0.001).

Conclusion: Aprotinin inhibited the survival and local invasion of human breast cancer cell lines MDA-MB231 and SK-BR-3. Although the growth inhibitory effect was significant in both cell lines, inhibition of local invasion was more pronounced in case of MDA-MB231. Future molecular studies could shed further lights on mechanisms underlying anti-neoplastic effects of aprotinin and its potential therapeutic effects.

Citation Format: Saeed Soleyman-Jahi, Kazem Zendehdel, Fatemeh Sadeghi, Ziba Afshari, Tahereh Barati, Samad Mohammadnezhad, Sevil Ghasemi, Saeed Amanpour. Inhibitory effects of aprotinin on survival and local invasion of human breast cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4. doi:10.1158/1538-7445.AM2017-4
Abstract 4172: Activation of AR signaling by mifepristone enhances prostate cancer growth and impairs enzalutamide response
Authors: Zhou Haiying , Jachan Nadine , Singh Mallika , Tran Chris , McWeeney Dan , Balbas Minna , Schenkein Emily , Zovorotinskaya Tatiana , Jackson Erica L. , Medina Julio , Sun Daqing , Rew Yosup , Du Xiaohui , Eksterowicz John , Yan Xuelei , Zhu Liusheng , Ye Qiuping , Fantin Valeria
Published: July 2017

Androgen receptor (AR) signaling is crucial for normal development and homeostasis of the prostate, and is a key driver of prostate cancer initiation and progression. Hormone therapies that deprive the cancer of androgen have long been a mainstay of prostate cancer treatment. More recently, anti-androgens, such as abiraterone and enzalutamide, have been approved for use in metastatic castration resistant prostate cancer (mCRPC). Evidence also suggests that AR may play an oncogenic role in certain breast cancers. Several recent publications have demonstrated that activation of the Glucocorticoid Receptor (GR) can confer resistance to enzalutamide, and GR has also been shown to provide protection from conventional chemotherapies in other solid tumor indications. Mifepristone, is a synthetic steroidal antagonist of progesterone receptor, and to a lesser extent of GR and AR. It is currently being tested in clinical trials in combination with enzalutamide in mCRPC, and in combination with chemotherapy in triple negative breast cancer (TNBC). We sought to characterize the effect of mifepristone in pre-clinical models of prostate and breast cancer. Here we show that in the absence of androgen, mifepristone acts as a partial AR agonist, and this agonism can only be partially overcome by enzalutamide. We find that in low androgen conditions, mifepristone promotes the growth of prostate cancer cells in vitro and accelerates the growth of prostate tumors in xenograft models. Moreover, when given in combination, mifepristone significantly reduces the efficacy of enzalutamide in the LN-AR xenograft model. We are currently assessing the effects of mifepristone treatment in TNBC. Our findings suggest that partial AR agonist activity of mifepristone may have a negative impact in prostate and other AR positive cancers. We have developed a GR antagonist that is devoid of AR agonism to circumvent undesired effects on proliferation and drug response.

Citation Format: Haiying Zhou, Nadine Jachan, Mallika Singh, Chris Tran, Dan McWeeney, Minna Balbas, Emily Schenkein, Tatiana Zovorotinskaya, Erica L. Jackson, Julio Medina, Daqing Sun, Yosup Rew, Xiaohui Du, John Eksterowicz, Xuelei Yan, Liusheng Zhu, Qiuping Ye, Valeria Fantin. Activation of AR signaling by mifepristone enhances prostate cancer growth and impairs enzalutamide response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4172. doi:10.1158/1538-7445.AM2017-4172
Abstract 4182: The two novel BTK-inhibitors M2951 and M7583 show in vivo anti-tumor activity in pre-clinical models of B cell lymphoma
Authors: Gaudio Eugenio , Tarantelli Chiara , Zucca Emanuele , Rossi Davide , Stathis Anastasios , Bertoni Francesco
Published: July 2017

Background. Diffuse large B cell lymphoma of the activated B cell like type (ABC DLBCL) is an aggressive lymphoma in which the inhibition of the Bruton’s tyrosine kinase (BTK) has shown biologic and clinical relevance. M7583 and M2951 are two novel highly selective irreversible BTK inhibitors (Fest C et al, 2016). M7583 has shown activity in a mouse model of lupus erythematosus (Bender et al, 2016). Here, we evaluated the anti-tumor activity of M2951 and M7583 in an in vivo ABC DLBCL model.

Methods. Xenografts were established by s.c. injecting TMD8 lymphoma cells (100 µL of PBS, 107 cells/mouse) into the left flanks of female NOD-SCID mice (6 weeks of age and around 20g of body weight). Treatments (QD, po) were started with an average tumor volume of 100mm3 on groups of 10 mice each.

Results. NOD-SCID mice engrafted with the ABC DLBCL TMD8 were treated with three different concentrations of M7583 (1 mg/Kg; 3 mg/Kg; 10mg/Kg) or M2951 (5 mg/Kg; 15 mg/Kg; 25mg/Kg) or vehicle, as control group. M2951 slowed tumor growth at all doses starting after 10 days of treatment. M7583 at 3 and 10 mg/Kg showed stronger anti-tumor activity than M2951, already evident after 5 days of treatment (P<0.05) and improving with time (after 10 days, P< 0.005). Moreover, while most groups had to be stopped after 2 weeks of treatment (D30 after engraftment) because of the reaching of the maximum tumor volume, allowed by the ethical veterinary committee, mice treated with M7583 at the two highest doses were able to continue treatment for additional six (3 mg/Kg) or ten days (10mg/Kg), when tumors became bigger than 2000 mm3, indicating a high activity of M7583. No body weight losses and sign of toxicity were detected.

Conclusion. The two novel BTK inhibitors M2951 and M7583 showed promising in vivo anti-tumor activity in an ABC-DLBCL model. These data provide the rational for further investigating these compounds.

Citation Format: Eugenio Gaudio, Chiara Tarantelli, Emanuele Zucca, Davide Rossi, Anastasios Stathis, Francesco Bertoni. The two novel BTK-inhibitors M2951 and M7583 show in vivo anti-tumor activity in pre-clinical models of B cell lymphoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4182. doi:10.1158/1538-7445.AM2017-4182
Abstract 4183: A novel selective DNA-PK inhibitor, M3814, as a potential combination partner of Etoposide and Cisplatin in the therapy of lung cancer
Authors: Sirrenberg Christian , Zimmermann Astrid , Grombacher Thomas , Vassilev Lyubomir T. , Damstrup Lars , Zenke Frank T. , Blaukat Andree
Published: July 2017

M3814 is a potent and selective inhibitor of DNA-PK, one of the key cellular regulators of DNA damage induced by ionizing radiation or certain cytostatics used in the treatment malignant disease. One of these drugs, Etoposide, induces double strand breaks (DSB) in cellular DNA. DSBs are most difficult to repair and, if left unrepaired, can lead to induction of cell cycle arrest and/or apoptosis and ultimately cell death. DNA-PK plays a critical role in the repair of DSB via the non-homologous end-joining pathway. M3814 was tested for activity in combination with Etoposide in a panel of 98 cancer cell lines derived from lung cancer. A broad potentiation effect of DNA-PK inhibitor was observed in most cancer cell lines. As a rule, cell lines sensitive to Etoposide demonstrated increased sensitivity to the combination. M3814 did not show significant effect on cancer cell growth/viability in combination with Cisplatin compared to Cisplatin alone at concentrations that effectively inhibit DNA-PK activity. At the same time, M3814 did not negatively affect the antitumor activity of Cisplatin. The therapeutic effect of M3814 in combination with the standard of care (SoC) regimen of Etoposide and Cisplatin was tested in the human small cell lung cancer xenograft model, NCI-H520. Triple combination of M3814, Etoposide and Cisplatin resulted in increased efficacy compared to SoC treatment arm. Since myeloid and lymphoid suppression is one of the dose limiting toxicities of the SoC regimen in patients, the effect of the triple combination on myeloid and lymphoid blood cells was investigated in immunocompetent mice. Whereas the SoC treatment showed reduction of the myeloid and lymphoid compartments, the addition of M3814 did not additionally reduce these cells neither in the treatment nor the recovery phase. Our results warrant further investigations to explore the potential of the combination in the clinical setting.

Citation Format: Christian Sirrenberg, Astrid Zimmermann, Thomas Grombacher, Lyubomir T. Vassilev, Lars Damstrup, Frank T. Zenke, Andree Blaukat. A novel selective DNA-PK inhibitor, M3814, as a potential combination partner of Etoposide and Cisplatin in the therapy of lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4183. doi:10.1158/1538-7445.AM2017-4183
Abstract 4198: Highly potent and selective DNA-PK inhibitor M3814 with sustainable anti-tumor activity in combination with radiotherapy
Authors: Fuchss Thomas , Mederski Werner W. , Emde Ulrich , Buchstallter Hans-Peter , Zenke Frank , Zimmermann Astrid , Sirrenberg Christian , Vassilev Lubo , Damstrup Lars , Urbahns Klaus , Blaukat Andree
Published: July 2017

Physical or chemical agents that damage DNA such as ionizing radiation are among the most widely used classes of cancer therapeutics today. Double strand breaks (dsb) generated in DNA by radiation induce a multitude of cellular responses, including DNA repair, cell cycle arrest or cell death once the damage is left unrepaired. A complex set of molecular events are responsible for DNA repair mediated by two major repair mechanisms - homologous recombination (HR) or non-homologous end joining (NHEJ). DNA-PKcs together with its regulatory protein subunits Ku70 & Ku80, is an integral component of the NHEJ process and considered an attractive intervention point to inhibit DNA dsb repair. We have developed an orally bioavailable, highly potent & selective inhibitor of DNA-PK, M3814, for cancer therapy in combination with DNA damaging modalities such as radiation & radio-chemotherapy. Here, we present the optimization of initial hit matter & its structure-activity relationships leading to M3814, its chemical structure (first-time disclosure) & preclinical characterization using biochemical, cellular & human tumor xenograft models. M3814 sensitized multiple tumor cell lines to radiation therapy in vitro and strongly enhanced the anti-tumor activity of ionizing radiation in vivo with complete tumor regression by applying a clinically relevant fractionated radiation regimen. These effects are due to inhibition of DNA-PK protein kinase activity as demonstrated by the levels of DNA-PK autophosphorylation in human tumor cell lines & xenograft tumors. M3814 is currently being investigated in PhI/II clinical trials.

Citation Format: Thomas Fuchss, Werner W. Mederski, Ulrich Emde, Hans-Peter Buchstallter, Frank Zenke, Astrid Zimmermann, Christian Sirrenberg, Lubo Vassilev, Lars Damstrup, Klaus Urbahns, Andree Blaukat. Highly potent and selective DNA-PK inhibitor M3814 with sustainable anti-tumor activity in combination with radiotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4198. doi:10.1158/1538-7445.AM2017-4198
Abstract 4302: Disulfiram possesses antitumor activity and inhibits cancer stem-like properties via inhibition of STAT3 activation in triple-negative breast cancer cells
Authors: Kim Yoon-Jae , Lee Nahyun , Oh Eunhye , Sung Daeil , Cho Tae-Min , Kim Ji Young , Seo Jae Hong
Published: July 2017

Disulfiram (DSF), a clinically used treatment for alcohol dependence, is considered as a potential anti-cancer agent due to its cytotoxicity in many cancer cell types on copper (Cu)-dependent processes. However, the molecular mechanisms underlying DSF-induced antitumor activity in triple-negative breast cancer (TNBC) are still poorly understood. Here, we hypothesized that DSF induces antitumor effects by targeting cancer stem cells (CSCs) and STAT3 signaling in TNBC. We then examined the effects of DSF in combination with Cu on the cell viability and apoptosis, cell proliferation and survival, as well as CSC-like properties in MDA-MB-231 and 4T1 TNBC cell lines. DSF/Cu effectively induced cytotoxicity and apoptotic cell death, concomitant with caspase-3 activation and PARP cleavage. DSF/Cu also significantly suppressed cell migration and mammosphere formation, which are accompanied by a remarkable decrease in the CD44+/CD24- (or CD49f+/CD24+ in mouse 4T1 cells) stem-like population and ALDH1 activity. These effects were functionally associated with the inhibition of STAT3 phosphorylation (Tyr705). Furthermore, DSF/Cu treatment resulted in the downregulation of cyclin D1 and survivin expression, which are downstream targets of STAT3 signaling. Interestingly, phosphorylated STAT3 and STAT3 downstream signaling were highly upregulated in the mammospheres, and these upregulated molecules were significantly decreased by DSF/Cu treatment. In MDA-MB-231-derived xenograft model, DSF administration elicited a significant reduction in tumor growth with a noticeable downregulation of CD44, ALDH1 and phosphorylated STAT3 levels. Taken together, our results suggest that therapeutic treatment to DSF can effectively manifest antitumor activity by reducing CSC-like properties, and that STAT3 could be a key molecule of aggressive TNBC that is targeted by DSF.

Note: This abstract was not presented at the meeting.

Citation Format: Yoon-Jae Kim, Nahyun Lee, Eunhye Oh, Daeil Sung, Tae-Min Cho, Ji Young Kim, Jae Hong Seo. Disulfiram possesses antitumor activity and inhibits cancer stem-like properties via inhibition of STAT3 activation in triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4302. doi:10.1158/1538-7445.AM2017-4302
Abstract 4362: Coordinated regulation of several microRNAs in lung cancer patients
Authors: Ferrer Irene , Salinas Ana , Marrugal Angela , Zugazagoitia Jon , Carnero Amancio , Paz-Ares Luis , Molina-Pinelo Sonia
Published: July 2017

Background Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancers. NSCLCs are mainly classified adenocarcinoma and squamous cell carcinoma. It seems more and more obvious that specific molecular analyses are necessary to elucidate the complexity of clinically relevant phenotypes which determine the lung carcinogenesis. This includes the role of microRNAs (miRNAs) in the disease, involved in the complexity of gene expression regulation. Different modulator mechanisms are involved in the biogenesis and function of miRNAs, one of them is DNA methylation. Alterations in these mechanisms may contribute to the tumor genesis and progression. The purpose of this study was to assess miRNA methylation patterns in patients with NSCLC to study the potential of coordinate regulation of miRNAs as a relevant mechanism involved in this pathogenesis.

Materials and Methods DNA methylation of gene clusters was analyzed by Illumina 70 subjects. The samples were divided into two cohorts. A first cohort constituted from 47 patients who had undergone surgical resection for clinical early stage NSCLC. A second group from 23 subjects was used as health cohort. DNA was extracted using the QIAamp DNA Mini Kit. For each assay, 500 ng of DNA was treated with sodium bisulfate using EZ DNA Methylation™ Kit and cleaned with ZR-96 DNA Clean-up Kit™, before standard Illumina amplification, hybridization, and imaging steps. Methylation data were processed using the RnBeads R package.

Results We identified two clusters clearly corregulated by methylation in lung cancer, involving 49 miRNAs differentially hypomethylated in tumor samples respect to control tissue. Some of these miRNAs have been implicated in several pathways in cancer.

Conclusions Our results strongly imply hypomethylation of two miRNA clusters, which represent key targets in unravelling of the mechanism of lung tumorigenesis.

Citation Format: Irene Ferrer, Ana Salinas, Angela Marrugal, Jon Zugazagoitia, Amancio Carnero, Luis Paz-Ares, Sonia Molina-Pinelo. Coordinated regulation of several microRNAs in lung cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4362. doi:10.1158/1538-7445.AM2017-4362
Abstract 4455: Identification of novel oncogene, copine-7 (CPNE7), in colorectal cancer
Authors: JEONG DONGJUN , Ban Seona , Kim Hyungjoo , Oh Seunghyun , Ji Sanghee , Kim Han Jo , Bae Sang Byung , Ahn Tae Sung , Kim Chang-Jin , Lee Moon Soo , Baek Moo-Jun
Published: July 2017

Background: CPNE7, a member of copine family, is composed of calcium-dependent membrane-binding proteins. The encoded protein may play a role in calcium-mediated intracellular processes and membrane trafficking mainly. Although this protein was known as tumor suppressor in breast cancer, we identified the possibility as an oncogene in colorectal cancer. In this study, we performed to evaluate the oncogenic functions of CPNE7 in the colorectal cancer cell line and the clinical significance of CPNE7 expression in colorectal cancer patients.

Method: The colorectal cancer cell lines (SW480, SW620, HCT116 and HT29) were transfected with siRNA for knockdown CPNE7. The oncogenic functions were identified in the transfected cell lines and compared them with CPNE7 highly expressing control cell lines. In vitro functional studies included cell proliferation assay, migration assay, invasion assay and semisolid agar colony forming assay. The clinical significance of CPNE7 expression was evaluated in 250 cases of colorectal cancer tissue by immunohistochemistry.

Results: The knockdowned cell lines significantly showed lower oncogenic function (proliferation, migration, invasion and colony forming) compared to the CPNE7 highly expressing control cell lines (p<0.05). The overall survival rate was decreased in patients of CPNE7 high expression (p=0.03). Multivariate and univariate Cox-regression analysis showed that CPNE7 high expression was an independent prognostic factor in colorectal cancer (HR=1. 54, 95% CI=1. 00-2.38: p=0. 048 / HR=1. 58, 95% CI=1. 03-2.41: p=0. 032).

Conclusion: This study announces that CPNE7 is a novel oncogene, an independent prognostic factor and therapeutic target in colorectal cancer. In the future, studies on in vivo assay and identifying oncogenic signal pathways of CPNE7 in colorectal cancer are necessary.

Citation Format: DONGJUN JEONG, Seona Ban, Hyungjoo Kim, Seunghyun Oh, Sanghee Ji, Han Jo Kim, Sang Byung Bae, Tae Sung Ahn, Chang-Jin Kim, Moon Soo Lee, Moo-Jun Baek. Identification of novel oncogene, copine-7 (CPNE7), in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4455. doi:10.1158/1538-7445.AM2017-4455
Abstract 4511: HIF-1-dependent regulation of creatine kinase metabolism promotes breast cancer invasion and metastasis
Authors: Barch Hilaire , Brooks Danielle L. , Krutilina Raya , Schwab Luciana P. , Parke Deanna , Seagroves Tiffany N.
Published: July 2017

Dysregulated tumor cell metabolism is a hallmark of cancer progression and therapeutic resistance. In a screen for Hypoxia-Inducible Factor (HIF)-dependent genes regulating metabolism, we identified creatine kinase, brain isoform (CKB) as down-regulated in HIF-1 knockout mammary tumor cells. Creatine kinases (CKs) reversibly catalyze the transfer of a high-energy phosphoryl group from ATP to creatine, generating phosphocreatine in the forward reaction, and ATP in the reverse reaction. CKs are up-regulated in a variety of solid tumors, including ovarian, breast, colon, lung and brain. Knockdown of CKB in the polyoma middle T (PyMT) transgenic mouse model of metastatic breast cancer suppressed the production of intracellular ATP and invasion in vitro, and inhibited metastasis from the mammary gland to the lung in vivo. CK activity is known to be inhibited by cyclocreatine, a creatine kinase substrate that represses CK-dependent generation of ATP from phosphocreatine. When female FVB/Nj mice were injected with wild type PyMT cells in a tail vein assay and then treated with cCr (1g/kg/day, IP), lung metastasis was repressed to the same extent as Ckb gene knockdown. Moreover, when cCr therapy was administered 7 days after tail vein injection, cCr was effective in preventing the transition of lung micrometastases to macrometastases. To explore the role of CK activity in regulating cell proliferation, survival in suspension, cellular metabolism and invasion, we next created CKB loss- and gain-of-function models using human breast cancer cell lines, and compared phenotypes to cCr treatment. Whereas deletion of CKB had no effect on cell proliferation or survival in adherent conditions or in suspension, either deletion of CKB or cCr therapy potently reduced ATP levels and invasive potential in vitro. Preliminary data also indicate that pre-treatment of triple negative breast cancer cell lines with cCr sensitizes cells to doxorubicin. Together, these data suggest that inhibition of CK activity may be effective in treating stage IV breast cancer. We are currently testing whether cCr has anti-metastatic efficacy as a monotherapy, or in combination with conventional chemotherapies, using luciferase-labeled patient-derived xenograft (PDX) models.

This work was sponsored by the NIH (CA138488), the Dept. of Defense (BC150640), the METAvivor foundation, and the West Cancer Center in Memphis, TN.

Citation Format: Hilaire Barch, Danielle L. Brooks, Raya Krutilina, Luciana P. Schwab, Deanna Parke, Tiffany N. Seagroves. HIF-1-dependent regulation of creatine kinase metabolism promotes breast cancer invasion and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4511. doi:10.1158/1538-7445.AM2017-4511