bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4445: Post-transcriptional inhibition of PTPN23 by SND1: Potential mechanism for hepatocellular carcinoma
Authors: Jariwala Nidhi H. , Mendoza Rachel G. , Dawn Garcia , Zhao Lai , Subler Mark A. , Windle Jolene J. , Fisher Paul B. , Yidong Chen , Sarkar Devanand
Published: August 2018

Oncoprotein SND1 regulates gene expression at a post-transcriptional level in multiple cancers including hepatocellular carcinoma (HCC). Staphylococcal nuclease domains of SND1 function as an RNAse and tudor domain facilitates protein-oligonucleotide interaction. In the present study, we aimed to identify RNA interactome of SND1 to obtain better insights into gene regulation by SND1. RNA interactome was identified by immunoprecipitation of RNA using anti-SND1 antibody from human HCC cells followed by RNA-sequencing (RIP-Seq). With an adjusted p value of <0.01 and >2-fold enrichment over control, 282 mRNAs were identified to significantly associate with SND1 protein. We focused on the tumor suppressor PTPN23 because its regulation by SND1 and its role in HCC are not known. PTPN23 levels were downregulated in human HCC cells versus normal hepatocytes and in human HCC tissues versus normal adjacent liver as revealed by immunohistochemistry. In human HCC cells, knocking down SND1 increased and overexpression of SND1 decreased PTPN23 protein. RNA binding and degradation assays revealed that SND1 specifically binds to and degrades 3'-UTR of PTPN23 mRNA. Tetracycline-inducible PTPN23 overexpression in human HCC cells resulted in significant inhibition in proliferation, migration and invasion and in vivo tumorigenesis. PTPN23 induction caused inhibition in activation of c-Met, EGFR, Src and FAK, suggesting that as a phosphatase PTPN23 inhibits activation of these oncogenic kinases. The present study unravels a novel function of SND1 in targeting PTPN23 and identifies PTPN23 as a unique tumor suppressor for HCC. PTPN23 might function as a homeostatic regulator of multiple kinases restraining their activation.

Citation Format: Nidhi H. Jariwala, Rachel G. Mendoza, Garcia Dawn, Lai Zhao, Mark A. Subler, Jolene J. Windle, Paul B. Fisher, Chen Yidong, Devanand Sarkar. Post-transcriptional inhibition of PTPN23 by SND1: Potential mechanism for hepatocellular 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 4445.
Abstract 4490: Autocrine and paracrine effects of miR-29 in hPSCs and PDAC cells
Authors: Factora Tricia , Briscoe Elise , Kwon Jason , Kota Janaiah
Published: August 2018

Pancreatic ductal adenocarcinoma (PDAC) continues to be one of the most lethal malignancies worldwide and responds poorly to current therapeutic modalities. Due to its late stage diagnosis and limited response to current treatments, it is imperative to develop novel and effective therapies for PDAC. Dense fibrotic stroma associated with PDAC abrogates drug perfusion into the tumor and activated pancreatic stellate cells (PSCs) are the major stromal cells responsible for stromal reaction. Furthermore, activated PSCs are known to secrete various factors causing autocrine effects to elicit sustained PSC activation and paracrine effects on cancer cells to promote tumor progression. Our previous work has shown that loss of miR-29 is a common phenomenon of both activated PSCs and pancreatic cancer cells, and its restored expression reduces stromal accumulation and cancer growth. In addition, conditioned media from miR-29 overexpressing PSCs reduces cancer cell viability. To extend our work on the role of miR-29 in tumor-stromal interactions, we examined how autocrine and paracrine signaling is affected by miR-29 overexpression in PSCs and cancer cells. We collected the conditioned media from miR-29a overexpressing human PSCs (hPSC-CM) or cancer cells (CC-CM) and cultured nascent PSCs or cancer cells to evaluate autocrine and paracrine effects. hPSCs and PDAC cells cultured in miR-29 hPSC-CM resulted in a significant reduction of cell viability. Likewise, miR-29 overexpressing CC-CM reduced the viability of hPSCs and cancer cells. Additionally, cancer cell migration was reduced when treated with miR-29 overexpressing hPSC- and CC-CM. Taken together, our results demonstrate that conditioned media collected from miR-29 overexpressing hPSCs and cancer cells reduces hPSC/cancer cell viability and cancer cell migration. Moreover, our findings highlight the important role of miR-29 in the autocrine/paracrine interactions between PSCs and cancer cells. In our future studies, we expect to identify the key factors that are regulated by miR-29 in tumor-stromal interactions.

Citation Format: Tricia Factora, Elise Briscoe, Jason Kwon, Janaiah Kota. Autocrine and paracrine effects of miR-29 in hPSCs and PDAC 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 4490.
Abstract 4541: MET gene copy number gains evaluated by NGS is more predictive than other methods to enrich for papillary RCC patients sensitive to savolitinib, a selective MET inhibitor
Authors: Frigault Melanie M. , Signoretti Sabina , Markovets Aleksandra , Stetson Daniel , Howat William , Roudier Martine , Haddad Vincent , Dougherty Brian , Choueiri Toni K. , Barrett J. Carl
Published: August 2018

There is no approved therapy specifically for the treatment of papillary renal cell carcinoma (PRCC). Advances in molecular profiling of PRCC have identified a segment of PRCC with 10% MET mutation rates (Linehan et al 2016). Chromosome 7 gain is a hallmark of PRCC and thought to occur at 50% frequency in PRCC (Jiang et al 1998). We undertook a retrospective analysis of archival tumors to evaluate MET pathway aberrations for correlation with efficacy in a phase II study of savolitinib (volitinib, AZD6094, HMPL-504) in patients with PRCC (NCT02127710). Archival diagnostic tumor samples were mandated for central confirmation of PRCC diagnosis, histological subtyping, and for exploratory biomarker analysis. Eighty-four archival tumors were obtained and profiled using four methods: H&E stain for PRCC histological subtype, immunohistochemistry (IHC) for c-Met protein expression (Ventana, CONFIRM), fluorescent in situ hybridization (FISH) for MET gene amplification (Abbott, VYSIS), and Next Generation Sequencing (NGS) as an orthogonal method for confirmation of MET amplifications, detection of HGF gene amplifications, MET mutations, chromosome 7 ploidy, and other exploratory genomic biomarkers (Foundation Medicine Inc., T7 panel). Eight patients with a confirmed partial response (PR) to savolitinib were used to define MET-driven PRCC. Retrospective analysis demonstrated that only NGS, not IHC, FISH or histological subtype, identified all PRs. NGS detected any one of the following biomarkers alone or in combination in PRs: chromosome 7 copy number gain, MET or HGF gene focal amplification, MET kinase domain mutations. The frequency of these biomarkers in the phase II population were 31.5%, 10%, 1%, and 7.5%, respectively and captured all responders (18% ORR); however, 42% of non-responders (PD and SD) were also classified as MET-driven by NGS. Focal MET amplifications were confirmed with MET FISH testing, but FISH was unable to identify gains in chromosome 7. IHC was not as sensitive as NGS (57% vs 100%) but had a higher specificity. Molecular characterization of MET status was more predictive of response to savolitinib than a classification based on pathology.

Citation Format: Melanie M. Frigault, Sabina Signoretti, Aleksandra Markovets, Daniel Stetson, William Howat, Martine Roudier, Vincent Haddad, Brian Dougherty, Toni K. Choueiri, J. Carl Barrett. MET gene copy number gains evaluated by NGS is more predictive than other methods to enrich for papillary RCC patients sensitive to savolitinib, a selective MET inhibitor [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 4541.
Abstract 4573: Localized synchrotron radiation in mice induces persistent systemic genotoxic events mediated by the functional immune system
Authors: Ventura Jessica , Lobachevsky Pavel , Palazzolo Jason , Forrester Helen , Haynes Nicole , Ivashkevich Alesia , Stevenson Andrew , Hall Christopher , Gorgoulis Vassilis , Hamilton John , Georgakilas Alexandros , Sprung Calr , Martin Olga A.
Published: August 2018

The discovery of the radiation-induced bystander effect has expanded knowledge of radiobiological mechanisms of ionizing radiation. A counterpart in vivo phenomenon is the radiation-induced abscopal effect (RIAE). It is not known how radiation settings affect non-targeted normal tissues and therefore the risk of adverse RIAE. At the Australian Synchrotron we examined systemic effects of microbeam radiotherapy (MRT) and broad beam (BB) configurations, in mice that were locally exposed to a very short pulse of a high dose-rate synchrotron beam (49 Gy/sec). We determined how radiation volume and dose impact the RIAE. C57BL/6 mice were irradiated with 10 or 40 Gy incident dose of MRT or BB in an 8x8, 8x1, or 2x2-mm area of the right hind leg. Blood samples, irradiated skin and a variety of normal unirradiated tissues were collected for DNA damage analysis of double-strand breaks (DSBs) quantified as gamma-H2AX foci and oxidatitive clustered DNA lesions (OCDL). OCDLs elevated in a wide variety of unirradiated normal tissues. In out-of-field duodenum, a trend for elevated apoptotic cells was observed, however DSBs elevated only after exposure to lower doses. These genotoxic events were accompanied by changes in concentrations of MDC, CCL2/MCP1, Eotaxin, IL-10, TIMP-1, VEGF, TGFβ-1 and TGFβ-2 plasma cytokines and by changes in frequencies of macrophages, neutrophils and T-lymphocytes in duodenum. Overall, systemic radiation responses were dose-independent (1). The MRT irradiations was repeated in immune-deficient mice: NSG, CCL2 knock-outs, and in C57BL/6 mice treated with anti-CSF1R antibody which effectively depletes macrophages. Strikingly, these effects and the abscopal innate and adaptive immune effector responses were completely or partially abrogated in the mice with various immune deficiencies, highlighting the role of the functional immune system in propagation of systemic genotoxic effects of radiation. These findings have implications for the planning of therapeutic and diagnostic radiation to reduce the risk of radiation-related adverse systemic effects. 1. Ventura et al, Cancer Research, e-pub (2017).

Citation Format: Jessica Ventura, Pavel Lobachevsky, Jason Palazzolo, Helen Forrester, Nicole Haynes, Alesia Ivashkevich, Andrew Stevenson, Christopher Hall, Vassilis Gorgoulis, John Hamilton, Alexandros Georgakilas, Calr Sprung, Olga A. Martin. Localized synchrotron radiation in mice induces persistent systemic genotoxic events mediated by the functional immune system [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 4573.
Abstract 4760: Joint modeling of longitudinal tumor dynamics and survival in non-small cell lung cancer (NSCLC) patients
Authors: Tong Xiao , Dunyak James , Zhou Diansong , Carlile David , Tomkinson Helen , Helmlinger Gabriel , Al-Huniti Nidal , Xu Hongmei
Published: August 2018

Objectives: Tumor size dynamics and survival are traditionally analyzed in a 2-stage approach, without consideration of joint dependencies. The objective of this analysis was to develop a joint model which associates tumor size dynamics and progression-free survival (PFS), to predict time-to-progression. Methods: Phase 2 trial data from selumetinib (AZD6244; ARRY-142886, MEK inhibitor), in NSCLC patients (SELECT-2; NCT01750281) were used to develop a joint model for tumor size dynamics and PFS. The analysis was performed using JM package in R. Treatment arm, KRAS mutation and WHO performance status were evaluated as covariates. Model was evaluated by survival estimation based on early time (e.g., first 3 months) tumor size data. The final joint model based on SELECT-2 data was then used to predict PFS of selumetinib in SELECT-1 (NCT01933932) phase 3 trial. Results: The joint model developed on SELECT-2 data identified a significant association (p value <0.001) between the slope of the longitudinal tumor dynamic and PFS. WHO performance status was identified to be the only significant covariate. Furthermore, the model built on phase 2 data adequately predicted PFS of the SELECT-1 data, using SELECT-1 tumor size data within 4 months of treatment. Prediction confirmed no significant difference in PFS between active treatment arm and chemotherapy arm in SELECT-1 trial. Conclusions: Using selumetinib as an example, we showed that joint modeling of tumor size dynamics and PFS may provide a novel quantitative tool to predict long-term outcome in NSCLC based on early tumor size measurements.

Citation Format: Xiao Tong, James Dunyak, Diansong Zhou, David Carlile, Helen Tomkinson, Gabriel Helmlinger, Nidal Al-Huniti, Hongmei Xu. Joint modeling of longitudinal tumor dynamics and survival in non-small cell lung cancer (NSCLC) patients [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 4760.
Abstract 4780: A novel, potent and selective FGFR4 inhibitor, HM81422 in hepatocellular carcinoma with FGFR4-driven pathway activation
Authors: Lee JaeHo , Kang Hyunjeong , Koo Kyounghwa , Ha Youngeun , Lim Sun Young , Byun Joo-Yun , Yu Hyunkyung , Song Taehun , Lee Moonsub , Jung Seung Hyun , Kim Taewoo , Bang Hyojeong , Kim Eunyoung , Kang Jahoon , Lee Ho Jeong , Kim YoungHoon , Ahn YoungGil , Suh KweeHyun , Kim Sun-Jin
Published: August 2018

Introduction: Hepatocellular carcinoma (HCC) is the sixth most prevalent cancer and the second most frequent cause of cancer-related death, however, treatment options are very limited. In recent studies, aberrant signaling through FGFR4 and its ligand, FGF19 have been identified as the oncogenic driver in a subset of HCCs and reported to be associated with poor prognosis. About 30% of HCC patients have altered FGF19/FGFR4 pathway signaling, therefore, the treatment with FGFR4 inhibitor may produce benefit. Materials and Method: Using the structure-based design, we have generated a novel, potent and selective FGFR4 inhibitor, HM81422 with irreversible-covalent binding mode, and evaluated its anti-tumor activity in a variety of HCC cell lines, HCC cell line xenografts and orthotopic grafts. Results: Biochemical selectivity assays demonstrated that HM81422 is highly selective towards FGFR4 compared to other FGFR isotypes as well as a panel of several kinases. The treatment of HM81422 to FGF19 amplified and overexpressed HCC cell lines led to suppression of FGF19/FGFR4 signaling pathway and concomitant reduction in cell viability in dose-dependent manner. Oral administration of HM81422 to mice bearing FGF19 altered HCC cells showed dose-dependent pharmacokinetics, pharmacodynamic modulation of FGFR4 signaling and antitumor efficacy in xenograft models. And HM81422 demonstrated inhibition of tumor growth in an orthotopic liver xenograft model of FGF19 altered HCC in nude mice. Conclusion: In conclusion, the treatment of HCC patients with a potent and selective FGFR4 inhibitor, HM81422, can be an attractive approach targeting approximately 30% of HCC patients by inhibiting altered FGF19-mediated signaling cascade. Further preclinical studies with HM81422 will be performed and reported soon.

Citation Format: JaeHo Lee, Hyunjeong Kang, Kyounghwa Koo, Youngeun Ha, Sun Young Lim, Joo-Yun Byun, Hyunkyung Yu, Taehun Song, Moonsub Lee, Seung Hyun Jung, Taewoo Kim, Hyojeong Bang, Eunyoung Kim, Jahoon Kang, Ho Jeong Lee, YoungHoon Kim, YoungGil Ahn, KweeHyun Suh, Sun-Jin Kim. A novel, potent and selective FGFR4 inhibitor, HM81422 in hepatocellular carcinoma with FGFR4-driven pathway activation [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 4780.
Abstract 4849: Targeting the lethal pediatric atypical teratoid/rhabdoid tumors with the DNA minor-groove binding agent quinacrine
Authors: Kaur Harpreet , Guo Huizi , Eberhart Charles G. , Raabe Eric H.
Published: August 2018

Atypical teratoid/rhabdoid tumors (AT/RT) are rare, incurable, and highly proliferative pediatric brain tumors. Discovering new targets and developing novel therapeutics are urgently needed for this devastating tumor. We have previously shown that AT/RT tumors and cell lines express increased amounts of the epigenetic modifier high mobility group AT-hook 2 (HMGA2). HMGA2 is a DNA-binding oncoprotein that regulates transcription during normal embryogenesis and in cancer stem cells. Targeting HMGA2 using short hairpins significantly decreased AT/RT cell growth and increased survival of xenografted mice in our studies. We hypothesized that pharmacological inhibition of HMGA proteins using DNA minor-groove binding drugs will decrease growth of AT/RT cell lines due to displacement of HMGA proteins from the DNA. We used the minor-groove binding agent quinacrine to test our hypothesis in 3 different AT/RT cell lines (BT37, CHLA-06 and CHLA-04). Quinacrine has been used in millions of humans to treat malaria and other parasitic infections and has a well known safety profile. Quinacrine penetrates the brain, and we can achieve micromolar levels of quinacrine in brain after oral administration. Quinacrine causes a dose-dependent reduction in AT/RT cell growth (MTS assay) and proliferation (BrdU incorporation) compared to vehicle-treated cells (P<0.05). Additionally, treatment of AT/RT cells with quinacrine significantly increased apoptotic cell death (increased cleaved caspase-3 and cleaved PARP expression) in a dose-dependent manner compared to vehicle-treated cells (P<0.05). Our results suggest that minor groove binding drugs like quinacrine are a viable potential treatment strategy for AT/RT. Future studies are aimed at testing the in vivo efficacy and validating the mechanism of action of quinacrine in AT/RT.

Citation Format: Harpreet Kaur, Huizi Guo, Charles G. Eberhart, Eric H. Raabe. Targeting the lethal pediatric atypical teratoid/rhabdoid tumors with the DNA minor-groove binding agent quinacrine [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 4849.
Abstract 4878: RMC-4550, an allosteric inhibitor of SHP2: Synthesis, structure, and anti-tumor activity
Authors: Koltun Elena S. , Aay Naing , Buckl Andreas , Jogalekar Ashutosh S. , Kiss Gert , Marquez Abby , Mellem Kevin T. , Mordec Kasia , Saldajeno-Concar Mae , Semko Chris M. , Tibrewal Nidhi , Tzitzilonis Christos , Won Walter , Smith Jacqueline A. , Wilson Susan E. , Nichols Robert J. , Wang Zhengping , Wilds David , Singh Mallika , Gill Adrian L.
Published: August 2018

Genetic and pharmacologic evidence has shown that SHP2, a non-receptor protein tyrosine phosphatase (PTP) and scaffold protein encoded by the PTPN11 gene, is a convergent signal transduction node that integrates growth factor signals from multiple receptors to promote activation of RAS and its downstream effectors. Guided by structural insights from X-ray data, we describe a strategy aimed at the identification of a highly potent and selective allosteric SHP2 inhibitor series. Our efforts led to the discovery of RMC-4550, a potent and selective SHP2 inhibitor which exhibits a high quality, drug-like preclinical profile. RMC-4550 inhibits purified, activated full length human SHP2 with an IC50 of 1.55 nM, and has cellular IC50 of 39 nM in PC9 cells with a pERK readout. RMC-4550 has no detectable inhibitory activity up to 10 µM against the catalytic domain of SHP2, a panel of 14 additional protein phosphatases, and a panel of 468 protein kinases. RMC-4550 exhibits low to moderate cross species in vitro intrinsic clearance (3.6-24 µL/min/million cells) in hepatocytes, a high passive permeability (458 nm/s) and efflux ratio of 1. The ADME properties translate into favorable pharmacokinetic profiles in preclinical species. RMC-4550 has moderate to high bioavailability and has a half-life amenable for once daily oral administration. In the EGFR-driven KYSE-520 human esophageal cancer xenograft model, we observed a dose dependent efficacy consistent with target modulation, assessed by phospho-ERK inhibition in tumors. RMC-4550 is well tolerated at doses that achieved maximal and sustained efficacy in this model. RMC-4550 was synthesized in 5 linear (6 total) steps from the readily accessible or commercially available intermediates. The chemical structure and synthesis of RMC-4550, along with detailed structure-activity relationships will be presented. In summary, RMC-4550 exemplifies a novel class of potent allosteric inhibitors of SHP2 with an excellent drug like property profile.

Citation Format: Elena S. Koltun, Naing Aay, Andreas Buckl, Ashutosh S. Jogalekar, Gert Kiss, Abby Marquez, Kevin T. Mellem, Kasia Mordec, Mae Saldajeno-Concar, Chris M. Semko, Nidhi Tibrewal, Christos Tzitzilonis, Walter Won, Jacqueline A. Smith, Susan E. Wilson, Robert J. Nichols, Zhengping Wang, David Wilds, Mallika Singh, Adrian L. Gill. RMC-4550, an allosteric inhibitor of SHP2: Synthesis, structure, and anti-tumor 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 4878.
Abstract 4909: Adaptive oncology phase 1 study of first-in-class inhibitor of ataxia telangiectasia mutated protein kinase (ATM), in combination with olaparib
Authors: Chen Yingxue , Pass Martin , Bruna Nuria Bui , Stephens Christine , Pierce Andrew , Hanekom Wouter , Gabra Hani , Tomkinson Helen , Al-Huniti Nidal
Published: August 2018

Background: AZD0156 is a highly selective first oral ATM inhibitor in the clinic. Coordinating DNA single and double strand break repair, AZD0156 enhances the preclinical activity of olaparib, leading to its development as an antitumor agent in combination. We report here results from an ongoing phase 1 adaptive dose escalation study of AZD0156 in combination with olaparib.

Methods: This phase I study is to assess the safety, tolerability, pharmacokinetics (PK) and preliminary efficacy of escalating doses of AZD0156 in combination with olaparib in patients with advanced malignancies. AZD0156 is mainly metabolized by CYP3A and FMO, which undergo genetic polymorphism. The flexible language incorporated in the protocol also allow rapid dose escalation beyond subtherapeutic doses.

Results: PK data from the first 2 dose cohorts were lower than predicted and the AZD0156 dose was increased 4 X and dosing changed from QD (daily) to BD (twice daily). The mean terminal half-life of AZD0156 was 9-12 hours and PK was dose proportional. No significant drug-drug interactions (DDI) were observed with olaparib. Minor toxicities included nausea, vomiting and anemia in about 40% of patients in all cohorts. Grade 3 and 4 hematologic toxicities were observed in AZD0156 120mg BD with olaparib 200mg BD, and this dose level was considered intolerable. At doses of AZD0156 of 30 and 60mg BD, active target engagement was demonstrated. PK profile was similar across FMO genotype.

Conclusion: Hematologic toxicities, consistent with the mode of action, emerged as dose-limiting toxicities of AZD0156 in combination with olaparib. Pharmacologically active doses of AZD0156 with olaparib were achieved more quickly in patients by using real-time PK, real-time genotyping, safety, DDI assessment, and PK/PD modelling.

Citation Format: Yingxue Chen, Martin Pass, Nuria Bui Bruna, Christine Stephens, Andrew Pierce, Wouter Hanekom, Hani Gabra, Helen Tomkinson, Nidal Al-Huniti. Adaptive oncology phase 1 study of first-in-class inhibitor of ataxia telangiectasia mutated protein kinase (ATM), in combination with olaparib [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 4909.
Abstract 5022: Precision medicine: High-throughput 3D bioprinting of embedded multicellular cancer spheroids
Authors: Atapattu Lakmali , Utama Robert , O'Mahony Aiden , Fife Christopher , Baek Jongho , Allard Théophile , O'Mahony Kieran , Ribeiro Julio , Gaus Katharina , Gooding Justin , Kavallaris Maria
Published: August 2018

Three-dimensional (3D) cancer cell models, such as 3D multicellular spheroids, are superior models of in vivo systems than the more popular two-dimensional (2D) cell culture. 3D cancer spheroids provide a closer reflection of tumor gene expression and biology than 2D cultures. This is particularly relevant for drug development and precision medicine programs were cell culture conditions that better reflect the 3D tumor environments, including matrix stiffness, would be highly advantageous. A key challenge for the routine application of embedded 3D cancer spheroids in precision and drug programs is the lack of high throughput and uniform production of spheroids in a biocompatible matrix. To address this, we have developed a high-throughput method of producing 3D multicellular cancer spheroids embedded inside a tissue-like matrix using a custom-built drop-on-demand 3D bioprinter. The 3D bioprinter is capable of printing a cell-suspension ink with up to 300 million cells/mL and a cell viability greater than 95%. We have validated the 3D bioprinting using glioblastoma, neuroblastoma and lung cancer cells. The 3D bioprinted spheroids were confirmed to maintain all the biological characteristics typically found in a cancer spheroid. In particular, the 3D bioprinted cancer spheroids were shown to be viable and proliferating, preserved the apoptotic and cancer-stemness characteristics and using super-resolution lattice-light sheet microscopy, we showed the spheroids maintained the same structural integrity as manually formed spheroids. The potential application of the 3D bioprinted spheroids for high-throughput drug screening in 3D environments was demonstrated using doxorubicin as the model drug. 3D-bioprinting of patient-derived cancer cells for precision medicine applications is now underway. High-throughput embedded 3D bioprinting has enormous potential to accelerate precision medicine, drug discovery and cancer biology.

Citation Format: Lakmali Atapattu, Robert Utama, Aiden O'Mahony, Christopher Fife, Jongho Baek, Théophile Allard, Kieran O'Mahony, Julio Ribeiro, Katharina Gaus, Justin Gooding, Maria Kavallaris. Precision medicine: High-throughput 3D bioprinting of embedded multicellular cancer spheroids [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 5022.
Abstract 5133: Lipids and suppressive functions of MDSC in cancer
Authors: Veglia Filippo , Tyurin Vladimir A. , Kagan Valerian E. , Gabrilovich Dmitry
Published: August 2018

Myeloid derived suppressor cells (MDSC) play an important role in the regulation of anti-tumor responses and their presence in cancer patients correlates with a poor prognosis. MDSC contribute to the failure of cancer therapies and favor tumor progression and metastasis. However, despite the recent advances in understanding the biology of MDSC, the mechanisms driving their expansion and suppressive function are not yet well defined. Polymorphonuclear, PMN-MDSC is by far the largest population of MDSC. Here, we found that PMN-MDSC from tumor bearing mice and from cancer patients have higher amount of lipids than their counterpart (neutrophils) in tumor free mice and healthy donors, respectively. The content of lipids in PMN-MDSC was much higher in MDSC infiltrating the tumor, and the accumulation of lipids in PMN-MDSC was caused by the uptake of extracellular lipids. Lipid profile of PMN-MDSC demonstrated accumulation of oxidized fatty acids as part of triglycerides and phospholipids. We demonstrated that fatty acids accumulated by PMN-MDSC are oxidized by myeloperoxidase (MPO) and reactive oxygen species (ROS) generated by Nox2. Genetic ablation of MPO or ROS in MDSC results in a drastic reduction of lipid oxidation and in an impaired ability of MDSC to suppress CD8 T cell responses. In line with these findings, unsaturated fatty acids, highly susceptible to oxidation, favor the expansion of highly suppressive MDSC from hematopoietic progenitors in vitro, contrary to non-oxidizable saturated fatty acids. Together, our data suggest that MDSC have increased ability to pick up lipids from the microenvironment and to oxidize them via their potent oxidative machinery. In turn oxygenated lipids contribute to the suppressive activity of MDSC. In conclusion, the regulation of the oxidation of lipids in MDSC could be an attractive therapeutic strategy in cancer.

Citation Format: Filippo Veglia, Vladimir A. Tyurin, Valerian E. Kagan, Dmitry Gabrilovich. Lipids and suppressive functions of MDSC in 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 5133.
Abstract 5236: ARV-110: An androgen receptor PROTAC degrader for prostate cancer
Authors: Neklesa Taavi , Snyder Lawrence B. , Willard Ryan R. , Vitale Nicholas , Raina Kanak , Pizzano Jennifer , Gordon Deborah , Bookbinder Mark , Macaluso Jennifer , Dong Hanqing , Liu Zheng , Ferraro Caterina , Wang Gan , Wang Jing , Crews Craig M. , Houston John , Crew Andrew P. , Taylor Ian
Published: August 2018

The Androgen Receptor (AR) remains the principal driver of castration-resistant prostate cancer during the transition from a localized to metastatic disease. Most patients initially respond to inhibitors of the AR pathway, but the response is often short-lived. The majority of patients progressing on enzalutamide or abiraterone exhibit genetic alterations in the AR locus, either in the form of amplifications or point mutations in the AR gene. Given these mechanisms of resistance, our goal is to eliminate the AR protein using the PROteolysis TArgeting Chimera (PROTAC) technology. Here we report an orally bioavailable small molecule ARV-110 that leads to ubiquitination and degradation of AR. ARV-110 completely degrades AR in all cell lines tested, with an observed 50% degradation concentration (DC50) < 1 nM. PROTAC-mediated AR degradation suppresses the expression of the AR-target genes PSA and FKBP5, inhibits AR-dependent cell proliferation, and induces potent apoptosis in VCaP cells. ARV-110 degrades clinically relevant mutant AR proteins and retains activity in a high androgen environment. In mouse xenograft studies, greater than 90% AR degradation is observed at a 1 mg/kg PO QD dose. Significant inhibition of tumor growth and AR signaling can be achieved in both an intact and castrate setting. Further, ARV-110 demonstrates in vivo efficacy and reduction of oncogenic Erg protein in a long term, castrate, enzalutamide-resistant VCaP tumor model. DMPK and exploratory toxicology studies show robust oral, dose proportional drug exposure in rodent and non-rodent species. In summary, we report preclinical data on ARV-110, an orally bioavailable androgen receptor PROTAC degrader that demonstrates efficacy in enzalutamide-resistant prostate cancer.

Citation Format: Taavi Neklesa, Lawrence B. Snyder, Ryan R. Willard, Nicholas Vitale, Kanak Raina, Jennifer Pizzano, Deborah Gordon, Mark Bookbinder, Jennifer Macaluso, Hanqing Dong, Zheng Liu, Caterina Ferraro, Gan Wang, Jing Wang, Craig M. Crews, John Houston, Andrew P. Crew, Ian Taylor. ARV-110: An androgen receptor PROTAC degrader 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 5236.
Abstract 5395: MicroRNA-196b-3p identified from serum microRNA expression profiling is a potential diagnostic biomarker for gastritis and early gastric cancer
Authors: Fujii Atsushi , Nambara Sho , Kuroda Yosuke , Ito Shuhei , Eguchi Hidetoshi , Masuda Takaaki , Otsuka Takao , Nakamura Masafumi , Mimori Koshi
Published: August 2018

Background: Gastric cancer is one of the most common forms of cancer worldwide. The progression of chronic atrophic gastritis (CAG) is involved in gastric carcinogenesis. With the progression of chronic atrophic gastritis, the risk of gastric cancer increased in a stepwise fashion. Non-invasive tools for early gastric cancer are required to improve the prognosis. MicroRNAs (miRNAs), a class of small non-coding RNAs of 19-22 nucleotides in length, function as post-transcriptional regulators by directly cleaving target messenger RNA or translational repression. MiRNAs in serum have been shown to be potential biomarkers in various type of cancers and several miRNAs have been reported in gastric cancer. The purpose of present study was to identify a serum miRNA expression profile that can serve as a novel diagnostic biomarker for early gastric cancer.

Materials and Method: We defined a normal gastric mucosa as grade0, CAG classified C-1, 2 in Kimura-Takemoto classification as grade1, CAG classified C-3, O-1 in Kimura-Takemoto classification as grade2, and CAG classified O-2, 3 in Kimura-Takemoto classification as grade3. We obtained 344 serum samples, which gathered from 19 health people (grade0), 125 patients with chronic atrophic gastritis (grade1: 54, grade2: 36, grade3: 35), and 200 patients with early gastric cancer. The miRNAs were extracted from serums and miRNA microarray analysis was performed. In the discovery phase, we analyzed 71 cases (grade0: 11, grade1: 20, grade2: 20, grade3: 20). Next, in the training phase, we analyzed 73 cases (grade0: 8, grade1: 34, grade2: 16, grade3: 15). Then, we analyzed 200 early gastric cancer serum samples.

Result: The microarray analysis identified a profile of miRNA (miRNA-196b-3p) as a biomarker for early gastric cancer. The analysis results showed that the expression level of the serum miRNA was correlated to degree of CAG and carcinogenesis. When compared with grade0,1,2 and grade3, the area under the receiver operating characteristic (ROC) curve of this serum miRNA signature was 0.730 and 0.763 for the two sets of serum samples. There was significant difference in serum value of miRNA-196b-3p between grade1-3 and early gastric cancer (p=0.003). Conclusion: Serum miRNA-196b-3p is good biomarker for early gastric cancer.

Citation Format: Atsushi Fujii, Sho Nambara, Yosuke Kuroda, Shuhei Ito, Hidetoshi Eguchi, Takaaki Masuda, Takao Otsuka, Masafumi Nakamura, Koshi Mimori. MicroRNA-196b-3p identified from serum microRNA expression profiling is a potential diagnostic biomarker for gastritis and early gastric 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 5395.
Abstract 5629: Preclinical characterization of Sym023 a human anti-TIM3 antibody with a novel mechanism of action
Authors: Lindsted Trine , Gad Monika , Grandal Michael V. , Frölich Camilla , Bhatia Vikram K. , Gjetting Torben , Lantto Johan , Horak Ivan D. , Kragh Michael , Koefoed Klaus , Pedersen Mikkel W.
Published: August 2018

Immunotherapy has become a major focus of research in oncology and blockade of immune checkpoints such as cytotoxic T-Lymphocyte Associated Protein 4 (CTLA4) and programmed cell death protein 1 (PD1) has been some of the most successful immunotherapies. The next wave of immunomodulatory targets that are being explored for cancer therapy include T cell immunoglobulin and mucin domain protein 3 (TIM3). TIM3 is constitutively expressed on cells of myeloid origin whereas the TIM3 expression is induced on T-cells upon activation. The exact function of TIM3 on the different immune cells is not clear and may be context dependent suggesting that TIM3 is not a classical immune check-point.

Sym023 is a human anti-TIM3 antibody, which binds human TIM3 and cross-reacts with cynomolgus monkey TIM3. Sym023 blocks binding of phosphatidyl serine but not galectin 9 and stimulates T-cell proliferation in mixed lymphocyte reactions and tumor growth inhibition in vivo. Here, we present data demonstrating that ligation of TIM3 by Sym023 increase cytokine production and T cell proliferation in vitro through a novel mechanism of action.

Citation Format: Trine Lindsted, Monika Gad, Michael V. Grandal, Camilla Frölich, Vikram K. Bhatia, Torben Gjetting, Johan Lantto, Ivan D. Horak, Michael Kragh, Klaus Koefoed, Mikkel W. Pedersen. Preclinical characterization of Sym023 a human anti-TIM3 antibody with a novel mechanism of action [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 5629.
Abstract 5754: Hypoxia-induced tumor plasticity and immune resistance involves an alteration of target recognition by a mechanism involving TGF-beta signaling
Authors: Terry Stephane , Buart Stephanie , Thiery Jean-Paul , Mami-Chouaib Fathia , Chouaib Salem
Published: August 2018

Backgound: Hypoxia is known to shape the tumor microenvironment of developing tumors and contributes to both tumor progression and escape to immune surveillance. Previous work using non-small cell lung carcinoma cells (NSCLC) showed that hypoxia promotes epithelial-mesenchymal transition (EMT) in a fraction of carcinoma cells and that acquisition of a more mesenchymal (Mes) phenotype by certain cancer subclones correlates with an increased propensity to resist cytotoxic T lymphocytes (CTL) attacks as compared to more Epithelial (Epi) cancer subclones. Moreover, numerous studies identified alterations or loss of major histocompatibility complex (MHC) class I as a potential mechanism of tumor escape in cancer. Here, we asked whether hypoxia-induced EMT could regulate MHC class I expression and whether that could impact on resistance to CTL-mediated killing.

Methods and Results : By comparing CMH class I in various NSCLC cancer subclones that have experienced hypoxic stress, we found that cancer clones with enhanced Mes features have a reduced expression of CMH class I molecules, as compared to those with a more Epi phenotype. Transient siRNA knockdown of hypoxia effectors HIF-1α or HIF-2α did not affect the expression of CMH class I molecules suggesting that EMT effectors, rather that HIFs, directly regulate CMH class I expression. Among EMT effectors, TGF-β expression was found to be highly upregulated in Mes cancer clones. Moreover, treatment of these cells with TGF-βR inhibitors markedly upregulated CMH class I expression. and as target cells, increases their susceptibility to CTL-mediated killing. However, blocking of TGF-β using Ab to TGF-β during the coculture time had no significant effects on CTL-mediated killing. This suggests that the observed changes in target cell susceptibility occurs through TGF-β-mediated repression of MHC class I rather than by cancer cell production of TGF-β affecting CTL effector functions. Conclusions: These results suggest that the increase in TGF-β signaling, especially found in EMTed Mes carcinoma cells in response to hypoxic stress, contributes to downregulate CMH class I expression in these cells while coinciding with a reduced susceptibilty to CTL attacks. The development of agents targeting hypoxia or EMT components should provide new therapeutic opportunities in combination with current approaches.

Citation Format: Stephane Terry, Stephanie Buart, Jean-Paul Thiery, Fathia Mami-Chouaib, Salem Chouaib. Hypoxia-induced tumor plasticity and immune resistance involves an alteration of target recognition by a mechanism involving TGF-beta signaling [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 5754.
Abstract 5811: Evaluation of antitumor efficacy of CA102N, H-Nim and hyaluronic acid (HA) conjugate, alone or in combination with 5-fluorouracil (5-FU) in in vivo human colorectal cancer models
Authors: Tchaparian Eskouhie , Lin Louis
Published: August 2018

CA102N is a conjugate of HA and H-Nim (N-(4-Amino-2-phenoxyphenyl, methanesulfonamide), a derivative of cyclooxygenase-2 inhibitor Nimesulide, and a CD44 receptor targeted system currently in preclinical development for metastatic colorectal cancer. The in vivo antitumor activities of CA102N was evaluated singly or combined with 5-FU in CR5038 (Patient Derived Xenograft (PDX) and HT29 Xenograft models. Inhibitory activity was assessed according to tumor-volume changes. CA102N (i.v. 200mg/kg, BIW) monotherapy did not show significant tumor growth inhibition at either the end of the treatment (21 days) or prolonged observational (45 days) period, in CR5038 PDX-SCID metastatic model in comparison to vehicle control or 5-FU (i.v. 50mg/kg, QW). Notably, the combination therapy displayed a marked reduction in tumor growth (>70%) compared to 5-FU alone (p=0.0003) or vehicle control (p<0.0001) in addition to prolonged median survival (p<0.05). Alternatively, HT29 xenografts nude mice were found to have significantly reduced tumor burden treated with either single agents CA102N (400 mg/kg, QW) or 5-FU (50 mg/kg, QW), (p<0.012) compared to control vehicle. The drug combination further displayed a synergistic antitumoral effect and greater median survival (p <0.05) in HT29 model. CA102N was well tolerated alone or in the combination therapy. Based on the results of this study, combined treatment of CA102N and 5-FU resulted in a complete response at the regimen given. The combination of CA102N and 5-FU represents a promising potential therapeutic strategy and a new approach to reverse drug resistant in the treatment of colorectal cancer. Studies presently are ongoing to further evaluate its efficacy in CT26 murine colorectal models.

Citation Format: Eskouhie Tchaparian, Louis Lin. Evaluation of antitumor efficacy of CA102N, H-Nim and hyaluronic acid (HA) conjugate, alone or in combination with 5-fluorouracil (5-FU) in in vivo human colorectal cancer models [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 5811.
Abstract 5857: Isorhapontigenin inhibits triple-negative breast cancer via activating NRF2-mediated pathway
Authors: Zhu Yusha , Murphy Anthony , Kluz Thomas , Li Bo , Costa Max , Huang Chuanshu , Sun Hong
Published: August 2018

Breast cancer is the most prevalent cancer among women worldwide, and is the second leading cause of cancer mortality among women. Triple-negative breast cancers (TNBCs; ER-/PR-/HER2-) are highly aggressive tumors with poor prognosis. Due to lack of these specific receptors, TNBCs cannot be treated with hormone therapies or anti-HER2 targeted therapies. Chemotherapy, combined with surgery and radiation therapy, is the primary strategy for treating TNBCs. Thus, discovery and evaluation of new alternative medications targeting triple-negative breast cancer is of tremendous importance for reducing breast cancer mortality. Isorhapontigenin (ISO), a new derivative of stilbene isolated from Chinese herb Gnetum Cleistostachyun, exhibits a strong inhibitory effect on human bladder cancers. However, its anticancer activity with respect to breast cancer remains unclear. In the present study, we investigated the potential anticancer effect of ISO on TNBCs. ISO treatment inhibited the proliferation and anchorage-independent growth of 4T1 cells, a murine triple-negative breast cancer cell line, in a dose-dependent manner. In addition, the results from wound healing and Transwell invasion assay indicated significantly reduced cell migration and invasion in ISO-treated cells. Moreover, we performed the whole transcriptome analysis using RNA-seq to elucidate the molecules and pathways that mediate ISO's anticancer activity. A total of 1972 differentially expressed genes were identified in ISO-treated cells. IPA analysis revealed that the top canonical pathway altered by ISO is NRF2-mediated pathway. ISO-induced NRF2 activation was further confirmed by nuclear translocation of NRF2 protein and upregulation of NRF2 downstream target genes (GSTA1, GSTM1, HMOX1, and NQO1). In summary, our study is the first to demonstrate that ISO inhibits breast cancer cell growth and invasion, in part, by activating NRF2-mediated signaling pathway. Our results provide a novel mechanism for ISO-induced anticancer effect, and strongly support ISO as a promising therapeutic agent for triple-negative breast cancers.

Citation Format: Yusha Zhu, Anthony Murphy, Thomas Kluz, Bo Li, Max Costa, Chuanshu Huang, Hong Sun. Isorhapontigenin inhibits triple-negative breast cancer via activating NRF2-mediated pathway [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 5857.