bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4522: Hypoxia-induced alteration of mitochondrial dynamics is linked to chemoresistance in ovarian cancer
Authors: Han Youngjin , Kim Boyun , Song Yong Sang
Published: July 2017

Hypoxia, commonly observed in the tumor microenvironment, has been shown to be related to the cancer hallmarks. Hypoxia generates reactive oxygen species (ROS), inducing changes in mitochondrial function and structure in various diseases. Mitochondria, dynamic intracellular organelles, go through incessant processes of fission and fusion for efficient generation of energy. We investigated alterations in mitochondrial networks under hypoxic conditions (< 1 % O2) and subsequent effects of the mitochondrial alteration on drug response in ovarian cancer cells. Mitochondrial fission was prevalent in cancer cells under hypoxic conditions. Translocation of phospho-Drp1 (Ser616), a biochemically active mitochondrial fission protein, to mitochondria was enhanced under hypoxic conditions. An increase in ROS production was accompanied under hypoxic conditions. ROS scavenging reversed the hypoxia-elicited mitochondrial fission, while exogenous H2O2 treatment promoted mitochondrial fission. These results strongly implicate that ROS are important modulators of mitochondrial dynamics under hypoxia. Interestingly, hypoxia-induced mitochondrial fission altered the response of cancer cells to cisplatin (CDDP). Ovarian cancer cells in hypoxic and normoxic conditions were differentially affected by CDDP when mitochondrial fission was inhibited. Taken together, the alteration of mitochondrial dynamics, a cellular adaptation to hypoxia, is associated with chemoresistance. Mitochondria adapted to the hypoxic tumor microenvironment could be a potential target for anti-cancer therapy.

Citation Format: Youngjin Han, Boyun Kim, Yong Sang Song. Hypoxia-induced alteration of mitochondrial dynamics is linked to chemoresistance in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4522. doi:10.1158/1538-7445.AM2017-4522
Abstract 4526: Attenuation of hypoxia in solid tumor with multifunctional upconversion nanoparticles to enhance photodynamic therapy
Authors: Zhang Yan , Li Jingqiu , Hu Jun , Yang Xiangliang
Published: July 2017

As a non-invasive therapeutic technique, photodynamic therapy (PDT) requires sufficient oxygen in order to generate adequate singlet oxygen upon the excitation of photosensitizer using an appropriate light wavelength. It has limited efficacy in preexisting hypoxia in tumor and PDT-induced hypoxia, owing to the limit oxygen level in the tissue and the consumption of the oxygen during the PDT treatment. Herein, taking advantage of hyperbaric oxygen (HBO), multifunctional upconversion nanovehicle is constructed to overcome the above-mentioned problems. HBO provides timely oxygen supply during the treatment, which greatly enhances the photodynamic effect of the loaded nanoparticles, as demonstrated by the accelerated generation of 1O2 and elevated cytotoxicity. Tumor growth is greatly regressed in the HBO-PDT treated mice. Our finding provides new insights into the modulation of unfavourable tumor microenvironment with the perfect marriage between HBO and NIR-light sensitized upconversion nanoparticles to overcome the current limitation of cancer photodynamic therapies.

Note: This abstract was not presented at the meeting.

Citation Format: Yan Zhang, Jingqiu Li, Jun Hu, Xiangliang Yang. Attenuation of hypoxia in solid tumor with multifunctional upconversion nanoparticles to enhance photodynamic therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4526. doi:10.1158/1538-7445.AM2017-4526
Abstract 4538: UniConSig: A new algorithm for genome wide quantification of gene functions and disease associations
Authors: Chi Xu , Anurag Meenakshi , Maureen Sartor A. , Wang Xiaosong
Published: July 2017

One of the greatest hurdles for cancer biologists is to identify the cancer-causal genes from thousands of candidate genes suggested by large-scale genomics or deep sequencing studies. In our previous study, we discovered that cancer genes possess a complicated yet distinct “gene concept signature.” Concept Signatures include cancer-related signaling pathways, molecular interactions, transcriptional motifs, protein domains, and gene ontologies. We developed a Concept Signature (or ConSig) analysis that prioritizes the biological importance of candidate genes underlying cancer by computing their strength of association with those cancer-related signature concepts. The ConSig analysis has facilitated the discovery of a recurrent ESR1-CCDC170 gene fusion in more aggressive Luminal B breast cancers (Nat. Commun. 2014) as well as TLK2, MAP3K3, and MYST3 amplifications in aggressive luminal breast cancer (Nat. Commun. 2016, J. Pathol. 2014, Oncogene In press). Nevertheless, current candidate gene prioritization methods, including ConSig, are subject to bias from redundancy in the compiled knowledgebase (also known as gene concept database). This leads to variation of the gene ranking and jeopardizes the reliability of the priority methods. In light of these problems, we developed an innovative, universal algorithm called uniConSig. By penalizing overlapping concepts with a stable parameter, “effective concept number”, we reduced the fluctuation in uniConSig scores, and stabilized the ranking of the genes even with the randomly duplicated gene concept databases. We tested the uniConSig algorithm by identifying known cancer genes based on a cancer gene list, and found that the uniConSig algorithm demonstrated significantly enhanced prioritization of known cancer genes compared to the ConSig algorithm, and the results are stable even in the presence of randomly duplicated gene concept databases. In addition, we used calculations based on the dominant/recessive cancer gene lists, and were able to provide a quantitative measure of the potential dominant/recessive functions of human genes underlying cancer. As an example application of this algorithm, we show that the uniConSig scores can directly reveal the primary oncogene targets from genomic amplicons in breast cancer.To our knowledge, UniConSig is the first tool for genome-wide quantification of gene functions and disease associations. UniConSig has broad applications on gene prioritization for genomic-based studies to discover new disease causal genes or new gene functions.

Citation Format: Xu Chi, Meenakshi Anurag, Sartor A. Maureen, Xiaosong Wang. UniConSig: A new algorithm for genome wide quantification of gene functions and disease associations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4538. doi:10.1158/1538-7445.AM2017-4538
Abstract 4547: Structural, thermodynamics and kinetics role of novel hot-spot mutations of BCR-ABL1 in resistance towards "Ibs" inhibitors
Authors: Laurini Erik , Marson Domenico , Boccardo Silvia , Fermeglia Maurizio , Pricl Sabrina
Published: July 2017

The present contribution integrates structural, computational, and molecular biology techniques to understand the eventual role of our newly reported hot-spot mutations of the BCR-ABL1 KD in TKI resistance observed in CML patients (1) using a "two-tier level" investigation: isolated KDs and SH2-liker-SH3-KD BCR-ABL1 constructs.

Specifically, in silico experiments will disclose the effects exerted by the “hot-spot” BCR-ABL1 mutations on the protein structure, thermodynamic stability, and ability to interfere with BCR-ABL1 binding to specific TKIs (e.g., imatinib, dasatinib, and ponatinib).

Structural biology (e.g., SAXS) evidences of wild type and mutant isolated KDs and SH2-linker-SH3-KD (SSK) constructs (per se and in complex with TKIs) will be discussed to support in silico predictions. Direct drug binding thermodynamics and kinetics to both KD and SSK constructs will be measured using isothermal titration calorimetry and surface plasmon resonance.

In vitro kinase assays will monitor the ability of these mutant isoforms to auto-phosphorylate and to phosphorylate a substrate peptide.

(1) Gibbons DL, Pricl S, Posocco P, et al., Molecular dynamics reveal BCR-ABL1 polymutants as a unique mechanism of resistance to PAN-BCR-ABL1 kinase inhibitor therapy. Proc Natl Acad Sci U S A. 2014 Mar 4;111(9):3550-5. doi: 10.1073/pnas.1321173111

Note: This abstract was not presented at the meeting.

Citation Format: Erik Laurini, Domenico Marson, Silvia Boccardo, Maurizio Fermeglia, Sabrina Pricl. Structural, thermodynamics and kinetics role of novel hot-spot mutations of BCR-ABL1 in resistance towards "Ibs" inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4547. doi:10.1158/1538-7445.AM2017-4547
Abstract 4549: Identification of clinically relevant patterns in complex genomic interactions of glioblastoma multiforme
Authors: Dimitrova Nevenka , Volyanskyy Konstantin , Menkovski Vlado , Cheung Yee Him , Zhong Minghao , Fallon John T. , Vail Eric
Published: July 2017

Background: In clinical practice of complex diseases such as cancer it is a challenging task to correctly identify driving factors and specific interactions between multitudes of genomic and clinical variables. In cancer where patterns are heterogeneous and involve highly complex interactions of biological pathways, this is by far a non-trivial task. Using novel approaches in machine learning, we demonstrate the ability to quantitatively and accurately reveal and describe patterns in genomic and clinical data.

Method: We developed a computational framework based on a number of interacting learners performing extensive pattern search and data characterization. This framework is multilevel and involves steps of relevant data transformations. We formulate and apply contextually relevant criteria to distinguish and rank iteratively obtained findings and identify hidden relations between features and characteristic connections to the clinical outcome.

Summary: We successfully tested our method on the public data of Glioblastoma Multiforme (GBM) from The Cancer Genome Atlas (TCGA). We identify interacting sets of genomic variables to represent clinically relevant subgroups of samples. We observe a dense presence of interacting known cellular processes in GBM such as EGFR, FGFR, ERK, RAS, MAPK, P53, as well as novel such as calcium signaling, kinesin motor and protein kinase pathways, tubulin and microtubule binding, phorbol esters/diacylglycerol binding domain (C1 domain), purine ribonucleoside triphosphate binding, positive regulation of phospholipase activity and others. The top ranking pathways recap the well-known and new biology. In addition, compared to known genes in the literature, we also observe involvement of novel genes in GBM subtype characterization. Finally, we propose two-fold solution: identification of distinctive patterns in the input genomic and output clinical data, and specific format of their interactions.

Citation Format: Nevenka Dimitrova, Konstantin Volyanskyy, Vlado Menkovski, Yee Him Cheung, Minghao Zhong, John T. Fallon, Eric Vail. Identification of clinically relevant patterns in complex genomic interactions of glioblastoma multiforme [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4549. doi:10.1158/1538-7445.AM2017-4549
Abstract 4550: IODNE: an integrated optimization method for identifying the deregulated sub-network for precision medicine in cancer
Authors: Cheng Lijun , Inavolu Sai Mounika , Radovich Milan , Vasudevaraja Varshini
Published: July 2017

Background and Purpose: Sub-network analysis can explore complex patterns of the entire molecular pathways and networks for the drug target identification, and therefore drug selection. Computational algorithms are needed to optimize the sub-network search for molecular pathways and networks under certain conditions.

Materials and Methods: In the paper, the gene expression profile of a cohort breast cancer patients and protein-protein interaction (PPI) network are integrated with edge score and node score simultaneously. A novel optimization algorithm, IODNE, is developed to search for the optimal de-regulated subnetwork of merging gene and protein network. The application of IODNE is demonstrated for a sub-network of drug target selection in breast cancer subtypes Luminal-A/Luminal-B data from Cancer Genome Atlas (TCGA). Where, the PPI networks data selected from Pathway Commons.

Results: IODNE selects sub-networks, which contain ER1/PR and HER2, the well-known targets in clinic Luminal-A/Luminal-B breast cancer subtypes. This validates the utility of IODNE. When applying IODNE to the triple-negative breast cancer subtype data, we identify sub-networks that contain genes such as ERBB2, HRAS, PGR, CAD, POLE and SLC2A1.

Conclusion: Using the known drug targets in breast cancer subtypes, we demonstrate that IODNE is a robust and powerful algorithm to identify sub-networks for drug target identification. Based on the gene expression profiles and protein interaction networks, it detects the optimum deregulated subnetwork for the candidate drug target selection. The IODNE is promising tool for the precision medicine.

Citation Format: Lijun Cheng, Sai Mounika Inavolu, Milan Radovich, Varshini Vasudevaraja. IODNE: an integrated optimization method for identifying the deregulated sub-network for precision medicine in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4550. doi:10.1158/1538-7445.AM2017-4550
Abstract 4554: Unified theoretical algorithms for graphics dynamic multiple transformations of dose-effect relationships with computer simulation
Authors: Chou Ting-Chao
Published: July 2017

Based on physico-chemical principle of the mass-action law, bio-system analysis (arrangement & combinatorial) several hundred rate equations have been derived and published. Mathematical induction allows the deduction of them into the unified general median-effect equation (MEE) that makes dose & effect “interchangeable” for the 1st order and higher order dynamics. Based on the MEE plot, all dose-effect curves can be “linealized” which allows the quantitative determination of x-intercept (log Dm which signifies potency) and slope (m which signifies shape or dynamic order). Extension of MEE to multi-drug combinations resulted the combination index (CI) theorem which quantifies synergism (CI<1), additive effect (CI=1), and antagonism (CI>1). This unprecedented approach and multi-step logics have been summarized in [Chou TC. Pharmacol Rev 58: 621-681, 2006] which has been cited 1,261 times in 485 bio-medical journals (http://www.researcherid.com/rid/B4111-2009 as of 10.19.2016). Dose-effect curves (single & multi-drugs) can be quantitatively transformed into the MEE plot, Fa-CI plot, isobologram, Fa-DRI Plot for dose-reduction index, and when n>2, the polygonogram. This presentation will illustrate of combinations (1:5) of docetaxel (TXT) with T-compound against HCT-116 colon carcinoma xenograft in nude mice. In all, only 66 nude mice have been used, i.e. TXT 3 doses, T 3 doses and TXT+T 4 doses, plus a control group, each N=6. The MEE “Linearization” allows the “Econo-Green Bio-research”, with small experimental size and small number of data points, using small number of animals. The algorithms based computer simulation using CompuSyn software can be completed within one second after the dose and effect data entries. The CompuSyn has been offered for free download upon registration via www.combosyn.com.

Note: This abstract was not presented at the meeting.

Citation Format: Ting-Chao Chou. Unified theoretical algorithms for graphics dynamic multiple transformations of dose-effect relationships with computer simulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4554. doi:10.1158/1538-7445.AM2017-4554
Abstract 4563: Local and abscopal effects in oncolytic virotherapy are boosted by immune checkpoint blockade, immunogenic chemotherapy, or IFNAR blockade
Authors: Fend Laetitia , Yamazaki Takahiro , Préville Xavier , Quéméneur Eric , Kepp Oliver , Adam Julien , Marabelle Aurélien , Pitt Jonathan , Kroemer Guido , Zitvogel Laurence
Published: July 2017

Athough the clinical efficacy of oncolytic viruses has been demonstrated for local treatment, the ability to induce immune-mediated regression of distant metastases is still poorly documented. We here report that an engineered oncolytic Vaccinia Virus, VVWR-TK-RR--Fcu1, is able to induce an immunogenic cell death and thus to generate a systemic immune response. Effect on tumor growth and survival is largely driven by CD8+ T-cells, and we could demonstrate that the immune cell infiltrate in the tumor could be reprogrammed towards a higher ratio of effector T-cells to regulatory CD4+ T-cells. The key role of the type 1-IFN pathway in oncolytic virotherapy was also highlighted, and we could show a strong abscopal response in Ifnar-/- tumors. In this model, the single administration of the virus directly into the tumors, on one flank, led to regression in the contralateral flank (i.e. opposite to the virus injection site). Moreover, we observed that these effects are further enhanced when the oncolytic treatment is combined with either immunogenic chemotherapy such as oxaliplatin, or with immune checkpoint blockers such as anti-PD-1 or anti-CTLA-4. Altogether, these data suggest that local oncolytic virotherapy in patients with tumors altered in IFNAR signaling could increase immune-mediated abscopal regression of distant metastases.

Citation Format: Laetitia Fend, Takahiro Yamazaki, Xavier Préville, Eric Quéméneur, Oliver Kepp, Julien Adam, Aurélien Marabelle, Jonathan Pitt, Guido Kroemer, Laurence Zitvogel. Local and abscopal effects in oncolytic virotherapy are boosted by immune checkpoint blockade, immunogenic chemotherapy, or IFNAR blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4563. doi:10.1158/1538-7445.AM2017-4563
Abstract 4567: Immunoclassification of gastric cancer in the context of clinical outcome
Authors: Wang Weili , Liao Ping , Xie Shangchen , Shen Dongya , Zhou Chengfang , Mcleod Howard , He Yijing
Published: July 2017

Background: Gastric cancer (GC) plays a leading role in all cancer deaths especially in Eastern Asia. Current classifications including WHO, Lauren, and TCGA defined molecular classification have illuminated the clinicopathological characteristics or genetic profile of GC. However, these classifications were lack of association with clinical outcome and guidance for medication selection.

Objective: We aimed to identify a new immunoclassification for GC to predict patient’s prognosis and provide evidence for choosing proper medication.

Methods: Formalin-fixed and paraffin-embedded (FFPE) samples of GC were obtained along with the clinical outcome of the patient. Epstein-Barr virus (EBV) infection was measured by RT-PCR. Immune markers including CD3, CD8 and PD-L1 were measured by immunohistochemistry (IHC) at tumor infiltration area (TI) and invasive margin area (IM). The expression of PD-L1 in tumor microenvironment (TME) was assessed by immune reactive score (IRS) system. For immunoclassification, patients were classified into two subgroups: strong immunoreaction (SIR) and weak immunoreaction (WIR) defined by the number of CD8+ T cells and PD-L1 expression at TI.

Results: EBV infection was associated with the number of CD3+T cells and PD-L1 expression in TME. EBV+ patients also showed a poor overall survival (OS) compared with EBV- patients. Importantly, WIR patients lived significantly shorter than SIR patients. Moreover, patients who were treated by taxanes in WIR group had a shorter OS compared with those not using taxanes.

Conclusion: In this study, we suggest a new immunoclassification for gastric cancer which is associated with patients’ outcome and may provide a way to guide chemotherapy.

Citation Format: Weili Wang, Ping Liao, Shangchen Xie, Dongya Shen, Chengfang Zhou, Howard Mcleod, Yijing He. Immunoclassification of gastric cancer in the context of clinical outcome [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4567. doi:10.1158/1538-7445.AM2017-4567
Abstract 4578: Does poor prognosis in African Americans with MSI-H colorectal cancer associate with altered immune markers
Authors: Golconda Umamaheshwari , Sokol Lena , Shokrani Babak , Lee Edward , Hansel Donna , Fadare Oluwole , Patel Sandip , Nouraie Mehdi , Soleimani Hooman , Sharif Zaki , Afsari Ali , Daremipouran Fareed , Brim Hassan , Ashktorab Hassan
Published: July 2017

BACKGROUND: Microsatellite instability high (MSI-H) of sporadic colorectal carcinomas (CRC) is usually associated with improved prognosis and a high density of tumor-infiltrating lymphocytes. However, African Americans with MSI-H have poor prognosis.

AIM: To evaluate whether or not expression of different immune and tumor markers individually or in combination in African American MSI-H CRC associate with the prognosis status.

METHODS: Tissues Microarray (TMA) were prepared by microdissection from FFPE (Formalin Fixed Paraffin Embedded) blocks of 15 MSI-H patients. Immune markers (CD8+, Granzymes, Perforins, CD4+, STAT1, IRF1 and IRF5) and PD-L1 status were analyzed by immunohistochemistry (IHC). Demography and clinical data including TNM, tumor grading (WHO standard), histological type of the tumor, date of diagnosis, date of the last follow-up examination, treatment, comorbidities, metastasis, recurrence, 5-year disease-free survival and death were collected. RESULTS: There were 4 patients with improved prognosis (27%) with relatively high CD4 density (2 with >50% and 2 with 11-50%) regardless of PD-L1 status (3+/1-), stage (II,III), and other immune markers (CD8, CD4, Granzymes, Perforins, IRF1, IRF5 and STAT1). One of the improved prognosis patients with KRAS mutation has elevated expression of all considered immune markers besides CD4. There were 9 patients (6 dead, 3 alive) with poor prognosis, with different immune and tumor markers level (3 were PD-L1+ and 5 have relatively high CD4 count; but with low level of STAT1. There were also 2 patients with expected improved prognosis based on their immune markers level (stage II), however, they died within the 5 year period post-diagnosis, due to their age (98 and 87 years old).

Conclusion: MSI-H Colorectal cancer from African American patients has poor prognosis which may correlate with the nature of tumor-associated immune response. Other factors such as MSH3 defects might cancel the positive prognosis of MSI-H status. Nonetheless, low STAT1 and low CD4 may be indicators of poorer prognosis.

Citation Format: Umamaheshwari Golconda, Lena Sokol, Babak Shokrani, Edward Lee, Donna Hansel, Oluwole Fadare, Sandip Patel, Mehdi Nouraie, Hooman Soleimani, Zaki Sharif, Ali Afsari, Fareed Daremipouran, Hassan Brim, Hassan Ashktorab. Does poor prognosis in African Americans with MSI-H colorectal cancer associate with altered immune markers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4578. doi:10.1158/1538-7445.AM2017-4578
Abstract 458: Choline metabolism in cancer cells modifies PD-L1 expression
Authors: Krishnamachary Balaji , Penet Marie-France , Mironchik Yelena , Bhujwalla Zaver M.
Published: July 2017

Increased expression of choline kinase-α (Chk-α), the enzyme that converts choline to phosphocholine has been observed in most cancer cells including breast (1). This increase of choline kinase results in increased production of phosphocholine that is used in the synthesis of membrane phosphatidylcholine. We investigated the relationship between Chk-α and the programmed death ligand-1 (PD-L1) expression in triple negative MDA-MB-231 human breast cancer cells. Expression of PD-L1 in response to the secretion of interferon gamma (IFNγ) by cytotoxic T cells and its binding to the immune checkpoint PD-1 on T cells results in cancer cells escaping destruction by the immune system (2). Quantitative real-time PCR analysis (q-RT-PCR) did not reveal any change in PD-L1 mRNA expression between MDA-MB-231 expressing an empty vector control plasmid (231-EV) and MDA-MB-23 stably expressing shRNA against Chk (231-Chk). However, there was a difference in the mean intensity fluorescence (MIF) between 231-EV (325 ±11.5, n=2) and 231-Chk (527±18.5, n=2) in the FACS analysis. Upon induction with IFNγ (10ng/ml) for 24h, there was 1.23 fold increase in PD-L1 mRNA in 231-Chk compared to 231-EV. Interestingly, the MIP for cell surface expression of PD-L1 was 600±21 (n=2) for 231-EV compared to 872±16 (n=2) for 231-Chk cells. Values represent mean + SE. These data suggest that the cell surface increase of PD-L1 in 231-Chk cells could be due to translocation. Increased PD-L1 expression creates an opportunity to deliver PD-L1 targeted nanoparticles carrying therapeutic cargo. Since most conventional treatments result in a decrease of Chk-α and phosphocholine, our data suggest that these treatments may also result in cancer cells escaping immune surveillance through expression of PD-L1.

Acknowledgements: This work was supported by NIH R01CA136576 and P50 CA103175.

Reference: 1) Glunde K et al, Cancer Res 65, 11034 (2005)., 2) Zou W, Chen L, Nat Rev Immunol 8(6), 467-77 (2008).

Citation Format: Balaji Krishnamachary, Marie-France Penet, Yelena Mironchik, Zaver M. Bhujwalla. Choline metabolism in cancer cells modifies PD-L1 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 458. doi:10.1158/1538-7445.AM2017-458
Abstract 4606: Preclinical characterization of a novel fully human IgG1 anti-PD-L1 mAb CK-301
Authors: Gorelik Leonid , Avgerinos George , Kunes Yune , Marasco Wayne A.
Published: July 2017

Antibodies targeting Programmed Death-1 (PD-1), or its ligand, PD-L1, have demonstrated remarkable efficacy in subsets of cancer patients, with inhibition of the interaction between PD-1 on T-cells and PD-L1 on tumor cells leading to the recovery of anti-tumor immune response and immune-mediated eradication of tumors. However, not all patients respond to existing PD-1 and PD-L1 targeting agents and relapses to therapy still occur. Therefore, there exists a need to identify additional therapeutics and approaches to engage the immune system to enhance the efficacy of current anticancer therapies. Using phage and yeast display approaches, we have discovered and optimized a novel, fully human, PD-L1 specific IgG1 antibody, CK-301, which exhibits subnanomolar binding affinity for PD-L1. CK-301 blocks binding of PD-L1 to both PD-1 and B7-1 in enzyme-linked immunosorbent assays (ELISA) and cell-based competition assays. Using an assay measuring inhibition of a nuclear factor of activated T-cells (NFAT) reporter caused by PD-L1 binding to PD-1, we demonstrate that CK-301 completely reverses reporter inhibition at concentration of less than 1 µg/ml, IC50 of the dose response curve is 80ng/ml. CK-301 enhances IFN-gamma secretion in allogeneic mixed lymphocyte reaction (MLR) using primary human T-cells and immature dendritic cells. CK-301 can also trigger antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) mediated killing of PD-L1+ cell lines, including lymphoma cells. CK-301 has similar subnanomolar affinity for cynomolgus monkey PD-L1 as for human PD-L1, hence we chose Macaca fascicularis for pre-clinical toxicology and safety pharmacology studies. Single-dose administration of CK-301 to monkeys up to the highest tested dose of 100 mg/kg was shown to be safe and demonstrated linear dose-dependent pharmacokinetic (PK) properties over the dose range from 1 to 100 mg/kg with a half-life of 15 days at 100 mg/kg. A first-in-human Phase 1 study of CK-301 is planned to commence in mid-2017.

Citation Format: Leonid Gorelik, George Avgerinos, Yune Kunes, Wayne A. Marasco. Preclinical characterization of a novel fully human IgG1 anti-PD-L1 mAb CK-301 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4606. doi:10.1158/1538-7445.AM2017-4606
Abstract 4612: Development and validation of a screening platform for the identification of novel immuno-oncology targets
Authors: Scoumanne Ariane , Rabolli Virginie , Legrand Lea , Martini Murielle , Letellier marie-claire , Crosignani Stefano , Quéva Christophe , Detheux Michel , Cauwenberghs Sandra , Swiercz Jakub
Published: July 2017

Although immune therapy of cancer, including immune-checkpoint blockade have demonstrated therapeutic benefit in patients with various advanced cancers, further understanding of human immune pathology triggered by the tumor microenvironment, is essential to improve these therapeutic approaches.

In order to shed light on novel immune suppressive mechanisms in tumor, iTeos Therapeutics developed a target discovery and drug repurposing platform based on phenotypic screening assays. We established a co-culture assay combining tumor immune suppressive cells and T-cells. This assay is flexible to allow the screening of chemicogenomics, shRNA and cDNA libraries. Multi-parameter readouts are combined to assess both T cell activation and proliferation, through high content imaging of T cell clusters formation, complemented with detection of IFNγ secretion and tumor cell death, as assessed using a cytotoxicity assay. The 96-well format of the assay allows medium-throughput testing of up to 3000 samples/screen. From the technical point of view we were able to adapt the assay to low level of automation, making it affordable to the biotech start-ups and academic laboratories.

As a proof-of-concept we evaluated the assay for its ability to detect metabolic immune-oncology targets in A549 cells, a lung cancer immune suppressive cell line. A549 express indoleamine-2,3-dioxygenase 1 (IDO1), an enzyme expressed in many cancers that mediates local T-cell suppression through depleting the essential amino acid tryptophan. The assay conditions were validated with an IDO1 inhibitor as positive control and subsequently scaled up for automation. A commercially available small molecule library of 1900 compounds, with a high percentage of clinically tested drugs was screened. The library was tested at two different concentrations (0.3μM and 3μM), with two independent T-cell donors and spiked with IDO1 inhibitor as control. Combined analysis of T-cell activity and tumor killing led to the identification of 42 compounds with activity on multiple, potential immune suppressive pathways, including metabolism, epigenetics, autophagy, TGFβ, Wnt/β-catenin and TNFα/NF-κB signaling.

Citation Format: Ariane Scoumanne, Virginie Rabolli, Lea Legrand, Murielle Martini, marie-claire Letellier, Stefano Crosignani, Christophe Quéva, Michel Detheux, Sandra Cauwenberghs, Jakub Swiercz. Development and validation of a screening platform for the identification of novel immuno-oncology targets [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4612. doi:10.1158/1538-7445.AM2017-4612
Abstract 4613: Cytotoxicity of selenium trastuzumab and bevacizumab immunoconjugates against triple negative breast cancer cells
Authors: Khandelwal Soni , Gollahon Lauren , Spallholz Julian , Boylan Mallory , Garcia-Hernandez Maria Del Mar
Published: July 2017

Breast cancer (BC) is the second leading cause of death among women with an estimated 246,660 new cases of invasive breast cancer expected to be diagnosed this year. Of the different subtypes, 15-20% are classified as triple negative breast cancer (TNBC) because they lack three key receptors; progesterone, estrogen and HER2. These receptors are considered essential for “targeted therapies”. Thus, systemic chemotherapy is the only TNBC drug treatment option. In this study, two clinical monoclonal antibodies; Herceptin® and Avastin® were covalently labeled with redox selenium and used to treat two TNBC cell lines (MDA-MB-231 and MDA-MB-468) in culture. TNBC cell lines were treated with vehicle control, increasing concentrations of selenium as Selenite (ranging from 2 to 20 µg); Se-Herceptin® or Se-Avastin® and compared to native antibodies with equal concentrations of protein. Cell counts and viability were analyzed using Beckman ViCell Counter and Trypan Blue exclusion over a seven day period. Selenite, Se-Avastin® and Se-Herceptin® were observed to be more cytotoxic over dose and time to both TNBC cell lines in comparison to control cells and cells treated with native antibodies alone. Morphological changes were observed under phase contrast microscopy in all selenium treated cell lines. In contrast, visible alterations in morphology were absent in native Herceptin® and Avastin® treated cells compared to control cells. We believe this is the first report of selenium Antibody-Drug Conjugates (ADCs) being demonstrated as cytotoxic to these TNBC cell lines, suggesting a potential strategy to design more effective treatments of TNBC resistant to chemotherapy.

Citation Format: Soni Khandelwal, Lauren Gollahon, Julian Spallholz, Mallory Boylan, Maria Del Mar Garcia-Hernandez. Cytotoxicity of selenium trastuzumab and bevacizumab immunoconjugates against triple negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4613. doi:10.1158/1538-7445.AM2017-4613
Abstract 462: The role of miR-24 as a race-related genetic factor in prostate cancer
Authors: Hashimoto Yutaka , Shiina Marisa , Kato Taku , Yamamura Soichiro , Tanaka Yuichiro , Majid Shahana , Saini Sharanjot , Shahryari Varahram , Kulkarni Priyanka , Dasgupta Pritha , Mitsui Yozo , Sumida Mitsuho , Deng Guoren , Tabatabai Laura , Kumar Deepak , Dahiya Rajvir
Published: July 2017

The incidence of prostate cancer (PCa) among African-Americans (AfA) is significantly higher than Caucasian-Americans (CaA) but the genetic basis for this disparity is not known. To address this problem, we analyzed miRNA expression in AfA (n=81) and CaA (n=51) PCa patients. Here, we found that miR-24 is differentially expressed in AfA and CaA PCa patients and attempt to clarify its role in AfA patients. Also, the public sequencing data of the miR-24 promoter confirmed that it was highly methylated and down-regulated in PCa patients. Utilizing a VAMCSF and NDRI patient cohorts, we discovered that miR-24 expression was linked to a racial difference between AfA/CaA PCa patients. Interestingly, miR-24 was restored after treatment of PCa cells with 5Aza-CdR in an AfA cell line (MDA-PCa-2b), while restoration of miR-24 was not observed in CaA cells, DU-145. Ectopic expression of miR-24 showed decreased growth and induced apoptosis, though the effect was less in the CaA cell line compared to the AfA cell line. Finally, we found unique changes in biological pathways and processes associated with miR-24 transfected AfA cells by quantitative PCR-based gene expression array. Evaluation of the altered pathways showed that AR, PDPK1, IGF1, and IGFBP5 were markedly decreased in the AfA derived cell line compared with CaA cells, and there was a reciprocal regulatory relationship of miR-24/target expression in prostate cancer patients. These results demonstrate that miR-24 may be a central regulator of key events that contribute to race-related tumorigenesis and has potential to be a therapeutic agent for PCa treatment.

Citation Format: Yutaka Hashimoto, Marisa Shiina, Taku Kato, Soichiro Yamamura, Yuichiro Tanaka, Shahana Majid, Sharanjot Saini, Varahram Shahryari, Priyanka Kulkarni, Pritha Dasgupta, Yozo Mitsui, Mitsuho Sumida, Guoren Deng, Laura Tabatabai, Deepak Kumar, Rajvir Dahiya. The role of miR-24 as a race-related genetic factor in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 462. doi:10.1158/1538-7445.AM2017-462
Abstract 4621: Metabolic activation of macrophages by CpG stimulates anti-tumor activity and overcomes CD47-independent inhibition by tumor cells in pancreatic ductal adenocarcinoma
Authors: Liu Mingen , Beatty Gregory
Published: July 2017

Macrophages dominate the immune infiltrate present in pancreatic ductal adenocarcinoma (PDAC). Although tumor-infiltrating macrophages have the potential to mediate anti-tumor activity, they most commonly acquire an immunosuppressive role in cancer. We have found that CpG oligonucleotides - containing unmethylated cytosine-guanine motifs - potently induce macrophages to phagocytose PDAC cells and mediate anti-tumor effects. When delivered in vivo, in a fully immunocompetent murine model of PDAC, CpG increased the presence of macrophages within PDAC tumors and suppressed tumor outgrowth in a macrophage-dependent manner. Though CpG did not polarize macrophages toward classical anti-tumor (M1) or pro-tumor (M2) phenotypes, CpG treatment increased the basal rate of oxygen consumption in macrophages. Reversing this metabolic shift by inhibiting fatty acid oxidation abolished pro-phagocytic and anti-tumor effects by macrophages in vitro and in vivo. We investigated the impact of CpG in overcoming anti-phagocytic signals from CD47, a membrane protein overexpressed in multiple malignancies, including PDAC. While targeted knockout of CD47 in PDAC cells was not sufficient to activate in vitro phagocytosis by macrophages or anti-tumor effects in vivo, delivery of CpG potentiated anti-tumor and phagocytic activity that was independent of CD47 expression by tumor cells. However, the combination of CD47 loss in tumor cells and systemic CpG treatment enhanced the survival of tumor-bearing mice, indicating that disruption of CD47 can sensitize tumors to macrophage-directed immunotherapy. Together, our findings demonstrate a key role for cellular metabolism in directing the anti-tumor functions of macrophages and overcoming negative regulatory signals imposed by malignant cells.

Citation Format: Mingen Liu, Gregory Beatty. Metabolic activation of macrophages by CpG stimulates anti-tumor activity and overcomes CD47-independent inhibition by tumor cells in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4621. doi:10.1158/1538-7445.AM2017-4621
Abstract 4631: Enhanced glycolysis promotes function of tumor associated macrophages in mouse breast cancer model
Authors: Liu Yan , Zhou Wei , Wang Wei , Bai Lipeng , He Huiwen , Guo Jian , Luo Yunping
Published: July 2017

Tumor associated macrophages(TAMs) represent a major part of immune cells in tumor microenvironment(TME) and play crucial roles in promoting tumor progression by secreting cytokines or enzymes that could remodel TME or enhance envision or migration capabilities of tumor cells. Given their indispensable effect on tumor, the mechanisms underlying TAMs’ tumor-promoting phenotype and function remain unclear. Glucose metabolism in tumor cells has been intensively studied; however, the status of glucose metabolism of TAMs is still unclear. In this study, we aim to identify glucose metabolism status in TAMs, and explore how glucose metabolism regulates phenotype and function of TAMs. In the begining, we isolated TAMs from 4T1 mouse breast tumor, spleen macrophages from normal mouse as a control, and performed RNA sequencing to analyze the expression pattern in glucose metabolism related genes. The results from transcriptomics showed that TAM shown up regulation of glycolysis genes pattern. Then we analyzed and found that HK’s activity was increased while IDH’s activity was decreased in mouse breast cancer moel. Furthermore, In vitro, we found that up regulation of impotent glycolysis genes including HK2, PFK-L and LDHA in mouse peritoneal macrophages (CMPM) and mouse bone marrow derived macrophages (CMBMDM) conditionally cultured with 4T1 supernatants. Glucose consumption or production of lactate and ATP were also increased in CMPM or CMBMDM. Then we monitored lactate producing rate with seahorse energy metabolism detector, and found that either CMPM or CMBMDM produced lactate faster than that of control. Moreover, inhibition of LDHA activity by oxamate ( inhibitor) also decreased glucose consumption and lactate production. Importantly, the expression of Arg1 and CD206, positively correlated with glycolysis and function of TAMs. Finally, we search the underneath mechanism and found PI3K/AKT pathway was activated in CMPM or CMBMDM. In detail it was found that p-p85 and p-AKT was upregulated at 8 hour after 4T1 supernatants stimulation. Inhibition of PI3K/AKT pathway by LY294002 (PI3K inhibitor), notably inhibited the expression of glycolysis genes and markers of TAMs, which indicated that PI3K/AKT pathway maybe involved in glycolysis of TAMs. Together, we identified that TAMs adopted glycolysis as a major way to consume glucose and glycolysis was directly contributed to the phenotype and function of TAMs, which was regulated by PI3K/AKT signaling pathway.

Citation Format: Yan Liu, Wei Zhou, Wei Wang, Lipeng Bai, Huiwen He, Jian Guo, Yunping Luo. Enhanced glycolysis promotes function of tumor associated macrophages in mouse breast cancer model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4631. doi:10.1158/1538-7445.AM2017-4631
Abstract 4637: Activation of toll-like receptor-2 promotes proliferation in human lung adenocarcinoma cells
Authors: Kohtz Patrick
Published: July 2017

Objective: Toll-like receptors (TLR) have been implicated in tumor progression by affecting the immune response in the tumor microenvironment. Toll like receptors have been described to be present in human cancer cells but the presence and the role of these receptors in the growth and proliferation of human lung cancer cells is not known. We analyzed human lung cancer cells for the presence of TLR-2 receptors and evaluated the proliferation response to a specific TLR-2 agonist.

Methods: Human non-small cell cancer cell (NSCLC) lines A549, (adenocarcinoma) H125 adenosquamous, and H1650 adenocarcinoma (American Type Culture Collection), were cultured using standard techniques. Cells were treated with Pam3CSK4, a specific agonist of TLR2, at doses of 0ug/ml, 5ug/ml, and 10ug/ml for 72 hours. Baseline protein levels of TLR2 were assessed using standard western blotting techniques. Proliferation assays were completed using the MTS assay in response to treatment with the agonist

Results: Protein analysis demonstrated varying but consistently detectable levels of TLR2 protein in each cell line. After treatment with Pam3CSK4 we found that proliferation was significantly promoted in the human lung adenocarcinoma cell lines (p=.03), but not in the adenosquamous cell line (p=0.001).

Conclusion: Toll-like receptor -2 is consistently present and detectable on human lung adenocarcinoma cells. Toll Like receptor -2 activation results in increased proliferation in human lung adenocarcinoma cells and this effect appears to be specific to adenocarcinoma cells. Further study is necessary to elucidate the downstream pathways responsible for the increased proliferation we see in these human lung adenocarcinoma cells, however these findings may suggest TLR-2 to be a possible therapeutic target in human lung cancer.

Citation Format: Patrick Kohtz. Activation of toll-like receptor-2 promotes proliferation in human lung adenocarcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4637. doi:10.1158/1538-7445.AM2017-4637
Abstract 4641: Wild type APC is an independent marker of poor prognosis in late stage colorectal cancer
Authors: Siraj Abdul K. , Bu Rong , Beg Shaham , Masoodi Tariq , Al-Kuraya Khawla S.
Published: July 2017

Background: Colorectal cancer (CRC) is one of the commonest cancer worldwide and ranks as the most common cancer in males and the third most common among females in Saudi Arabia. Wnt pathway alteration is well known in colorectal cancer. In this study we aimed to comprehensively study Wnt pathway alterations in a large cohort of Middle Eastern colorectal patients and to know its role in this unique ethnic group.

Methods: 426 CRC cases were analyzed for APC, AXIN2 and CTNNB1 somatic mutations by targeted capture sequencing and protein expression status by immunohistochemistry in tissue microarray format. Results were analyzed for association with any clinicopathological parameters and for prognostic significance.

Results: The incidence of APC, AXIN2 and CTNNB1 mutations in our patient cohort were 58.2%, 4.2% and 3.8% respectively. APC mutation was associated with grade 2 tumors (p=0.0494) and male sex (p=0.0349). AXIN2 mutation were associated with grade 3 tumors (p=0.0231), MSI-high tumors (p<0.0001) and AXIN2 protein expression loss (p=0.0056). CTNNB1 mutations were associated with MSI-high tumors (p=0.0076). AXIN2 and CTNNB1 mutation were significantly associated with each other (p=0.0263). Wild type APC was associated with poor overall survival and was an independent marker of poor prognosis in late stage CRC (p=0.0200).

Conclusion: We reported the incidence of Wnt pathway alterations in this unique ethnic group on which such data is lacking. APC mutation plays major and other genes minor role in Wnt pathway activation. Incidence of APC mutation in this group is comparatively high compared to reported incidence worldwide. Wild type APC independently predicts poor prognosis in Middle Eastern late stage colorectal patients.

Citation Format: Abdul K. Siraj, Rong Bu, Shaham Beg, Tariq Masoodi, Khawla S. Al-Kuraya. Wild type APC is an independent marker of poor prognosis in late stage colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4641. doi:10.1158/1538-7445.AM2017-4641