bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1393: JARID1 family inhibitor recovers sensitivity of drug-tolerant subpopulation in lung cancer cell lines
Authors: Ariga Shin , Kinoshita Ichiro , Kikuchi Junko , Shimizu Yasushi , Dosaka-Akita Hirotoshi
Published: August 2018

Epigenetic alteration contributes to tumor initiation, progression and invasion. Epigenetic machinery has become a major focus for the new development of molecular targeted cancer therapeutics. H3K4 trimethylation (H3K4me3) is an epigenetic mark which exist surrounding the transcription start site and have been presumed to be associated with cancer stem like cells. JARID1 family is demethylase of H3K4me2/me3. Especially, JARID1a and JARID1b have been suggested to have oncogenic properties in several cancers, including lung cancer. Here we examined the effects of a JARID1 family inhibitor against human lung cancer cell lines. 2-4(4-methylphenyl)-1,2-benzisothiazol-3(2H)-one (PBIT) was screened from more than 15000 small molecules as a JARID1 family inhibitor by Sayegh et al. First, we confirmed that PBIT increased H3K4me3 levels in time-dependent and dose-dependent manners in non-small cell lung cancer cell lines. PBIT inhibited cell growth in a dose-dependent manner with the IC50 value of about 10 µM to 30 µM. Next, we detected drug-tolerant persisters (DTPs) from each cell line by exposing them to very high concentration of the drug that is effective to each of the cell lines (gefitinib 10µ for PC9, PC3 and HCC827; AZD9291 5µM for H1975; CDDP 100µM for H226; paclitaxel 500nM for H460, A549 and H1299). DTPs detected were less than 1% in each cell line after 9-day exposure. These small subpopulations showed increased expression of JARID1a and/or JARID1b. Likewise, side population cells, which are considered to contain cancer stem like cells, showed increased expression of JARID1a and decreased H3K4me3 levels. PBIT in combination with anticancer drugs reduced colony formation of DTPs from all cell lines whose DTPs formed colonies. The DTPs showed decreased H3K4me3 levels and increased expression of cancer stem cell marker Oct-4, while these changes were abrogated by subsequent PBIT treatment. These results indicate that PBIT, a JARID1 family inhibitor, recovers sensitivity of drug-resistant lung cancer cells.

Citation Format: Shin Ariga, Ichiro Kinoshita, Junko Kikuchi, Yasushi Shimizu, Hirotoshi Dosaka-Akita. JARID1 family inhibitor recovers sensitivity of drug-tolerant subpopulation in lung cancer cell lines [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 1393.
Abstract 1394: Resistance to BET inhibitor leads to new therapeutic vulnerabilities in castration resistant prostate cancer
Authors: Pawar Aishwarya S. , Asangani Irfan A.
Published: August 2018

BRD4 plays a major role in the transcription networks orchestrated by androgen receptor (AR) in castration resistant prostate cancer (CRPC) cells. Bromodomain and extraterminal protein (BET) inhibitors displace BRD4 protein from chromatin, resulting in the inhibition of oncogenic transcriptional programs. Several BET inhibitors (BETi) are in phase I clinical trials for a variety of malignancies including CRPC. Here we describe a general mechanism of acquired resistance to BETi due to modulation of cellular pathways that are amenable to targeted therapies in CRPC cells. BETi resistant CRPC cells displayed cross resistance to a variety of BETi in the absence of gatekeeper mutations or persistent drug pump activation. Resistant cells exhibited reduced chromatin bound BRD4, and were less sensitive to Proteolysis Targeting Chimeric (PROTAC) -mediated degradation or genetic knockdown, suggesting a BRD4-independent transcription program. Transcriptomic analysis revealed reactivation of AR-signaling due to CDK9-mediated serine-81 phosphorylation of AR, with a consequent increase in sensitivity to CDK9 inhibitors and anti-androgens in BETi resistant cells. Additionally, increased DNA damage associated with PRC2-mediated transcriptional silencing of DNA damage response (DDR) genes was observed due to the loss of BRD4 from their proximal promoter regions in the resistant cells, leading to PARP inhibitor sensitivity. Collectively, our results identify the therapeutic limitation of BETi as a monotherapy in CRPC. However, data showing the reactivation of AR-signaling and increased DNA damage in the BETi resistant cells provide unique opportunities for combination therapies in managing CRPC.

Citation Format: Aishwarya S. Pawar, Irfan A. Asangani. Resistance to BET inhibitor leads to new therapeutic vulnerabilities in castration resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1394.
Abstract 1397: The regulation of DNMT3B expression by lncRNA-NCRMS in cancer
Authors: Peng Wan-xin , Mo Yin-yuan
Published: August 2018

5-aza-2'-deoxycytidine (decitabine; DAC) is an inhibitor of DNA methyltransferases which catalyze the transfer of a methyl group to DNA. It is well known that aberrant hypermethylation occurs in many types of tumors and often contributes to suppression of important genes such as tumor suppressor genes. Reactivation of the methylated genes by DAC has been shown to induce cytotoxicity and cell cycle dynamics. Increasing evidence indicates that lncRNAs may play critical roles in cancer development through epigenetic regulation of gene expression. However, little is known whether and how lncRNAs are involved in regulation of methylation pathways. In present study, we applies the CRISPR/Cas9-based synergistic activation mediator (SAM) system to a DAC-induced cytotoxicity model to identify potential lncRNAs capable of conferring the resistance to DAC in breast cancer. This screen identified several potential lncRNA candidates and among them is NCRMS which is capable of promoting resistance to DAC. We further confirmed that ectopic expression of NCRMS causes resistance to DAC whereas knockout of NCRMS increases the sensitivity to DAC. Mechanistically, NCRMS functions as a positive regulator of DNA (cytosine-5-)-methyltransferase 3 beta (DNMT3B). Further characterizations demonstrated that NCRMS promotes the stability of DNMT3B mRNA. Together, these results suggest that NCRMS may serve as a potential target for cancer therapy. Studies are underway to delineate the detailed mechanism of how NCRMS regulates the stability of DNMT3B mRNA.

Citation Format: Wan-xin Peng, Yin-yuan Mo. The regulation of DNMT3B expression by lncRNA-NCRMS in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1397.
Abstract 1399: Passenger deletion vulnerability in glioblastoma
Authors: Oatman Nicole , Komurov Kakajan , Arora Priyanka , Desai Pankaj , Dasgupta Nupur , Choi Kwangmin , Dasgupta Biplab
Published: August 2018

Glioblastoma multiforme (GBM) is the most common, aggressive and lethal primary brain tumor in humans. Despite aggressive therapy the median survival of GBM patients is approximately 15 months. Monoallelic loss of PTEN occurs in ~70% of GBM. Through experimental validation studies in patient-derived primary GBM cell lines, we discovered that the lipogenic gene SCD undergoes unintended co-deletion as a passenger to PTEN, resulting in hemizygous loss of SCD in a subset of GBM. Additional in silico, genetic and biochemical analysis lead to the identification of two distinct PTEN-deleted subgroups: one with hemizygous co-deletion of PTEN and SCD (hereafter SCD-expressing), and another with very little or no PTEN/SCD expression due to PTEN mutation/homozygous loss and epigenetic suppression of the remaining SCD allele (hereafter SCD-non-expressing). SCD is an integral membrane protein of the endoplasmic reticulum (ER), involved in mediating the rate-limiting step of unsaturated fatty acid biosynthesis. SCD catalyzes the desaturation of the saturated fatty acids stearic (C18:0) and palmitic acids (C16:0) to the monounsaturated fatty acids oleic (C18:1) and palmitoleic acids (C16:1), respectively. We showed that the SCD-expressing lines are highly sensitive to SCD inhibition in the absence of dietary oleate and that the SCD-non-expressing lines are resistant to SCD inhibition. In vivo, we showed that mice treated with the SCD inhibitor survive longer than mice treated with vehicle. We also showed that SCD-expressing GBM lines acquire inhibitor resistance upon long-term exposure to the inhibitor in vitro. To understand the mechanism of resistance, we performed Reverse Phase Protein Array (RRPA) and RNA-Seq. Results illustrated the up-regulation of SCD among other genes. To identify the gene/s that provide SCD inhibitor resistance, we are currently performing a targeted shRNA screen of the highly expressed genes that we determined through RPPA and RNA-Seq.

Citation Format: Nicole Oatman, Kakajan Komurov, Priyanka Arora, Pankaj Desai, Nupur Dasgupta, Kwangmin Choi, Biplab Dasgupta. Passenger deletion vulnerability in glioblastoma [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 1399.
Abstract 1404: Role of catechol-O-methyltransferase gene in prostate cancer
Authors: Maekawa Shigekatsu , Kato Taku , Hashimoto Yutaka , Shiina Marisa , Wong Ryan K. , Shahryari Varahram , Yamamura Soichiro , Majid Sahana , Saini Sharanjot , Tabatabai Laura Z. , Homma Yukio , Dahiya Rajvir , Tanaka Yuichiro
Published: August 2018

Prostate cancer is one of the most common malignancies and ranks the second most common cause of cancer-related deaths in men in the United States. Catechol-O-methyltransferase (COMT) is a phase II enzyme that detoxifies various catechol compounds that are reactive toward DNA and damaging to the cell. Studies have shown COMT to play a protective role against cancers such as renal and breast, but their effect on prostate is not well understood. In this study, the biological properties and function of COMT in prostate cancer were studied. Expression of COMT was initially measured in normal/benign and cancerous prostate tissues by immunohistochemistry, and cell lines by real-time PCR and western blotting. Cancerous cells displaying the lowest levels was then transfected with COMT. Gene effect on various cellular properties such as cell proliferation, migration, invasion and apoptosis, as well as growth in athymic nude mice were determined. COMT protein expression was lower in cancer regions compared to benign and normal regions of prostate tissues. Cancerous DU145 and DuPro cells also had reduced mRNA levels of COMT but with undetectable protein levels. Interestingly, re-expressing COMT in DU145 and DuPro cells led to decreased cell proliferation, migration, wound healing ability and invasion, and increased apoptosis compared to vector control. COMT also inhibited cell tumor formation in animal models. As a possible target, the TNFRSF11B gene was upregulated due to COMT. These results demonstrate COMT to protect against prostate cancer progression and to have a functional role by affecting apoptosis. COMT may thus be a potential biomarker or therapeutic target for prostate cancer.

Citation Format: Shigekatsu Maekawa, Taku Kato, Yutaka Hashimoto, Marisa Shiina, Ryan K. Wong, Varahram Shahryari, Soichiro Yamamura, Sahana Majid, Sharanjot Saini, Laura Z. Tabatabai, Yukio Homma, Rajvir Dahiya, Yuichiro Tanaka. Role of catechol-O-methyltransferase gene in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1404.
Abstract 1412: Comprehensive characterization of clock genes in human cancers
Authors: Ye Youqiong , Xiang Yu , Takahashi Joseph , Mills Gordon , Yoo Seung-Hee , Han Leng
Published: August 2018

Cancer chronotherapy has been considered to optimize the balance of anti-tumor effects and toxicity. To develop cancer chronotherapy, the first crucial step is to understand the significant roles of clock genes in cancer. Circadian machinery is controlled by two main transcription auto-regulatory feedback loops, which including 14 core circadian genes. Core clock genes have at least one paralog gene may be functionally redundant in circadian regulation. Dysregulation of circadian genes promotes tumorigenesis through mechanisms that include the cell cycle, DNA damage and metabolism. However, there has been no comprehensive analysis to characterize the roles of circadian genes and circadian rhythms in tumorigenesis and to assess their potential utility in cancer chronotherapy. We determined genetic and epigenetic alterations of clock genes in human cancer, and explore clock gene clinical effect through integrative analysis, based on The Cancer Genome Atlas (TCGA) profiles contained DNA, RNA, protein, epigenetic levels and clinical data. Our results suggested several clock genes may function as oncogene, such as ARNTL2, NR1D1, and NPAS2, while several other clock genes may function as tumor suppressors, such as PERs, CRYs, RORs. We revealed disruption and reprogramming of circadian rhythms across patient samples by showing significantly reduced number of genes correlated to clock genes. We further showed that transcriptional dysregulation of clock genes is strongly associated with patient survival, tumor stage and subtype. Taken together, our results provide the comprehensive analysis of clock system across different cancer types and expected outcomes will lay the groundwork for the future development of cancer chronotherapy.

Citation Format: Youqiong Ye, Yu Xiang, Joseph Takahashi, Gordon Mills, Seung-Hee Yoo, Leng Han. Comprehensive characterization of clock genes in human 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 1412.
Abstract 1414: Germline cancer predisposition gene mutations among patients with early onset rectal cancer
Authors: Beltrami Caroline , Alvim Luisa Matos , Kupper Bruna Elisa , Petersen Annabeth , Jørgensen Mads , Junior Samuel Aguiar , Rogatto Silvia Regina
Published: August 2018

Background: Although the incidence rates of rectal cancer (ReCa) have been increasing, the mechanisms or mutations involved in the development of the disease are poorly explored, especially in younger patients. To date, rectal tumors from young patients show molecular, clinical and pathological differences compared with colorectal cancer from older patients. Few studies have reported the presence of a hereditary factor involved in the risk of developing ReCa in a set of patients even in the absence of family history of cancer.

Patients and Method: Twenty-seven ReCa younger patients (<40 years old) were screened for germline alterations in DNA from blood samples. The family history and clinical-pathological findings were assessed from the medical records from all subjects. DNA library was performed from SureSelectQXT Library Prep kit (Agilent, Santa Clara, CA) and next-generation sequencing was applied using a multigenic panel with 105 cancer-related genes (SureSelectXT Custom Panel; Agilent, Santa Clara). The generated sequences were aligned to human genome (hg19) and reported as pathogenic, likely pathogenic, variant of uncertain significance (VUS), benign and likely benign according to the American College of Medical Genetics and Genomics guidelines. Sanger sequencing was used to confirm the pathogenic variants.

Results: Two patients carried out high penetrance genes associated with colorectal cancer, MUTYH biallelic and BMPR1A. MTHFR and MUTYH variants were found in two patients. POT1 (melanoma predisposition), MEN1 (high penetrance) and ATM (moderate penetrance) variants were found in three additional ReCa patients. Three patients carried out likely pathogenic varaints (BMPR1A, CHEK2 and RAD51D genes). Four of ten patients with pathogenic variants had no family history of cancer.

Conclusion: Pathogenic (7/27) and likely pathogenic (3/27) variants were found in 37% of ReCa patients younger than 40 years old affecting genes with high or moderate penetrance. Our data suggest that genetic testing using a multigene panel and genetic counseling should be offered for younger patients with rectal cancer.

Citation Format: Caroline Beltrami, Luisa Matos Alvim, Bruna Elisa Kupper, Annabeth Petersen, Mads Jørgensen, Samuel Aguiar Junior, Silvia Regina Rogatto. Germline cancer predisposition gene mutations among patients with early onset rectal 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 1414.
Abstract 1415: Target sequencing of tumor driver genes in lung adenocarcinoma tissues
Authors: Zhu Jiawei , Feng Lin , Xiao Ting
Published: August 2018

Background and objective As the main histological subtype of lung cancer, lung adenocarcinoma (ADC) contributes to a cancer-related death in the word. With the development of high-throughput genomic analysis, whole-genome sequencing provides a new opportunity to identify genetic mutations that drive the origination, growth, and spread of cancer. The aim of this study is to detect the mutated genes in lung adenocarcinoma tissues and to analyze its clinical significance, based on 483 tumor driver gene sequence of ADC samples.

Methods We obtained 40 pairs of samples from ADC tumors and matched normal adjacent tissue, which included female (58%), male (43%), people over 60 years old (63%), stage I (40%), stage II (20%), stage III (38%), and stage IV (3%). Based on the Illumina HiSeq2500 sequencing technology platform, small fragment library was built, high coverage of the target area was captured by NovoP products (Agilent sure select XT Custom 0.5-2.9Mb kit, contains 483 tumor related target gene probe). DNA samples was examined by Paired-End sequencing; sequencing data of 40 cases of tumor samples was obtained. We analyzed the sequencing results of lung adenocarcinoma tumor and matched normal adjacent tissues by using bioinformatics analysis methods, and sought for somatic mutations in lung adenocarcinoma tumor and matched normal adjacent tissues.

Results There were 31 ADC samples had reliable exons mutations in the target area, totally 142 (102 genes) mutation positions, including non-synonymous mutations (65%), synonymous mutations (26%), and terminating translation mutation (8%). Totally 116 mutation positions (81.69%) were located in the target capture area. Gene mutations were more frequent in smoking and aged patients. Top ten mutation frequency genes were TP53, EGFR, PIK3CA, ATN1, ATXN1, ABL1, ALK, EGF, EPHA3, and KRAS. And compared to The Cancer Genome Atlas (TCGA) data, the mutation frequency of EGFR was significantly higher in our research.

Conclusion These data provided evidence for the further determination of the molecular mechanism and molecular typing of lung adenocarcinoma. Mutation frequency of TP53, EGFR and PIK3CA in lung adenocarcinoma tissue was the highest.

Citation Format: Jiawei Zhu, Lin Feng, Ting Xiao. Target sequencing of tumor driver genes in lung adenocarcinoma tissues [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 1415.
Abstract 1416: Pan-cancer mutual vulnerability and viability
Authors: Yeerna Huwate , Kim William , Hahn William C. , Mesirov Jill , Tamayo Pablo
Published: August 2018

We used a skew-distribution-based statistical methodology to fit parametric models of cell-viability profiles from genome-wide RNAi-gene knockdowns. We identify the subset of genes with the highest degree of skewness that represent most oncogenes and tumor suppressors. We decomposed those genes using matrix decomposition in order to define mutual vulnerabilities. The resulting matrix decomposition components represent coordinated patterns of gene dependency shared by many genes in many cell lines across cancer types. We identify the most salient mutual vulnerabilities, generate a comprehensive catalog of them, and annotate them based on their association with a variety of genomic features including: gene, protein, and pathway expression, mutational landscape, copy number alterations, promoter methylations, metabolites, miRNAs, and drug sensitivities. We also generate an Onco-GPS (Oncogenic Positioning System) Map that provides a summary and visual representation of the mutual vulnerabilities and their components, and allow us to compare them with their counterparts in Pan-Cancer transcriptional and drug sensitivity spaces.

Citation Format: Huwate Yeerna, William Kim, William C. Hahn, Jill Mesirov, Pablo Tamayo. Pan-cancer mutual vulnerability and viability [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 1416.
Abstract 1422: NEBNext Direct® Custom Ready myeloid genes for NGS target enrichment
Authors: Adams Scott M. , Patel Kruti M. , Emerman Amy B. , Bowman Sarah K. , Elfe Charles D. , Mauceli Evan , Barry Andrew , Davis Theodore , Hendrickson Cynthia L.
Published: August 2018

Comprehensive genetic evaluation of patients with blood cancer can be imperative for identifying and classifying myeloid malignancies in a patient. Since liquid tumors comprise of highly variable percentages of normal cells or multiple tumor clones, this type of screening is critical for early stage disease patients where a cytogenetic profile informs diagnosis or disease stratification, prognosis, and treatment options. For patients with late-stage disease, genetic testing can assist in choosing options for alternative treatments, including targeted therapies. Thus, next generation sequencing technologies are needed that can provide the sensitivity and specificity to identify these genetic complexities. Here we introduce a selection of over 50 genes from the NEBNext Direct® Custom Ready gene catalog that are known to be involved in myeloid malignancies. The NEBNext Direct® technology uses a novel approach to selectively enrich nucleic acid targets, and rapid customized panels can be generated by combining one to several hundred genes from the Custom Ready catalog. Unlike alternative hybridization methods, NEBNext Direct® does not require upfront library preparation. The approach rapidly hybridizes both strands of genomic DNA to biotinylated baits, captures the targets on streptavidin beads, enzymatically removes off-target sequence, and directly converts captured molecules into Illumina-compatible sequencing libraries in a single day protocol. A 12-base unique molecular identifier (UMI) tags each individual molecule prior to the final PCR amplification to enable identification of PCR duplicates, and an 8-base barcode is added to each library during PCR for high-throughput pooling. With 100ng of DNA input the panel provides highly uniform coverage and approximately 95% of sequence reads map to the targeted myeloid genes. The NEBNext Direct® Custom Ready myeloid genes demonstrate a high-throughput and cost-effective approach for the rapid and sensitive analysis of exon coding regions (~200kb) for over 50 oncogenic genes important in the pathogenesis of these types of blood disorders. These gene mutations are powerful prognostic markers that can be used as a guide for effective patient management.

Citation Format: Scott M. Adams, Kruti M. Patel, Amy B. Emerman, Sarah K. Bowman, Charles D. Elfe, Evan Mauceli, Andrew Barry, Theodore Davis, Cynthia L. Hendrickson. NEBNext Direct® Custom Ready myeloid genes for NGS target enrichment [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 1422.
Abstract 1433: Estrogen-related receptor alpha (ERRα) functions to promote prostate cancer cell stemness via its transcriptional regulation of ZIP1 and ACON2 to enhance oxidative phosphorylation
Authors: Ma Taiyang , Xu Zhenyu , Wang Yuliang , Wang Zhu , Gao Weijie , You Wenxin , Chan Leung Franky
Published: August 2018

Accumulating evidences suggest that cancer stem-like cells (CSCs) play critical roles in cancer relapse and metastasis, likely due to their strong self-renewal capacity and chemo-radiotherapy resistance. Recent advances also indicate that the energy metabolism status of the CSCs differs greatly from the bulk cancer cells, and that such energy metabolism shift may favor the maintenance of cancer cell stemness. Estrogen-related receptor alpha (ERRα, NR3B1) is a key energy metabolism regulator. In this study, we aim to characterize its role in the energy metabolism regulation in prostate cancer stem-like cells (PCSCs). By analyzing the oxygen consumption and extracellular acidification rates using the Seahorse XF Analyzer, we revealed that the PCSCs, isolated from prostate cancer cell lines (LNCaP, DU145 and PC3) by low attachment 3D-culture, exhibited lower aerobic glycolysis but higher oxidative phosphorylation status. Expression analyses by qPCR and Western blot showed that the isolated PCSCs expressed higher levels of ERRα. Further functional studies showed that overexpression of ERRα could promote the oxidative phosphorylation but lower the glycolysis status, and also enhance the 3-D spheroid formation capacity (stemness) of prostate cancer cells (LNCaP, DU145 and PC3), whereas its knockdown could reverse the effects. Furthermore, we identified by combined chromatin immunoprecipitation (ChIP) and PCR analysis that ERRα could directly regulate two key energy metabolic genes ACON2 (a rate limiting enzyme in TCA cycle) and ZIP1 (a zinc transporter) in PCSCs. Western blot analysis showed that ERRα could up-regulate the ACON2 but repress the ZIP1 expression, whereas its knockdown reverse the expressions of these two proteins in prostate cancer cells. Together, our results show that ERRα can promote the stemness of prostate cancer cells and enhance the cellular energy metabolism towards oxidative phosphorylation in PCSCs via its transcriptional regulation of ACON2 and ZIP1 genes.This study is supported by a General Research Fund from the Research Grants Council of Hong Kong (project code 14107116).

Citation Format: Taiyang Ma, Zhenyu Xu, Yuliang Wang, Zhu Wang, Weijie Gao, Wenxin You, Leung Franky Chan. Estrogen-related receptor alpha (ERRα) functions to promote prostate cancer cell stemness via its transcriptional regulation of ZIP1 and ACON2 to enhance oxidative phosphorylation [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 1433.
Abstract 1435: Investigation of phosphoserine aminotransferase 1: Its role in breast cancer progression
Authors: Metcalf Stephanie , Kruer Traci , Wittliff James , Klinge Carolyn , Clem Brian F.
Published: August 2018

Metastasis and endocrine resistance are two factors that complicate therapeutic intervention in breast cancer patients and lead to poorer overall survival. Metastasis is known to be responsible for 90% of cancer related deaths, and is especially prevalent in triple negative breast cancer (TNBC); while endocrine resistance can affect up to 50% of patients diagnosed with estrogen receptor positive breast cancer (ER+BC). Phosphoserine aminotransferase 1 (PSAT1) catalyzes the second step within de novo serine biosynthesis and increased expression of enzymes in this pathway have been linked to progression of breast cancer and poor clinical outcomes. Within our preliminary retrospective analysis of human breast cancer patients, we identified an inverse association with elevated transcript levels and poorer distant metastasis free survival, which, coupled to a previously reported correlation of PSAT1 with response to endocrine therapy in patients with ER+BC, we postulate that PSAT1 contributes to breast cancer progression through promotion of metastasis and/or endocrine resistance. To initially determine relevance for PSAT1, immunohistochemistry was performed to determine PSAT1 expression in human breast cancer patients. We found that PSAT1 expression is increased through breast cancer progression, with highest levels observed within metastatic conditions. To investigate the metastatic contribution of PSAT1, we silenced PSAT1 expression within the triple negative breast cancer cell line (TNBC), MDA-MB-231. While suppression of PSAT1 had no effect on proliferation, there was a significant decrease in the motility and invasion of these cells. In addition, decreased PSAT1 substantially inhibited tumor nodule formation following tail-vein injections of MDA-MB-231 cells in vivo. To investigate PSAT1's role in endocrine resistance, we used parental MCF-7 cells and an endocrine-resistant derivative cell lines and found that resistant cells exhibited higher PSAT1 expression compared to parental MCF-7 cells. Lastly, suppression of PSAT1 trended to sensitize the resistant cells to 4-hydroxytamoxifen treatment. Taken together, these data indicate that PSAT1 may contribute to the progression of human breast cancer via either metastasis or endocrine resistance or both and could potentially serve as a viable target for new therapies.

Citation Format: Stephanie Metcalf, Traci Kruer, James Wittliff, Carolyn Klinge, Brian F. Clem. Investigation of phosphoserine aminotransferase 1: Its role in breast cancer progression [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 1435.
Abstract 1445: Metastasis-resistant fatty liver induced by a choline-deficient diet
Authors: Nakamura Miki , Suetsugu Atsushi , Satake Tomoyuki , Hasegawa Kosuke , Shimizu Masahito , Saji Shigetoyo , Moriwaki Hisataka , Hoffman Robert M.
Published: August 2018

Background: Fatty liver disease is increasing worldwide. The differences of the liver metastasis between fatty liver and normal liver remain unclear. We established the EL4-RFP lymphoma models in mice with fatty liver induced by a choline-deficient-diet (CDD) comparing the frequency of liver-metastasis of this tumor in mice with fatty and normal livers.

Material and method: C57BL/6-GFP transgenic mice were fed with a CDD in order to establish a fatty liver model. EL4-RFP malignant lymphoma cells were injected in the spleen of fatty-liver mice and normal mice. Using the Olympus SZX7 microscope and the Olympus FV1000 confocal microscope, metastases in mice with fatty liver or normal liver were imaged.

Results: EL4-RFP tumors were observed to metastasize to the liver and bone marrow as well as form ascites. Fewer metastases in the fatty liver were observed than in the normal liver. Similarly, fewer cancer-associated fibroblasts (CAFs) were observed in the fatty liver.

Conclusion: The relative metastatic resistance of the fatty liver may be due to the reduced number of CAFs in the fatty liver. The fatty liver may not be a suitable environment for CAFs which may be due to re-programmed methionine metabolism which may have occurred during the time the animals were on the CDD.

Citation Format: Miki Nakamura, Atsushi Suetsugu, Tomoyuki Satake, Kosuke Hasegawa, Masahito Shimizu, Shigetoyo Saji, Hisataka Moriwaki, Robert M. Hoffman. Metastasis-resistant fatty liver induced by a choline-deficient diet [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 1445.
Abstract 1446: Glycogen accumulation in aggressive breast cancers under hypoxia
Authors: Altemus Megan Ann , Yates Joel A. , Wu ZhiFen , Bao LiWei , Merajver Sofia D.
Published: August 2018

Cancers utilize diverse metabolic pathways to increase survival and proliferation; therefore, the identification of specific metabolic alterations in aggressive triple-negative and inflammatory breast cancers is a compelling avenue for the development of new treatments. The ability to survive and grow in hypoxia is necessary for breast cancer cells in the center of the tumor mass or in the lymphatic system and other organ environments. Glucose reserves are typically stored in liver and muscle cells as the polysaccharide glycogen. Elevated levels of glycogen have been observed in various cancers, and recent studies have suggested that glycogen metabolism is important for promoting cancer cell survival. We hypothesize that aggressive breast cancers modulate glycogen metabolism to provide nutritional flexibility for proliferation and invasion during cancer progression. To test our hypothesis, we investigated the regulation of glycogen levels in breast cancer and normal-like cells in atmospheric and 1% oxygen. The glycogen content of six breast cancer and one normal-like cell line was measured under normoxic and hypoxic conditions. All cell lines had increased glycogen accumulation under hypoxia, but to varying degrees that did not correlate with receptor status. Expression levels of various genes involved in glycogen synthesis and degradation were determined by qRT-PCR, with no consensus expression pattern between all cell lines. Several key alterations were noted in synthesis and degradation enzymes. We will present an integrative picture of glycogen synthesis and degradation in breast cancer cells. Studies are ongoing to determine the mechanisms through which these changes occur and whether the accumulation of glycogen confers an advantage for continued survival and growth of cancer.

Citation Format: Megan Ann Altemus, Joel A. Yates, ZhiFen Wu, LiWei Bao, Sofia D. Merajver. Glycogen accumulation in aggressive breast cancers under hypoxia [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 1446.
Abstract 145: Pharmacologic inhibition of SIAH2 stabilizes DYRK2 and inhibits survival and self-renewal in chronic myeloid leukemia (CML) leukemic stem cells
Authors: Park Chun Shik , Shen Ye , Lewis Andrew , Suppipat Koramit , Puppi Monica , Tomolonis Julie , Chen Taylor , Pang Paul , Mistretta Toni-Ann , Ma Leyuan , Green Michael , Rau Rachel , Lacorazza Daniel
Published: August 2018

Leukemia stem cells (LSCs) are a rare population able to sustain and recapitulate leukemia through a poorly understood mechanism of self-renewal. Because more than half of patients relapse after the cessation of TKI therapy, it is clear that a cure is not possible with TKIs alone, and LSC-specific drugs are urgently needed to simultaneously eliminate bulk leukemia with TKIs and LSCs. Here we report that KLF4 promotes disease progression in chronic myeloid leukemia (CML) by repressing an inhibitory mechanism in LSCs that can be activated with small molecules. Deletion of the Klf4 gene severely abrogated maintenance of BCR-ABL(p210)-induced CML by impairing survival and self-renewal in LSCs whereas increased self-renewal was observed in hematopoietic stem cells during serial transplantation. Mechanistically, KLF4 represses the Dyrk2 gene and thus loss-of-KLF4 resulted in elevated levels of the DYRK2 kinase in LSCs, which was associated with p53-mediated apoptosis and inhibition of self-renewal through depletion of c-Myc protein. Supporting this model, stabilization of DYRK2 protein, by inhibiting the ubiquitin E3 ligase SIAH2 with vitamin K3, promoted apoptosis in a panel of CML cell lines (K562, KU-812, and KCL-22) by inducing DYRK2 expression. Knocking out the DYRK2 gene in K562 cells by Cas9/CRISPR abrogated cytotoxicity induced by vitamin K3 and the presence of p53 significantly lowered IC50. In vivo treatment of CML mice diminished the number of LSCs evaluated in secondary transplants. In humans, vitamin K3 induced apoptosis in bone marrow cells from CML patients but not in healthy individuals by inducing DYRK2, p53 phosphorylation, and c-Myc depletion; furthermore, vitamin K3 abrogated capacity of CD34+ cells to generate colonies in methylcellulose only in CML. Altogether, our results suggest that DYRK2 is a molecular checkpoint controlling both p53 and c-Myc mediated regulation of survival and self-renewal in CML LSCs that can be activated pharmacologically.

Citation Format: Chun Shik Park, Ye Shen, Andrew Lewis, Koramit Suppipat, Monica Puppi, Julie Tomolonis, Taylor Chen, Paul Pang, Toni-Ann Mistretta, Leyuan Ma, Michael Green, Rachel Rau, Daniel Lacorazza. Pharmacologic inhibition of SIAH2 stabilizes DYRK2 and inhibits survival and self-renewal in chronic myeloid leukemia (CML) leukemic stem 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 145.
Abstract 1452: The receptor tyrosine kinase EphA2 promotes glutamine metabolism in tumors by activating the transcriptional coactivators YAP and TAZ
Authors: Edwards Deanna N. , Ngwa Verra M. , Wang Shan , Shiuan Eileen , Brantley-Sieders Dana , Kim Laura , Reynolds Albert B. , Chen Jin
Published: August 2018

Malignant tumors reprogram cellular metabolism to support cancer cell proliferation and survival. Although most cancers depend on a high rate of aerobic glycolysis, many cancer cells also display addiction to glutamine. Glutamine transporters and glutaminase activity are critical for glutamine metabolism in tumor cells. We found that the receptor tyrosine kinase EphA2 activated the TEAD family transcriptional coactivators YAP and TAZ (YAP/TAZ), likely in a ligand-independent manner, to promote glutamine metabolism in cells and mouse models of HER2-positive breast cancer. Overexpression of EphA2 induced the nuclear accumulation of YAP and TAZ and increased the expression of YAP/TAZ target genes. Inhibition of the GTPase Rho or the kinase ROCK abolished EphA2-dependent YAP/TAZ nuclear localization. Silencing YAP or TAZ substantially reduced the amount of intracellular glutamate through decreased expression of SLC1A5 and GLS, respectively, genes that encode proteins that promote glutamine uptake and metabolism. The regulatory DNA elements of both SLC1A5 and GLS contain TEAD binding sites and were bound by TEAD4 in an EphA2-dependent manner. In patient breast cancer tissues, EphA2 expression positively correlated with that of YAP and TAZ, as well as that of GLS and SLC1A5. Although high expression or EphA2 predicted enhanced metastatic potential and poor patient survival, it also rendered HER2-positive breast cancer cells more sensitive to glutaminase inhibition. The findings define a previously unknown mechanism of EphA2-mediated glutaminolysis through YAP/TAZ activation in HER2-positive breast cancer and identify therapeutic targets in patients.

Citation Format: Deanna N. Edwards, Verra M. Ngwa, Shan Wang, Eileen Shiuan, Dana Brantley-Sieders, Laura Kim, Albert B. Reynolds, Jin Chen. The receptor tyrosine kinase EphA2 promotes glutamine metabolism in tumors by activating the transcriptional coactivators YAP and TAZ [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 1452.
Abstract 1453: SLC25A32 sustains cancer cell proliferation by regulating flavin adenine dinucleotide (FAD) metabolism
Authors: Santoro Valeria , Kovalenko Ilya , Vriens Kim , Christen Stefan , Bernthaler Andreas , Haegebarth Andrea , Fendt Sarah-Maria , Christian Sven
Published: August 2018

SLC25A32 is a member of the solute carrier 25 family of mitochondrial transporters. SLC25A32 transports tetrahydrofolate (THF) as well as FAD into mitochondria and regulates mitochondrial one-carbon metabolism and redox balance. While it is known that cancer cells require one-carbon and FAD-dependent mitochondrial metabolism for generation of nucleotides, lipids and for maintenance of redox homeostasis to sustain growth, the role of SLC25A32 in cancer cell survival remains unexplored.

Our results indicate that the SLC25A32 gene is highly amplified in different tumors and that its amplification correlates with increased mRNA expression and with reduced patient survival. SiRNA-mediated knock-down and CRISPR-mediated knock-out of SLC25A32 in cancer cells of different origins, resulted in the identification of sensitive and resistant cells to SLC25A32 inhibition. Mechanistically, tracing of deuterated serine upon SLC25A32 knock-down did not affect the mitochondrial/cytosolic folate flux as measured by Liquid Chromatography coupled Mass Spectrometry (LC-MS). Instead, SLC25A32 inhibition resulted in respiratory chain dysfunction at the FAD-dependent complex II enzyme, induction of Reactive Oxygen

Species (ROS) and depletion of reduced glutathione (GSH), impairing cancer cell proliferation. Moreover, Buthionine sulfoximine (BSO) treatment further sensitized cells to ROS-mediated inhibition of cell proliferation upon SLC25A32 knock-down. Treatment of cells with the FAD precursor riboflavin and with GSH rescued cancer cell proliferation upon SLC25A32 down-regulation.

Our results indicate that inhibition of mitochondrial FAD transport by targeting SLC25A32 has potential clinical applications as a single agent or in combination with approved cancer drugs that lead to increased oxidative stress and reduced tumor growth.

Citation Format: Valeria Santoro, Ilya Kovalenko, Kim Vriens, Stefan Christen, Andreas Bernthaler, Andrea Haegebarth, Sarah-Maria Fendt, Sven Christian. SLC25A32 sustains cancer cell proliferation by regulating flavin adenine dinucleotide (FAD) metabolism [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 1453.
Abstract 1454: Glutamate dehydrogenase 1 mediates metabolic reprogramming and promotes tumor metastasis through CamKK2-AMPK signaling in LKB1-deficient lung cancer
Authors: Kang Sumin , Jin Lingtao , Pan Chaoyun , Chun Jaemoo
Published: August 2018

Although elevated glutaminolysis has been demonstrated in cancer cells, the precise mechanism by which glutaminolysis is exploited to promote tumor metastasis has not been elucidated. Lung cancer frequently metastasizes and is the leading cause of cancer-related death worldwide. Loss of LKB1 is associated with increased metastasis and poor prognosis in lung cancer, but the development of targeted agents in LKB1-deficient lung cancer is still in its infancy. Our recent study reveal that glutamate dehydrogenase 1, upregulated upon detachment via the transcription factor pleomorphic adenoma gene 1 (PLAG1), provides anti-anoikis and pro-metastatic signals in LKB1-deficient lung cancer. Mechanistically, the GDH1 product alpha-KG activates CamKK2 by enhancing its substrate AMPK binding and triggers CamKK2-mediated AMPK signaling, which contributes to energy production that confers anoikis resistance. The effect of GDH1 on AMPK is evident in LKB1-deficient lung cancer, where AMPK activation predominantly depends on CamKK2. Targeting GDH1 with the small molecule GDH1 inhibitor R162 led to attenuated anoikis resistance and tumor metastasis in an LKB1-deficient lung patient-derived xenograft model. Correlation studies between GDH1 signaling and metastatic cancer progression in lung cancer patients further validated the clinical relevance of our finding. Our study provides insight into the molecular mechanism by which GDH1-mediated metabolic reprogramming of glutaminolysis mediates lung cancer metastasis. This knowledge could be leveraged for the development of effective therapeutic strategies for patients with LKB1-deficient lung cancer.

Citation Format: Sumin Kang, Lingtao Jin, Chaoyun Pan, Jaemoo Chun. Glutamate dehydrogenase 1 mediates metabolic reprogramming and promotes tumor metastasis through CamKK2-AMPK signaling in LKB1-deficient lung 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 1454.
Abstract 1455: FAK overexpression upregulates glucose metabolism in glioblastoma
Authors: Velpula Kiran Kumar , Pajor Michael , Guda Maheedhara R. , Labak Collin M. , Park Simon , Tsung Andrew J.
Published: August 2018

Glioblastoma (GBM) is the most aggressive brain cancer, afflicting nearly 1 in 6 adult patients with brain tumors. Currently, no effective curative treatment for this CNS neoplasm exists. The high prevalence and dismal prognosis of GBM increases the need to identify novel therapeutic targets. Previous research has suggested that increased expression of focal adhesion kinase (FAK) may play a role in tumor metabolism. Our preliminary data using datamining approach of the TCGA and Oncomine™ databases showed that mRNA levels of FAK correlate with expression levels of the glucose transporter GLUT1. Subsequent mass-spectrometric and immunoprecipitation experiments provided evidence that FAK binds to GLUT1, while immunohistochemistry and immunocytochemistry experiments confirmed their interaction. Western blot analysis using shRNA to silence FAK demonstrated a significant decrease in GLUT1 expression. Cells treated with the same FAK shRNA showed decreased metabolites from anaerobic glycolysis, instead relying on oxygen-dependent metabolism utilizing oxidative phosphorylation for their energy supply. Collectively, we propose that FAK is a viable target in modulating glucose uptake and metabolism in GBM.

Citation Format: Kiran Kumar Velpula, Michael Pajor, Maheedhara R. Guda, Collin M. Labak, Simon Park, Andrew J. Tsung. FAK overexpression upregulates glucose metabolism in glioblastoma [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 1455.