bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 43: ASN004, a novel 5T4-targeted Dolaflexin ADC, achieves complete regressions and tumor-free survivors in a broad variety of solid tumor models
Authors: Smith Roger A. , Zammit David J. , Reddy Sanjeeva P.
Published: July 2017

ASN004 is an Antibody Drug Conjugate (ADC) that targets the 5T4 oncofetal antigen (trophoblast glycoprotein). High expression of 5T4 is observed in a wide range of malignant tumors, while very limited expression is found in normal tissues. ASN004 incorporates a novel single-chain homo-dimer antibody, Fleximer® linker technology (Mersana Therapeutics), and several cytotoxic dolastatin (auristatin) analog warheads per ADC molecule with a drug/antibody ratio of ~15:1. ASN004 shows high affinity for the 5T4 antigen (Kd < 30 pM) and 5T4-expressing tumor cells; rapid cellular internalization; and potent, selective cytotoxicity. ASN004 provides complete tumor regressions and tumor-free survivors in multiple tumor xenograft models, derived from human tumor cell lines having both high and low 5T4 expression levels. Tumor-free survivors were achieved at doses that are well-tolerated, based on xenograft body weight measures and exploratory toxicology studies in pharmacologically-relevant non-human primates. As well, tumor-free survivors were achieved in xenograft tumor models following administration of a single dose of ASN004, as low as 1 mg/kg iv. The broad activity of ASN004 has been demonstrated by its superior efficacy in a head-to-head study against trastuzumab-DM1, in a low-5T4, high-HER2 expressing tumor xenograft model. Finally, tumor xenografts that initially responded to ASN004 and later showed tumor regrowth, had not developed resistance and responded well to subsequent treatment with ASN004. Overall, ASN004 is demonstrated to be an ADC with high therapeutic potential in multiple tumor types, encompassing a wide range of 5T4-expression levels. IND-enabling toxicology studies are ongoing, in preparation for advancement to clinical studies in patients with solid tumors.

Citation Format: Roger A. Smith, David J. Zammit, Sanjeeva P. Reddy. ASN004, a novel 5T4-targeted Dolaflexin ADC, achieves complete regressions and tumor-free survivors in a broad variety of solid tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 43. doi:10.1158/1538-7445.AM2017-43
Abstract 4332: Interactome between vascular endothelial cells and Proneural glioma stem cells protects seeds for GBM recurrence from radiation therapy
Authors: Bastola Soniya , Pavlyukov Marat S. , Yamaguchi Shinobu , Wang Jun , Komarova Svetlana , kornblum Harley I. , Nakano Ichiro
Published: July 2017

Glioblastoma (GBM) is the most aggressive form of intracranial tumors. Despite multimodal treatment strategies that include surgical resection, radiation and chemotherapy, most patients survive for less than two years. GBM cells tend to invade into adjacent normal brain tissues and therefore, cannot be completely resected by surgery. These remaining tumor cells are the “seeds” to escape post-surgical therapies, thereby giving rise to recurrent tumors during or after treatment. The exact mechanisms by which these seeds contain pre-existing resistant cells, and/or gain resistance to therapy remain elusive. Recent findings demonstrated that communication between normal and GBM cells play physiological roles in tumor initiation and progression. Here, we aim to determine how vascular endothelial (VE) cells affect GBM recurrence in response to radiotherapy. To this end, we first assessed the effect of secreted factors from VE cells on patient-derived GBM neurospheres in vitro by using their conditioned medium (CM). GBM neurospheres promoted their growth with CM from VE cells. qPCR and FACS exhibited that the CM treatment preferentially protected CD133+ population in GBM neurospheres that otherwise were depleted by irradiation treatment. Furthermore, RNA sequencing of VE cells and A2B5-expressing GBM stem-like cells directly isolated from GBM and epilepsy patients demonstrated that only tumor-derived VE cells and GBM stem-like cells express a high level of a secreted factor Endocan (ESM1) and its receptor CD11a, respectively. Preferential expression of CD11a in perivascular GBM cells was also confirmed by immunocytochemistry. Further, we found that recombinant Endocan is able to promote GBM neurosphere growth in vitro after irradiation and protected them from irradiation-induced aggressive mesenchymal transition of these cells. Taken together, these results demonstrate a potential role of a crosstalk between peri-vascular GBM stem-like cells and their adjacent VE cells, via the ligand-receptor interaction mechanism. This interactome network may help Proneural GBM stem cells to escape from radiation therapy, thereby reinitiating the tumor as recurrence via an Endocan-CD11a signaling. Hence, targeting Endocan in endothelial cells or CD11a in GBM cells may have potential as a novel efficacious strategy for preventing recurrence.

Citation Format: Soniya Bastola, Marat S. Pavlyukov, Shinobu Yamaguchi, Jun Wang, Svetlana Komarova, Harley I. kornblum, Ichiro Nakano. Interactome between vascular endothelial cells and Proneural glioma stem cells protects seeds for GBM recurrence from radiation therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4332. doi:10.1158/1538-7445.AM2017-4332
Abstract 4334: Interleukin-6 is a key player in stroma-induced resistance to chemotherapy for gastric carcinomas
Authors: Ham In-Hye , Lee Dakeun , Jin Hyejin , Son Sang Yong , Kim Yong Bae , Han Sang-Uk , Hur Hoon
Published: July 2017

Introduction Gastric carcinomas (GCs) are known to progress through the interaction between cancer cells and the tumor stroma. Moreover, recent molecular analysis revealed that the expression of stroma-related genes was significantly associated with poor response to chemotherapy in GCs. However, the specific targets to inhibit the interaction between stroma and cancer cells and to reverse stroma-induced chemoresistance have never been suggested in GCs.

Experimental procedures The gene expression in the biopsied GCs tissues from 10 patients, who were treated with preoperative chemotherapy, was analyzed using Nanostring analyzer. We compared the expression pattern of genes involved in the cancer progression panel between chemotherapy response and non-response groups. In addition, we investigated the level of serum interleukin-6 (IL-6) in 39 GCs patients, who underwent curative gastrectomy and postoperative adjuvant chemotherapy. GCs patients were subclassified into recurrence and non-recurrence groups, and the level of IL-6 was compared between two groups. We performed cell proliferation assay, qRT-PCR, ELISA and Western blot to investigate the role of cancer-associated fibroblasts(CAFs)-produced IL-6 on the chemoresistance.

Summary of the new data For pretreated biopsied tissues of patients with preoperative chemotherapy, the expression of IL-6 gene in non-response group was significantly higher than response group. In addition, for patients with postoperative adjuvant chemotherapy, the serum level of IL-6 in GCs patients with recurrence was higher than patients without recurrence. Moreover, In vitro studies demonstrate that co-culture with CAFs increased chemoresistance of various GCs cell lines to 5-fluouracil (5-FU) and CAFs-derived IL-6 activated the membrane gp130 and the STAT3 signal pathway in GCs cell lines. When anti-IL-6R monoclonal antibody was added to 5-FU for GCs cell lines in co-culture with CAFs, activation of STAT3 was significantly reduced and apoptosis markers efficiently induced.

Conclusion Our study showed that the stroma-induced IL-6 is a key player in the resistance to chemotherapy in GCs. We convinced that the interaction between stroma cells and cancer cells could be inhibited by blockade of IL-6 signal pathway. We suggest the use of IL-6 inhibitor as therapeutic agent to enhance the response to chemotherapy in GCs.

Citation Format: In-Hye Ham, Dakeun Lee, Hyejin Jin, Sang Yong Son, Yong Bae Kim, Sang-Uk Han, Hoon Hur. Interleukin-6 is a key player in stroma-induced resistance to chemotherapy for gastric carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4334. doi:10.1158/1538-7445.AM2017-4334
Abstract 4340: RHBDF2 in stromal fibroblasts mediates TGF-β signaling and enhances gastric cancer cell invasion via intercellular crosstalk
Authors: Ishimoto Takatsugu , Miyake Keisuke , Yashiro Masakazu , Eto Tsugio , Izumi Daisuke , Arima Kota , Baba Yoshifumi , Watanabe Masayuki , Hirakawa Kosei , Baba Hideo , Tan Patrick
Published: July 2017

Background: Cancer-associated fibroblasts (CAFs) have been reported to promote various types of tumor through secretion of soluble factors. However, there have been no systematic studies of CAFs in diffuse-type gastric cancers (DGCs). We investigated the characteristics and functional roles of CAFs in DGCs using comprehensive genomic approach.

Methods: We established primary fibroblasts, normal fibroblasts (NFs) and CAFs from more than 100 tissue samples from GC patients. NFs/CAFs were subjected to Exome and RNA sequencing, and the possible candidates for functional assay were selected from among the acquired comprehensive data. The candidate molecules were examined the functional roles for GC tumor progression by in vitro assays.

Results: CAFs exhibited an invasive molecular pattern and acquired motility in extracellular matrix (ECM). We identified RHBDF2 as a mediator of TGF-β signaling and an enhancer of CAF motility. RHBDF2 silencing in CAFs significantly decreased their motility stimulated by TGF-β1 in ECM, whereas NFs transfected with the RHBDF2 expression vector displayed a greater motility in ECM than did control NFs. RHBDF2 silencing also decreased type I TGF-β receptor (TβRI) cleavage through tumor necrosis factor (TNF)-α converting enzyme (TACE) activity and attenuated the invasive molecular pattern of CAFs. Consequently, high-motility CAFs confer on DGC cells the ability to invade the ECM. Furthermore, simultaneous treatment with interleukin (IL)-1α, IL-1β and TNF-α resulted in a strong induction of RHBDF2 expression in NFs. The expression of these cytokines in GC tissues was associated with poor prognosis in GC patients.

Conclusion: These findings highlight the underlying mechanism whereby DGC cells take advantage of CAFs to acquire invasiveness.

Citation Format: Takatsugu Ishimoto, Keisuke Miyake, Masakazu Yashiro, Tsugio Eto, Daisuke Izumi, Kota Arima, Yoshifumi Baba, Masayuki Watanabe, Kosei Hirakawa, Hideo Baba, Patrick Tan. RHBDF2 in stromal fibroblasts mediates TGF-β signaling and enhances gastric cancer cell invasion via intercellular crosstalk [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4340. doi:10.1158/1538-7445.AM2017-4340
Abstract 4342: Loss of MSLN impairs pancreatic cancer growth in the peritoneal cavity
Authors: Rudloff Michael W. , Arons Danielle , El-Behaedi Salma , Albalawy Rakan , Alewine Christine
Published: July 2017

Mesothelin (MSLN) is a cell surface glycoprotein that is expressed in at least 85% of pancreatic adenocarcinoma but not on cells of the healthy pancreas nor in the parenchyma of other vital organs. Due to this differential expression, MSLN has been used as a target for various antibody based treatments and cancer vaccines. The physiologic role of MSLN is unknown. Previous reports have suggested that overexpression of MSLN may increase tumorigenicity and metastatic potential of pancreatic adenocarcinoma. To further define the role of MSLN in this disease, we deleted MSLN from the human pancreatic cancer cell line, KLM1, using CRISPR/Cas9 gene editing. Successful deletion of MSLN was confirmed via flow cytometry and immunoblotting. Furthermore, KLMΔMSLN cells were rendered invulnerable to the MSLN-targeted immunotoxin, RG7787, which depends on the presence of surface MSLN for cytotoxicity. In cell culture, KLMΔMSLN cells grew at the same rate as control cells. KLMΔMSLN formed subcutaneous tumors in nude mice with the same frequency as control cells and these tumors grew at the same rate. However, when nude mice were inoculated with KLMΔMSLN or control cells intraperitoneally, a marked decrease in tumor burden was observed in cells lacking MSLN. In summary, we have engineered a pancreatic cancer cell line that lacks MSLN and demonstrated that loss of MSLN impairs tumor growth and spread specifically within the peritoneal cavity. Further experiments are in progress to identify the factors contributing to this phenotype.

Citation Format: Michael W. Rudloff, Danielle Arons, Salma El-Behaedi, Rakan Albalawy, Christine Alewine. Loss of MSLN impairs pancreatic cancer growth in the peritoneal cavity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4342. doi:10.1158/1538-7445.AM2017-4342
Abstract 4350: Cell-of-origin differentiation stages define methylation-based subtypes of human colorectal cancer
Authors: Bormann Felix , Rodríguez-Paredes Manuel , Bergman Yehudit , Linhart Heinz G. , Lyko Frank
Published: July 2017

Introduction: Colorectal cancer accounts for a significant fraction of cancer-related mortalities, but has proven to be surprisingly refractory to consensus pathological and molecular subclassification. Similarly, little is known about the molecular profiles of colorectal cancer cells-of-origin. DNA methylation is an important epigenomic marker of cellular identity and can therefore be used to infer cancer cells-of-origin.

Methods: Infinium 450k data from colorectal cancer samples provided by “The Cancer Genome Atlas” was applied to a consensus clustering algorithm. The identified epigenetic subgroups were tested for features including well established clinical parameters or subgroup specific gene expression. Additionally, the subgroups were compared with Infinium 450k data of non-malignant colorectal adenomas. Finally, clinical significance was addressed by testing for subgroup specific overall survival rates.

Results: Analysis of the TCGA dataset defined 5 distinct epigenetic subtypes of human colorectal cancer. These subgroups showed an overlap to the microsatellite instability phenotype and the CpG island methylator phenotype. Gene expression based analysis revealed that the subtypes also form a continuum of epigenetic programs reflecting various intestinal crypt cell differentiation stages. Patient survival correlated with the differentiation stage, with a particular poor prognosis for patients with stem cell-related signatures. Interestingly, its prognostic potential outperformed a recently established gene expression-based classifier. Finally, non-malignant colorectal adenomas could be classified into the same methylation subtypes, reflecting their shared cell-types of origin with carcinomas

Conclusion: Our results establish a novel and clinically relevant approach for colorectal cancer classification and illustrate how differences in the cell-type of origin shape the tumor methylome.

Citation Format: Felix Bormann, Manuel Rodríguez-Paredes, Yehudit Bergman, Heinz G. Linhart, Frank Lyko. Cell-of-origin differentiation stages define methylation-based subtypes of human colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4350. doi:10.1158/1538-7445.AM2017-4350
Abstract 4354: Methylation-mediated silenced PYCARD plays a key role in human prostate cancer
Authors: Fukushige Shinichi , Miyauchi Toshiya , Okubo Teppei , Mitsuzuka Koji , Arai Yoichi , Horii Akira
Published: July 2017

Epigenetic gene silencing by aberrant DNA methylation leads to loss of key cellular pathways in tumorigenesis. In order to analyze effects of DNA methylation in prostate cancer, we constructed LNCaP-derived human prostate cancer cells that can induce global reactivation of hypermethylated genes by the methyl-CpG targeted transcriptional activation (MeTA) method. In MeTA, a cassette construct of MBD2-derived methyl-CpG binding domain (MBD) with NFκB-derived transcriptional activation domain is transfected into cancer cells. Then expressed MBD domain specifically binds to the hypermethylated promoter regions, and NFκB transcriptional activation domain recruits p300/CREB-binding protein (CBP) and reactivates hypermethylation-mediated silenced genes. A cell proliferation assay indicated that MeTA suppressed the growth of LNCaP cells. Furthermore, both flow cytometry and TUNEL assays demonstrated that MeTA induced apoptosis. In order to search genes responsible for apoptosis, we performed gene expression microarray analysis of MeTA-uninduced and -induced LNCaP cells: Five genes encoding apoptosis-inducing factors upregulated two-fold or more in accordance with the induction of MeTA; PYCARD (PYD and CARD domain containing), TNFRSF25 (tumor necrosis factor receptor superfamily 25), HRK (harakiri, BCL2 interacting protein), BIK (BCL2 interacting killer), and CIDEA (cell death-inducing DFFA-like effector a). These genes also contained CpG islands (CGIs) within ± 1,000-bp of the transcription start site. We focused on PYCARD and TNFRSF25 because these two genes upregulated 10-fold or more in accordance with the induction of MeTA. We found that both genes were hypermethylated and showed low expression levels in LNCaP prostate cancer cells, whereas only PYCARD was unmethylated in RWPE-1 normal prostate epithelial cells. We further analyzed primary prostate cancer tissues and found tumor-specific promoter hypermethylation of PYCARD in 53.8% (7/13; P < 0.01). These results suggest that methylation-mediated silencing of PYCARD contributes to escape from apoptosis in human prostate cancer, and MeTA may provide important clues to analyze changes in cancer cell phenotypes by DNA methylation alterations.

Citation Format: Shinichi Fukushige, Toshiya Miyauchi, Teppei Okubo, Koji Mitsuzuka, Yoichi Arai, Akira Horii. Methylation-mediated silenced PYCARD plays a key role in human prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4354. doi:10.1158/1538-7445.AM2017-4354
Abstract 4373: SOX2-related gene signature predicting the prognosis in head and neck squamous cell cancer
Authors: Eun Young-Gyu , Lee Young Chan , Hwang Jun Eul
Published: July 2017

Purpose: Numerous studies have indicated that Sex-determining region Y-box 2 (SOX2) is involved in many squamous cell carcinomas. However, the role of SOX2 in head and neck squamous cell carcinoma (HNSCC) remains unclear. Therefore, we investigated whether activation of SOX2 is significantly associated with prognosis in HNSCC.

Methods: Gene expression signature reflecting SOX2 activation was identified in HNSCC cohort, and patients were stratified into two groups according to this signature: SOX2-high group or SOX2-low group. Validation of the signature was sought in two independent patient groups. The association between the signature and prognosis of patients was assessed.

Results: The SOX2-low group was associated with poor prognosis for HNSCC in three independent patient cohorts. In a multivariate analysis, the impact of the SOX2 signature on overall survival (OS) was independent of other clinical variables [hazard ratio (HR), 1.45; 95% confidence interval (CI), 1.09 - 1.92; P=0.01]. In patients who received radiotherapy (RT), SOX2-low group had significantly poor OS than those in SOX2-high group.

Conclusions: SOX2-low signature is associated with poor prognosis in patients with HNSCC and could be used to predict patients who would benefit from RT.

Note: This abstract was not presented at the meeting.

Citation Format: Young-Gyu Eun, Young Chan Lee, Jun Eul Hwang. SOX2-related gene signature predicting the prognosis in head and neck squamous cell cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4373. doi:10.1158/1538-7445.AM2017-4373
Abstract 4375: Complex interactions between genomic and epigenomic alterations in esophageal adenocarcinomas
Authors: Peng Dunfa , Guo Yan , Chen Heidi , Zhao Shilin , Washington Kay , Hu Tianling , Shyr Yu , El-Rafai Wael
Published: July 2017

Background: The incidence of Barrett’s-related esophageal adenocarcinoma (EAC) is rapidly rising in the USA and the Western countries. However, the interaction between genomic and epigenomic alterations in regulating gene expression networks in EAC is poorly understood.

Results and Discussion: We carried out an integrative analytical approach to identify DNA copy number, promoter DNA methylation and gene expression in a panel of esophageal adenocarcinomas, using chip-based technologies. Our analysis displayed significant differences between EAC and normal samples in DNA copy numbers, gene expression, and DNA methylation profiles. Our results identified remarkable incidence of DNA copy number (CN) alterations showing that 90% of probes had a copy number aberration (gain or loss) in at least one tumor, suggesting a vast chromosomal instability in EAC. Integrative analysis indicated that altered expression of 1755 genes was associated with changes in CN or methylation. We found that expression alterations in 84 genes (56 overexpressed and 28 down regulated) were associated with changes in both CN and methylation. These data suggest a strong interaction between genetic and epigenetic events to modulate gene expression in EAC. Of note, we detected a prominent K-RAS signature in EAC. Network analysis predicted activation of several important transcription factor networks in EAC, including β-catenin, MYB, TWIST1, SOX7, GATA3 and GATA6. 56 genes were overexpressed in EAC with simultaneous promoter hypomethylation and gene amplification, including CDH17 and GATA6. Notably, several genes associated with inflammation such as COX2, IL8 and IL23R were overexpressed and hypomethylated in EAC, a reaffirmation of the underlying inflammatory process in Barrett’s tumorigenesis and a suggestion of an epigenetic regulation of these genes.

Conclusions: In summary, our integrative analysis of genetic and epigenetic alterations in EAC demonstrates a complex interaction between molecular mechanisms in EAC and provides several novel insights for future studies.

Citation Format: Dunfa Peng, Yan Guo, Heidi Chen, Shilin Zhao, Kay Washington, Tianling Hu, Yu Shyr, Wael El-Rafai. Complex interactions between genomic and epigenomic alterations in esophageal adenocarcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4375. doi:10.1158/1538-7445.AM2017-4375
Abstract 4377: EBV associated gastric cancer in Korean
Authors: Lee Woochan
Published: July 2017

Gastric cancer was the third leading cause of death worldwide and most prevalent malignant cancer in Korea, comprising 20.8% of all cancers. The Cancer Genome Atlas (TCGA) divided gastric cancer to four subgroups; EBV (Epstein-Barr Virus infected, 9%), MSI (Microsatellite instability, 22%), GS (Genomically stable, 20%), CIN (Chromosomal instability, 50%). We sequenced transcriptomes of 11 gastric cancers associated with EBV infection (EBVaGC) tissues, 11 gastric cancers not associated with EBV infection (EBVnGC) tissues and 22 matched normal gastric mucosa for find unique features of Korean EBVaGC. We compared RNA expression, EBV gene expression, germline mutations, and somatic mutation between EBVaGC and EBVnGC. We found IKBKE gene, which is taking an essential role in regulating antiviral signaling pathways, expressed higher on EBVaGC tumor than EBVnGC tumor (p-value=1.73 x 10-4). While most EBV genes appeared to be similar, BMRF1,2 genes made distinctive patterns compared to previous TCGA report in expression of EBV which integrated into human cells. The BMRF-2 gene is associated with cell-to-cell spread of EBV, which is major infection mechanism of EBV. We found novel germline mutations on PABPC3, PGA3, and C4A,B in almost every samples, which are candidate for Korean specific mutations.

Citation Format: Woochan Lee. EBV associated gastric cancer in Korean [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4377. doi:10.1158/1538-7445.AM2017-4377
Abstract 4392: Genetic diagnosis of multiple hepatocellular carcinoma
Authors: Midorikawa Yutaka , Yamamoto Shogo , Tatsuno Kenji , Ueda Hiroki , Tsuji Shingo , Nagae Genta , Takayama Tadatoshi , Aburatani Hiroyuki
Published: July 2017

Multiple hepatocellular carcinoma (HCC) is categorized into two types; multicentric hepatocarcinogenesis (MC) and intrahepatic metastasis (IM). Although discrimination of the types of multiple HCC is of clinical importance, it is quite difficult to determine it correctly even after histological diagnosis. In order to determine multiple HCC as MC or IM, we compared pairs of multiple HCC samples from 40 patients with multiple HCC and 2 patients with pairs of primary HCC and extrahepatic metastasis such as adrenal grand and lung metastasis with regard to molecular aberrations. Exome sequence showed that more than 70% of somatic mutations were common in the pairs of IM, which were consistent with the result of a pair of primary HCC and extrahepatic metastasis, while no common somatic mutations was detected in genomic-MC (gMC) pairs. Notably, more than 60% of clinically metachronous-MC cases were diagnosed as genomic-IM (gIM) due to the concordance of mutation and shows significantly frequent tumor thrombus, suggesting that gIM pairs developed from the common ancestor. In addition, cell-free DNA was analyzed using exome sequence in five patients with recurrent HCC, and found that somatic mutations of cell free DNA were similar to the primary HCC only in gIM but not in gMC cases. Taken together, comparison of mutations in a pair of HCCs makes it possible to classify multiple HCCs into MC and IM, which is difficult to be correctly determined in clinical practice. Furthermore liquid biopsy for mutation analysis is available to make treatment plans for multiple HCC patients.

Citation Format: Yutaka Midorikawa, Shogo Yamamoto, Kenji Tatsuno, Hiroki Ueda, Shingo Tsuji, Genta Nagae, Tadatoshi Takayama, Hiroyuki Aburatani. Genetic diagnosis of multiple hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4392. doi:10.1158/1538-7445.AM2017-4392
Abstract 4415: Fatty acid synthase-mediated palmitate production impacts epidermal growth factor receptor signaling to regulate specificity protein 1 in breast cancer cells
Authors: Barlow Lincoln J. , Zhang Jian-Ting
Published: July 2017

Fatty acid synthase (FASN), the sole cytosolic enzyme responsible for de novo palmitate synthesis, is critical for cancer cell survival and is a marker of poor prognosis. FASN overexpression has been shown to confer resistance to both drug and radiation resistance by inhibiting drug-induced ceramide production and by increasing DNA repair activities. Recently, it was found that FASN upregulation of non-homologous end joining (NHEJ) repair of double strand DNA breaks was due to FASN up-regulation of Poly (ADP-ribose) polymerase 1 (PARP1) by increasing expression of specificity protein 1 (SP1). However, how FASN regulates the expression of SP1 remains unknown. In this study, we tested the hypothesis that, palmitate, the end product of FASN catalysis, plays an important role in regulating SP1 expression by posttranslationally modifying epidermal growth factor receptor (EGFR) and affecting its activity. Using paired isogenic cell lines with FASN overexpression and FASN knockdown as well as exogenous supplementation of palmitate, we found that depleting palmitate in culture reduced SP1 expression while supplementing palmitate rescues this effect. Furthermore, FASN overexpression and knockdown increased and decreased, respectively, phosphorylation and activation of EGFR. Treatment with EGFR inhibitor, Erlotinib, reduced SP1 expression. These findings suggest that FASN regulation of SP1 and NHEJ repair may be via regulating EGFR activity. We are in the process of determining the potential effect of palmitoylation of EGFR on its activation and regulation of SP1 expression.

Citation Format: Lincoln J. Barlow, Jian-Ting Zhang. Fatty acid synthase-mediated palmitate production impacts epidermal growth factor receptor signaling to regulate specificity protein 1 in breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4415. doi:10.1158/1538-7445.AM2017-4415
Abstract 4422: Capsaicin suppresses pancreatic tumor growth by inhibiting tumor cell metabolism
Authors: Ramachandran Sharavan , Srivastava Sanjay K.
Published: July 2017

Tumor cell metabolism is considered to be a hallmark of tumorigenic progression. Pancreatic cancer cells are shown to develop addiction towards the metabolic pathways like glycolysis and mitochondrial respiration to meet energy demands. Pancreatic tumor cells predominantly utilize cytosolic aerobic glycolysis for energy production. In our previous studies, we have reported that capsaicin treatment inhibits the survival of AsPC-1 and BxPC-3 pancreatic cancer cell lines. In this study, we evaluated the effect of capsaicin on glycolysis and mitochondrial respiration in pancreatic cancer cells. Our results showed that capsaicin treatment reduced the extra-cellular acidification rate (glycolysis) and oxygen consumption rate (mitochondrial respiration) in AsPC-1 and BxPC-3 cells in a concentration-dependent manner after 24h of treatment. Capsaicin treatment caused 86% and 55% down-regulation of glycolysis process in AsPC-1and BxPC-3 cells respectively. Our results also indicate the inhibition of glycolytic capacity and glycolytic reserve by capsaicin treatment indicating the potential of capsaicin to inhibit glycolysis. In addition, capsaicin treatment reduced 98% of basal oxygen consumption rate and ATP production in AsPC-1 cells. Furthermore, capsaicin treatment inhibited spare respiratory capacity & proton leak in a concentration dependent manner in AsPC-1 cells. Treatment of AsPC-1 and BxPC-3 with capsaicin for 48h also inhibited the expression of LDH-A and its upstream regulators such as HIF-1α, pSTAT3 (Y705) and EGFR as evaluated by Western blot. Our study thus indicate that capsaicin suppresses pancreatic tumor growth by down-regulating glycolysis and mitochondrial respiration. Further mechanistic studies are in progress. [Supported in part by R01 grant CA129038, awarded to (S.K.S.) by the National Cancer Institute].

Citation Format: Sharavan Ramachandran, Sanjay K. Srivastava. Capsaicin suppresses pancreatic tumor growth by inhibiting tumor cell metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4422. doi:10.1158/1538-7445.AM2017-4422
Abstract 4436: Novel microRNA signature score to predict bone metastasis and prognosis of breast cancer
Authors: Kawaguchi Tstutomu , Yan Li , Qi Qianya , Liu Song , Takabe Kazuaki
Published: July 2017

Backgrounds: 40,000 US women still die with breast cancer every year. Vast majority of death occur after they develop metastasis, where bone is the most frequent site for breast cancer. New measures to identify the patients who develop metastasis allow us to intervene early, which is expected to prolong survival. The aim of this study is to establish a microRNA (miRNA) signature scoring system that can predict bone metastasis and survival utilizing integrated transcriptomics analyses in breast cancer.

Materials and Methods: Both clinical and RNA expression data, including microRNA and mRNA, of 1051 patients were retrieved from The Cancer Genome Atlas (TCGA). 1) Multivariate Cox proportional hazard model and Kaplan-Meier for overall survival were performed to construct and identify novel models of miRNAs signature for predicting patient survival. 2) Competing risk analysis using the miRNAs signature was conducted to clarify its association with metastatic distributions. 3) Gene Set Enrichment Analysis (GSEA) was performed to identify the genome-epigenome significance of the miRNAs signature.

Results: 1) Utilizing Cox model on TCGA cohort, we established a novel risk scoring system with three miRNAs signatures (miR-19a, miR-93, and miR-106a) that identified the patients population with extremely poor overall survival (p = 0.0004; 5-y survival rate, 49.2%). This result was reproduced in two other completely independent cohorts with microarray datasets (GSE19536, p = 0.0009; GSE22220, p = 0.0003, respectively). 2) Utilizing competing risk analysis for each metastatic sites of breast cancer, we found that the patients with bone metastasis demonstrated significantly high scores (p = 0.0052). The evaluation also showed a statistical tendency of association with lung metastasis (p = 0.0854). 3) We found that high score is associated with several critical gene sets such as angiogenesis (p < 0.0001) or epithelial mesenchymal transition (p = 0.0155) by GSEA that suggests that high signature score is associated with enhanced metastasis in breast cancer patients.

Conclusions: We established a miRNAs signature scoring system to predict bone metastasis and poor overall survival in breast cancer using novel integrated transcriptomics concept.

Citation Format: Tstutomu Kawaguchi, Li Yan, Qianya Qi, Song Liu, Kazuaki Takabe. Novel microRNA signature score to predict bone metastasis and prognosis of breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4436. doi:10.1158/1538-7445.AM2017-4436
Abstract 4439: The urine miRNA transcriptome of prostate cancer is temporally stable and predicts disease aggressivity
Authors: Jeon Jouhyun
Published: July 2017

The clinical management of prostate cancer is the most common non-skin male malignancy in the world is hindered by the limitations of current diagnostic and prognostic tests, such as the low specificity of prostate-specific antigen (PSA) testing, low sensitivity of digital rectal examination (DRE) and complications of biopsies. To try to resolve these issues, we evaluated miRNA abundance in the urine of prostate cancer patients. Alterations in miRNA abundance levels have been reported as playing essential roles in the pathogenesis of cancer and is occurred in a tumor phenotype-specific manner (e.g. aggressive and non-aggressive). miRNAs are stable under diverse analytical conditions and can be detected in various types of body fluids including urine. These characteristics make them as promising non-invasive biomarkers. Here, we examined the intra- and inter-individual variance of urine miRNA abundance by investigating longitudinal changes over months to years in a cohort of patients with localized prostate cancer. We observed a large dynamic range of intra-individual variance in miRNA abundances and identified a set of miRNAs that is stable within individuals, and is biased toward specific biological functions including regulation of transmembrane channel activity. We combined this observation with machine-learning techniques to create a predictive model that can identify aggressive prostate cancer. This four miRNAs predictive model was validated in an independent prostate cancer cohort to non-invasively predict high-risk disease. Remarkably it showed comparable performance to the best existing tissue-based prognostic markers. These results demonstrated that non-invasive biomarkers can be developed to precede or supplement tissue-based tests by understanding the intra- and inter-tumoural heterogeneity of the urine miRNA transcriptome.

Citation Format: Jouhyun Jeon. The urine miRNA transcriptome of prostate cancer is temporally stable and predicts disease aggressivity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4439. doi:10.1158/1538-7445.AM2017-4439
Abstract 4440: New predictive and preventive biomarkers for preclinical hepatocellular carcinoma in Egyptian population by detection of serum miRNAs
Authors: Ghazy Sara Salama , Fyala Amira S. , Sultan Ahmed S.
Published: July 2017

The hepatocellular carcinoma (HCC) is one of the most common malignant tumors and carries a poor survival rate. Prognosis, survival and management of patients at risk for developing HCC remain challenging in Egypt and worldwide. Furthermore, poor prognosis of patients with symptomatic (HCC) diagnosed clinically at advanced stages suggests an urgent need for new biomarkers detection that can be used for pre-clinical screening for early detection of premalignant lesions and tumors in high risk to hepatitis C infection.

In the present study we used novel formulated anticancer compound(s), combined with or without Sorafenib treatment to exert their antiproliferative effects by eliciting concomitant expression of known or novel biomarkers, including different serum miRNAs, which can be quantitatively analyzed for differences in the levels of metabolites and proteins in the blood and liver tissues of treated HCC xenografts and liver cirrhosis animal models. In addition, circulating miRNAs levels were measured as promising markers in 12 month interval-serum samples that were collected from normal, chronic hepatitis C virus infected patient cohorts at the pre-malignant or pre-clinical stages and liver cirrhotic patients that did not develop cancer. Different serum miRNAs, including miR-21, miR-142, let-7a, let-7b, miR-429 and miR-34a were detected in pre-clinical HCC patients and have the potential to screen for hepatitis C virus infected patients at high risk to develop HCC 12 months after miRNAs detection. Our data of detected serum miRNAs from animal models and patient samples can be used to confirm and correlate histological and histochemical results at various time points with serum or imaging biomarkers, which may have a great promise for the prediction and prevention of HCC in high risk Egyptian populations.

Citation Format: Sara Salama Ghazy, Amira S. Fyala, Ahmed S. Sultan. New predictive and preventive biomarkers for preclinical hepatocellular carcinoma in Egyptian population by detection of serum miRNAs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4440. doi:10.1158/1538-7445.AM2017-4440
Abstract 4443: SPARC overexpression alters microRNA expression profiles involved in tumor progression
Authors: Ahir Bhavesh K. , Lakka Sajani
Published: July 2017

Medulloblastoma is the most common of malignant brain tumor in children. SPARC (secreted protein acidic and rich in cysteine), a multicellular non-structural glycoprotein is known to be involved in multiple processes in various cancers. Previously, we reported that SPARC expression significantly impairs medulloblastoma tumor growth in vivo and alters chemo sensitivity. MicroRNAs are a class of post-transcriptional gene regulators with critical functions in tumor progression. In addition, they also play role in regulation of vital genes that control mechanisms of chemo-resistance. Herein, we assessed microRNA (miRNA) profiling to identify the functional network and biological pathways altered in SPARC-overexpressed medulloblastoma cells. A total of 27 differentially expressed miRNAs were identified between the control and SPARC-overexpressed samples. Potential messenger RNA (mRNA) targets of the differentially expressed miRNA were identified using Ingenuity Pathway Analysis (IPA). Network-based functional analyses were performed on the available human protein interaction and miRNA-gene association data to highlight versatile miRNAs among the significantly deregulated miRNAs using the IPA, and the biological pathway analysis using the PANTHER web-based tool. We identified six miRNAs (miR-125b1*, miR-146a-5p, miR-181a-5p, miR-204-5p, miR-219-5p and miR-509-3p) which are differentially regulated with SPARC expression in medulloblastoma cells. Furthermore, pathway enrichment analysis outline that these six microRNAs mainly belong to biological processes related to cancer related signaling pathways. Collectively, these results suggest that a perturbed expression for these six miRNAs and their complex regulatory loops contribute to tumor suppressive effects in SPARC overexpressed medulloblastoma cells.

Citation Format: Bhavesh K. Ahir, Sajani Lakka. SPARC overexpression alters microRNA expression profiles involved in tumor progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4443. doi:10.1158/1538-7445.AM2017-4443
Abstract 4448: Elevated levels of circulating miR-34a-5p and miR-181c-3p during cancer initiation by aflatoxin B1
Authors: Livingstone Merricka C. , Roebuck Bill D. , Johnson Natalie M. , Kensler Thomas W. , Groopman John D.
Published: July 2017

MicroRNAs are small non-coding RNAs that affect gene expression. They are essential in normal physiological processes and have been implicated in carcinogenesis. Hepatocellular carcinoma (HCC) is the second leading cause of cancer death globally, with the risk factor aflatoxin (AFB1) estimated to play a causal role in up to 28% of all HCC cases. In this study, we analyzed circulating miRNAs in a quantitative rat cancer model that were previously found to be associated with AFB1 exposure in liver. Sera were examined from 3 groups of animals: rats dosed with AFB1 (200 µg) for 28 consecutive days receiving vehicle only or AFB1 plus the chemopreventive agent CDDO-Im (30 µmol), and controls (Johnson et al., CaPR, 2014). Small RNAs were isolated from these serum samples at the end of the dosing period and analyzed by RT-qPCR. Quantitative PCR results at the end of the carcinogenic 28-day dosing regimen revealed notable increased expression of miR-34a-5p and miR-181c-3p (13 fold and 170 fold, respectively) in the AFB1 treated samples compared to controls. This expression was reduced in animals that received complete protection from developing HCC by concurrent intervention with CDDO-Im. Additionally, miR-122-5p displayed increased expression (5 fold) and miR-541-5p was downregulated (0.08 fold) in the sera of AFB1-treated animals. Overall, the data suggest that serum expression of miR-34a-5p and 181c-3p during liver cancer initiation could give insight into increased risk of developing HCC. Supported by T32ES007141-31A1 and CA197222.

Citation Format: Merricka C. Livingstone, Bill D. Roebuck, Natalie M. Johnson, Thomas W. Kensler, John D. Groopman. Elevated levels of circulating miR-34a-5p and miR-181c-3p during cancer initiation by aflatoxin B1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4448. doi:10.1158/1538-7445.AM2017-4448
Abstract 4453: Identification and characterization of oncogenic kinase LASK2 as a diagnostic and therapeutic target for lung cancer
Authors: Daigo Yataro , Takano Atsushi , Nakamura Yusuke
Published: July 2017

Lung cancer is the leading cause of cancer deaths worldwide. Since the number of patients responding well to standard therapies is still limited, further development of new anti-cancer agents with minimum risk of adverse event is urgently awaited. To screen oncoantigens which could be used for the development of new diagnostic biomarkers and molecular targeted drugs, we have established a screening system as follows; i) To identify overexpressed genes in 120 lung cancers by genome-wide screening using the microarray representing 27,648 genes and pure populations of tumor cells taken from cancer tissues by laser microdissection, ii) To verify the genes for their very low or absent expression in normal tissues by northern-blotting, iii) To validate the clinicopathological significance of their expression with tissue microarray covering hundreds of archived lung cancers, iv) To verify whether the target genes are essential for the growth or survival of lung cancer cells by RNAi and cell growth assays. During this process, we selected dozens of druggable oncogenes with various enzymatic activities, and identified overexpression of protein kinase LASK2 (lung cancer-associated kinase 2) in the majority of lung cancers, but not in normal tissues except testis. Immunocytochemical analysis found LASK2 to be mainly localized in the cytoplasm and nucleus of lung cancer cells. Immunohistochemical analysis revealed that strong LASK2 expression was an independent prognostic factor for non-small cell lung cancer (NSCLC) (P<0.0001). Suppression of LASK2 expression with its specific siRNAs caused growth inhibition of lung cancer cells. The enhanced cellular proliferation by introduction of LASK2 in mammalian cells also supported its oncogenic function. Since LASK2 plays a significant role in the cell proliferation and malignant nature of lung cancers, LASK2 is likely to be a prognostic biomarker and a potential therapeutic target for developing new anti-cancer drugs.

Citation Format: Yataro Daigo, Atsushi Takano, Yusuke Nakamura. Identification and characterization of oncogenic kinase LASK2 as a diagnostic and therapeutic target for lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4453. doi:10.1158/1538-7445.AM2017-4453