bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3827: Bispecific antibodies with an anti-PD-1 backbone for cancer therapy generate enhanced immune activity
Authors: Li Baiyong , Huang Zhaoliang , Pang Xinghua , Zhong Tingting , Chen Na , Wang Max , Jin Xiaoping , Xia Dennis , Zhang Peng , Xia Michelle Yu
Published: August 2018

PD-1 antibody combination therapy shows great promise to further enhance clinical efficacy of PD-1 antibody. Bispecific antibody represents an emerging platform that allows simultaneous targeting of multiple pathways employing a single agent. Here we present a series of bispecific antibodies using anti-PD-1 antibody as backbone, including PD-1/CTLA-4 and PD-1/VEGF bispecific antibodies, which target additional immune suppressive pathways to further enhance cancer immunity. These include complementary check point pathway and suppression of dendritic cell maturation and antigen presentation. Clinical candidates for these projects have been selected demonstrating superior immune enhancing activity in preclinical study. These bispecific antibodes are designed based on Akeso Tetrabody platform, which has demonstrated robust developability properties. Akeso PD-1/CTLA-4 bispecific antibody AK104 has entered clinical trial for advanced solid tumors (ClinicalTrials.gov Identifier: NCT03261011).

Citation Format: Baiyong Li, Zhaoliang Huang, Xinghua Pang, Tingting Zhong, Na Chen, Max Wang, Xiaoping Jin, Dennis Xia, Peng Zhang, Michelle Yu Xia. Bispecific antibodies with an anti-PD-1 backbone for cancer therapy generate enhanced immune activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3827.
Abstract 3839: A small-molecule inhibitor of the β-catenin-TCF4 interaction suppresses colorectal cancer growth in vitro and in vivo
Authors: Shin Seung Ho , Lim Do Young , Reddy Kanamata , Malakhova Margarita , Liu Fangfang , Wang Ting , Song Mengqiu , Chen Hanyong , Bae Ki Beom , Ryu Joohyun , Liu Kangdong , Lee Mee-Hyun , Bode Ann M. , Dong Zigang
Published: August 2018

Colorectal cancer is associated with aberrant activation of the Wnt pathway. β-Catenin plays essential roles in the Wnt pathway by interacting with T-cell factor 4 (TCF4) to transcribe oncogenes. We synthesized a small molecule, referred to as HI-B1, and evaluated signaling changes and biologic consequences induced by the compound. HI-B1 inhibited β-catenin/TCF4 luciferase activity and preferentially caused apoptosis of cancer cells in which the survival is dependent on β-catenin. The formation of the β-catenin/TCF4 complex was disrupted by HI-B1 due to the direct interaction of HI-B1 with β-catenin. Colon cancer patient-derived xenograft (PDX) studies showed that a tumor with higher levels of β-catenin expression was more sensitive to HI-B1 treatment, compared to a tumor with lower expression levels of β-catenin. The different sensitivities of PDX tumors to HI-B1 were dependent on the β-catenin expression level and potentially could be further exploited for biomarker development and therapeutic applications against colon cancer.

Citation Format: Seung Ho Shin, Do Young Lim, Kanamata Reddy, Margarita Malakhova, Fangfang Liu, Ting Wang, Mengqiu Song, Hanyong Chen, Ki Beom Bae, Joohyun Ryu, Kangdong Liu, Mee-Hyun Lee, Ann M. Bode, Zigang Dong. A small-molecule inhibitor of the β-catenin-TCF4 interaction suppresses colorectal cancer growth in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3839.
Abstract 3853: AXL inhibitor promotes anti-tumor immunity through modulation of macrophage polarization
Authors: Lai Shenshen , Xu Qin , Yan Jun , Zhang Hong , Zhang Zaihui
Published: August 2018

Polarization of tumor-associated macrophages (TAMs) to classic pro-inflammatory M1 or alternatively activated M2 types plays an important role in establishing tumor microenvironment and determining therapeutic responses. It has become increasingly clear that M2 macrophages contribute to tumor progression by producing anti-inflammatory cytokines and suppressing anti-tumor immunity. AXL receptor tyrosine kinase has recently emerged as a dual therapeutic target in oncology, due to its function in tumor growth, survival and metastasis, as well as immunosuppressive activity. AXL and its ligand Gas6 are both involved in attenuating anti-tumor immune response through modulation of TAM polarization. Thus, targeting the AXL signaling pathway is expected to promote a pro-inflammatory tumor microenvironment, in addition to direct inhibition of tumor growth. SLC-391, a selective small molecule inhibitor for AXL, displays high potency against numerous cancer cell lines through inhibition of AXL/PI3K/AKT-dependent cell proliferation and survival in vitro. Additionally, this compound was also found to alter the cytokine profile expressed in THP1-derived M2 macrophages, including upregulating pro-inflammatory CXCL10 and downregulating anti-inflammatory IL-10. Noticeably, the expression level of AXL was significantly upregulated in THP1-derived macrophages in presence of tumor cell-conditioned medium, which indicated the involvement of AXL in M2 polarization, and therefore strengthening the notion of AXL as a dual therapeutic target. In a co-culture system consisting of THP1-derived macrophages and A549 non-small cell lung cancer cells, SLC-391 targeted AXL activity in both cell types for overall tumor cell suppression through direct and macrophage-mediated inhibition. This observation is supported by inhibition of tumor growth and the increased ratio of M1/M2-polarized TAMs from mice treated with SLC-391 in a CT-26 murine colon carcinoma syngeneic model, considering CT26 cells are not sensitive to SLC-391 in cell-based proliferation assay. In summary, in addition to direct inhibition of tumor cells, SLC-391 also appears to promote anti-tumor immunity through modulation of M2 to M1 transition.

Citation Format: Shenshen Lai, Qin Xu, Jun Yan, Hong Zhang, Zaihui Zhang. AXL inhibitor promotes anti-tumor immunity through modulation of macrophage polarization [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3853.
Abstract 3854: Antizyme inhibitor editing modulates nuclear localization but not antizyme binding: Implications for prostate cancer
Authors: Rice James M. , Ghalali Aram , Wang Liangzhe , Kusztos Amanda , Jernigan Finith , Wu Chin Lee , Zetter Bruce , Rogers Michael S.
Published: August 2018

The endogenous antizyme inhibitor (AZIN) binds to, and is a key regulator of, the cell cycle suppressor antizyme. Antizyme itself is a key regulator of ornithine decarboxylase (ODC), the rate-limiting enzyme in the polyamine synthesis. We have measured the expression and localization of AZIN in 202 prostate cancer specimens, along with 26 adjacent benign samples, and found that nuclear localization of AZIN is dramatically associated with worse outcome. Upregulation and nuclear localization of AZIN have been observed in several cancers, as has editing of the AZIN1 mRNA. Other have hypothesized that the RNA- edited AZIN (edAZIN) may have an increased affinity to antizyme and that could explain the association of edAZIN to the various cancers. We have studied the mechanism behind the nuclear localization of AZIN and found the single base pair substitution caused by RNA editing is sufficient to result in nuclear localization of the protein in all cell types tested. To determine if the nuclear localization might result from increased antizyme-edAZIN affinity, we developed fluorescent protein FRET sensor for protein-protein interaction using Clover-AZIN and antizyme-mRuby2 fusion proteins. Unexpectedly, we found that the editing event decreases edAZIN affinity for antizyme, notwithstanding increased interaction in vivo. Thus, the data indicate that the change in protein localization to the nucleus may be more important to oncogenic function than the actual degree of binding to antizyme. Other functional differences between edAZIN and AZIN might be explained by altered kinetics of binding, by the contribution of an additional adapter protein that modulates the intracellular antizyme:AZIN complex, or by competition for AZIN binding by other partners whose interaction is affected by the editing event. Therefore, we plan to identify AZIN and edAZIN interacting proteins by using proteomic analysis and later study the interaction between AZIN or edAZIN with their binding partners in more detail. Furthermore, our validated FRET assays can be used to identify antagonists of AZIN-antizyme binding (no antagonists are currently known). Given the important role of AZIN in tumor growth, discovery of small-molecule inhibitors of AZIN should lead to antitumor therapies.

Citation Format: James M. Rice, Aram Ghalali, Liangzhe Wang, Amanda Kusztos, Finith Jernigan, Chin Lee Wu, Bruce Zetter, Michael S. Rogers. Antizyme inhibitor editing modulates nuclear localization but not antizyme binding: Implications for prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3854.
Abstract 3907: Birinapant co-treatments of colon cancer cell lines show consensus molecular subtype-specific synergistic effects
Authors: Fichtner Michael , Bozkurt Emir , McAllister Katherine , McCann Christopher , Longley Daniel , Prehn Jochen H.
Published: August 2018

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. Despite recent advances, the treatment success differs strongly between colorectal cancer patients and new treatment options are urgently needed. Since most tumors avoid apoptosis by a dysregulation of Inhibitor of apoptosis proteins (IAPs), IAP inhibitors such as the SMAC mimetic birinapant are promising new drugs for the treatment of CRC. However, it is currently not possible to stratify for patients who might benefit from adjuvant therapy and those who need new treatments. In 2015, an international consortium unified six independent subtyping approaches and defined 4 distinct, so-called consensus molecular subtypes (CMS). The CMSs are the current gold standard for CRC subtyping and have some prognostic values. Nevertheless, whether the CMS classification can be used to estimate the treatment success is currently under investigation. This study aimed to evaluate the efficacy of treatments with the chemotherapeutics oxaliplatin and 5-fluoruracil (5-FU), TNFalpha, the SMAC-mimetic birinapant and a combination of these drugs. We selected 10 cell lines, representing the four CMSs, and treated them for 24h and 48h with the respective drugs. At both time points, we assessed the cell death by staining with Annexin V/PI followed by flow cytometric analysis of the cells. We were able to show that birinapant has very strong synergistic effects if used in combination with either oxaliplatin/5-FU or TNFalpha, but does not induce apoptosis alone. Importantly, the efficacy of the various co-treatments differed between the cell lines. These differences can, to some extent, be linked to the corresponding CMS of the cells. Cells belonging to the CMS1 subtype showed strong synergistic effects of birinapant and oxaliplatin/5-FU. In contrast, birinapant and TNFalpha co-treatments showed synergistic effects in CMS2 cells, but not in CMS1 cells. Cells belonging to the CMS3 subtype, however, showed a mixed but generally weak response to the co-treatments. Our results highlight birinapant as a promising drug for the treatment of CRC. Our data also suggest that CMSs not only have prognostic values but may also be used as patient stratification tools.

Citation Format: Michael Fichtner, Emir Bozkurt, Katherine McAllister, Christopher McCann, Daniel Longley, Jochen H. Prehn. Birinapant co-treatments of colon cancer cell lines show consensus molecular subtype-specific synergistic effects [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3907.
Abstract 3914: ZW49, a HER2-targeted biparatopic antibody-drug conjugate for the treatment of HER2-expressing cancers
Authors: Hamblett Kevin J. , Hammond Phil W. , Barnscher Stuart D. , Fung Vincent K. , Davies Rupert H. , Wickman Grant R. , Hernandez Andrea , Ding Tong , Galey Adam S. , Winters Geoffrey C. , Rich Jamie R. , Babcook John S.
Published: August 2018

Therapies targeting HER2 have transformed the treatment of patients with HER2-expressing breast and gastric cancers. Unfortunately, many patients recur following HER2-targeted treatments and new therapies are needed. Multiple antibody-drug conjugate (ADC) technologies are being explored in this setting, some of which utilize the anti-HER2 antibody trastuzumab. Here we present the preclinical characterization of a new anti-HER2 biparatopic ADC, ZW49, which is generated from the conjugation of a novel N-acyl sulfonamide auristatin payload to the inter-chain disulfide bond cysteines of the bispecific anti-HER2 IgG1 antibody ZW25, via a protease cleavable linker. A series of in vitro and in vivo experiments were performed to characterize ZW49 as a potential therapeutic candidate. In cellular binding assays, it was confirmed that the payload conjugation to ZW25 did not affect the antibody's binding to HER2-expressing cells. ZW49 displayed potent in vitro cytotoxicity in multiple cancer cell lines expressing HER2 and was efficacious in multiple patient-derived xenograft (PDX) models. In mice bearing the HBCx-13b HER2 3+ PDX, two doses of ZW49 administered two weeks apart generated tumor regressions. Furthermore, preliminary results from PDX models with lower levels of HER2 expression treated with ZW49 also generated regressions. In nonhuman primates ZW49 administered intravenously every two weeks for three doses was well tolerated. Based on these findings, we are proceeding with further development of ZW49 as a therapeutic candidate in HER2-expressing cancers.

Citation Format: Kevin J. Hamblett, Phil W. Hammond, Stuart D. Barnscher, Vincent K. Fung, Rupert H. Davies, Grant R. Wickman, Andrea Hernandez, Tong Ding, Adam S. Galey, Geoffrey C. Winters, Jamie R. Rich, John S. Babcook. ZW49, a HER2-targeted biparatopic antibody-drug conjugate for the treatment of HER2-expressing cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3914.
Abstract 393: Dynamic networks of transcriptome, DNA methylome, and DNA hydroxymethylome during T-cell lineage commitment
Authors: Yoon Byoung-Ha
Published: August 2018

Immunotherapy is a treatment that uses a part of a personal immune system to overcome diseases such as cancer. T cell therapy has been used in a variety of cancer types, including lymphoma, melanoma and colon cancer. There are several strategies used to take advantage of cell-mediated anti-tumor properties of T cells. Cellular immune responses to prevent tumors are typically caused by CD4 and CD8 T cells. The stepwise development of T cells from a multipotent precursor is guided by diverse mechanisms including interactions of lineage specific transcription factors and epigenetic changes and acquires lineage-specific trait. However, in this T-cell development process, a greater understanding of the regulation of genes involved in immunotherapy is needed.

To elucidate the transcriptional networks and epigenetic mechanisms underlying T-cell therapy and T cell lineage commitment, we investigated genome-wide changes in gene expression, DNA methylation and hydroxymethylation among five successive populations of T cell development (DN3, DN4, DP, CD4+, and CD8+) using RNA-seq, MBD-seq and hMeDIP-seq, respectively. And investigated interactive networks of transcription factors, chromatin modifiers, and DNA methylation. We found stage specific differentially methylated regions and some of them occurred near the specific differentially expressed genes related to immune cells or cancer immunotherapy such as Cd4, Ccr9 and Pdcd1. Dynamic changes of DNA methylation and hydroxymethylation is associated with the recruitment of the stage specific transcription factors.

This study may provide a frame work for understanding the complexity of T cell lineage commitment and immunotherapy-related genes were regulated during T-cell differentiation.

Citation Format: Byoung-Ha Yoon. Dynamic networks of transcriptome, DNA methylome, and DNA hydroxymethylome during T-cell lineage commitment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 393.
Abstract 3941: Heparan sulfate, a new target for platinum in metastatic TNBC
Authors: Katner Samantha J. , Hampton James D. , Peterson Erica J. , Katsuta Eriko , Sayyad Megan R. , Takabe Kazuaki , Koblinski Jennifer , Farrell Nicholas P.
Published: August 2018

Heparan sulfate (HS) are linear polysaccharides conjugated to proteins, heparan sulfate proteoglycans (HSPGs), located on the cell surface and extracellular matrix. The HS chains display varying degrees of sulfation. These sulfate clusters mediate the interaction of polynuclear platinum complexes (PPCs) with HSPG through a “sulfate clamp.” Such PPC-HS interactions can be conceptualized as “polyarginine” mimics. Strong HS-PPC binding protects the oligosaccharide against sulfate loss through metalloshielding. The biological consequences of PPCs metalloshielding HS will in principle affect HS interactions with relevant enzymes and proteins such as heparanase and growth factors, similar in concept to inhibition of DNA-protein binding through modification of DNA structure and conformation. HSPGs, associated growth factors, and heparanase promote tumor progression by facilitating invasion, angiogenesis, and metastasis. High heparanase expression correlates with increased metastatic potential and poor clinical prognosis. PPC-HS interactions inhibit cleavage of a model pentasaccharide by heparanase and further modulated bFGF binding to HS, and bFGF-induced migration and signaling in colon cancer cells. The end-point of functional modulation of HS interactions is inhibition of angiogenesis and metastasis. We report proof-of-principle of strong in vivo anti-metastatic activity of PPCs in triple negative breast cancer (TNBC) models. Further, we examine the anti-metastatic and anti-angiogenic effects of PPC-HS metalloshielding in these in vivo models through multiple growth factor signaling pathways (bFGF, HB-EGF, VEGF), and heparanase activity in breast cancer and endothelial cells. PPCs represent a novel class of intrinsically dual-function agents combining platinum cytotoxicity through DNA targeting with anti-angiogenic effects through glycan targeting.

Chicago 14-17 April 2018

Citation Format: Samantha J. Katner, James D. Hampton, Erica J. Peterson, Eriko Katsuta, Megan R. Sayyad, Kazuaki Takabe, Jennifer Koblinski, Nicholas P. Farrell. Heparan sulfate, a new target for platinum in metastatic TNBC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3941.
Abstract 3942: The fraction of garlic protein(s) affect HCC via regulation of angiogenesis
Authors: Chatterjee Nabanita , Das Subhadip , Pal Prajnamoy , Saha Krishna Das
Published: August 2018

Cancers are leading cause of human mortality. Most of deaths are due cancer because of its high resistance to the chemotherapeutic drugs. Thus, there is an ongoing demand for generation of new potent and more effective therapies for cancer treatment. Hepatocellular carcinoma (HCC), is the most leading cause of cancer-related death in all over the world (seems to more than 80% of death cases in USA). It is frequently linked with continuous hepatocytes death, inflammatory cell infiltration, and compensatory liver regeneration with higher angiogenic rate. Considering the different signaling process during this tumorigenesis with the involvement of new ‘lead' identification of various sources, desire to be less toxic and higher bioavailability. The use of natural or its active metabolic component(s) has begun to emerge as a potential innovative approach in medicinal research to deliver bioactive entities. Thus, garlic has already proven as the therapeutic implication on immune system and cancer due to presence of sulfur compound. But the role of active protein fraction present in fresh garlic is still not revealed yet. In our study, we are isolated an active garlic protein from fresh garlic aqueous extract with molecular weight about 40-45kD. This Garlic protein (GP) having non-cytotoxic to human/murine normal cells, could induce apoptosis in human HepG2 cells, as characterized by phosphatydylserine serine exposure, DNA fragmentation, alteration with loss of mitochondrial membrane potential and cytochrome C release, activation of series of caspases, other physiochemical changes lead to induce cancer cell death. This study also focused that the GP effectively suppressed DEN induced liver inflammation and hyperplasia in murine also decline in lipid peroxidation and changes in enzymic antioxidants and biochemical parameter with improvement of their survival rate. It reduced the elevated level of cytokines and growth factors in hepatocytes from liver fibrosis bearing mice. Thus, finding indicated that, GP may induce apoptosis and in hyper proliferating cancer cells via altering cytokines-chemokines level with tumor growth regulation in carcinogen exposed rodent system. Future direction, we are proposing to execute the experiments on p53-/- and cMyc-/- deficient mice to find out the role of GP in these circumstances.

Citation Format: Nabanita Chatterjee, Subhadip Das, Prajnamoy Pal, Krishna Das Saha. The fraction of garlic protein(s) affect HCC via regulation of angiogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3942.
Abstract 3944: Blockage of EGFR signaling repurposes tumor metabolism through suppression of glycolysis and Kreb cycle in head and neck cancer
Authors: Kuo Ching-Chuan , Chang Jang-Yang , Hsieh Hsing-Pang , Kung Hsing-Jien , Chang Hsih-Huei , Huang Chih-Hsiang , Kuo Cheng-Chin , Ke Yi-Yu
Published: August 2018

Head and neck squamous cell carcinoma (HNSCC), the most common malignant neoplasm arising in the mucosa of the upper aerodigestive tract, remains a significant cause of morbidity worldwide. With respect to the cancer treatment, HNSCC has a moderately good survival rate, however, the disease often recurs, leading to a poor prognostic disease course and tends to fail in treatment. The metabolic properties of cancer cells diverge significantly from those of normal cells. Emerging evidence suggests these metabolic alterations are also linked to therapeutic resistance in cancer treatment. EGFR targeted agents currently approved or under investigation for HNSCC. We recently identified a novel EGFR tyrosine kinase inhibitor (EGFR-TKI), BPR3K007S0, and found that this compound significantly inhibited EGFR phosphorylation in EGFR-overexpressing HNSCC cells in vitro and in vivo than that of gefitinib. BPR3K007S0 significantly suppressed the expression of a wide range of metabolic genes, including metabolic-related transcription factors, glycolysis, TCA cycle, and pentose phosphate pathway genes. Collectively, we also found that BPR3K007S0 significantly decreased glycolytic and mitochondria respiratory capacity. Pharmacological and genetic manipulation demonstrated that c-Myc/hexokinase 2 axis was one of downstream effector in response to EGFR inhibition. Furthermore, we found that EGFR-TKIs were able to suppress succinate dehydrogenase A leading to reduce fumarate, an oncometabolite generates from Kreb cycle, and contributed to EGFR-TKIs mediated antitumor effect. Taken together, these results revealed that blockage of EGFR signaling repurposes tumor metabolism through suppression of hexokinase 2 and succinate dehydrogenase A, in glycolysis and Kreb cycle, respectively, and demonstrated added benefits to treatment of HNSCCs.

Citation Format: Ching-Chuan Kuo, Jang-Yang Chang, Hsing-Pang Hsieh, Hsing-Jien Kung, Hsih-Huei Chang, Chih-Hsiang Huang, Cheng-Chin Kuo, Yi-Yu Ke. Blockage of EGFR signaling repurposes tumor metabolism through suppression of glycolysis and Kreb cycle in head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3944.
Abstract 3952: Selective high affinity MYC-binding compound inhibits MYC-MAX interaction and MYC-dependent tumor cell growth
Authors: Castell Alina , Yan Qinzi , Fawkner Karin , Hydbring Per , Zhang Fan , Verschut Vasiliki , Franco Marcela , Zinzalla Giovanna , Larsson Lars-Gunnar
Published: August 2018

The MYC family oncoproteins/transcription factors MYC, MYCN and MYCL (here referred to as MYC) are key players in tumor development and are particularly associated with aggressive disease and poor prognosis. Efficient and specific MYC-targeting drugs are therefore highly warranted, but no such drugs are available in the clinic at present. MYC is strictly dependent on heterodimerization with MAX for activation of transcription. In a cell-based Bimolecular Fluorescence Complementation protein-protein interaction screen for small molecule inhibitors we identified a molecule that exhibits strong selective inhibition of MYC-MAX interaction in cells as validated by Gaussia luciferase protein complementation assay, coimmunoprecipitation and in situ proximity ligation (isPLA) assay, reaching an IC50 at single-digit micromolar concentrations. The molecule was shown to inhibit MYC-MAX interactions in a biochemical FRET assay and binds selectively to the MYC bHLHZip domain with affinity in the single digit micromolar range as demonstrated by Microscale Thermophoresis and Surface Plasmon Resonance. Further, within the same concentration range, this molecule blocks MYC-driven transcription and efficiently inhibits tumor cell growth in a MYC-dependent manner, but spares normal cells. Moreover, the growth inhibitory responses to the molecule correlated significantly with MYC expression levels in a cohort of 60 human tumor cell lines. Importantly, utilizing a mouse tumor model of MYCN-amplified neuroblastoma, treatment with the molecule resulted in significant inhibition of the MYC-MAX interaction in tumor tissue, as shown by isPLA, and massive induction of apoptosis in the tumors. Since this molecule, unlike many experimental MYC inhibitors, is selective, has high affinity for MYC, has high efficacy in cells, reaches its target in vivo and does not affect MYC expression levels, it can be used as a chemical tool to specifically study the role of the MYC-MAX complex in MYC biology in normal and cancerous cells, and it has potential for drug development.

Citation Format: Alina Castell, Qinzi Yan, Karin Fawkner, Per Hydbring, Fan Zhang, Vasiliki Verschut, Marcela Franco, Giovanna Zinzalla, Lars-Gunnar Larsson. Selective high affinity MYC-binding compound inhibits MYC-MAX interaction and MYC-dependent tumor cell growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3952.
Abstract 3958: Inhibiting the inhibitor: Exploring the p53-Rbm38 loop as a potential cancer therapeutic target
Authors: Lucchesi Chris A.
Published: August 2018

Modulating Rbm38 negative regulation of p53 translation may be an effective therapeutic target in human cancers. As a focal regulatory component of RNA metabolism, RNA-binding proteins (RBPs) modulate all facets of RNA biogenesis. Given the fact that RBPs control RNA metabolism, defects in the function of RBPs can have far reaching implications including neurodegenerative disorders and cancer. Our group discovered that Rbm38 suppresses p53 mRNA translation by interacting with eukaryotic translation initiation factor 4E (eIF4E) on p53 mRNA. Conceptually, blocking the Rbm38 interaction with eIF4E employing synthetic peptides corresponding to their respective binding interface should relieve the translational repression caused by Rbm38. Indeed, treating cancer cells with synthetic peptides corresponding to the binding interface between Rbm38 and eIF4E resulted in robust p53 protein expression. Moreover, addition of peptide and low dose doxorubicin caused further accumulation of p53. Consequently, treatment of cancer cells with peptide alone or in combination with low dose doxorubicin leads to a decrease in colony formation as well as tumor sphere formation. Of significance, RKO xenografts with wild-type p53 were highly sensitive to treatment with a synthetic peptide comprising just 8 amino acids (Pep8) compared to the control group. Together, these data support that p53 expression can be modulated by using small peptides to inhibit Rbm38 from interacting with eIF4E, which may be used as a therapeutic approach in cancers carrying wild-type p53.

Citation Format: Chris A. Lucchesi. Inhibiting the inhibitor: Exploring the p53-Rbm38 loop as a potential cancer therapeutic target [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3958.
Abstract 3990: Clinicopathologic findings and in vitro efficacy of MYC-targeting agentsin NUT midline carcinoma
Authors: Kim Soyeon , Jung Minsun , Park Heae Surng , Yoon Sun Och , Hong Soon Won , Park Weon-Seo , Keam Bhumsuk , Kim Hyun Jik , Kim Dong-Wan , Kim Young Tae , Heo Dae Seong , Kim Tae Min , Jeon Yoon Kyung
Published: August 2018

Purpose: NUT midline carcinoma (NMC) is an aggressive and poorly differentiated carcinoma defined by chromosomal rearrangements of NUT gene, which is fused to the BET family genes (BRD3/4). Although NMC is extremely rare and mostly reported in a Western population, clinico-pathologic findings of Asian NMC patients are not well studied. Here, we performed the clinicopathologic analysis of Korean NMC cases and in vitro efficacy of MYC-targeting agents against patient-derived NMC cell lines.

Experimental Design: A total of 12 NMC patients (3 thoracic and 9 head and neck origins) were collected in Korean multi-centers by immunohistochemistry (IHC) and fluorescence in situ hybridization for NUT. IHC staining was performed for c-MYC, p53, EGFR, HER2, and PD-L1. NMC cell lines (SNU-2972-1, SNU-3178S, HCC2429, and Ty-82) were exposed to MYC-targeting agents including BET (I-BET, OTX-015, and AZD5153) and HDAC (vorinostat, romidepsin, panobinostat, and CUDC-907) inhibitors. Cell viability assays were performed and IC50 values were determined.

Results: Primary sites of tumor included sinonasal area (n=7), salivary gland (n=2), and thorax/lung (n=3). The median age was 47.5 years (range, 8-73 years) with equal male-to-female ratio. NUT immunostaining showed diffuse positivity in all except for one who had focal NUT positivity. C-MYC and p53 were expressed in all evaluated cases. PD-L1 expression was observed only in a patient (8.3%). Seven of 12 patients died at 3-24 months (median, 9 months) after diagnosis. Long-term survivor was observed in one patient with stage IV NMC who received mass excision plus metastasectomy followed by 6 cycles of docetaxel plus cisplatin and radiotherapy (survival > 27 months). Although BET and HDAC inhibitors showed limited in vitro efficacies against NMC cells, a dual HDAC/PI3K inhibitor, CDUC-907 exhibited the most potent efficacy with the IC50 of 6.2 to 9 pmol/L.

Conclusions: Korean NMC patients pursued grave prognosis like Western patients except one long-term survivor who received tri-modality treatments. Dual HDAC/PI3K inhibitor might be promising in NMC treatment.

Citation Format: Soyeon Kim, Minsun Jung, Heae Surng Park, Sun Och Yoon, Soon Won Hong, Weon-Seo Park, Bhumsuk Keam, Hyun Jik Kim, Dong-Wan Kim, Young Tae Kim, Dae Seong Heo, Tae Min Kim, Yoon Kyung Jeon. Clinicopathologic findings and in vitro efficacy of MYC-targeting agentsin NUT midline carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3990.
Abstract 4010: Identification of signaling pathways involved in colorectal adenoma-to-carcinoma progression using phosphoproteomics
Authors: Kemp Sanne Martens-de , Henneman Alex , Haas Richard de Goeij-de , Piersma Sander , Pham Thang , Meijer Gerrit , Carvalho Beatriz , Jimenez Connie
Published: August 2018

Background Colorectal cancer (CRC) develops in a multi-step-process from normal epithelium, through a pre-malignant lesion (adenoma), into a malignant lesion (adenocarcinoma). A minority of about 5% of adenomas will ultimately progress into cancer. To improve clinical practice it is important to identify the subgroup of colorectal adenomas that is at risk of progressing to cancer. By extensive genomic analysis we have shown that specific copy number alterations are associated with risk of progression. An example of such an alteration is the gain of chromosome arm 20q and we identified AURKA and TPX2 as major drivers of this amplicon. Yet, the activation status of these and other genes during colorectal carcinogenesis and the downstream signaling pathways affected with gene (in)activation are not fully known. In order to better understand the biology of adenoma to carcinoma transition, we performed a comprehensive analysis of phosphoproteomes at different stages of colorectal carcinogenesis.

Methods Phosphotyrosine containing peptides were immunoprecipitated from 5 mg of colorectal adenoma (n=81) and colorectal carcinoma tissues (n=50) using agarose bead-coupled phosphotyrosine antibody P-Tyr-1000. A Q Exactive HF mass spectrometer was used to perform NanoLC-MS/MS. Spectral counts of phosphoproteins and ion intensities of phosphopeptides were defined by MaxQuant for relative quantitation of protein phosphorylation.

Results Phosphotyrosine-based phosphoproteomics of the 131 colorectal tissue samples yielded 6056 phosphopeptides, corresponding to 2745 unique phosphoproteins including 183 phosphokinases. Data analysis to identify regulated phosphorylation states and inference of kinase activity is ongoing.

Conclusions and future plans This is the first large phosphoproteomics dataset of colorectal adenomas and carcinomas. Preliminary data analysis shows promising differences in kinase activities between adenomas and carcinomas. After further analysis, we will manipulate the activity of the driver kinases in adenoma-derived organoids and monitor tumor outgrowth in mice.

Citation Format: Sanne Martens-de Kemp, Alex Henneman, Richard de Goeij-de Haas, Sander Piersma, Thang Pham, Gerrit Meijer, Beatriz Carvalho, Connie Jimenez. Identification of signaling pathways involved in colorectal adenoma-to-carcinoma progression using phosphoproteomics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4010.
Abstract 4012: SAG/RBX2/ROC2 regulates pancreatic cancer initiation, cystic formation in mice driven by KrasG12D
Authors: Tan Mingjia , Zhang Qiang , Sun Yi
Published: August 2018

BACKGROUND & AIMS: SAG, a small ring finger protein, a component of CRLs, might be involved in pancreatic tumorigenesis. SAG overexpression in human pancreatic ductal adenocarcinomas indicating that SAG could be promotes pancreatic tumorigenesis. We investigated how manipulated SAG (overexpression or disruption of SAG) cooperate KRAS to regulate pancreatic cancer progression in mice.

METHODS: We disrupted or overexpressed SAG specifically in pancreata of mice (Sagfl/fl;P48-Cre or SagTG;P48-Cre mice) and crossed them with LSL-KRASG12D mice, which express an activated form of KRAS and develop spontaneous pancreatic tumors. The resulting Sagfl/fl;LSL-KRASG12D;P48-Cre or SagTG;LSL-KRASG12D;P48-Cre mice were monitored; Pancreatic tissues were collected and analyzed by histology and immunohistochemical analyses. Protein was isolated from mouse tissues and primary acinar cells were prepared and cultured in 3D condition. Human PDA TMA was analyzed by immunohistochemistry.

RESULTS: Overexpression of Sag with KRASG12D in pancreata of mice increased the proliferation of pancreatic epithelial cells, lead to formation of acinar to ductal metaplasia and pancreatic intraepithelial neoplasias (mPanINs) and shorten mice survival. Disruption of Sag in LSL-KRASG12D;P48-Cre mice accelerated the growth of pancreatic cystic lesions compared to LSL-KRASG12D;P48-Cre mice, but did not change mouse survival. In mechanism, Sag promotes pancreatic tumorigenesis by regulating Deptor/mTOR/S6K axis and EGFR signaling pathway.

CONCLUSION: Sag is an oncogenic cooperator of activated KRAS for pancreatic tumorigenesis. Targeting SAG E3 ligase may, therefore, have therapeutic value for the treatment of prostate cancer associated with mutant KRAS.

Citation Format: Mingjia Tan, Qiang Zhang, Yi Sun. SAG/RBX2/ROC2 regulates pancreatic cancer initiation, cystic formation in mice driven by KrasG12D [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4012.
Abstract 4019: The chronic intermittent hypoxia enhances cancer progression in lung cancer mouse model
Authors: Lee SangHaak , Kim In Kyoung , Kang Hyeon Hui , Kwon Hee Young , Kang Hye Sun , Yeo Chang Dong
Published: August 2018

Background: Obstructive sleep apnoea (OSA) is a very prevalent disorder charaterized by chronic intermittent hypoxia (CIH) and some reports suggested that OSA is related to increased incidence of cancer as well as cancer progression. The purpose of this study was to evaluate whether intermittent hypoxia with a time course similar to that found in OSA influence lung cancer development and progression in metastatic lung cancer animal model.

Methods: C57/BL6 mice were divided into two groups; control condition (room air) or CIH. The mice of CIH group were exposed to IH for 2 weeks before and immediately for 2 weeks after tumour induction by tail vein injection of carcinoma cells. At day 14, the mice were sacrificed and measured cancer growth. Hypoxia inducible factor (HIF-1α), vascular endothelial growth factor (VEGF) and metastasis related matrix metalloproteinases (MMP) were measured by ELISA and gelatin zymography in lung tissue and serum, respectively. The mRNA expressions of epithelial mesenchymal transition markers were measured by real-time PCR in lung tissue.

Results: The marked increase in tumor number and volume was found in CIH group (p < 0.05). The accumulation of total cell and neutrophils in BAL fluids and the activation of MMP -2 and -9 in serum were also increased by CIH. The level of HIF-1α in the nuclear fraction of pneumocytes and the expression of VEGF were increased significantly in CIH group compared to the control (p < 0.05). The mRNA expression of E-cadherin was decreased in CIH group compared to the control group while vimentin was increased.

Conclusions: Our data shows that CIH is associated with enhanced proliferative and migratory properties of cancer. These results suggest evidence for the causal link between OSA and lung cancer progression.

Key words: chronic intermittent hypoxia, obstructive sleep apnea, lung cancer

Citation Format: SangHaak Lee, In Kyoung Kim, Hyeon Hui Kang, Hee Young Kwon, Hye Sun Kang, Chang Dong Yeo. The chronic intermittent hypoxia enhances cancer progression in lung cancer mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4019.
Abstract 4028: Molecular mechanisms of magnolin resistance in ovarian cancer cells
Authors: Yoo Sun-Mi , Lee Cheol-Jung , Kim Seung-Min , Cho Seon-Yeon , Park Juhee , Cho Yong-Yeon
Published: August 2018

Mitogen-activated protein kinase (MAPK) and PKB signaling pathways are highly activated in many human solid cancers and play a key role in carcinogenesis, tumor growth and cancer cell survival. Although regulation mechanisms of each signaling pathway are well-elucidated, interrelationship between both signaling pathways in cancer growth has not been understood. In this study, we found that magnolin, ERK1 and ERK2 inhibitor with 16 nM and 68 nM of IC50 value, suppressed cell proliferation and colony growth of ovarian cancer cells. In signaling pathway analysis, we found that TOV-112 and SKOV3 showed differential responsiveness by EGF stimulation. The differential responsiveness was caused by differential activation signaling through MAPK and PKB signaling pathway. Notably, TOV-112D sensitized by magnolin treatment in cell proliferation and colony growth in soft agar by alteration of cell cycle regulation, but not in SKOV3 cells. Notably, phosphorylation of ATF-1, a downstream target of RSK2, at Ser63 was inhibited in TOV-112 by magnolin treatment, resulted in cellular senescence. However, SKOV3 was vice versa. We further found that magnolin fraction suppressed TOV-112D cancer growth in xenograft animal model. Taken together, we concluded that signaling alteration might involve in magnolin resistance in SKOV3.

Keywords: MAP kinase, molecular targets, ovarian cancer, cancer growth

Citation Format: Sun-Mi Yoo, Cheol-Jung Lee, Seung-Min Kim, Seon-Yeon Cho, Juhee Park, Yong-Yeon Cho. Molecular mechanisms of magnolin resistance in ovarian cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4028.
Abstract 4035: Cetuximab combined with dasatinib reduces drug resistance in KRAS/BRAF mutated colorectal carcinoma cells
Authors: Chiu Hsiang-Ling , Wu Yi-Che , Wei Wan-Chen , Kao Ling-Yi , Chao Wei-Ting
Published: August 2018

Colorectal carcinoma is the most common cancer worldwide. Cetuximab is the first-line target drug for late-stage metastatic colon cancer treatment; however, resistance to it was found in 45% patients having KRAS mutation. Therefore, the alternative signal axis to target mutated cancer cells is urgent and investigated. The role and the importance of Src has emerged in colon cancer; this study is to investigate the drug combined efficacy of cetuximab and dasatinib, the Src inhibitor and the underling molecular mechanism in KRAS/BRAF mutated cells. In this study, KRAS and BRAF mutated colon cancer cell lines SW480 and HT-29 were treated with cetuximab, dasatinib and fluorouracil (5-FU) solitary or combined. Cell viability and migration ability were monitored, and the molecular mechanisms were analyzed by Western blot for signal protein expressions. The in vivo study was carried with subcutaneous xenograft mouse model to evaluate the drug efficacy. The result showed that cetuximab treatment induced Src activation dose dependently, and induced matrix-dependent cell migration in SW480 cells. Cetuximab, dasatinib and 5-FU combined treatment compared to solitary treatment can reduce cell viability in either wild-type or KRAS/BRAF mutated colon cells. In the signal protein expressions, Src expression was suppressed sufficiently with dasatinib treatment; the phosphorylation of MAPK in SW480 cells and the phosphorylation of PI3K in HT-29 cells were downregulated after treated with cetuximab together with dasatinib and 5-FU. The drug combination was also demonstrated to suppress subcutaneous tumor growth as well. These findings demonstrate the combination of cetuximab, dasatinib and 5-FU can be the potential treating strategy for colon cancer cells with KRAS/BRAF mutation.

Citation Format: Hsiang-Ling Chiu, Yi-Che Wu, Wan-Chen Wei, Ling-Yi Kao, Wei-Ting Chao. Cetuximab combined with dasatinib reduces drug resistance in KRAS/BRAF mutated colorectal carcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4035.
Abstract 4041: K-ras transformation leads to increased HS-degradation
Authors: Ju Donghong , Kosir Mary A.
Published: August 2018

INTRODUCTION: Degradation of glycosaminoglycans in the extracellular matrix impacts cell movement, angiogenesis and other cancer cell activities. The purpose of study was to detect and measure extracellular heparan sulfate (HS)-degrading enzyme in malignant ras-transformed cells where HS degradation occurred at attachment sites. METHODS: Serum free conditioned medium (CM) and lysate from subconfluent Balb/c3T3 and KiMSV (Ki-ras-transformed 3T3) cells were collected and protease inhibitors added. CM was concentrated by ultracentrifugal filters using 3K and 30K MWCO. Heparan degrading enzyme assay kit was use with lysate and CM for each cell line. Western blot analysis of lysate and CM used anti-HPSE, anti-NAP2, and anti-PPBP antibodies with actin control. mRNA for HPSE and PPBP was determined by RTPCR. RESULTS: The heparan degrading activity of CM from KiMSV cells was 12 fold higher than from Balb/c3T3 cells with 30K MWCO concentration. Further, HPSE was detected in the lysate, but not CM. Anti-NAP2 and anti-PPBP antibody detected protein greater than 30kD in CM and lysate for KiMSV cells. However, mRNA expression of PPBP/NAP2 was less in KiMSV than Balb/c3T3 cells. CONCLUSION: HS degradation in the CM of KiMSV cells is greater than for Balb/c3T3 cells from which they are derived after Ki-ras-transformation. The HPSE is not detected in the CM, but PPBP-like proteins at higher molecular weight are detected which may be responsible for the extracellular HS degradation.

Citation Format: Donghong Ju, Mary A. Kosir. K-ras transformation leads to increased HS-degradation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4041.