bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4744: Disruption of Clever-1 in macrophages activates the innate immune response and mediates tumor rejection
Authors: Viitala Miro , Virtakoivu Reetta , Tadayon Sina , Jalkanen Sirpa , Hollmén Maija
Published: August 2018

Macrophages are considered the main obstacles for successful cancer treatment since they can dampen anti-tumor immune responses and promote tumor progression and resistance formation. Macrophages are highly eligible candidates for targeted therapies since these cells are abundantly present in various tumors, they are very plastic and can be converted into pro-inflammatory macrophages supporting T cell activation and tumor rejection. Clever-1 (also known as Stabilin-1) is a multifunctional molecule conferring scavenging ability on a subset of alternatively activated macrophages. The aim of this study is to uncover previously unknown functions of Clever-1 in the regulation of macrophage-mediated anti-tumor immune responses and develop anti-Clever-1 therapy into clinical use for those suffering from refractory, untreatable solid tumors.

By using in vivo tumor models and sophisticated immunological assays, our data show that mice carrying a macrophage-specific genetic deletion of Clever-1 (Lyz2-Cre/Stab1fl/fl) reject implanted Lewis lung carcinoma and EL4 lymphoma tumors. Macrophage Clever-1 deficiency induced proliferation of Ly6Chi monocytes in the tumor microenvironment but impaired their maturation and polarization into tumorigenic Ly6Clo macrophages. Tumors in Lyz2-Cre/Stab1fl/fl mice consisted almost completely of proliferating CD8 T cells with an exhausted PD-1+Lag3+ phenotype two weeks after implantation. Functionally, Clever-1 depleted macrophages showed enhanced pro-inflammatory responses following TLR activation with a shift in IL-12/IL-10 secretion levels. This was due to enhanced activation of mTORC1 and NFκB signaling, which resulted in increased secretion of TNFα in steady state and after LPS induction.

In conclusion, our data show an important function of Clever-1 in controlling macrophage mediated local immune responses. Moreover, our data supports Clever-1 targeting as a novel approach to increase host defenses against immune compromised tumors and promotes anti-Clever-1 immunotherapy into clinical trials where it may have benefits in comparison with currently available immune activating drugs.

Citation Format: Miro Viitala, Reetta Virtakoivu, Sina Tadayon, Sirpa Jalkanen, Maija Hollmén. Disruption of Clever-1 in macrophages activates the innate immune response and mediates tumor rejection [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 4744.
Abstract 4835: UD-017, a novel highly selective and orally active CDK7 inhibitor, shows a significant anticancer activity in patient-derived cancers
Authors: Matsushita Takashi , Ogi Sayaka , Onuma Kazuhiro , Sunamoto Hidetoshi , Ogawa Ayumi , Hasegawa Toru , Tokunaga Yasunori , Aga Yasuhiro , Ushiyama Shigeru
Published: August 2018

Background: Cyclin dependent kinase 7 (CDK7) is an attractive target for anticancer drugs due to its dual roles, cell cycle regulation and gene transcription/RNA processing. We synthesized UD-017, a small-molecule, highly selective, orally active CDK7 inhibitor with a novel chemotype. In this study, we characterized anticancer activities of UD-017 in vitro and in vivo using patient-derived (PD) cancer cells.

Methods: We first evaluated an antiproliferative activity of UD-017 broadly in a panel of cancer cell assay including patient-derived 3-dimensional tumor clonogenic assay. To verify the potential of c-Myc expression as a biomarker, we investigated the correlation between c-Myc expression levels and anticancer activity in vitro and in vivo using a colorectal cancer cell. In the main part, anticancer efficacy of UD-017 was investigated in vivo in PD-xenograft (PDX) models using representative PD cancer cells.

Results: In antiproliferative panel assays using over 100 cancer cell lines, UD-017 broadly inhibited a wide range of cancer cells from colon, breast, lung, kidney, blood, pancreas, osteosarcoma, sarcoma and urinary bladder cancers. Especially, UD-017 showed high sensitivity in solid tumor such as non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), gastric, osteosarcoma and sarcoma cells with IC50s of 10-200 nM. UD-017 showed a good correlation of c-Myc expression levels with antiproliferative activity using colorectal cancer cells in vitro, and UD-017 reduced the intratumoral MYC mRNA levels by an administration at 100 mg/kg in a HCT-116 xenograft model in mice. In PDX models, UD-017 showed strong inhibition and even regression of tumor growth, and the tumors were almost disappeared on day 14 without body weight loss in non-small cell lung cancer (LXFL1121). In a pleuramesothelioma (PXF541) PDX model, UD-017 also showed regressive effect, and in gastric cancer (GXA3067) and sarcoma (SXFS117) PDX models, UD-017 completely inhibited the tumor growth.

Conclusions: We propose UD-017 as a novel type of anticancer drug that shows complete antitumor responses in patient-derived xenograft models of diverse cancer types. c-Myc expression in cancer cells may be a biomarker for the antitumor effect of UD-017. These data support the rationale for further advancing towards clinical development.

Citation Format: Takashi Matsushita, Sayaka Ogi, Kazuhiro Onuma, Hidetoshi Sunamoto, Ayumi Ogawa, Toru Hasegawa, Yasunori Tokunaga, Yasuhiro Aga, Shigeru Ushiyama. UD-017, a novel highly selective and orally active CDK7 inhibitor, shows a significant anticancer activity in patient-derived cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4835.
Abstract 4857: Discovery of PF-06855800, a SAM competitive PRMT5 inhibitor with potent antitumor activity
Authors: Mcalpine Indrawan J. , Tatlock John , Billitti Joseph , Braganza John , Brooun Alexei , Ya-Li Deng , Hirakawa Brad , Jensen-Pergakes Kristen , Kumpf Robert , Liu Wei , Maegley Karen , McTigue Michele , Patman Ryan , Rui Eugene , Scales Stephanie , Spiegel Noah , Tran-Dubé Michelle , Wang Fen , Wang Zhenxiong , Yamazaki Shinji , Zhang Tao , Wythes Martin
Published: August 2018

Protein arginine methyltransferase 5 (PRMT5) is the primary type II arginine methyltransferase responsible for symmetric dimethylation of protein arginine residues. PRMT5 utilizes S-adenosylmethionine (SAM) to methylate a number of cytoplasmic and nuclear substrates that are involved in tumorigenesis. A number of these substrates affect a variety of biologic functions including RNA splicing, transcriptional regulation and translation, all of which are dysregulated in cancer. The discovery of adenosine as an efficient inhibitor of PRMT5 catalyzed a medicinal chemistry effort aimed at nucleoside optimization. Protein structure-based and ligand property-based design were combined to deliver molecules with desirable potency, selectivity and ADME properties. Here we will describe a series of deazapurine nucleoside analogues that culminate in the identification of PF-06855800, a SAM competitive PRMT5 inhibitor with antiproliferative activity in both in vitro and in vivo models.

Citation Format: Indrawan J. Mcalpine, John Tatlock, Joseph Billitti, John Braganza, Alexei Brooun, Deng Ya-Li, Brad Hirakawa, Kristen Jensen-Pergakes, Robert Kumpf, Wei Liu, Karen Maegley, Michele McTigue, Ryan Patman, Eugene Rui, Stephanie Scales, Noah Spiegel, Michelle Tran-Dubé, Fen Wang, Zhenxiong Wang, Shinji Yamazaki, Tao Zhang, Martin Wythes. Discovery of PF-06855800, a SAM competitive PRMT5 inhibitor with potent antitumor 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 4857.
Abstract 4859: JNJ-64619178, a selective and pseudo-irreversible PRMT5 inhibitor with potent in vitro and in vivo activity, demonstrated in several lung cancer models
Authors: Wu Tongfei , Millar Hillary , Gaffney Dana , Beke Lijs , Mannens Geert , Vinken Petra , Sommers Ivan , Thuring Jan-Willem , Sun Weimei , Moy Christopher , Pande Vineet , Zhou Junguo , Haddish-Berhane Nahor , Salvati Mark , Laquerre Sylvie , Lorenzi Matthew V. , Brehmer Dirk
Published: August 2018

PRMT5 is a type II methyltransferase that symmetrically di-methylates arginine residues on proteins involved in signal transduction and cellular transcription. For example, PRMT5 acts as the enzymatic machinery of the methylosome complex, crucial for spliceosome assembly and activity. Although not frequently mutated or amplified in tumors, an elevated PRMT5 protein level that leads to higher methylosome activity and promotes epithelial–mesenchymal transition, has recently been correlated with a poor survival of cancer patients.

The PRMT5 inhibitor JNJ-64619178 has been selected as a clinical candidate based on its high selectivity and potency, paired with favorable oral pharmacokinetics and safety properties.

JNJ-64619178 binds simultaneously to the SAM- and protein substrate- binding pockets of the PRMT5/MEP50 complex with a pseudo-irreversible mode-of-action. Chemical proteomics, methylomics and RNA-sequencing analyses of PRMT5 inhibitor treated cell line samples support the current biological understanding of PRMT5 as a regulator of alternative splicing events.

JNJ-64619178 showed potent and broad inhibition of cellular growth, observed in several cell line panels that represent diverse cancer histologies. Ongoing investigations will explore the potential synthetic lethal correlation between PRMT5 inhibition and cancer driver pathways, including those addicted to altered splicing.

Oral administration of JNJ-64619178 resulted in efficient inhibition of di-methylation of SMD1/3 proteins, components of the splicing machinery and direct substrates of the methylosome, in several human NSCLC and SCLC cancer mouse xenograft models. JNJ-64619178 demonstrated dose-dependent tumor growth inhibition and regression in several human NSCLC and SCLC cancer mouse xenograft models with sustained blockage of tumor re-growth after dosing cessation.

In summary, JNJ-64619178 has a favorable pre-clinical profile supporting clinical testing in patients with lung cancer and other malignancies.

Citation Format: Tongfei Wu, Hillary Millar, Dana Gaffney, Lijs Beke, Geert Mannens, Petra Vinken, Ivan Sommers, Jan-Willem Thuring, Weimei Sun, Christopher Moy, Vineet Pande, Junguo Zhou, Nahor Haddish-Berhane, Mark Salvati, Sylvie Laquerre, Matthew V. Lorenzi, Dirk Brehmer. JNJ-64619178, a selective and pseudo-irreversible PRMT5 inhibitor with potent in vitro and in vivo activity, demonstrated in several lung 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 4859.
Abstract 5038: Ready to use 3D spheroid cultures as a standard tool to screen for compounds targeting cancer
Authors: Martin Angel Garcia , Leis Olatz , Gumuzio Juan , Prieto Inés , Ruiz Estibaliz
Published: August 2018

The cancer stem cell (CSC) concept has important implications not only for our understanding of carcinogenesis, but also for the development of cancer therapeutics. There is a growing body of preclinical evidence showing that cancer stem cells contribute to chemotherapy and radiation resistance in breast cancer. The use of drugs that interfere with stem cell self-renewal represents the strategy of choice for novel effective anti-cancer treatments, but also a great challenge because cancer stem cells and their normal counterparts share many pathways. The biology of cancer stem cells has proven complex and difficult to translate into effective therapeutic strategies. The question arises as: how do we test compounds for anti-cancer stem cell activity? The answer is: phenotypic screening. There are indeed several functional assays well validated in the scientific literature that have been used for years associated to the ability of cancer cells to demonstrate stem cell behavior. The most relevant is the 3D tumor spheroid assay. This assay has been used to uncover and culture stem cells from many tissues as well as from tumors. There are multiple reports now that show that spheroid derived cells are enriched in tumor initiating or cancer stem cells, derived from cell lines and from natural fresh tumors as well. Here we describe the use of 3D spheroid models to profile compound activity against cancer stem cells. Furthermore, a case of compounds preventing hypoxia-inducible transcription factor (HIFs) activity is presented. Recently, HIF transcription factor biology has been linked to pathways that regulate stem cell self-renewal and pluripotency, suggesting a new mechanism whereby HIF proteins may drive tumor growth, through the generation of tumour-initiating cells or cancer stem cells. Therefore, targeting the HIF pathway may provide a novel therapeutic avenue to target cancer stem cells. We demonstrate that interfering with HIF pathway activation prevents mammosphere formation, validated through indenpendent confirmation through Sox2 promoter activation, Aldefluor® assay and in vivo proof-of concept experiments targeting tumor initiation.

Citation Format: Angel Garcia Martin, Olatz Leis, Juan Gumuzio, Inés Prieto, Estibaliz Ruiz. Ready to use 3D spheroid cultures as a standard tool to screen for compounds targeting 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 5038.
Abstract 5309: Determining EGFR and STK11 mutational status in lung adenocarcinoma histopathology images using deep learning
Authors: Coudray Nicolas , Moreira Andre L. , Sakellaropoulos Theodore , Fenyö David , Razavian Narges , Tsirigos Aristotelis
Published: August 2018

To characterize a tumor and suggest appropriate treatments, pathologists rely on the visual assessment of histopathology slides as well as molecular profiling of the tumors. This is especially true in lung adenocarcinoma cases where targeted therapies exist for tumors with EGFR mutations. In this work, we studied the use of deep learning approaches to classify lung cancer histopathology images and predict the mutational status of frequently mutated genes.

To achieve these tasks, more than 1600 scanned histopathology slides, obtained from TCGA, were separated into independent training, validation and test sets. They were then tiled into 512x512 pixel windows, and the training set was used to feed the inception v3 convolutional neural network developed by Google. The resulting model was evaluated on the test set, and the generated per-tile probabilities were aggregated to generate a per-slide classification score. We based our study on whole-slide images of adenocarcinoma, squamous cells carcinoma and normal lung tissues and the neural network was trained on several tasks: first, the goal was to identify normal from tumor regions, and then, once the tumor regions were identified, it was trained to identify adenocarcinoma versus squamous cell carcinoma. Finally, the neural network was trained predict the mutational status of the most frequently mutated genes in lung adenocarcinoma.

After verifying that the trained network properly identifies tumor slides compared to normal tissue (AUC=0.99) and distinguishes adenocarcinomas from squamous cell carcinomas (AUC=0.95), the network was trained to simultaneously identify different mutations. It achieved AUC scores of 0.82 and 0.86 for STK11 and EGFR respectively, showing that mutations in these two genes may confer specific macroscopic features, visible in histopathology images, and that a deep convolutional neural network can be trained to recognize those features.

Our results demonstrate that, given the availability of sufficient data, convolutional neural networks learn to perform complex diagnoses and thus assist pathologists in their work.

Citation Format: Nicolas Coudray, Andre L. Moreira, Theodore Sakellaropoulos, David Fenyö, Narges Razavian, Aristotelis Tsirigos. Determining EGFR and STK11 mutational status in lung adenocarcinoma histopathology images using deep learning [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 5309.
Abstract 5327: DNA methylation loss in late-replicating domains is linked to mitotic cell division
Authors: Zhou Wanding , Dinh Huy Q. , Ramjan Zachary , Weisenberger Daniel J. , Nicolet Charles M. , Shen Hui , Laird Peter W. , Berman Benjamin P.
Published: August 2018

DNA methylation loss occurs frequently in cancer genomes, primarily within lamina-associated, late-replicating regions termed Partially Methylated Domains (PMDs). We profiled 39 diverse primary tumors and 8 matched adjacent tissues using Whole-Genome Bisulfite Sequencing (WGBS), and analyzed them alongside 343 additional human and 206 mouse WGBS datasets. We identified a local CpG sequence context associated with preferential hypomethylation in PMDs. Analysis of CpGs in this context (“Solo-WCGWs”) revealed previously undetected PMD hypomethylation in almost all healthy tissue types. PMD hypomethylation increases with age in both fetal and postnatal tissues, appearing to track accumulation of cell divisions beginning early in life. In cancer, the degree of PMD hypomethylation correlates with somatic mutation density and expression of cell-cycle dependent genes, reinforcing its mitotic clock-like role. We propose that late replication leads to progressive methylation loss throughout an individual's lifespan, potentially contributing to the onset of cancer and acting as a biomarker for cellular aging.

Citation Format: Wanding Zhou, Huy Q. Dinh, Zachary Ramjan, Daniel J. Weisenberger, Charles M. Nicolet, Hui Shen, Peter W. Laird, Benjamin P. Berman. DNA methylation loss in late-replicating domains is linked to mitotic cell division [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 5327.
Abstract 5450: Anticancer effects of picrasidine Q from Angelica keiski on esophageal squamous cell carcinoma by downregulation of FGFR2 signaling pathways
Authors: Liu Xuejiao , Shi Yuanyuan , Song Mengqiu , Zhao Ran , Chen Hanyong , Shim Jung Hyun , Dong Zigang , Lee Mee-Hyun
Published: August 2018

Fibroblast growth factor receptor (FGFR) 2 and its downstream signaling cascades, PI3 K/AKT/mTOR is playing an important role in cell survival and proliferations. In this study, we firstly found that picrasidine Q (PQ), an alkaloid component extracted from Angelica keiskei species, has the capacity of anti-cell transformation and anticancer. After ligand shape similarity approach of PQ, we found that PQ targeted FGFR 2 and verified by FGFR2 kinase assay as well as computational docking model. FGFR2 highly expressed in esophageal cancer tissues and PQ inhibited fibroblast growth factor (FGF)-induced cell transformation. Furthermore, PQ inhibited cell proliferation and induced G1 phase arrest of cell cycle and apoptosis in KYSE30, KYSE410, and KYSE450 esophageal squamous cell carcinoma (ESCC) cells. It was confirmed by detecting of biological markers such as cyclinD1, cyclinD3 and cyclinB1 for cell cycle or cleaved caspase-7, caspase-3, and PARP for apoptosis. PQ targeting of FGFR2 kinase activities suppressed downstream target proteins including phosphorylation of AKT and mTOR but not MEK/ERK signaling pathways. Taken together, our results are the first to identify that PQ might be a chemopreventive and chemotherapeutic agent by direct targeting FGFR2 and inhibiting cell proliferation of ESCC cells.

Citation Format: Xuejiao Liu, Yuanyuan Shi, Mengqiu Song, Ran Zhao, Hanyong Chen, Jung Hyun Shim, Zigang Dong, Mee-Hyun Lee. Anticancer effects of picrasidine Q from Angelica keiski on esophageal squamous cell carcinoma by downregulation of FGFR2 signaling pathways [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 5450.
Abstract 5463: Free fatty acids augment radiation-induced cytotoxicity in cervical cancer cells via mTOR/Akt//PPARg/p53-mediated pathway
Authors: Muhammad Naoshad , Rashmi Ramachandran , Ruiz Fiona , Zahner Michael C. , Schwarz Julie K.
Published: August 2018

Purposes of the study: Our clinical results show that obese patients with cervical cancer who are treated with definitive chemoradiation have favorable outcomes, and this effect is independent of diabetes or metformin use. PI3K/AKT pathway signaling is decreased in cervical tumors from obese hosts, even in the context of genomic mutations that would otherwise activate PI3K signaling, such as PTEN deletion. The objective of the present study is to determine whether other factors related to obesity, such as circulating free fatty acids, influence radiation responses in vitro.

Experimental procedures: Radiation sensitivity experiments were performed in cervical cancer cells with and without PI3K pathway mutations (CaSki (mutant), ME180 (mutant), SiHa (WT). To determine whether combination treatment of free fatty acids (palmitic, oleic and linoleic) plus radiation increases cytotoxicity in cervical cells, we performed cell viability assay, long term colony formation assays (CFA) and morphological examination by vertical microscope. To determine whether the addition of free fatty acids to radiation influences cell survival signaling, a series of Western blots were performed.

Summary: We observed that combination treatment of free fatty acids and radiation significantly increased cytotoxicity in all the cervical cancer cell lines tested. Mechanistic investigations revealed that free fatty acids induced potentiation of radiation-induced cytotoxicity through downregulation of p-mTOR, p-Akt and PTEN. Furthermore, we observed that this combination regimen upregulates the expression PPARγ and p53.

Conclusions: Free fatty acids potentiated the effect of radiation in PI3K-activated cervical cancer cell lines through mTOR/Akt//PPARg/p53 mediated pathway. Future studies will characterize the mechanism of free fatty acid potentiation of radiation-induced tumor cell death in detail, including the potential contribution of lipid peroxidation and alterations in glucose metabolism.

Citation Format: Naoshad Muhammad, Ramachandran Rashmi, Fiona Ruiz, Michael C. Zahner, Julie K. Schwarz. Free fatty acids augment radiation-induced cytotoxicity in cervical cancer cells via mTOR/Akt//PPARg/p53-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 5463.
Abstract 5566: LYC-55716, a first-in-class RORγ agonist: Rationale and preclinical data to support clinical combinations with established immunotherapies
Authors: Hu Xiao , Liu Xikui , Li Hongxiu , Weems Garry , Zawidzka Elizabeth , Gao Yilin , Wilkins H. Jeffrey , Carter Laura
Published: August 2018

The retinoic acid-related orphan receptor γt (RORγt) is a nuclear receptor transcription factor that acts as an immune cell master control switch driving the generation and function of Th17 (helper) and Tc17 (cytotoxic) T cells. Preclinical data show that synthetic RORγ agonists modulate immune cell gene expression to both increase immune activity and decrease immunosuppressive mechanisms in the tumor microenvironment. Because RORγ agonists impact multiple antitumor mechanisms, these compounds have the potential to combine with other agents to enhance tumor immunity. In addition to preclinical data supporting a biologic rationale, bioinformatic and machine learning approaches based on the RORγ pathway were undertaken to prioritize possible clinical combinations. This work highlighted other immuno-oncology agents as well as targeted therapies and more conventional treatments. Preclinical, syngeneic mouse cancer models were used to test some of the hypotheses. Based on the finding that RORγ agonists decrease PD-1 expression and that anti-CTLA4 induces ICOS, which stabilizes RORγ expression, anti-PD1 or anti-CTLA4 agents were combined with the RORγ agonist LYC-54143 in 12 syngeneic murine models. Compared with individual agents, combination treatments elicited superior tumor growth inhibition in 6 of the models (Table). RORγ agonist treatment was also able to augment antitumor activity of doxorubicin. In preclinical and phase 1 clinical testing, the first-in-class, investigational oral small-molecule RORγ agonist LYC-55716 has demonstrated a favorable safety profile supporting combination with other immuno-oncology agents. Ongoing clinical trials include a phase 2a trial of LYC-55716 in patients with select solid tumors (NCT02929862) and a phase 1b trial of LYC-55716 combined with pembrolizumab in patients with non-small cell lung cancer.

Percentage of tumor growth inhibition induced in syngeneic murine models of cancer.Murine modelAnti-PD1 aloneAnti-CTLA4 aloneRORγ agonist aloneRORγ agonist + anti-PD1RORγ agonist + anti-CTLA4H22 (Liver)30-50%>50%*10-30%>50%*>50%*Pan02 (Pancreas)<10%10-30%<10%10-30%*10-30%*CT26 (Colon)10-30%<10%<10%<10%30-50%B16F10 (Melanoma)10-30%10%10-30%10-30%30-50%A230 (Lymphoma)<10%<10%10-30%30-50%10%Renca (Renal)<10%10-30%30-50%*30-50%*30-50%*P<.05 vs vehicle.

Citation Format: Xiao Hu, Xikui Liu, Hongxiu Li, Garry Weems, Elizabeth Zawidzka, Yilin Gao, H. Jeffrey Wilkins, Laura Carter. LYC-55716, a first-in-class RORγ agonist: Rationale and preclinical data to support clinical combinations with established immunotherapies [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 5566.
Abstract 5700: IDO1 high expression is associated with a poor prognosis in patients with esophageal cancer
Authors: Kiyozumi Yuki
Published: August 2018

[Background] Esophageal cancer is the sixth most common cause of death from cancer worldwide. Although the development of multidisciplinary treatment for esophageal cancer, the prognosis of those patients remains poor. Recently, immunotherapy is expected as innovative therapy for several types of cancer. Indoleamine 2, 3- Dioxygenase (IDO1) is an enzyme responsible for the oxidation of tryptophan into kynurenine and implicated in immune modulation through its ability to limit T cell function and engage mechanisms of immune tolerance. IDO1 is focused as one of the immune subversion strategy and the therapy of targeting IDO1 is in clinical development for many types of cancer. IDO1 activation induces the downregulation of activating natural killer (NK) cell receptors on NK cells and inhibition of cytotoxic T cells by promotion of cell cycle arrest and apoptosis. However, the significance of IDO1 expression in esophageal cancer remains still unknown. The aim of this study is to examine the relationship between IDO1 expression and tumoral immune status, and its prognostic impact in esophageal cancer.

[Methods] IDO1 expression, cluster of differentiation (CD8) expression as a marker of cytotoxic T cells, forkhead Box P3 (FOXP3) as a maker of regulatory T cells were evaluated by immunostaining in 305 clinical samples of curative resected esophageal cancer.

[Result] In 305 clinical samples, high IDO1 high expression (n=71) was significantly associated with poor prognosis in overall survival(OS)(p=0.0034). In uni-and multivariate analysis, IDO1 high expression was an independent prognostic factor for OS. In those patients, CD8 low expression was also associated with poor prognosis in OS (p = 0.025). Furthermore, double immunostaning with IDO1 and CD8 revealed that CD8 expression levels was inverse correlated with IDO1 expression levels in those patients. In combination analysis of IDO1 and CD8 expression, IDO1 high expression and CD8 low expression group experienced worse overall survival (p=0.0024). Conversely, IDO1 low expression and CD8 high expression group was shown the best overall survival. In addition, IDO1 high expression group is correlate with the high counts of FOXP3 positive cells (p = 0.02).

[Conclusion] IDO1 expression was associated with an unfavorable clinical outcomes by induced immune tolerance and the classification of IDO1 and CD8 expression levels might serve as a predictive biomarker in esophageal cancer. The correlation between IDO1 and FOXP3 expression suggested that IDO1 involved in immune tolerance in esophageal cancer. These findings indicate that IDO1 would be a regulator of, and candidate therapeutic target in esophageal cancer.

Citation Format: Yuki Kiyozumi. IDO1 high expression is associated with a poor prognosis in patients with esophageal 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 5700.
Abstract 681: Targeting diacylglycerol kinases for immuno-oncology
Authors: Hsu Ku-Lung ,   Hsu Lab at the University of Virginia
Published: August 2018

Diacylglycerol kinases (DGKs) are integral components of signal transduction cascades that regulate cell biology through phosphorylation of the ubiquitous secondary messenger diacylglycerol. Emerging clinical evidence supports DGK-alpha (DGKA) as a promising target for cancer immunotherapy because of its critical role in regulating lipid signaling necessary for proper T cell activation. Methods for direct evaluation of DGK activity in native biologic systems are lacking and needed to develop chemical probes for studying isoform-specific functions. Here, I will discuss efforts from my group to utilize ATP acyl phosphate activity-based probes and quantitative mass spectrometry to map previously undefined ATP- and small molecule-binding sites of representative members from all subtypes of the DGK family. We use our chemical proteomics strategy to discover an unusual binding mode for DGKA inhibitors, including interactions at a novel binding site remote from the ATP binding pocket. I will also describe our efforts towards discovery of new lead inhibitors for future development of highly potent and selective DGKA inhibitors. Collectively, our studies illustrate the power of chemical proteomics to site-specifically profile protein-small molecule interactions and reveal key ligand binding sites for selective inactivation of the DGK family of lipid kinases.

Citation Format: Ku-Lung Hsu, Hsu Lab at the University of Virginia. Targeting diacylglycerol kinases for immuno-oncology [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 681.
Abstract 79: Stromal regulation of prostate cancer cell proliferation by mevalonate pathway enzymes HMGCS1 and HMGCR
Authors: Ashida Shingo , Kawada Chiaki , Inoue Keiji
Published: August 2018

It has been suggested that the tumor microenvironment plays an important role in tumor progression, acquisition of androgen independence, and distant metastasis in prostate cancer (PC). However, little is known about the transcriptional basis of cellular interactions in the human PC microenvironment. To clarify the mechanism of PC progression and metastasis, we investigated the interaction of PC, epithelial, and stromal cells by analyzing genome-wide gene expression profiles. Our hypothesis is that PC cells could induce stromal cells to differentiate into so-called cancer-associated fibroblasts (CAFs), which might contribute to cancer invasion and metastasis. We identified the up-regulated genes in normal human prostate stromal cells (PrSC) co-cultured with human PC cells (LNCaP), which included the mevalonate pathway enzymes 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR). Knockdown of endogenous HMGCS1 or HMGCR in PC cells by shRNA resulted in a significant reduction of PC cell viability. Importantly, exogenous overexpression of HMGCS1 or HMGCR in either PC cells or prostate stromal cells stimulated PC cell growth, suggesting a possible autocrine/paracrine regulation. Immunohistochemical analysis confirmed that HMGCS1 and HMGCR were overexpressed in PC stroma, especially in the early stage of PC. These results provide clues to the molecular mechanisms underlying PC invasion and metastasis, and suggest that HMGCS1 and HMGCR in PC, as well as in PC stroma, might serve as molecular targets for the treatment of PC.

Citation Format: Shingo Ashida, Chiaki Kawada, Keiji Inoue. Stromal regulation of prostate cancer cell proliferation by mevalonate pathway enzymes HMGCS1 and HMGCR [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 79.
Abstract 886: Metformin effect on multidrug resistant leukemia cells: metalloproteinases 2 and 9
Authors: Oliveira Ligia P. , Perea Camila , Baldi Caroline , Prata Rodrigo F. , Renó Débora L. , Lima Luana P. , Curvello Rodrigo , Souza Ana Carolina S.
Published: August 2018

Besides the progress in studies of cancer, the rates of morbidity and mortality due to this disease still considerably elevated. Chronic myeloid leukemia (CML) is a hematological neoplasm, therefore chemotherapy is the main therapeutic approach used, however most patients develop resistance to chemotherapy. Several factors contribute to this scenario, among them the lack of selective treatment, the emergence of tumor cells resistant to the wide variety of cytotoxic agents during treatment and the formation of metastases, which accounts for the majority of cancer deaths. Therefore, application of new drugs in antitumor therapies has been constantly studied. Studies have shown that metformin, an oral euglycemic used in the treatment of type II diabetes, has potential use in the treatment of cancer, potentializing the effects of standard chemotherapy used in treatment. In this work the effects of metformin on the metastatic capacity of K562 (non-resistant leukemic cell), Lucena and FEPS (leukemic cells resistant to multiple drugs) were evaluated through the study of MMP2 and MMP9 metalloproteinases, enzymes essential for the metastatic process. Thus, the present project aimed to evaluate the gene expression and enzymatic activity of MMP2 and MMP9 metalloproteinases in CML cells, before and after treatment with metformin, seeking to verify the effects of metformin on the expression and activity of these enzymes. Gene expression was evaluated using quantitative PCR and the quantification of enzymatic activity was performed by the substrate degradation using the zymography method. The results showed that metformin has the potential to modulate the expression and activity of the studied metalloproteinases.

Citation Format: Ligia P. Oliveira, Camila Perea, Caroline Baldi, Rodrigo F. Prata, Débora L. Renó, Luana P. Lima, Rodrigo Curvello, Ana Carolina S. Souza. Metformin effect on multidrug resistant leukemia cells: metalloproteinases 2 and 9 [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 886.
Abstract 889: Metformin treatment inhibits proliferation and induces apoptosis in medulloblastoma cells
Authors: Payne Kristen C. , Hunter Abigail , Bowman W Paul , Murray Jeffrey C. , Basha Riyaz , Sankpal Umesh T.
Published: August 2018

Medulloblastoma is the most common malignant brain tumor in children that typically requires a patient specific combination of surgical resection, chemotherapy, and radiation. Up to 80% of medulloblastoma patients respond to treatment and survive long-term. However, survivors often suffer from severe long term sequelae secondary to treatment that affects their neurocognitive and growth potential. Therefore, there is an urgency to develop less toxic, more focused treatment strategies that target specific pathways or molecules involved in medulloblastoma development and progression. Metformin is a widely used non-toxic anti-diabetic drug that has mild, generally well-tolerated side effects. Studies have shown that it can reduce cancer risk and improve the prognosis of certain malignancies. However, the mechanism underlying its anti-cancer effect is still unclear and varies depending on the type of cancer. There are reports that metformin can target the specificity protein 1 (Sp1) transcription factor. Sp1 regulates the expression of a variety of proteins involved in regulation of cancer cell proliferation including survivin, an Inhibitor of Apoptosis Protein family member. Expression of survivin is increased in medulloblastoma cells and often correlates with poor prognosis. We studied the anti-cancer activity of metformin on medulloblastoma using DAOY and D283 cells. Metformin treatment resulted in a dose and time dependent reduction in cell proliferation in both cell lines. This growth inhibition was accompanied by the induction of apoptosis as determined by annexin-V staining, PARP cleavage, and activation of caspase3/7. Metformin also induced cell cycle arrest in G0/G1 phase. Western blot analysis using extracts from cells treated with metformin revealed that the anti-proliferative and apoptotic effects of metformin in medulloblastoma were accompanied by a reduction in the expression of survivin protein. Overexpression of survivin is reported to be associated with chemo- and radio-resistance. This study suggests the use of metformin as a novel anticancer agent in combination with standard therapy for the treatment of medulloblastoma.

Citation Format: Kristen C. Payne, Abigail Hunter, W Paul Bowman, Jeffrey C. Murray, Riyaz Basha, Umesh T. Sankpal. Metformin treatment inhibits proliferation and induces apoptosis in medulloblastoma 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 889.
Abstract 95: Heparan sulfate and glypican-1 mediate renal carcinoma metastasis
Authors: Qazi Henry , Moran Heriberto , Cancel Limary M. , Mayer Mariya , Roberge Sylvie , Huang Peigen , Munn Lance L. , Tarbell John M.
Published: August 2018

The surface proteoglycan/glycoprotein layer (glycocalyx) on tumor cells has been associated with cellular functions that can potentially enable invasion and metastasis. In addition, aggressive renal carcinoma cells (SN12L1) with high metastatic potential have enhanced invasion rates compared to low metastatic (SN12C) cells in response to interstitial flow stimuli in vitro. Our previous studies suggest that heparan sulfate (HS) and hyaluronic acid (HA) in the glycocalyx play an important role in this flow mediated mechanotransduction and upregulation of invasive and metastatic potential. In our recent study, SN12L1 cells were genetically modified to suppress HS production by knocking down its synthetic enzyme NDST1. Using modified Boyden chambers with defined interstitial flow, we showed that flow-enhanced invasion is suppressed in HS deficient cells. We also examined two prominent HSPGs on renal carcinoma cells, glypican-1 and syndecan-1, and one prominent HA receptor, CD44. We observed higher glypican-1 levels in flow dependent SN12L1 cells when compared to SN12C cells. Caki-1 (highly metastatic) cells did not display flow-dependent invasion in vitro and did not display elevated glypican-1 compared to low metastatic Caki-2 cells. However, we did observe significantly increased HS, HA, syndecan-1 and CD44 in Caki-1 compared to Caki-2 cells suggesting an alternative mechanism for reported higher metastatic rates in these cells. All of our data are consistent with the hypothesis that glypican-1 is the core protein responsible for flow sensing in metastatic cancer cells. This is also consistent with observations in endothelial cells. To assess the ability of tumor cells to metastasize in vivo, parental or HS knockdown SN12L1 cells expressing fluorescent reporters were injected into kidney capsules in SCID mice. Histological analysis confirmed that there was a large reduction (95%) in metastasis to distant organs by tumors formed from knockdown cells compared to control cells with intact HS. The reduction was even greater (98%) in lungs where most of the metastases from the control cells were observed. The ability of these knockdown cells to invade surrounding tissue was also impaired. The substantial inhibition of metastasis and invasion upon reduction of HS suggests an active role for the tumor cell glycocalyx and glypican-1 in tumor progression.

Citation Format: Henry Qazi, Heriberto Moran, Limary M. Cancel, Mariya Mayer, Sylvie Roberge, Peigen Huang, Lance L. Munn, John M. Tarbell. Heparan sulfate and glypican-1 mediate renal carcinoma metastasis [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 95.
Abstract 964: Genome-wide CRISPR screen identifies potential therapeutic combination of EGFR and FGFR inhibitors in oral cancer
Authors: Ludwig Megan , Birkeland Andrew , Nimmagadda Sai , Foltin Sue , Kulkarni Aditi , Jiang Hui , Carey Thomas , Brenner John Chad
Published: August 2018

Oral squamous cell carcinoma (OSCC) has remained a disease with poor survival for decades. Current treatment options such as targeting EGFR has had limited success. We hypothesize that inhibiting compensatory pathways in combination with EGFR signaling may result in greater efficacy. To identify synergistic combinations with EGFR-targeted therapy, we introduced Genome-scale CRISPR-Cas9 KnockOut (GeCKO) libraries into OSCC cell lines. CRISPR-Cas9 generates individual genetic knockouts through targeted gene editing. We used a pool of CRISPRs targeting over 18,000 genes to perform genome-scale screening for drivers of sensitivity to EGFR-targeted therapy. Upon selection of the OSCC GeCKO pool, we identified gene knockouts in the FGFR pathway that increased sensitivity to the EGFR inhibitor gefitinib. Using resazurin viability assays we tested combinations of EGFR and FGFR inhibitors in 14 OSCC cell lines. Six/14 (43%) of the cell lines were responsive to the combination, indicating that the FGFR pathway is an alternate mechanism of resistance to EGFR-targeted therapy in some tumors. We advanced an OSCC cell line to test in vivo by establishing subcutaneous cell line xenografts in mice. After treatment for 21 days, the mice receiving the combination of EGFR and FGFR inhibitors, gefitinib and BGJ398, had significantly smaller tumors than mice receiving vehicle or either monotherapy. These results suggest targeting EGFR in combination with its compensatory pathway such as FGFR signaling could be effective in vivo. While OSCC remains a common and frequently lethal cancer, there is great potential for the development of novel personalized targeted therapies. Here, we describe the use of genome-wide CRISPR-Cas9 library to discover the synergistic combination of EGFR and FGFR inhibition. Further investigation suggests the FGFR pathway is a common compensatory mechanism to EGFR inhibition, and that targeting EGFR and FGFR in combination has efficacy in an in vivo model. Overall, targeting compensatory mechanisms in combination with EGFR-targeted therapy could generate novel, beneficial treatment plans for patients with OSCC.

Citation Format: Megan Ludwig, Andrew Birkeland, Sai Nimmagadda, Sue Foltin, Aditi Kulkarni, Hui Jiang, Thomas Carey, John Chad Brenner. Genome-wide CRISPR screen identifies potential therapeutic combination of EGFR and FGFR inhibitors in oral 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 964.
Abstract CT104: Evaluation of peripheral T cell subset proliferation as a pharmacodynamic assay to guide the development of anti-CTLA-4 and PD-1 antibody combinations in patients with solid tumors
Authors: Souza Paul De , Malczewski Agnieszka , Proscurshim Igor , Yuan Guojun , Coart Elisabeth , Dupont Christopher , Manrique Mariana , Drouin Elise , Gonzalez Ana , Carini Meg , Shebanova Olga , Cuillerot Jean-Marie
Published: August 2018

The combination of anti-CTLA-4 and anti-PD-1 antibodies has demonstrated significant clinical efficacy in metastatic melanoma, but also significant toxicities. Pharmacodynamic (PD) markers are important tools in optimizing the safety and efficacy of anti-CTLA-4/anti-PD-1 combinations, but have remained elusive to date. To test the hypothesis that blockade of both CTLA-4 and PD-1 together results in increased expression of the proliferation marker Ki67 in CD4+ T cells, we studied two dose regimens of ipilimumab in combination with nivolumab in patients with advanced or metastatic solid tumors, using a validated assay. Subjects received either 1.0 mg or 0.3 mg /kg of ipilimumab every 6 weeks in combination with nivolumab 3mg/kg every 2 weeks for a maximum of 12 weeks (2 cycles). A total of 9 subjects were enrolled at the 1.0 mg/kg of ipilimumab cohort and 12 subjects in the 0.3 mg/kg cohort. Highly statistically significant increases of 4.56 fold (range 3.35-6.20) from baseline at cycle 1 day 8 post-treatment (p < 0.001) in cohort 1, and 3.13 fold (range 2.31-4.33) from baseline at cycle 1 day 8 post-treatment (p < 0.001) in cohort 2, in Ki67 expression were observed in CD4+ T cells. The absolute increase in Ki67 expression was also observed to be dose dependent numerically. Significant increases in Ki67 expression were also observed in CD8+ T cells, regulatory T cells, and natural killer T (NKT) cells following treatment. These results identify and support Ki67 expression as a PD biomarker in the context of CTLA-4 and PD-1 blockade. More work is required to correlate these finding with clinical outcomes in order to establish a benchmark biomarker assay for anti-CTLA-4 and anti-PD-1 combinations. (ACTRN12617001059358)

Citation Format: Paul De Souza, Agnieszka Malczewski, Igor Proscurshim, Guojun Yuan, Elisabeth Coart, Christopher Dupont, Mariana Manrique, Elise Drouin, Ana Gonzalez, Meg Carini, Olga Shebanova, Jean-Marie Cuillerot. Evaluation of peripheral T cell subset proliferation as a pharmacodynamic assay to guide the development of anti-CTLA-4 and PD-1 antibody combinations in patients with solid tumors [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 CT104.