bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2543: Circulating s100a9 myeloid-derived suppressor cells are predictive for poor pfs in patients with egfr mutation positive lung adenocarcinoma treated with gefitinib
Authors: Feng Po-Hao , Yu Chih-Teng , Lee Kang-Yun
Published: May 2015

Introduction: Around 10-30% of patients with EGFR-mutation positive lung adenocarcinoma do not have objective response to EGFR-TKI, whilst the majority of the EGFR-mutation positive patients eventually develop acquired resistance. We have recently demonstrated that the distinct S100A9+ myeloid derived suppressor cells (MDSC) predicted poor outcome in response to cisplatin-based chemotherapy in patients with advanced NSCLC. In this study, we tested whether numbers of circulating S100A9+ MDSC, among other potential factors, could be a predictor for treatment response to EGFR-TKI.

Methods: Between 2010-2012, thirty-four patients with pathologically proved, stage IIIB-IV lung adenocarcinoma with sensitizing EGFR-mutations, were prospectively enrolled. Gefitinib was used as the first line treatment. Pretreatment S100A9 MDSC was determined by FACS analysis and serum HGF by ELISA.

Results:

Among the patients, 47.1% (16/34) had Ex19Del and 50.0% (17/34) had L858R mutations. The objective response rate was 84.4% and the median progression-free survival (PFS) 10.16 months (95% CI 8.18-12.14 months). In univariate analysis, body weight loss, brain metastasis and S100A9 MDSC (p = 0.008, 0.022 and 0.014, respectively), but not gender, smoking, type of EGFR mutation, or serum HGF, were significant factors for PFS. Multivariate Cox regression analysis revealed brain metastasis (HR=3.924, 95% CI 1.449-10.627, p=0.007) and S100A9+ MDSC (HR=4.944, 95% CI 1.578-15.490, p=0.006) were both independent factors for PFS. When the medium number of S100A9 MDSC, e.g. 17.9% of the PBMC, was chosen as a cut off level, patients with high MDSC had shorter PFS compared with those with low MDSC (median 7.2 months (95% CI, 5.5-8.9 months) vs. 12.7 months (95% CI, 8.1-17.2 months), respectively, p=0.014 by Log-Rank Test).

Conclusions: Pretreatment S100A9+ MDSC and brain metastasis are independent predictors for shorter PFS to EGFR TKI in EGFR-mutation positive patients. This MDSC might potentially play some roles in treatment response to EGFR-TKI.

Keyword: NSCLC, S100A9+ MDSC, EGFR mutation, EGFR-TKI

Citation Format: Po-Hao Feng, Chih-Teng Yu, Kang-Yun Lee. Circulating s100a9 myeloid-derived suppressor cells are predictive for poor pfs in patients with egfr mutation positive lung adenocarcinoma treated with gefitinib. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2543. doi:10.1158/1538-7445.AM2014-2543
Abstract 3117: BET bromodomain inhibition triggers apoptosis of NF1-associated malignant peripheral nerve sheath tumors through Bim induction
Authors: Patel Amish J. , Liao Chung-Ping , Chen Zhiguo , Liu Chiachi , Wang Yong , Le Lu Q.
Published: May 2015

Malignant Peripheral Nerve Sheath Tumors (MPNSTs) are highly aggressive sarcomas that develop sporadically or in Neurofibromatosis type 1 (NF1) patients. MPNSTs are typically fatal, and effective treatments remain limited. To gain insights into MPNST pathogenesis, we utilized a novel MPNST mouse model that allowed us to study the evolution of these tumors at the transcriptome level. Strikingly, in MPNSTs we found upregulation of chromatin regulator Brd4, and show that BRD4 inhibition profoundly suppresses both growth and tumorigenesis. Our findings reveal new roles for BET bromodomains in MPNST development, and report a novel mechanism by which bromodomain inhibition induces apoptosis through induction of pro-apoptotic Bim, which may represent a paradigm shift in therapy for MPNST patients. Moreover, these findings indicate novel epigenetic mechanisms underlying the balance of anti-/pro-apoptotic molecules, and suggest that bromodomain inhibition can shift this balance in favor of cancer cell apoptosis.

Citation Format: Amish J. Patel, Chung-Ping Liao, Zhiguo Chen, Chiachi Liu, Yong Wang, Lu Q. Le. BET bromodomain inhibition triggers apoptosis of NF1-associated malignant peripheral nerve sheath tumors through Bim induction. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3117. doi:10.1158/1538-7445.AM2014-3117
Abstract 3166: Rab11 regulates E-cadherin expression and induces cell transformation in colorectal carcinoma
Authors: Chung Yuan-Chiang , Wei Wan-Chen , Huang Shin-Han , Shih Chi-Min , Hsu Chih-Ping , Chang King-Jen , Chao Wei-Ting
Published: May 2015

In the process of epithelial mesenchymal transition EMT, the disassembly of junctional adhesion complexes such as E-cadherin is a remarkable sign during changes in cell morphology and polarity. However, E-cadherin expression is dynamic, and is regulated by the cellular endocytic system; it is also involved in cell signaling mechanisms. In this study, we investigated the role of E-cadherin in colorectal tumors and the relationship with recycling endosome protein Rab11 in colon cell transformation. For tissue screening, the expressions of E-cadherin and Rab11 in colorectal tumors were identified by immunohistochemistry in 113 patients with colorectal carcinoma. For the in vitro cell experiment, GFP-tagged Rab11 plasmid was transfected into HT29 colon cells, E-cadherin expression and cell transformation were monitored by Western blot and confocal microscopy. In immunohistochemistry, the mean score of E-cadherin in tumor and normal tissues was 1.41 ± 0.06 and 1.08 ± 0.06 (p < 0.05). The mean score of Rab11 in tumor and normal tissues was 0.51 ± 0.05 and 0.18 ± 0.02 (p < 0.05). Synchronous overexpression of E-cadherin and Rab11 was noted in 74 patients (66.5%) with colorectal carcinoma. When GFP-tagged Rab11 plasmid was overexpressed in cultured colon cell line HT-29, the E-cadherin expression was up-regulated, and a fibroblast-like cell polarity was induced, which resulted in cell transformation. This study demonstrated the importance of the overexpression of Rab11 and E-cadherin in colorectal cancer. The results indicated that Rab11 together with E-cadherin might be potential markers for colorectal cancer progression and treatment.

Citation Format: Yuan-Chiang Chung, Wan-Chen Wei, Shin-Han Huang, Chi-Min Shih, Chih-Ping Hsu, King-Jen Chang, Wei-Ting Chao. Rab11 regulates E-cadherin expression and induces cell transformation in colorectal carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3166. doi:10.1158/1538-7445.AM2014-3166
Abstract 3177: Hepatitis C virus transcriptionally downregulates miR-181c for promotion of hepatocyte growth towards hepatocellular carcinoma
Authors: Ray Ratna B. , Mukherjee Anupam , Shrivastava Shubham , Ray Ranjit
Published: May 2015

Hepatitis C virus (HCV) induced chronic liver disease is one of the leading causes for hepatocellular carcinoma (HCC). The molecular events leading to HCC following HCV infection remains poorly defined. MicroRNAs (miRNAs) have been implicated in the control of many biological processes and their deregulation associate with different viral infections. In this study, we have identified a mechanism by which HCV suppresses miR-181c in human hepatocytes and target pathways involved in cell proliferation. Our results suggested that HCV infection transcriptionally downregulates miR-181c expression by modulating C/EBP-β. In silico prediction suggests that HOXA1 is a direct target of miR-181c. Homeobox A1 (HOXA1) is a member of homeodomain containing transcription factor family, and possesses pivotal roles in normal growth, development and differentiation of mammalian tissues. We have observed that HOXA1 expression is enhanced in HCV infected hepatocytes, and overexpression of mimic miR-181c inhibits HOXA1. Subsequently, exogenous expression of miR-181c inhibited HOXA1 downstream molecules STAT3 and STAT5, important transcription factors involved in HCV mediated cell survival and proliferation. Taken together, our results suggested that miR-181c is downregulated during HCV infection, which in turn, enhanced HOXA1 signaling pathway and promoted hepatocyte growth towards HCC.

Citation Format: Ratna B. Ray, Anupam Mukherjee, Shubham Shrivastava, Ranjit Ray. Hepatitis C virus transcriptionally downregulates miR-181c for promotion of hepatocyte growth towards hepatocellular carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3177. doi:10.1158/1538-7445.AM2014-3177
Abstract 3200: The effect of a sesquiterpenoid from Tussilago farfara on cellular proliferation via the Wnt/β-catenin signaling pathway in colon cancer
Authors: Ryu Jae-Ha , Li Hua , Lee Hwa Jin , Kim Woo-Young
Published: May 2015

The β-catenin, a major transcriptional modulator in Wnt signaling, plays a key role in embryogenesis, stem cell renewal and organ regeneration.

Abnormal activation of the Wnt/β-catenin signaling pathway frequently induces colon cancer progression. Therefore, the modulation of Wnt/β-catenin signaling pathway has been regarded as a guaranteed target for the treatment of cancer. In the past decade, various pharmacological inhibitors of β-catenin have been identified from medicinal plants. Most of the plant-derived inhibitors are flavonoids such as quercetin, fisetin, silibene and so on. In the present study, we identified tussilagone (TSL), a sesquiterpenoid isolated from the flower buds of medicinal plant Tussilago farfara, as an inhibitor on β-catenin dependent Wnt pathway. TSL suppressed β-catenin/T-cell factor transcriptional activity in a dose dependent manner (2, 10, 30 μM), and also down-regulated β-catenin level both in cytoplasm and nuclei of HEK293 reporter cells when they were activated by Wnt3a or 20 mM LiCl, an inhibitor of glycogen synthase kinase-3β. Since the mRNA level was not affected by TSL up to 30 μM, proteasomal degradation might be responsible for the decreased level of β-catenin. In SW480 and HCT116 colon cancer cell lines, the levels of cytosolic and nuclear β-catenin were decreased significantly by the treatment with 10 μM TSL in Western blot and immunofluorescence analysis. Moreover, TSL also decreased the expression of cyclin D1 and c-myc, the representative target genes of the Wnt/β-catenin signaling pathway, and consequently inhibited the proliferation of colon cancer cells. Taken together, tussilagone from T. farfara may be a new lead compound for the development of therapeutic agents against colon cancer through modulating Wnt/β-catenin signaling pathway. Acknowledgements: This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science, ICT & Future Planning (Project No 2011-0030699 and 2012R1A1A3013645).

Citation Format: Jae-Ha Ryu, Hua Li, Hwa Jin Lee, Woo-Young Kim. The effect of a sesquiterpenoid from Tussilago farfara on cellular proliferation via the Wnt/β-catenin signaling pathway in colon cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3200. doi:10.1158/1538-7445.AM2014-3200
Abstract 3339: The regulatory subunit of PI3K, p85β, induces cellular transformation, enhanced cell proliferation and increased PI3K signaling
Authors: Ito Yoshihiro , Hart Jonathan R. , Ueno Lynn , Vogt Peter K.
Published: May 2015

We have expressed the human p85β in chicken embryo fibroblasts (CEF) using the RCAS retroviral vector. At low concentrations, RCAS-p85β induces the formation of distinct foci of transformed cells. CEF transfected with high concentrations of RCAS-p85β show enhanced cell proliferation and PI3K signaling. RCAS-p85β shares this growth promoting potential with the cancer-derived mutants of p85α, but the wild type p85α protein expressed by the RCAS vector lacks this potential. The transforming and signaling activities of p85β depend on the PI3K catalytic subunit p110. We have used p110 isoform-specific inhibitors to show that the transforming and signaling activities of p85β can be mediated by p110α or p110β or both. This is in contrast to the transforming mutants of p85α which depend almost exclusively on p110α for their growth-promoting effect. p85β can be phosphorylated at Y655. Mutations of the phosphorylation site Y655A or Y655E lose almost all of their transforming and enhanced signaling activity. This result indicates that p85β could be subject to control by tyrosine kinases. Our data document important differences between p85α and p85β and suggest that p85β could carry out special functions in signaling and in cancer.

This work was supported by NIH Grant R01 CA078230. This is abstract 26049 of The Scripps Research Institute.

Citation Format: Yoshihiro Ito, Jonathan R. Hart, Lynn Ueno, Peter K. Vogt. The regulatory subunit of PI3K, p85β, induces cellular transformation, enhanced cell proliferation and increased PI3K signaling. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3339. doi:10.1158/1538-7445.AM2014-3339
Abstract 3348: Netrin-4/Integrin beta-4 interaction promotes glioblastoma cell proliferation and protects from temozolomide induced cellular senescence via activating PI3K/AKT pathway
Authors: Hu Yizhou , Ylivinkka Irene , Li Li , Chen Ping , Hautaniemi Sampsa , Nyman Tuula A. , Keski-Oja Jorma Keski-Oja , Hyytiäinen Marko
Published: May 2015

Netrin-4 (NTN4) is a secreted laminin-related protein, which was originally observed to guide neuronal axons during neuronal development. Recently, it has been found to be expressed in many other tissues and tumor types, and to contribute to the regulation of cell adhesion, migration, proliferation, and apoptosis. Glioblastoma multiforme (GBM) is the most common primary tumor of central nervous system (CNS). Although there is no remedy for this fatal disease, the efficacy of temozolomide (TMZ) -an orally taken alkylating agent- has been demonstrated in the treatment of glioblastoma. However, the sensitivity of GBM cells to TMZ is interfered by many factors such as MGMT expression, AKT activation, etc. Here we have explored the functions and molecular mechanisms of NTN4 in GBM.

The suppression of NTN4 expression in GBM cell lines significantly reduced cell proliferation and motility and increased serum deprivation-induced apoptosis. By using tandem affinity purification and mass spectrometric analysis, we identified the physical interaction between integrin beta-4 (ITGB4) and NTN4. This interaction mediates AKT phosphorylation and concomitant mitogenic effects. Furthermore, silencing either NTN4 or ITGB4 in GBM cell induced cellular senescence. Temozolomide induces cellular senescence in MGMT devoid glioma cell lines. Addition of exogenous recombinant NTN4 protein rescued TMZ induced senescence and AKT dephosphorylation in GBM cells in an ITGB4 dependent manner.

Current data suggest that the NTN4-ITGB4 interaction promotes GBM cell proliferation and protects GBM cell from TMZ triggered cellular senescence via activating PI3K/AKT pathway.

Citation Format: Yizhou Hu, Irene Ylivinkka, Li Li, Ping Chen, Sampsa Hautaniemi, Tuula A. Nyman, Jorma Keski-Oja Keski-Oja, Marko Hyytiäinen. Netrin-4/Integrin beta-4 interaction promotes glioblastoma cell proliferation and protects from temozolomide induced cellular senescence via activating PI3K/AKT pathway. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3348. doi:10.1158/1538-7445.AM2014-3348
Abstract 3380: Cancer usurps skeletal muscle as an energy repository
Authors: Luo Yi , Yoneda Junya , Ohmori Hitoshi , Sasaki Takamitsu , Kuniyasu Hiroki
Published: May 2015

Cancer cells produce energy through aerobic glycolysis, but contributions of host tissues to cancer energy metabolism is unclear. In this study, we aimed to elucidate the cancer-host energy production relationship, in particular, between cancer energy production and host muscle. During the development and progression of colorectal cancer (CRC), expression of the secreted autophagy-inducing stress protein HMGB1 increased in the muscle of tumor-bearing animals. This effect was associated with decreased expression of pyruvate kinase PKM1 and pyruvate kinase activity in muscle via the HMGB1 receptor RAGE. However, muscle mitochondrial energy production was maintained. In contrast, HMGB1 addition to CRC cells increased lactate fermentation. In the muscle, HMGB1 addition induced autophagy by decreasing levels of active mTOR and increasing autophagy-associated proteins, plasma glutamate and 13C-glutamine incorporation into acetyl-CoA. In a mouse model of colon carcinogenesis, a temporal increase in HMGB1 occurred in serum and colonic mucosa with an increase in autophagy associated with altered plasma free amino acids levels, increased glutamine and decreased PKM1 levels. These differences were abolished by administration of an HMGB1 neutralizing antibody. Similar results were obtained in a mouse xenograft model of human CRC. Taken together, our findings suggest that HMGB1 released during tumorigenesis recruits muscle to supply glutamine to cancer cells as an energy source.

Citation Format: Yi Luo, Junya Yoneda, Hitoshi Ohmori, Takamitsu Sasaki, Hiroki Kuniyasu. Cancer usurps skeletal muscle as an energy repository. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3380. doi:10.1158/1538-7445.AM2014-3380
Abstract 4441: Ube2e inhibits the ubiquitination and degradation of EGFR mediated by Cbl and Ube2d
Authors: Ma Ke , Lipkowitz Stanley , Klevit Rachel , Liyasova Mariya Sergeyevna
Published: May 2015

Receptor tyrosine kinases (RTKs) have a demonstrated role as drivers of malignant transformation. Many RTKs (e.g., EGFR, MET, FLT3) are negatively regulated by ubiquitination and degradation mediated by Cbl proteins a family of RING finger (RF) ubiquitin ligases (E3s). There are three mammalian family members Cbl, Cbl-b and Cbl-c. All Cbl proteins have a highly conserved N-terminal tyrosine kinase binding domain and a C3HC4 (RF) domain. Loss of Cbl protein function has be associated with malignant transformation drive by increased RTK activity. E3s such as the Cbl proteins confer specificity to the ubiquitination process and direct the conjugation of ubiquitin to one or more lysines on the specific target proteins in collaboration with ubiquitin-conjugating enzymes (E2s). The E2s bind to the E3 and together they mediate ubiquitination of specific substrates. There are approximately 30 human E2s. All of the E2s have a highly conserved catalytic domain (UBC domain). In this study, we characterized the interaction of Cbl with E2s. Using an in vitro E3 assay, we found that only the Ube2d family (Ube2d1, Ube2d2, and Ube2d3) of E2s mediates autoubiquitination of the three Cbl proteins. Subsequently, using the yeast two-hybrid system, we found that Ube2e1, Ube2e2, Ube2e3, Ube2L3, Ube2u, Ube2w and Ube2z can interact with Cbl even though they do not support autoubiquitination of Cbl in vitro E3 assay. Among these E2s, three Ube2e family members and Ube2w bind to the RF domain of Cbl as demonstrated by the loss of interaction when the RF domain is disrupted. Further, we demonstrated that Ube2e family members are involved in regulation of EGFR activity mediated by Cbl in HeLa cells. Knockdown of Ube2e increases down-regulation and ubiquitination of EGFR in HeLa cells. We further showed that three Ube2e inhibit autoubiquitination of Cbl mediated by Ube2d2 in vitro. Together these data suggest that RTK degradation can be modulated by competition between E2s for the Cbl E3s.

Citation Format: Ke Ma, Stanley Lipkowitz, Rachel Klevit, Mariya Sergeyevna Liyasova. Ube2e inhibits the ubiquitination and degradation of EGFR mediated by Cbl and Ube2d. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4441. doi:10.1158/1538-7445.AM2014-4441
Abstract 4463: Targeting SN38 delivery to gastrointestinal cancer cells using a fucose-bound nanoparticle approach
Authors: Kato Junji , Takimoto Rishu , Osuga Takahiro , Ono Michihiro , Hirakawa Masahiro , Yoshida Makoto , Sato Yasushi , Tamura Fumito
Published: May 2015

Background & Aims: Prognosis of patients with metastatic colorectal cancer (CRC) has been improving by introduction of molecular targeting therapy combined with chemotherapeutic agents. However, metastatic CRC still needs new strategies to improve treatment efficacy and survival rate. Increased expression of fucosyltransferases (FUTs) is common in CRC cells. FUTs are key enzymes accelerating malignant transformation through the fucosylation of different sialylated precursors, suggesting a crucial requirement for fucose by metastatic CRC cells. With this in mind, we developed fucose-bound nanoparticles as vehicles for delivery of anticancer drugs specifically to CRC cells. Methods: L-fucose-bound liposomes were produced by the modified cholate dialysis method as vehicles for delivery of Cy5.5, FAM and SN38. The production of CA19-9 was measured by enzyme-linked immunosorbent assay. The CRC cell lines were exposed to SN38 or L-fucose-bound liposomes carrying SN38, and cytotoxicity was monitored with BrdU assay. Introduction of Cy5.5 or FAM by L-fucose-bound liposomes was analyzed by flow cytometry or fluorescence microscopy. The effect of L-fucose-bound liposome carrying SN38 in vivo was analyzed by intravenous injection in mice engrafted with CRC cell lines. All statistical tests were two-sided. Results: L-fucose-bound liposomes containing Cy5.5 or SN38 were effectively delivered into CA19-9 expressing CRC cancer cells. Excess L-fucose decreased the efficiency of Cy5.5 introduction by L-fucose-bound liposomes, suggesting L-fucose-receptor dependency. Intravenously injected L-fucose-bound liposomes carrying SN38 were successfully delivered to CRC cells, mediating efficient tumor growth inhibition as well as prolonging survival in mouse xenograft models. Conclusions: Fucose-bound liposomes carrying anti-cancer drugs should provide a new strategy for patients with colorectal cancer.

Citation Format: Junji Kato, Rishu Takimoto, Takahiro Osuga, Michihiro Ono, Masahiro Hirakawa, Makoto Yoshida, Yasushi Sato, Fumito Tamura. Targeting SN38 delivery to gastrointestinal cancer cells using a fucose-bound nanoparticle approach. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4463. doi:10.1158/1538-7445.AM2014-4463
Abstract 4491: FGFR2-ADC potently and selectively inhibits growth of gastric and breast cancer xenograft models
Authors: Sommer Anette , Nising Carl F. , Mahlert Christoph , Kopitz Charlotte C. , Lerchen Hans-Georg , Greven Simone , Stelte-Ludwig Beatrix , Schuhmacher Joachim , Zierz Ruprecht , Wittemer-Rump Sabine , Schatz Christoph , Reetz Frank , Apeler Heiner , Jautelat Rolf , Kreft Bertolt , Ziegelbauer Karl
Published: May 2015

Antibody-drug conjugates (ADCs) represent a promising therapeutic approach for treatment of cancer. We have developed a novel ADC directed against fibroblast growth factor receptor 2 (FGFR2). FGFR2 is overexpressed in several cancer indications, such as gastric cancer and breast cancer, thus representing an interesting therapeutic target for the treatment of FGFR2 positive cancer patients with an ADC-based therapy. The FGFR2-ADC consists of the fully human anti-FGFR2-mAb BAY 1179470 conjugated via a stable linker to a novel auristatin cytotoxic agent (technology licensed from Seattle Genetics). The FGFR2-mAb BAY 1179470, which is cross-reactive with human, mouse, rat and monkey, induces internalization of FGFR2. Quantitative data on FGFR2 antibody bound per cell (ABC) were determined with the QuantiBrite assay using BAY 1179470. FGFR2-ADC has a potency in the single digit nM to sub nM range in a panel of FGFR2-positive cells lines (e.g., SNU-16, KatoIII, SUM52-PE, MFM-223) and shows more than 100-fold selectivity versus FGFR2-negative cell lines. High levels of FGFR2 on cancer cells correlate with internalization efficacy and cytotoxic activity in vitro. FGFR2-ADC is highly efficacious in monotherapy and results in tumor growth inhibition in the gastric cancer xenograft model SNU-16 and tumor regression in the breast cancer xenograft model MFM-223. At doses efficacious in mice, FGFR2-ADC is well tolerated. The pre-clinical efficacy and tolerability data obtained for FGFR2-ADC suggest a therapeutic index and support clinical testing.

Citation Format: Anette Sommer, Carl F. Nising, Christoph Mahlert, Charlotte C. Kopitz, Hans-Georg Lerchen, Simone Greven, Beatrix Stelte-Ludwig, Joachim Schuhmacher, Ruprecht Zierz, Sabine Wittemer-Rump, Christoph Schatz, Frank Reetz, Heiner Apeler, Rolf Jautelat, Bertolt Kreft, Karl Ziegelbauer. FGFR2-ADC potently and selectively inhibits growth of gastric and breast cancer xenograft models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4491. doi:10.1158/1538-7445.AM2014-4491
Abstract 450: Head and neck cancer and HPV infection: A potential role for peritumoral neurogenesis in HPV-associated malignancy
Authors: Graves Christian A. , Tomar Swati , Altomare Diego , Wells James R. , Creek Kim E. , Pirisi Lucia
Published: May 2015

Background: Head and neck cancers account for 5% of all cancer related deaths and present with complex molecular pathology and clinical features owing to their functional anatomical location. The molecular etiology of head and neck squamous cancer (HNSC) has gained extensive attention recently revealing key mutations, viral infection, and an emerging landscape of varying disease sub-types. While perineural invasion has long been associated with poor outcome in HNSC, the association of neurogenesis-related genes with cancer progression has only recently come into focus in HNSC. Using prospectively acquired patient samples and TCGA HNSC mutational analyses we further characterized the sequelae of molecular changes in neurogenesis-related genes and propose that these genes play a significant role in HNSC development.

Methods: All clinical specimens were collected under approval by Institutional Review Board. Prospectively collected samples were analyzed for HPV DNA by linear arrays (InnoLipa) and q-PCR for E7-RNA. Gene expression arrays were conducted to determine differences in HPV activity and neuronal associated gene ontologies (Agilent). Gene expression profiles were compared in oral HPV+ and HPV- cultured oral keratinocytes as well as the TCGA Kandoth & Tang datasets. Gene targets were validated by Immunohistochemistry and qPCR.

Results: Initial unsupervised hierarchical clustering analysis of our HNSC cohort revealed a separation of HPV+ HPV+E7-(HPV-inactive: HPVin), and HPV- tumors with changes in the expression of key neurogenesis-related genes. For example, POU4F1, NGFR, and GRIN2C were significantly up-regulated (P<0.05) in HPV+ vs HPVin cancers. Additional analysis of TCGA data revealed divergence of the mutational spectra in HNSC with and without perineural invasion.

Conclusion: Our preliminary findings indicate the emerging importance of neurogenesis-related genes and perineural involvement in HNSC and support a divergence of HPV+, HPVin, and HPV- HNSC sub-types.

Citation Format: Christian A. Graves, Swati Tomar, Diego Altomare, James R. Wells, Kim E. Creek, Lucia Pirisi. Head and neck cancer and HPV infection: A potential role for peritumoral neurogenesis in HPV-associated malignancy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 450. doi:10.1158/1538-7445.AM2014-450
Abstract 4522: Akt2 and acid ceramidase cooperate to induce malignant transformation
Authors: Sebti Said M. , Berndt Norbert , Patel Ronil , Yang Hua , Balasis Maria
Published: May 2015

The ability of Akt 2 and acid ceramidase (ASAH1) to cooperate and induce malignant transformation is not known. Here we show that in immortalized, non-transformed cells, ectopic co-expression of Akt2 and ASAH1 is significantly more effective than expression of each gene alone at inducing cell invasion and at conferring resistance to apoptosis. Consistent with these observations, siRNA-mediated depletion of both Akt2 and ASAH1 is much more potent than depleting each alone at inhibiting cell viability/proliferation and cell invasion. Furthermore, pharmacological inhibitors of Akt (TCN or MK-2206) and ASAH1 (B13) synergize to inhibit cell viability/proliferation, and combinations of these drugs are more effective than single agent treatments at inhibiting cell invasion. Taken together, the results suggest that these two enzymes cooperate to induce malignant transformation and warrant further preclinical studies to evaluate the potential of combining inhibitors of Akt and ASAH1 to treat cancer.

Citation Format: Said M. Sebti, Norbert Berndt, Ronil Patel, Hua Yang, Maria Balasis. Akt2 and acid ceramidase cooperate to induce malignant transformation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4522. doi:10.1158/1538-7445.AM2014-4522
Abstract 4524: Combined inhibition of PIM and PI3 kinases shows an enhanced efficacy in a number of solid tumour cell lines
Authors: O'Neil Michael , Aparicio C. Blanco , Jiang S. , Martinez S. , McKenzie A. , Page Martin , Pastor J.
Published: May 2015

Introduction: PIM kinases play an important role in a number of aspects of the regulation of cell cycle and proliferation. Over expression of PIM kinases is associated with oncogenicity in a number of haematological and solid tumours. Recent evidencesuggests that PIM expression is upregulated by inhibition of signalling on the AKT/PI3K/mTOR pathway. We set out to examine the effects of inhibition of PIM kinases in a number of solid tumor cell lines and to explore the effects of concomitant inhibition of the AKT/PI3K/mTOR pathway with novel agents having dual PIM kinase and PI3k inhibition. Representative cell lines for colorectal, lung, oesophageal and prostate tumors (HCT116, A549 and H460, OE21, PC3 respectively) were chosen for testing in a cytotoxicity assay. Compounds tested were GDC-0941, a prototypical PI3K inhibitor, AZD 1208, a pan-PIM inhibitor (i.e. equi-active on all 3 PIM isoforms), IBL- 201 and IBL-202 (dual-acting PI3K/PIM inhibitors) and IBL-301 and IBL-302 (triple-acting PI3K/PIM/mTOR inhibitors). Rapamycin the prototypical mTOR inhibitor and cisplatin were also tested as reference compounds.

Results: The pan-PIM compound AZD1208 showed efficacy in all cell lines in the micromolar range. Rapamycin was also active across similar concentrations. GDC-0941, the PI3K inhibitor exhibited moderate efficacy across the cell line panel . The dual and triple inhibitors showed cytotoxic effects at sub micromolar concentrations, with the OE21 and PC3 oesophageal and prostate lines respectively showing the most sensitive responses.

Conclusion: These results suggest that in selected solid tumor cell lines combined inhibition of PIM and PI3 kinases can lead to an increased cytotoxic effect. Further studies are under way to determine if this observation is corroborated in vivo. Combined inhibition of PIM and PI3 kinases with a single molecule may offer significant opportunities in the clinic for efficacy in key tumour subsets and to circumvent acquired resistance via feedback mechanisms.

Citation Format: Michael O'Neil, C. Blanco Aparicio, S. Jiang, S. Martinez, A. McKenzie, Martin Page, J. Pastor. Combined inhibition of PIM and PI3 kinases shows an enhanced efficacy in a number of solid tumour cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4524. doi:10.1158/1538-7445.AM2014-4524
Abstract 4549: CDK2 inhibition causes anaphase catastrophe through the centrosomal protein CP110
Authors: Hu Shanhu , Danilov Alexey V. , Godek Kristina M. , Orr Bernardo , Tafe Laura J. , Memoli Vincent A. , Galimberti Fabrizio , Ravi Saranya , DeCastro Andrew J. , Lu Yun , Mustachio Lisa Maria , Sekula David J. , Andrew Angeline S. , Freemantle Sarah J. , Compton Duane A. , Dmitrovsky Ethan
Published: May 2015

Aneuploidy is frequently detected in human cancers and is implicated in carcinogenesis. Whether aneuploidy is an anti-neoplastic target remains to be learnt. We previously uncovered an unrecognized consequence of Cdk2 inhibition that causes lung cancer cells to undergo anaphase catastrophe, growth inhibition and apoptosis. Cells with activating KRAS mutations were especially sensitive to Cdk2 inhibition. The mechanism of Cdk2-mediated anaphase catastrophe and how activated KRAS promotes this effect was the subject of this study. Live-cell imaging provided direct evidence that after Cdk2 inhibition, lung cancer cells develop multipolar anaphase, undergo multipolar cell division, and the fate of the progeny of these divisions is apoptosis and death. The centrosomal protein CP110, a Cdk2 target protein, was found to be a critical mediator of Cdk2-inhibition-driven anaphase catastrophe. Intriguingly, siRNA-mediated CP110 repression induced anaphase catastrophe of lung cancer cells. By contrast, overexpression of CP110 antagonized anaphase catastrophe caused by Cdk2 inhibition. Furthermore, we found that activated KRAS mutations sensitized lung cancer cells to Cdk2 inhibition by deregulating CP110 expression. Immunohistochemical examination of a paired normal-malignant lung cancer tissue array revealed a statistically significant upregulation of CP110 in lung cancers as compared with normal lung tissues. Notably, CP110 expression was reduced in smokers where KRAS mutations are frequently present. Taken together, this study presents a CP110-dependent pathway that engages anaphase catastrophe in lung cancer cells. It also provides a mechanistic explanation that lung cancers and potentially other cancers with activated KRAS mutation are more sensitive to Cdk2 inhibition due to CP110 deregulation.

Citation Format: Shanhu Hu, Alexey V. Danilov, Kristina M. Godek, Bernardo Orr, Laura J. Tafe, Vincent A. Memoli, Fabrizio Galimberti, Saranya Ravi, Andrew J. DeCastro, Yun Lu, Lisa Maria Mustachio, David J. Sekula, Angeline S. Andrew, Sarah J. Freemantle, Duane A. Compton, Ethan Dmitrovsky. CDK2 inhibition causes anaphase catastrophe through the centrosomal protein CP110. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4549. doi:10.1158/1538-7445.AM2014-4549
Abstract 4619: Protein palmitoylation in non-small cell lung cancer (NSCLC): DHHC5 palmitoyltransferase as a potential therapeutic target
Authors: Tian Hui , Lu Jui-Yun , Shao Chunli , Huffman Kenneth , Carstens Ryan , Minna John D. , Hofmann Sandra L.
Published: May 2015

Post-translational palmitoylation of intracellular proteins is mediated by protein palmitoyltransferases (PAT) belonging to the DHHC family, which share a common catalytic Asp-His-His-Cys (DHHC) motif. According to a genome-wide siRNA screen scoring for growth suppression in over 50 non-small cell lung cancer (NSCLC) cell lines, DHHC family members are strikingly over-represented. Among the tested DHHC palmitoyltransferases, knockdown of DHHC5 showed cell growth inhibition in many cell lines in the initial screen.

To further investigate the growth suppression in response to DHHC5 depletion, transient and stable knockdowns of DHHC5 were undertaken in a dozen of NSCLC cell lines identified in the genome-wide screen using siRNAs and lentiviral shRNAs. Such knockdown was also performed in three immortalized human lung bronchial epithelial cell lines (HBECs). The gene expression and protein levels of DHHC5 were analyzed by RT-PCR and immunoblotting, both of which showed efficient knockdown levels. Cell proliferation and anchorage-dependent/independent colony formation assays indicated that DHHC5 knockdown led to cell growth arrest and decreased cell clonogenicity in most of the tested NSCLC cell lines in vitro, but not in the HBECs. H1299, H358 and H2009 cell lines were particularly dependent on DHHC5 expression and the growth-suppression phenotype of DHHC5 stable knockdown H1299/H358/H2009 cells was rescued by overexpression of DHHC5. The catalytically inactive DHHC5 mutant was not able to provide rescue. In vivo tumor formation assays were then carried out by injecting H1299 and H358 control cells and DHHC5 stable knockdown cells subcutaneously into NOD/SCID mice. DHHC5 knockdown led to absence or dramatically decreased tumor xenograft formation. Furthermore, a Tet-On inducible system with TRIPZ shRNA lentiviral delivery was established in H1299. Mouse xenografts using this H1299 Tet-on inducible cell line showed similar results.

As DHHC5 function is needed for optimal lung cancer cell growth, it will be important to identify physiologically relevant substrates in order to develop inhibitors of DHHC5 function. Future plans include microarray and protein kinase array analysis to distinguish possible signaling pathways in which DHHC5 may be involved, as well as palmitoyl-protein profiling of normal and knockdown DHHC5 cell lines by mass spectrometry.

Citation Format: Hui Tian, Jui-Yun Lu, Chunli Shao, Kenneth Huffman, Ryan Carstens, John D. Minna, Sandra L. Hofmann. Protein palmitoylation in non-small cell lung cancer (NSCLC): DHHC5 palmitoyltransferase as a potential therapeutic target. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4619. doi:10.1158/1538-7445.AM2014-4619
Abstract 4657: Evaluation of genomic profiling in the GALAXY-1 (NCT01348126), a randomized Phase 2b study of ganetespib in combination with docetaxel versus docetaxel alone as second line therapy in patients with advanced NSCLC
Authors: Fennell Dean , Antonov Alexey , Martins Miguel L. , Popat Sanjay , Ramalingam Suresh S. , Spicer James , Vukovic Vojislav M. , El-Hariry Iman , Reichert Vienna , Rosell Rafael
Published: May 2015

Background: Inhibition of Hsp90, a key molecular chaperone required for activation of many oncoproteins, can lead to cancer cell death. Ganetespib (G) is a 2nd generation Hsp90 inhibitor (Hsp90i) that has single agent clinical activity in patients with ALK, KRAS, HER2, and BRAF mutations. G also inhibits pathways implicated in resistance to taxanes, including hypoxia pathways (HIF-1α) as well as cell-cycle and DNA repair pathways. Combination of G with docetaxel (D) has shown synergy in NSCLC xenografts and prolongation of progression-free survival (PFS) and overall survival (OS) of patients with NSCLC adenocarcinoma in interim analyses of the GALAXY-1 trial. Methods: Patients enrolled in GALAXY-1 (n=253) receive D 75 mg/m2 on D1 of a 3-week treatment cycle; combination arm patients receive G 150 mg/m2 on D1 and D15 in addition. The co-primary endpoints are PFS in patients with elevated baseline level of serum LDH and PFS in the mutant KRAS population. PFS and OS in all adenocarcinoma patients are key secondary endpoints. Genomic profiling using Affymetrix Oncoscan™ was performed on archival tissues obtained at baseline, with the goal of determining biomarkers predictive of ganetespib activity

Results: Tumor tissue samples from 65 patients and 26 matched normal tissues were processed on the Affymetrix OncoScan™ FFPE Express 2.0 Services assay. The OncoScan™ FFPE Express 2.0 assay contains 332941 copy number probes. After removing samples with average call rates lower than 90%, 296522 markers on chromosome 1-22 were selected for copy number analyses. For preliminary evaluation of interactions between copy number aberrations and treatment response in the G arm, a genome scale Kaplan-Meier Estimates PFS analysis at each of 296522 markers was performed.

Conclusions: Genomic profiling using OncoScan™ is utilized in the GALAXY-1 trial to identify biomarkers of response/resistance to ganetespib administered in combination with docetaxel. Complete analyses will be presented at the meeting.

Note: This abstract was not presented at the meeting.

Citation Format: Dean Fennell, Alexey Antonov, Miguel L. Martins, Sanjay Popat, Suresh S. Ramalingam, James Spicer, Vojislav M. Vukovic, Iman El-Hariry, Vienna Reichert, Rafael Rosell. Evaluation of genomic profiling in the GALAXY-1 (NCT01348126), a randomized Phase 2b study of ganetespib in combination with docetaxel versus docetaxel alone as second line therapy in patients with advanced NSCLC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4657. doi:10.1158/1538-7445.AM2014-4657
Abstract 4660: Temporal interrogation of EGFR signaling in head and neck cancer reveals highly distinct phosphorylation waves for individual EGFR inhibitors
Authors: Iyer Gopal
Published: May 2015

Targeting the epidermal growth factor receptor (EGFR/HER) family is an effective target for cancer therapeutics with thousands of cancer patients treated annually with EGFR inhibitors. However, we are only beginning to appreciate the rapid and dynamic signaling impact of various EGFR inhibitors on cancer cell phosphorylation events. The purpose of this study was to characterize the temporal phosphoprotein kinetics in head and neck cancer cells treated with the anti-EGFR antibodies cetuximab and Sym004. For the first time, perturbation data were generated using reverse phase protein array probed with 183 phosphorylated proteins involved in various signaling pathways at 11 different time points starting as early as 5 min and up to 24 hours. A novel signaling module comprised of phospho-ATM at Ser-1981, p-53(Ser15), p-IkB-α (Ser32/36) and p-Bad (Ser112) was found to be highly upregulated following exposure to Sym004. Concomitantly, high resolution imaging also revealed rapid EGFR degradation and increased endosomal trafficking with Sym004 treatment. Correlation matrix on a temporal scale revealed distinct network topologies suggesting that phosphorylation occurs in distinct waves over time. These response patterns were highly distinct when comparing cetuximab with Sym004. Using this platform, we are able to quantify the strength of various signaling networks triggered by EGFR inhibition and are able to model new pathways. These findings may have significant implications for developing more personalized treatment strategies including logical combination with other molecular target inhibitors.

Note: This abstract was not presented at the meeting.

Citation Format: Gopal Iyer. Temporal interrogation of EGFR signaling in head and neck cancer reveals highly distinct phosphorylation waves for individual EGFR inhibitors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4660. doi:10.1158/1538-7445.AM2014-4660
Abstract 4678: Application of the EGFR in squamous lung cancer and its relationship between p63
Authors: Han Yiping , Wang Bibo , Huang Ling , Qiang Li
Published: May 2015

OBJECTIVE: Epidermal growth factor receptor (EGFR) mutations are rarely found in squamous cell carcinoma (SQC).

METHODS: Between J February 2010 and March 2013, we investigated the incidence of EGFR activating mutations in SQC of the lung using the automated direct sequencing of polymerase chain reaction products method. The efficacies of formal therapy, observation, targeting therapy (EGFR-tyrosine kinase inhibitors [TKIs]) were retrospectively evaluated in patients with EGFR-mutated SQC. Further pathologic analyses were performed using immunohistochemistry.

RESULTS: Sixteen of the 282 patients with SQC (5.67%) had EGFR mutations, including exon 19 deletion (7 of 16 patients, 44%), L858R point mutation in exon 21 (9 of 16, 56%). The patients' median progression-free survival (PFS) and median overall survival (OS) in formal therapy, observation and targeting therapy were 6.28 and 9.03, 5.23 and 6.17, and 4.20 and 10.57 months. The median OS of 4 EGFR-mutated patients received p63 (partly +) was 11.28 months (95% confidence interval [CI], 6.33-16.23months) verse that of 11 EGFR-mutated patients received p63 (+) was 3.57 months (95%CI, 1.75- 5.39 months) (p=0.039).

CONCLUSION: EGFR-TKIs for EGFR-mutated SQC are generally inferior to that for EGFR-mutated ADC. Chemotherapy or radiation therapy seemed to be less inferior to TKI therapy considering the PFS and OS results and the economy when patients are in good status. Patients presenting partly + of p63 had longer OS than p63 + patients. EGFR and p63 might be two separate factors which affected the proliferation of squamous tumor cells and result remained to be confirmed by large samples.

KEYWORDS: squamous lung cancer, SQC,epidermal growth factor receptor, EGFR, P63

Note: This abstract was not presented at the meeting.

Citation Format: Yiping Han, Bibo Wang, Ling Huang, Li Qiang. Application of the EGFR in squamous lung cancer and its relationship between p63. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4678. doi:10.1158/1538-7445.AM2014-4678