bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2693: Targeting the Hippo-YAP pathway with novel small-molecule inhibitors of the YAP-TEAD transcription activity
Authors: Tang Tracy T. , Konradi Andrei W. , Feng Ying , Peng Xiao , Qiao Sofie , Post Leonard
Published: August 2019

We have discovered and are developing multiple novel small molecule medicines targeting the Hippo-YAP pathway. Consisting of upstream regulatory components, the Hippo core kinase components (MST1/2, MAP4K1-7, and LATS1/2), and the downstream transcriptional machinery (YAP, TAZ, and TEAD), the mammalian Hippo-YAP pathway is involved in the regulation of cell proliferation, survival, and cell migration. It functions normally to control tissue regeneration and homeostasis and limit organ size/shape. Genetic alterations of pathway components have been reported in a variety of human malignancies. YAP activity is also linked to resistance to targeted therapies and cancer relapse. Through high-throughput phenotypic screens, we have identified novel classes of small molecule compounds that inhibit the YAP-TEAD transcription activity. Medicinal chemistry efforts on lead optimization resulted in highly potent inhibitors that block YAP-TEAD transcription activity at single digit nM. The classes of inhibitors that were demonstrated to interact directly with the TEAD transcription factors by thermal shift assay, co-crystal structures, and inhibition of autopalmitoylation of purified recombinant TEAD proteins will be presented. In cell-based assays, these inhibitors block TEAD palmitoylation in cells and selectively inhibit proliferation of NF2-deficient mesothelioma. In vivo, complete growth inhibition of NF2 mutant xenograft tumors can be achieved at doses that are well tolerated in mice without adverse effect on body weight. With the discovery and development of potent TEAD inhibitors, we bring forth a new generation of cancer therapeutics targeting a thus far undruggable pathway that is dysregulated in many cancers and critically associated with targeted therapy resistance.

Citation Format: Tracy T. Tang, Andrei W. Konradi, Ying Feng, Xiao Peng, Sofie Qiao, Leonard Post. Targeting the Hippo-YAP pathway with novel small-molecule inhibitors of the YAP-TEAD transcription activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2693.
Abstract 2697: YPN005, an oral CDK7 inhibitor, exhibits a significant antitumor activity in Myc-driven cancers
Authors: Lee Kwang-Ok , Yoo Jakyung , Lee Mi Jung , Lee Kang Woo , Min Ji Eun , Kim Jinhwan , Min Ki-Nam , Roh Tae Chul , Seo Kang-Sik , Rhee Hae In , Lee Jun Hee , Jeon Da-Hye , Lim Dae Seong
Published: August 2019

Background: CDK7 plays an important role in regulating cell cycle progression and gene via activation of cell cycle kinases (CDK1, CDK2, CDK4 and CDK6) and RNA polymerase II (PolII). Recent studies indicate that the inhibition of CDK7 is an attractive strategy for the treatment of cancer by down-regulation of c-Myc expression. (Wang et al 2018) We have investigated the therapeutic efficacy of YPN005, a novel oral CDK7 inhibitor, in triple negative breast cancer (TNBC) and hepatocellular carcinoma (HCC).

Methods: We evaluated antiproliferative activity of YPN005 on TNBC and HCC cells and the expression of RNA PolII and c-Myc was studied by Western Blot analyses to investigate the mechanism of action. To identify a biomarker, we examined the correlation between the level of c-Myc expression and anticancer activity of YPN005 in vitro using HCC cells. The therapeutic efficacy of YPN005 was evaluated in TNBC xenograft mouse model.

Results: YPN005 significantly inhibited the proliferation of TNBC and HCC cells and IC50s was determined as 10-20 nM and 5-30 nM range, respectively. Inhibition of cell proliferation was accompanied by a decrease in the levels of RNA PolII phosphorylation and c-Myc expression. Western Blot analyses revealed that the sensitivity of HCC cells to YPN005 was correlated with the level of c-Myc expression. In vivo xenograft model of TNBC showed that oral daily administration of YPN005 for 21 days (once, and 10mg/kg) efficiently inhibited the growth of tumor. All mice survived during the dosing period without significant changes of the hematologic profiles.

Conclusion: We propose that oral administration of YPN005, an orally available CDK7 inhibitor, could be a potent and attractive approach to treat the Myc overexpressing cancers.

Citation Format: Kwang-Ok Lee, Jakyung Yoo, Mi Jung Lee, Kang Woo Lee, Ji Eun Min, Jinhwan Kim, Ki-Nam Min, Tae Chul Roh, Kang-Sik Seo, Hae In Rhee, Jun Hee Lee, Da-Hye Jeon, Dae Seong Lim. YPN005, an oral CDK7 inhibitor, exhibits a significant antitumor activity in Myc-driven cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2697.
Abstract 27: Identification and characterization of a novel dual FLT3/microtubule polymerization inhibitor through cell-based screening
Authors: Malik Haleema Sadia , Bilal Aishah , Ullah Rahim , Iqbal Maheen , Saleem Rahman Shah , Hussain Hidayat , Faisal Amir
Published: August 2019

Activating mutations in FLT3 receptor tyrosine kinase are found in one-third of acute myeloid leukemia (AML) patients and are associated with disease relapse and bad prognosis. Majority of these mutations are Internal tandem duplications (ITD) in the juxtamembrane domain of FLT3 and have been validated as a therapeutic target. Clinical success of selective inhibitors targeting oncogenic FLT3, however, has been limited due to the acquisition of drug resistance. Here we report the identification of a dual FLT3/microtubule polymerization inhibitor, HH-IA-208, through cell-based screenings for the differential killing of leukemia cell lines expressing FLT3-ITD (MV411) or BCR-ABL (K562) using an MTS proliferation assay and for mitotic inducers using a dot-blot assay for histone H3 phosphorylation in HCT116 cell line. HH-IA-208, a chalcone derivative, inhibits FLT3 in vitro and treatment of FLT3-ITD+ cell lines with HH-IA-208 results in reduced FLT3 signaling. Similarly, HH-IA-208 inhibits tubulin polymerization in a biochemical assay and in cells that results in a mitotic arrest. HH-IA-208 is selectively more potent in leukemia cell lines expressing FLT3-ITD and treatment for 24 and 48 hours with the inhibitor results in apoptotic cell death. Furthermore, HH-IA-208 is able to overcome TKD mutation-mediated acquired resistance to FLT3 inhibitors in a MOLM13 cell line expressing FLT3-ITD with the D835Y mutation. HH-IA-208, therefore, is a promising lead for the discovery of dual target FLT3 inhibitors that can overcome resistance to selective FLT3 inhibitors.

Citation Format: Haleema Sadia Malik, Aishah Bilal, Rahim Ullah, Maheen Iqbal, Rahman Shah Saleem, Hidayat Hussain, Amir Faisal. Identification and characterization of a novel dual FLT3/microtubule polymerization inhibitor through cell-based screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 27.
Abstract 2734: Poly-ligand profiling of plasma samples as a potential predictor of prostate biopsy results
Authors: Wei Xixi , Liu Xiaowei , Santhanam Radhika , Magee Daniel , Helmstetter Anthony , Miglarese Mark , Spetzler David , Wei Xixi
Published: August 2019

Prostate cancer is the most common cancer in men, excluding skin cancer, and is also one of the leading causes of cancer death. Currently the prostate-specific antigen (PSA) blood test is employed as an initial screening method in men without symptoms, and patients with blood PSA level higher than 4 ng/mL are commonly recommended for prostate biopsy for definitive diagnosis. However, other non-cancerous conditions may also cause elevation in PSA, such as prostatitis, benign prostatic hyperplasia and prostate intraepithelial neoplasia. Among men with PSA levels of 4-10 ng/mL, only about 1 in 4 will show positive (cancer) biopsy, and for those with PSA levels higher than 10 ng/mL, the chance of having prostate cancer increases to over 50%. Moreover, approximately 15% of men with a PSA levels below 4 ng/mL have a positive prostate biopsy. Because of these scenarios, as well as significant risks associated with prostate biopsies, there is a need for diagnostic adjuncts that provide additional data to inform decisions on whether or not to perform biopsies.

Here we describe a method using single stranded oligonucleotide (ssODN) libraries to perform Poly-Ligand Profiling on plasma exosomes from men scheduled for biopsy to classify those with or without a subsequent diagnosis of prostate cancer. Through alternating rounds of positive and negative selection in binding assays, we isolated ssODNs that selectively bound to exosomes from prostate cancer patients (PSA level of at least 4 ng/mL; Gleason Score range 7-8, and at least 3 positive cores from at least 10 biopsy cores) and not to benign cases (all negative cores from at least 10 biopsy cores). An independent test set of 29 prostate cancer cases and 28 benign cases was then evaluated in binding assays. Based on 100 runs of 10-fold cross validation with multiple statistic models, an average successful classification rate of 82% was achieved with an area under the ROC curve of 0.89. While further refinement and testing on larger cohorts are needed, these results suggest that Poly-Ligand Profiling may provide an additional approach to stratify men with suspicious PSA levels for prostate biopsies.

Citation Format: Xixi Wei, Xiaowei Liu, Radhika Santhanam, Daniel Magee, Anthony Helmstetter, Mark Miglarese, David Spetzler, Xixi Wei. Poly-ligand profiling of plasma samples as a potential predictor of prostate biopsy results [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2734.
Abstract 2741: Comprehensive molecular profiling of primary tumors and paired distant metastases in non-small cell lung cancer
Authors: Lee Won-Chul , Gomez Daniel , Zhang Jianhua , Reuben Alexandre , Jalali Ali , Roh Whijae , Wu Chia Chin , Lu Wei , Chow Chi-Wan , Fujimoto Junya , Antonoff Mara , Moran Cesar , Sulman Erik , Rao Ganesh , Swisher Stephen , Heymach John , Wistuba Ignacio I. , Futreal Andrew , Zhang Jianjun
Published: August 2019

Despite complete resection, many non-small cell lung cancer (NSCLC) patients still develop and succumb to distant metastases, which is a major cause of cancer-related death worldwide. However, our understanding of the molecular mechanisms driving metastasis is rudimentary. We performed whole exome sequencing, RNA sequencing (RNA-seq), methylation microarray, and immunohistochemistry using multiple immune markers on 8 pairs of NSCLC primary tumors and matched distant metastases including 7 metachronous brain and 1 synchronous liver metastases. On average, 60% of all somatic mutations (22% to 90%) and 86% of canonical cancer gene mutations were shared between primary tumors and paired distant metastases. Metastases also resembled paired primary tumors closely in regard to their mutational spectrum, copy number aberrations, allelic imbalance, and methylation profiles. Genomic aberrations unique to metastases were rather patient-specific than commonly observed across multiple patients. Of particular interest, subclonal architecture analysis suggested that monoclonal metastatic seeding is a prevalent mode of metastasis in NSCLC. We validated these findings in a large published dataset consisting of 38 pairs of primary lung tumors and matched distant metastases. RNA-seq showed gene expression profiles in metastases were similar to those of paired primary tumors. Metastases commonly up-regulated metabolism-associated pathways and down-regulated immune-related pathways, consistent with the immunohistochemical staining data for CD3, CD4, CD8, CD20, CD68, PD1, and PD-L1. Our data suggest that distant metastasis is a late event during carcinogenesis and that a majority of genomic and epigenetic aberrations occur before metastatic outgrowth. While molecular mechanisms underlying postsurgical distant metastasis seem to be variable among NSCLC patients, immune suppression may be a common characteristic of cancer cells with metastatic plasticity.

Citation Format: Won-Chul Lee, Daniel Gomez, Jianhua Zhang, Alexandre Reuben, Ali Jalali, Whijae Roh, Chia Chin Wu, Wei Lu, Chi-Wan Chow, Junya Fujimoto, Mara Antonoff, Cesar Moran, Erik Sulman, Ganesh Rao, Stephen Swisher, John Heymach, Ignacio I. Wistuba, Andrew Futreal, Jianjun Zhang. Comprehensive molecular profiling of primary tumors and paired distant metastases in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2741.
Abstract 2742: Bone marrow-derived stromal cells in the tumor microenvironment might be associated with the progression of gastric cancer
Authors: Yashiro Masakazu , Kasashima Hiroaki , Kushiyama Syuhei , Nishimura Sadaaki , Kuroda Kenji , Togano Shingo , Okuno Tomohisa , Miki Yuichiro , Fukuoka Tatsunari , Nakamae Hirohisa , Hino Masayuki , Ohira Masaichi
Published: August 2019

Purpose: It has been reported that the interaction between cancer cells and cancer-associated fibroblasts (CAFs) is closely associated with the progression of various types of cancer. The aim of this study is to clarify the role of bone marrow-derived stromal cells (BM-SCs), which are supposed to be the origin of CAFs in the tumor microenvironment, on the development of gastric cancer.

Methods: The effect of human BM-SCs which frequently express CD271 on the proliferation and motility of six gastric cancer cell lines, OCUM-2M, OCUM-2MD3, OCUM-12, KATO-III, NUGC-3, and MKN-74, was examined. CD271 expression levels in BM-SCs were analyzed by flow cytometry. We also generated a gastric tumor model by orthotopic inoculation of OCUM-2MD3 cells in mice that had received transplantation of bone marrow from CAG-EGFP mice. The correlation between the clinicopathological features of 279 primary gastric carcinomas and CD271 expression in tumor stroma was examined by immunohistochemistry.

Results: Numerous BM-SCs infiltrated the gastric tumor microenvironment; CD271 expression was found approximately 25% of BM-SCs. Conditioned medium from BM-SCs significantly increased the proliferation of gastric cancer cell lines. Furthermore, conditioned medium from gastric cancer cells significantly increased the number of BM-SCs, while migration of OCUM-12 and NUGC-3 cells was significantly increased by conditioned medium from BM-SCs. CD271 expression in stromal cells was significantly associated with macroscopic type-4 cancers, diffuse-type tumors, and tumor invasion depth. The overall survival of patients (n=279) with CD271-positive stromal cells was significantly worse than that of patients with CD271-negative stromal cells. This is the first report of the significance of BM-SCs in gastric cancer progression.

Conclusions: BM-SCs were abundant in the tumor microenvironment of gastric cancer in vivo. In vitro study also indicated that the crosstalk between BM-SCs and gastric cancer cells might play an important role for the development of gastric cancer. CD271-positive tumor stromal cells might be a useful predictive prognostic factor for patients with gastric cancer. BM-SCs, therefore, might be one of therapeutic targets for cancer treatment.

Citation Format: Masakazu Yashiro, Hiroaki Kasashima, Syuhei Kushiyama, Sadaaki Nishimura, Kenji Kuroda, Shingo Togano, Tomohisa Okuno, Yuichiro Miki, Tatsunari Fukuoka, Hirohisa Nakamae, Masayuki Hino, Masaichi Ohira. Bone marrow-derived stromal cells in the tumor microenvironment might be associated with the progression of gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2742.
Abstract 2748: The role of TMPRSS2_ERG fusions in modulating tumor microenvironment in prostate cancer
Authors: Pakula Hubert , Ribeiro Caroline F. , Fanelli Giuseppe N. , Kirchner Rory , Syamala Sudeepa , Chowdhury Basudev , Zadra Giorgia , Chetta Paolo , Li Zhe , Loda Massimo
Published: August 2019

The control region of TMPRSS2 fuses with ETS transcription factor family members, particularly ERG. This occurs in ~20-50% of human prostate cancers depending on the ethnicity. TMPRSS2-ERG (T2E) fusions, however, do not display a notable phenotype in genetically engineered mouse models (GEMM) recapitulating the fusion expression and T2E also has no prognostic significance in humans, despite its high incidence. Wnt signaling has been implicated in prostate cancer. Here, we report a discovery of a role for T2E fusion that impacts the prostate stroma in pre-clinical models. The composition of stromal cells within the microenvironment in T2E mice was studied by single cell RNAseq analysis. We identified stromal cell clusters in T2E mice that differed from wild type mice in the expression of Pdgfrβ, Pdgfrα and Col1a1. Interestingly, these clusters showed an increase of expression of Wnt receptors such as Lgr5 and Fzd7 as well as Wnt ligands such as Sfrp2, Wnt2 and Wnt6. T2E GEMMs also exhibited upregulation of the Wnt-secretion regulator porcupine (PORCN-Protein-serine O-palmitoleoyltransferase) in the stroma adjacent to the T2E-expression prostate epithelial cells. Furthermore, lineage-tracing with the Wnt reporter and epithelial stem cell marker Lgr5 showed that Wnt-active cells were increased in prostatic intraepithelial neoplasia (PIN) lesions in T2E;Pten+/- mice. Strikingly, Lgr5+ cells were also found in the prostate stroma surrounding these PINs. Since, TMPRSS2/ERG fusions represent an early event in prostate tumorigenesis, here we provide a mechanism whereby induction of Wnt signaling in the stroma by T2E-expressing prostate epithelial cells increases the stem cell compartments in both epithelial and stromal cells. These data suggest that aberrant ERG-expressing prostate epithelial cells activate upregulation of Wnt signaling in stromal cells, which provides a possible route for enhancing prostate carcinogenesis.

Citation Format: Hubert Pakula, Caroline F. Ribeiro, Giuseppe N. Fanelli, Rory Kirchner, Sudeepa Syamala, Basudev Chowdhury, Giorgia Zadra, Paolo Chetta, Zhe Li, Massimo Loda. The role of TMPRSS2_ERG fusions in modulating tumor microenvironment in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2748.
Abstract 2760: Photoinducible detection of the oncometabolite fumarate
Authors: Briney Chloe , Bergholtz Sarah , Kulkarni Rhushikesh , Crooks Daniel , Mushti Chandrasekhar , Lockett Stephen , Swenson Rolf , Linehan W. Marston , Meier Jordan
Published: August 2019

Dysregulated metabolism is an important marker of many disease states, including cancer. For example, in hereditary leiomyomatosis and renal cell cancer (HLRCC), inactivating mutations in fumarate hydratase (FH) lead to accumulation of high levels of fumarate, a so-called “oncometabolite.” Substantial evidence indicates that fumarate stimulates various oncogenic signaling pathways, necessitating sensitive methods to detect the oncometabolite in order to more rapidly diagnose HLRCC as well as to identify new disease settings in which fumarate may play a signaling role. Here, we report development of novel photoactivatable, fluorogenic chemical probes for detection and profiling of fumarate in biological systems. These chemical probes, diaryl tetrazoles, are by themselves inert towards fumarate. However, upon irradiation with UV light, they release nitrileimines that can form fluorescent cycloadducts with fumarate. We have demonstrated that diaryl tetrazoles can sensitively detect FH activity as well as low micromolar levels of fumarate in complex biological samples. We have also shown that diaryl tetrazoles can be used to monitor changes in intracellular fumarate levels in biological samples by live-cell imaging and flow cytometry. Moreover, these compounds are capable of visualizing differences between patient-derived primary HLRCC tumors lacking FH activity and the adjacent normal kidney, highlighting their potential utility in clinical diagnostics. By offering new insights into fumarate reactivity, our studies provide the chemical basis for novel approaches to therapy and diagnosis in cancers driven by oncometabolite accumulation.

Citation Format: Chloe Briney, Sarah Bergholtz, Rhushikesh Kulkarni, Daniel Crooks, Chandrasekhar Mushti, Stephen Lockett, Rolf Swenson, W. Marston Linehan, Jordan Meier. Photoinducible detection of the oncometabolite fumarate [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2760.
Abstract 2764: Small activating RNA induced expression of VHL gene in renal cell carcinoma
Authors: Kang Jong Soon , Kang Moo RIm , Park Ki Hwan
Published: August 2019

Recent studies have reported that chemically synthesized double-stranded RNAs (dsRNAs), also known as small activating RNA (saRNAs), can specifically induce gene expression by targeting promoter sequences in a mechanism termed RNA activation (RNAa). In the present study, we designed four candidate dsRNAs targeting the Von Hippel-Lindau (VHL) gene promoter. Among these dsRNAs, dsVHL-821 significantly inhibited cell growth by up-regulating VHL at both the mRNA and protein levels in renal cell carcinoma 769-P cells. Functional analysis showed that dsVHL-821 induced apoptosis by increasing p53, decreasing Bcl-xL, activating caspase 3/7 and poly-ADP-ribose polymerase in a dose-dependent manner. Chromatin immunoprecipitation analysis revealed that dsVHL-821 increased the enrichment of Ago2 and RNA polymerase II at the dsVHL-821 target site. In addition, Ago2 depletion significantly suppressed dsVHL-821-induced up-regulation of VHL gene expression and related effects. Consistent with previous studies on RNAa of other genes, single transfection of dsVHL-821 caused long-lasting (14 days) VHL up-regulation. Furthermore, the activation of VHL by dsVHL-821 was accompanied by an increase in dimethylation of histone 3 at lysine 4 (H3K4me2) and acetylation of histone 4 (H4ac) and a decrease in dimethylation of histone 3 at lysine 9 (H3K9me2) and lysine 27 (H3K27me2) in the dsVHL-821 target region. Taken together, these results demonstrate that dsVHL-821, a novel saRNA for VHL, induces the expression of the VHL gene by epigenetic changes, resulting in inhibition of cell growth and induction of apoptosis. Targeted activation of VHL by dsVHL-821 may be explored as a novel treatment of renal cell carcinoma.

Citation Format: Jong Soon Kang, Moo RIm Kang, Ki Hwan Park. Small activating RNA induced expression of VHL gene in renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2764.
Abstract 2767: Expression of defensin-associated genes may be correlated with lymph node metastasis of early stage tongue cancer
Authors: Lee Doh Young
Published: August 2019

OBJECTIVES: Appropriate patient selection for elective neck dissection in N0 tongue cancer has been controversial, while there has been no genetic consideration so far. We aimed to analyze genetic difference between early and late cervical lymph node metastasis in oral tongue cancer patients using the Seoul National University Hospital (SNUH) cancer cohort and cancer genome atlas (TCGA) data, and to suggest genetic background for choosing eligible patient for elective neck dissection. METHODS: A total of 35 cases with RNAseq in SNUH cancer cohort with tongue cancer were also enrolled in this study. To investigate gene expression in early cervical lymph node metastasis, genomic data of following 2 groups was compared; 1) N0 group: T1/2 and N0, 2) N+ group: T1/2 and N+. Differentially expressed genes (DEGs) were extracted using R and limma package in bioconductor program. Gene ontology and pathway enrichment analysis were performed using DAVID online tool. Immune cell infiltration was analyzed using CIBERSORT online program. Additionally, 70 cases with matched RNAseq data of primary tumor and clinical TCGA data were analyzed to validate the role of DEGs. RESULTS: N0 and N+ groups showed no difference in age, gender, size and thickness of tumor, and nearest resection margin. In addition, there was no significant difference in 22 types of immune cell infiltration. A total of 51 DEGs were identified, and 14 genes were significantly upregulated and 37 genes were significantly downregulated (p<0.01, fold-change >2). Significant genes were PLA2G4D, NELL2, PCOLCE2, PI3, and PLA2G3. On KEGG pathway analysis, calcium, muscle contraction, and arachidonic acid metabolism-related pathway was significantly correlated. The most significant difference was found in 6 genes as follows; DEFB4A, SPRR2B, DEFB103B, SPRR2G, DEFB4B, FAM25A.DEFB which was the most significantly difference in expression was associated with ‘antibiotic’, ‘antimicrobial’, ‘beta defensing type’, defense reponse to bacterium’, and ‘defensin’. In TCGA data, DEFB4A and DEFB103B were more highly expressed in N0 group than N+ group, although it did not attain significant significance. CONCLUSIONS: Defensin (DEFB4A, DEFB103B, DEFB4B) may be a novel biomarker for early regional metastasis in T1/2 tongue cancer. Considering that defensin can be detected in saliva, simple saliva test for prediction of metastasis can be developed in the future.

Note: This abstract was not presented at the meeting.

Citation Format: Doh Young Lee. Expression of defensin-associated genes may be correlated with lymph node metastasis of early stage tongue cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2767.
Abstract 2771: Identification of novel genes associated with metastasis from TCGA transcriptomic data
Authors: Koyama Takahiko , Parida Laxmi
Published: August 2019

Introduction: Many cancer patients suffer from metastasis in bone, brain, liver and lung. However, understanding of metastasis in molecular level is quite limited. To improve survival rate and quality of life, preventing metastasis is crucial. In this study, we analyzed transcriptomic data with tumor node metastasis (TNM) classification from The Cancer Genome Atlas (TCGA) to identify genes associated with metastasis.

Method: TCGA samples in 20 solid tumor types with both transcriptome and TNM annotation data are used. To obtain the best cut point separating cohorts of high and low gene expression, Hothorn and Lausen’s method is used. A contingency table is created with expression separated by the the cut point and presence of staging T, N, M for each gene. Association between the staging category and the gene was calculated with Fisher’s exact test.

Results: In total, 13,220 distinct genes including miRNAs are associated with either lymph node or distant metastasis for some cancer types with a loose criteria of p-value < 1.0e-3. Among them, we identified 1,357 genes with p-value < 1.0e-8. ADAMTS12, AURKB, BUB1, C17orf53, CCNA2, CEP55, EPR1, KIF4A, KIF11, KRT80, MELK, MKI67, NCAPG, NEIL3, PBK, PLK1, RRM2, TPX2, miR139, miR30a, miR375, and miR379 appear most commonly in lymph node metastasis for many cancer types. Likewise, NOP56, PKD1L2, miR150, miR365, miR9, miR149, miR210, miR425, miR675, and miR937 are most commonly associated genes in distant metastasis. Many of these genes have been already reported in association with metastasis.

Conclusions: This study created comprehensive list of metastasis associated genes for the 20 solid tumors in TCGA. They would be useful as diagnostic and prognostic biomarkers; however, we found therapeutically relevant genes such as PLK1, AURKB, MET, MELK, CDK1, BUB1, and PBK. Some adhesion molecules associated with a particular cancer type might explain its organ destination preference. Among distant metastatic miRNAs discovered, metastasis promoting miRNAs in high abundance might be engaged in homing as exosomal miRNAs by reprogramming destination niche in favor of migrating tumor cells. Further work to reveal metastasis mechanism and to discover some pharmaceutical agents to prevent metastasis will be conducted.

Citation Format: Takahiko Koyama, Laxmi Parida. Identification of novel genes associated with metastasis from TCGA transcriptomic data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2771.
Abstract 2778: ADD3-loss in glioblastoma is associated with increased angiogenesis and malignancy
Authors: Kiang Mei-Yee , Leung Gilberto Ka-Kit
Published: August 2019

Introduction: Adducin 3 (ADD3) is one of the major functional component in the cytoskeleton. Dysregulation of ADD3 has been implicated in various diseases including cancer and participate in different signaling transductions. However, little is known about the functional properties of ADD3 in cancer, its role in cancer biology remains unclear and controversial. We aim to delineate the role of ADD3 dysregulation in GBM pathogenesis and to unravel the underlying molecular mechanisms.

Methods: Microarray datasets were used to analyze the gene expression of ADD3 in normal tissue and grade II, III, IV gliomas. Gene expression levels were verified by western blot, qPCR and immuno-histological analyses by using human clinical specimens. Luciferase-expressing ADD3 stable knockdown U87 malignant glioma cells were injected to immunocompromised mice subcutaneously and orthotopically for xenograft models. ADD3 stable knockdown U87 cells were co-cultured with human endothelial cells (HUVEC) to assess the functional changes.

Findings: Our study demonstrated significant downregulation of ADD3 in human GBM when compared with lower-grade gliomas. Downregulation of ADD3 was only shown in GBM (i.e., WHO grade IV lesion) but not in grade II and grade III tumor. Importantly, we found that ADD3 downregulation is associated with poor clinical outcome in GBM patients. And that ADD3 knock-down enhanced tumor growth in vivo, possibly mediated by promoting angiogenesis.

Conclusions: These findings suggest that ADD3 is associated with glioma malignancy, implying that cancer progression may occur along with the loss of ADD3. It may contribute towards tumor growth by enhancing angiogenesis.

Citation Format: Mei-Yee Kiang, Gilberto Ka-Kit Leung. ADD3-loss in glioblastoma is associated with increased angiogenesis and malignancy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2778.
Abstract 278: System xc- inhibition shares features of necroptosis and ferroptosis in hepatocellular carcinoma cells
Authors: Abdullah Md , Kim Do Hyung , Lee Seung Jin
Published: August 2019

Ferroptosis is characterized by iron-dependent non-apoptotic cell death involving the accumulation of lipid peroxides. Inhibition of system xc- which uptake L-cystine with the exchange of L-glutamate decreases glutathione synthesis, leading to ferroptosis. However, molecular interplay between ferroptosis and necroptosis has not been clearly identified. In this study, we investigated molecular process for cell death by SLC7A11 inhibition in hepatocellular carcinoma cell lines. The sensitivity for SLC7A11 inhibition by sulfasalazine was in the order of HepG2, Huh7 and Huh6, and then PLC/PRF/5. The decrease of cell viability by sulfasalazine and erastin was prevented by co-treatment of β-mercaptoethanol in all kinds of cell lines, indicating that the limited supply of L-cystine may contribute to cell death. Both ferrostatin and necrostatin, an inhibitor of ferroptosis and necroptosis, respectively, rescued sulfasalazine-induced growth inhibition by 60% in Huh7 and Huh6 and by 20% in HepG2. However, they failed to restore cell survival in PLC/PRF/5 cells. Sulfasalazine at the dose inducing equivalent level of cell death significantly increased lipid peroxidation in Huh7 and Huh6, but not in HepG2 and PLC/PRF/5 cells, as determined by C11-BODIPY dye. RIPK activation in necroptosis pathway by sulfasalazine was monitored in these cell lines. These results suggest that SLC7A11 inhibition triggers mixed-type of cell death via ferroptosis and necroptosis in a context-dependent manner in hepatocellular carcinoma cells (MD Abdullah and DH Kim are equally contributed to this work).

Citation Format: Md Abdullah, Do Hyung Kim, Seung Jin Lee. System xc- inhibition shares features of necroptosis and ferroptosis in hepatocellular carcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 278.
Abstract 2784: Driver genes network promotes mesenchymal to epithelial transition and organ-specific metastasis
Authors: Chang Ming , Huang Xin , Lu Songjian , Zhang Wei , Liang Weiping , Keller Evan , Lu Xinghua , Zhang Jian
Published: August 2019

Metastasis, a major characteristic of malignancy, causes high mortality. Accumulating evidence suggests that cancer intends to metastasize to specific organ, however the molecular mechanisms remain unknown. Others and us have demonstrated that epithelial to mesenchymal transition (EMT) plays key roles in initiation of primary tumor adherent, migration, and invasion, whereas mesenchymal to epithelial transition (MET) revokes “sleeping” disseminated tumor cell (DTC) to colonization at distant metastatic organ sites and facilitates the tumor cell proliferation. We have successfully generated the lung-specific metastatic subclones of human prostate cancer PC3 and murine RM1 cells. Through omic bench work and data analysis and comparison with TCGA database, we, for the first time, identified a preliminary network of “driver genes” that potentially promotes the reversible EMT/MET process We utilized cutting-edge tools of data analysis to identify the selected driver genes (LAMA, SPP1, ITGB3, S100A8, SERPINE, DKK2) and further characterized the gene functions via in vitro bioassay of cell proliferation, migration, invasion, cell cycle, and apoptotic testing etc, and in vivo with our unique cell and animal metastatic models. We observed that the rate of organ-specific metastasis was dramatically reduced when these gene expression levels were changed. Finally, we confirmed our findings using TMA slides, Our results may provide novel mechanisms of metastasis and clinical targets of cancer therapy. Supported by NSFC projects 81773146; Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research (2017B030301018); JCYJ20170412152943794, JCYJ20170412154619484, JCYJ20170307105128101, JCYJ2017030711041760

Note: This abstract was not presented at the meeting.

Citation Format: Ming Chang, Xin Huang, Songjian Lu, Wei Zhang, Weiping Liang, Evan Keller, Xinghua Lu, Jian Zhang. Driver genes network promotes mesenchymal to epithelial transition and organ-specific metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2784.
Abstract 2787: Artificial intelligence approach identifies IL2RB as a common prognostic and potential predictive biomarker associated with immune checkpoints in colorectal cancer
Authors: Alderdice Matthew , Craig Stephanie , Humphries Matt , Gilmore Alan , Bingham Victoria , Johnston Nicole , McQuaid Stephen , Salto-Tellez Manuel , Lawler Mark , McArt Darragh G.
Published: August 2019

Identifying robust predictive biomarkers to enable stratification of colorectal cancer (CRC) patients, based on their response to immune checkpoint therapy, is an area of unmet clinical need. Genetic algorithms represent an exciting branch of artificial intelligence which can be used to extract meaningful associations from ‘big data’ now emerging more frequently in oncological research. We have employed Atlas Correlation Explorer (ACE), a user-friendly workbench that utilises a genetic algorithm to mine data deposited in The Cancer Genome Atlas (TCGA). Our aim was to establish common intersections between gene expression analyses in ACE using nine well established immune checkpoint markers (CD274, PDCD1, CTLA4, LAG3, TIM3, TIGIT, ICOS, IDO1 and BTLA). We observed IL2RB to be the common gene associated with immune checkpoints in both microarray and RNA sequencing data from the TCGA (7/9 gene lists). Assessment of IL2RB indicates that it is highly expressed on CD56+ natural killer cells and is associated with an increased infiltration of cytotoxic lymphocytes and a decreased infiltration of fibroblasts. It is also significantly enriched in the immune consensus molecular subtype group CMS1. We next demonstrated that patients with high IL2RB gene expression have better relapse free survival in the TCGA CRC cohort (n = 322, log-rank p = 0.011) and an all stage CRC validation cohort GSE39582 (n = 519, log-rank p = 0.006). It is also an independent prognostic factor by multivariate analysis (p = 0.01). We next observed strong correlations between IL2RB gene expression in CRC and previously published predictive gene signatures for anti-PD1 therapies in other solid tumours (Pearson correlation, R = 0.88). Finally, we optimised assessment of IL2RB immunohistochemistry in a large CRC cohort (n=661) using a digital pathology approach with the open-source QuPath software. To conclude, we have validated IL2RB as prognostic biomarker and have provided evidence to demonstrate that IL2RB expression could be used for CRC patient stratification in future immunotherapy based clinical trials.

Citation Format: Matthew Alderdice, Stephanie Craig, Matt Humphries, Alan Gilmore, Victoria Bingham, Nicole Johnston, Stephen McQuaid, Manuel Salto-Tellez, Mark Lawler, Darragh G. McArt. Artificial intelligence approach identifies IL2RB as a common prognostic and potential predictive biomarker associated with immune checkpoints in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2787.
Abstract 2790: Characterization of the immune microenvironment in brain metastasis from non-small cell lung cancer
Authors: Kudo Yujin , Haymaker Cara , Zhang Jiexin , Reuben Alexandre , Duose Dzifa Yawa , Fujimoto Junya , Roy-Chowdhuri Sinchita , Solis Luisa Maren , Dejima Hitoshi , Cuentas Edwin Roger , Mino Barbara , Abraham Ronald , Ikeda Norihiko , Luthra Rajyalakshmi , Lee Jack J. , Kadara Humam , Huse Jason T. , Wistuba Ignacio I.
Published: August 2019

Purpose Brain metastases can induce life-threatening events and their immune microenvironment is still unrevealed. To elucidate the probability of immune-oncology therapy for brain metastases, we utilized immune profiling of resected brain metastases and paired non-small cell lung cancer (NSCLC).

Methods To compare pure tumor-infiltrating lymphocytes (TILs) or tumor tissues between NSCLC and brain metastases, macro-dissection was performed before extracting DNA and RNA from 39 patients who received surgical resection of local NSCLC tumor lesions and brain metastases from matched patients. We applied gene expression analysis of mRNA by using a 770 gene panel related with cancer immune cells. Targeted sequencing of 50 cancer hotspot genes on extracted DNA was performed for genotyping. TCRß sequencing of extracted DNA was utilized for profiling of TCR repertoire.

Results We identified 161 differentially expressed genes, which reflected inhibition of dendritic cell maturation pathway, Th1 pathway, and leukocyte extravasation signaling pathway in brain metastases (p < 0.01). VCAM1 was one of highly inter-connected network genes with low expression level in brain metastases. Immune cell profiling analysis illustrated low T-cell infiltration and higher ratio of macrophages against TILs in the brain metastases (p < 0.001). The detected mutations in genotyping analysis were completely shared in 28 patients (71%). TCR sequencing analysis revealed that T-cell clones were expanded in 64% of patients with brain metastases. The majority of TCR repertoire in brain metastases were principally shared with the paired NSCLC, while the density of T cells was sparse in the brain metastases.

Conclusion The microenvironment in brain metastases from NSCLC was revealed to be immunosuppressive, with the majority of T-cell clones in brain metastases being shared with the primary lesion.

Citation Format: Yujin Kudo, Cara Haymaker, Jiexin Zhang, Alexandre Reuben, Dzifa Yawa Duose, Junya Fujimoto, Sinchita Roy-Chowdhuri, Luisa Maren Solis, Hitoshi Dejima, Edwin Roger Cuentas, Barbara Mino, Ronald Abraham, Norihiko Ikeda, Rajyalakshmi Luthra, Jack J. Lee, Humam Kadara, Jason T. Huse, Ignacio I. Wistuba. Characterization of the immune microenvironment in brain metastasis from non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2790.
Abstract 2791: RelB upregulates PD-L1 in advanced prostate cancer: An insight into tumor immunoescape
Authors: Zhang Yanyan , Zhu Shuyi , Qian Peipei , Wang Xiumei , Xu Zhi , Sun Wenbo , Xu Yong
Published: August 2019

Interaction between programmed death (PD)-1 receptor and its ligand (PD-L1) is essential for suppression of activated T-lymphocytes. Thus, the high level of PD-L1 in tumors is critical for triggering the inhibitory effect on T cell responses leading to immunoescape. However, the mechanism underlying PD-L1 overexpression in cancers remains fully elucidated. Hence, we demonstrate that PD-L1 is uniquely expressed at a high level in advanced prostate cancer with high constitutive RelB, a member to drive the NF-κB alternative pathway. Clinical investigation indicated that the high levels of RelB/PD-L1 are correlated to the patient’s Gleason scores, which was confirmed by quantification of their levels in several prostate cancer cell lines vs. normal prostate epithelial cell. Knocked out RelB in androgen-independent PC-3 cells resulted in reducing PD-L1 expression. Consistently, ectopic expression of RelB in androgen-dependent 22Rv1 cells led to increasing PD-L1 level, implicating that RelB participates in PD-L1 regulation in prostate cancer. Cytokine IFN-γ is able to induce PD-L1 expression through promoting RelB nuclear translocation, revealing that RelB may directly regulate PD-L1 transcription. Accordingly, an NF-κB enhancer element was identified in the promoter region, which is responsible for RelB-mediated PD-L1 transcriptional activation. These results suggest that overexpression of PD-L1 in advanced prostate cancer is, at least in part, through RelB activation. The finding from this study provides a potential approach for enhancing immunotherapeutic efficiency by targeting RelB-activated NF-κB alternative pathway.

Note: This abstract was not presented at the meeting.

Citation Format: Yanyan Zhang, Shuyi Zhu, Peipei Qian, Xiumei Wang, Zhi Xu, Wenbo Sun, Yong Xu. RelB upregulates PD-L1 in advanced prostate cancer: An insight into tumor immunoescape [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2791.
Abstract 2793: Mismatch repair proficient colorectal cancer and adaptive immunosuppression of endogenous anti-tumor immune response: Implications for immunotherapy
Authors: Llosa Nicolas , Awan Anas H. , Tam Ada J. , Fan Hongni , Smith Kellie N. , Sears Cynthia L. , Wang Hao , Pardoll Drew M. , Anders Robert A. , Housseau Franck
Published: August 2019

The tumor immune microenvironment (TiME) of mismatch-repair deficient (MMRd) colorectal cancer (CRC) is characterized by a cytotoxic T cell immune signature and counter-expression of IFNγ -inducible T cell checkpoints, features underlining intratumoral adaptive immunosuppression and sensitivity to immune checkpoint blockade. Since single-agent checkpoint inhibitors have not demonstrated, so far, similar meaningful clinical activity for MMR proficient (MMRp) CRC, we compared the tumor immune microenvironment (TiME) in CRC patients that responded to anti-PD1 therapy with the TiME of patients that did not (NCT01876511) to identify immune signatures and biomarkers that could help deciding on combinatorial immunotherapies to treat patients with MMRp CRC. With this approach, we detected immunoreactive MMRp (irMMRp) tumors characterized by a dense infiltration with cytotoxic T cells (CTL) as well as high expression of IFNγ and CD274 (PD-L1), despite their low tumor mutation burden. We observed that these irMMRp colon tumors are characterized by a negative association between Indolamine 2,3 dioxygenase 1 (IDO1) expression and Th17 infiltration. Despite the high expression of CD8 and PD-L1 expression, only irMMRp CRC with high expression of IDO1 on tumor cells and low infiltration with Th17 cells have a TiME resembling the TiME of CRC responding to anti-PD1. Since IDO1-derived tryptophan metabolites, kynurenines, are known to repress Th17 differentiation, these results suggested that the use of IDO1 small molecule inhibitors may derepress Th17 infiltration in CRC and blunt the effect of anti-PD1. Combining therapy with IDO1/PD1 inhibitors with drugs inhibiting IL-17 signaling pathway may help to treat irMMRp CRC patients.

Citation Format: Nicolas Llosa, Anas H. Awan, Ada J. Tam, Hongni Fan, Kellie N. Smith, Cynthia L. Sears, Hao Wang, Drew M. Pardoll, Robert A. Anders, Franck Housseau. Mismatch repair proficient colorectal cancer and adaptive immunosuppression of endogenous anti-tumor immune response: Implications for immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2793.