bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4879: Genomic and epigenomic profiling of high-stage neuroblastoma
Authors: Ohira Miki , Tatsumi Yasutoshi , Nakamura Yohko , Tatsuno Kenji , Tsutsumi Shuichi , Yamamoto Shogo , Nagae Genta , Renard-Guillet Claire , Sugino Ryuichi , Nagase Hiroki , Kamijo Takehiko , Aburatani Hiroyuki , Nakagawara Akira
Published: July 2017

Neuroblastoma (NB) is the most common pediatric extracranial solid tumor with a wide range of clinical phenotypes from spontaneous regression to highly resistant to chemotherapy. Despite intense multi-modal therapy, NB remains the second most common cause of cancer deaths among children. MYCN amplification is the most prominent poor prognosis factor for NB, however, certain subgroup of high stage tumor without MYCN amplification also shows low survival rate in terms of long-term follow-up (10 years survival rate was 51%, n=161). To elucidate the molecular basis of malignant subtype of NB, integrated genomic and epigenomic analyses were performed. Array CGH of 537 tumors indicated that genomic subgroups of 17q gain, with 1p loss and/or 11q loss, exhibited aggressive characteristics even without MYCN amplification. Whole exome sequencing of 92 high stage, MYCN-non-amplified primary tumors (SureSelect XT Human All Exon V4, mean depth CDS:138, x20 coverage:96%) identified 1,735 non-silent somatic mutations, among which 53 were significant mutations observed more than once in the sample set. Aggressive tumors (died of disease, n=48) had several notable features, with mutations in chromatin remodeling/modulating genes (27%, cf. ATRX), in DNA-damage response (30%), in TP53 (28%) and RB (22%)-related gene networks, as well as with high TERT expression (38%). By hierarchical clustering of methylome signature (Infinium HumanMethylation450 BeadChip), the 92 tumors were subdivided into four clusters. Unfavorable cluster-2 and -4 exhibited hyper-methylation phenotypes in CpG islands and the former was enriched by MYCN-amplified tumors. These two clusters (53%) showed strong correlation with patient prognosis (p=0.0055). Thus, these prognosis-related genomic features could be useful markers for risk classification and help understanding molecular mechanism for aggressive phenotypes of MYCN-non-amplified NBs.

Citation Format: Miki Ohira, Yasutoshi Tatsumi, Yohko Nakamura, Kenji Tatsuno, Shuichi Tsutsumi, Shogo Yamamoto, Genta Nagae, Claire Renard-Guillet, Ryuichi Sugino, Hiroki Nagase, Takehiko Kamijo, Hiroyuki Aburatani, Akira Nakagawara. Genomic and epigenomic profiling of high-stage neuroblastoma [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 4879. doi:10.1158/1538-7445.AM2017-4879
Abstract 4889: Characterizing the viromes in pediatric cancers through whole genome sequencing
Authors: Chang Ti-Cheng , Easton John , Pinto Emilia M. , Zhang Jinghui , Wu Gang
Published: July 2017

Viral infections have been associated with the development of adult cancers such as cervical carcinomas and head/neck cancers. However, there is lack of clear understanding of the composition of virome in pediatric cancers. Here, we made the first attempt to identify differentially enriched viral communities in biological material from pediatric cancer patients, and to predict virus integration sites that may be associated with etiology or tumor progression of pediatric cancers.

The Pediatric Cancer Genome project (PCGP) has previously generated whole-genome sequencing (WGS) data of matched cancer-normal samples for over 600 pediatric patients from three classes of cancers: leukemia, solid tumors, and central nervous system (CNS) cancers. We re-constructed the virome composition profiles in each sample based on the unmapped and partially mapped Illumina reads extracted from the PCGP WGS data. Taxonomic assignments of the extracted read data were conducted using the Kraken metagenomic pipeline and virus integration sites were predicted by the VyPER pipeline. Custom scripts were used for downstream comparative analyses.

A comparison of the alpha diversity of each virome indicated that the tumor samples in general have a lower diversity compared to the normal samples across all tumor types. This trend holds true in leukemia, where both tumor and normal samples were blood derived, suggesting that this is not due to different origins of the tumor samples.

Ordination analyses based on the viral composition identified distinct clusters in the three classes of cancers. These clusters were formed mainly by samples of medulloblastoma for CNS, T-ALL and hypodiploid B-ALL for leukemia, retinoblastoma and rhabdomyosarcoma for solid tumors. Viruses, such as phages, herpesviraidae and several sarcoma viruses, were differentially enriched in these clusters.

Potential integration with retroviruses or herpesviruses was predicted at a high frequency. Less frequent integration events were also detected with known oncogenic viruses, such as simian virus 40 and human adenovirus C, which were under deep sequencing validation.

In summary, our comprehensive characterization of the virome in a wide spectrum of pediatric cancers revealed a potential connection between specific viral communities and integration sites in certain distinct types of pediatric cancers. Our findings highlight a possible mechanism underlying the tumor progression in pediatric patients.

Citation Format: Ti-Cheng Chang, John Easton, Emilia M. Pinto, Jinghui Zhang, Gang Wu. Characterizing the viromes in pediatric cancers through whole genome sequencing [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 4889. doi:10.1158/1538-7445.AM2017-4889
Abstract 4892: MOSPD2, a newly characterized protein, promotes breast cancer metastasis
Authors: Mendel Itzhak , Salem Yaniv , Yacov Niva , Propheta-Meiran Oshrat , Breitbart Eyal
Published: July 2017

Introduction: We have previously described motile sperm domain-containing protein 2 (MOSPD2), a protein which up until recently no function was ascribed to, as a key regulator of monocyte migration in-vitro. In this study, the role of MOSPD2 in promoting breast cancer migration and metastasis in-vitro and in-vivo was assessed.

Experimental procedure: The prevalence of MOSPD2 was evaluated by IHC in tissue microarray layered with cores, representing different stages of invasiveness in breast cancer. MOSPD2 abundance was scored according to the staining intensity on a scale from 0 to 3. MOSPD2 expression was silenced in MDA-231 breast cancer cell line using lentiviral particles for sh-RNA or CRISPR-CAS9 (CRISPR). Cells were then tested by a trans-well assay for migration towards EGF in vitro as well as for lung dissemination following orthotopic or systemic inoculation in vivo. For mechanism studies, EGF-induced signaling events were analyzed.

Results: Within normal adjacent tissue (NAT), 82% of the samples had no staining and the remaining 18% percent displayed an intensity of 1. In the tissue cores with in-situ carcinoma pathology, 79% of the samples had no staining intensity while the remaining 21% only scored 1 or 2. However, analysis of invasive and metastatic cancer demonstrated higher frequency in score of 2 and increased staining intensity up to score of 3 compared with NAT and in-situ carcinoma. Thus, the percent of combined scores 2 and 3 for invasive lobular carcinoma, invasive ductal carcinoma and metastatic invasive ductal carcinoma were 63%, 77% and 81%, respectively. MOSPD2-silenced cells, either by sh-RNA or by CRISPR, were severely impaired in their ability to migrate in-vitro towards EGF. Following EGF activation of CRISPR-Control MDA-231 cells, all tested proteins down-stream to the EGFR pathway, including AKT and p38, became phosphorylated. However, in CRISPR-MOSPD2 silenced cells, phosphorylation of all tested proteins excluding p38 were markedly inhibited. Moreover, activation with IGF of CRISPR-MOSPD2 MDA-231 cells did not hinder AKT phosphorylation. In vivo, SCID mice injected orthotopically or systemically with CRISPR-MOSPD2 silenced MDA-231 cells displayed significantly reduced number of metastases in the lungs relatively to mice injected with CRISPR-Control treated cells.

Conclusions: We suggest that MOSPD2 promotes metastasis of breast cancer cells and is a potential target for the treatment of metastatic breast cancer.

Citation Format: Itzhak Mendel, Yaniv Salem, Niva Yacov, Oshrat Propheta-Meiran, Eyal Breitbart. MOSPD2, a newly characterized protein, promotes breast cancer metastasis [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 4892. doi:10.1158/1538-7445.AM2017-4892
Abstract 4894: Identification of small molecules for the prevention and treatment of colon cancer
Authors: Pal Deeksha , Baby Becca Von , Chandrasekaran Balaji , Sharma Arun , Damodaran Chendil
Published: July 2017

Metastatic colon cancer or colorectal cancer (CRC) is associated with poor survival rate which is less than 5% due to chemo resistance. We previously showed that overexpression of AKT and Notch1 are key players for progression of metastasis of colon cancer cells. Withaferin-A (WA) is a potent herbal molecule that inhibited CRC growth by down regulating AKT and Notch1 signaling in both in vitro and in vivo colon cancer models. We found that WA binds to AKT with a predicted binding affinity of -10.6 kcal/mol and inhibited its activation. So, we designed and developed several analogs of WA, that exhibits a stronger AKT binding (binding energy higher than -10 kcal/mole) and more potent than the parental WA. UL16 is one such compound which inhibited the growth of colon cancer cells (HCT-116 and SW-620: IC50-300nM) at nanomolar concentration. Western blot analysis revealed that UL16 inhibits AKT/NF-κβ/ Bcl2- signaling axis in colon cancer cells. In addition, UL16 concurrently inhibited Notch1 signaling pathway in colon cancer cells. Then we systematically investigated the anti-migratory and anti-invasive effect of UL16by cell migration and invasion assays. Inhibition of both migration and invasion of colon cancer cells corresponds with downregulation of mesenchymal markers of Snail, Slug and β-catenin expression. We also examined the in vivo efficacy of UL16 in ApcMin/+ mouse model. Treatment of ApcMin/+ with UL16 (0.5 mg/kg/IP/twice a week) over 12 weeks significantly reduced the number of intestinal polyps (distal 84%; Middle 63% and Proximal 2%) as compared vehicle treated mice. In addition, we also observed 50% reduction in number of colon tumors in UL16 treated mice. Our ongoing immunohistochemistry studies may confirm the in vitro mechanistic results. In conclusion, our studies suggest that UL16 inhibits both AKT and Notch1 signaling and suppresses intestinal polyp formation in ApcMin/+ mice. Hence UL16could be a viable therapeutic agent for treating patients with metastatic colon cancer.

Citation Format: Deeksha Pal, Becca Von Baby, Balaji Chandrasekaran, Arun Sharma, Chendil Damodaran. Identification of small molecules for the prevention and treatment of colon 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 4894. doi:10.1158/1538-7445.AM2017-4894
Abstract 4899: The small molecule cathepsin L and K inhibitor KGP-94 impairs the metastatic phenotype of osteosarcoma cells
Authors: Fasanya Henrietta , Siemann Dietmar
Published: July 2017

Osteosarcoma (OS) is the most common primary malignant bone cancer and primarily affects adolescents between the ages 10-20 years old. Today, OS remains the second leading cause of cancer-related death for patients in this age group. With current therapy, the 5-year survival for patients with clinically metastatic disease has remained unchanged at 20% since the early 1970s. There is a significant need for novel therapies that will increase overall survival for patients with metastatic OS.

Proteases secreted within the tumor microenvironment enable primary tumor development and potentiate the ability of OS cells to metastasize. Typically, cysteine proteases under normal physiologic conditions function primarily within the lysosome to facilitate protein cell turnover. During tumorigenesis, Cathepsin L (CTSL) is secreted into the tumor environment by the tumor. CTSL secretion potentiates metastasis through its regulation of cell motility, extracellular matrix degradation, angiogenesis, and bone remodeling. Cathepsin K (CTSK) is overexpressed in OS and is secreted during bone remodeling by osteoclast to degrade the bone matrix. The bone environment is hypoxic which induces proteomic changes that enable OS cells to survive in a nutrient deprived environment and increase CTSL/CTSK secretion by the tumor cells.

The goal of these studies was to determine if CTSL/CTSK inhibition via KPG-94 (3-bromophenyl-3-hydroxyphenyl-ketone thiosemicarbazone) could decrease the metastatic phenotype of human osteosarcoma cells under normoxic and hypoxic conditions. The Hu09 cell line has been characterized as an osteoblastic cell line with the potential to grow orthotopically in vivo. Hu09 and its metastatic sublines Hu09-L6 and Hu09-M112 have been characterized to low and high metastatic capacity, respectively. Treatment with KGP-94 at 10 and 25 uM effectively decreases cell motility and invasion of Hu09 and its metastatic sublines under normoxic and hypoxic conditions.

Our results demonstrate that dual CTSL/CTSK inhibition is effective at decreasing two signature methods of measuring metastasis: 1) cell motility and 2) invasiveness. Targeting CTSL/CTSK may provide a novel therapeutic approach for patients with metastatic OS.

Citation Format: Henrietta Fasanya, Dietmar Siemann. The small molecule cathepsin L and K inhibitor KGP-94 impairs the metastatic phenotype of osteosarcoma 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 4899. doi:10.1158/1538-7445.AM2017-4899
Abstract 4901: Urokinase-derived peptide UP-7 effectively inhibits metastatic growth of breast cancer through suppression of FAK activation
Authors: Joe Young Ae , Kim Hyun-Kyung , Naidansuren Purevjargal , Lee Seung Woo , Kim Rae-Kwon , Lee Su-Jae , Lee Suk Keun , Hong Yong-Kil
Published: July 2017

The recombinant kringle domain of urokinase-type plasminogen activator (UK1) has been shown to inhibit angiogenesis in vitro and in vivo and suppress brain tumor in vivo. To avoid limitations in application due to mass production of recombinant proteins, we dissected UK1 sequences to seven peptides based on structure and amino acid characteristics, and examined the anti-angiogenic activities for the constructed peptides. UP-7 peptide derived from ß-sheets region of UK1 was the most potent inhibitor of the proliferation and migration of endothelial cells (ECs) in vitro, and it also inhibited in vivo angiogenesis in the mouse matrigel plug assay. Such anti-angiogenic activities were not exerted by the scrambled peptide. At molecular level, UP-7 inhibited the VEGF or bFGF-induced phosphorylation of FAK and ERK1/2 and suppressed formation of stress fibers and focal adhesions. It also inhibited the attachment and spreading of ECs onto immobilized fibronectin. In a lung cancer animal model xenografted with UP-7-non-sensitive NCI H460, systemic treatment of UP-7 effectively suppressed tumor growth through inhibition of angiogenesis. Interestingly, breast cancer cells such as MDA-MB231 were sensitive to UP-7 in proliferation differently from other cancer cells. It inhibited the migration and invasion of LM-2-MDA-MD231 and suppressed the phosphorylation of FAK. Accordingly, UP-7 potently inhibited lung metastatic growth of LM2-MDA-MB-231 in experimental metastasis model. Taken together, these results suggest that novel UK1-derived peptide, UP-7 can be effectively used for treatment of the metastatic growth of breast cancer. (NRF-2012R1A1A2007175, NRF-2012R1A5A2051476)

Citation Format: Young Ae Joe, Hyun-Kyung Kim, Purevjargal Naidansuren, Seung Woo Lee, Rae-Kwon Kim, Su-Jae Lee, Suk Keun Lee, Yong-Kil Hong. Urokinase-derived peptide UP-7 effectively inhibits metastatic growth of breast cancer through suppression of FAK activation [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 4901. doi:10.1158/1538-7445.AM2017-4901
Abstract 4903: Cationic polymer inhibits pancreatic cancer invasion in vitro and metastasis in vivo
Authors: Naqvi Ibtehaj A. , Gunaratne Ruwan , McDade Jessica , Rouse Douglas , Yeh Jen Jen , Pisetsky David , Lee Jaewoo , White Rebekah , Sullenger Bruce
Published: July 2017

Pancreatic cancer has the poorest prognosis of all major cancers with a 5-year survival of less than 5%. Cell-free DNA (cfDNA) has been well-validated as a biomarker that correlates with tumor burden. More recently, a role for cfDNA in facilitating tumor progression has also been identified. The latter occurs through activation of nucleic acid sensing receptors such as toll-like receptors (TLRs) in both tumor cells and the host environment, which can upregulate pro-metastatic signaling pathways. In addition, tumor-derived extracellular vesicles such as microparticles and exosomes have also been implicated in promoting metastasis by activating pro-invasive pathways in tumor cells and pre-conditioning secondary sites for metastatic establishment. Our laboratory has previously shown that cationic polymers can scavenge negatively charged nucleic acids like cfDNA and abrogate aberrant inflammation in disease models of autoimmunity and infection. Herein, we investigated the ability of the cationic polymer, PAMAM-G3, to bind and inhibit cfDNA as well as anionic tumor-derived extracellular vesicles using in vitro models of pancreatic cancer cell line invasion. We also evaluated the effect of PAMAM-G3 treatment in a bioluminescent syngeneic murine model of pancreatic cancer metastasis. PAMAM-G3 significantly inhibited in vitro invasion of pancreatic cancer cell lines in response to both cfDNA and tumor-derived extracellular vesicles. Moreover, biweekly intraperitoneal treatment with PAMAM-G3 (20 mg/kg) starting 48 hours after pancreatic tumor cell implantation led to a significant reduction in liver metastasis without affecting primary tumor growth. Thus, cationic polymers such as PAMAM-G3 may represent a novel class of therapeutics to combat pancreatic cancer metastasis.

Citation Format: Ibtehaj A. Naqvi, Ruwan Gunaratne, Jessica McDade, Douglas Rouse, Jen Jen Yeh, David Pisetsky, Jaewoo Lee, Rebekah White, Bruce Sullenger. Cationic polymer inhibits pancreatic cancer invasion in vitro and metastasis in vivo [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 4903. doi:10.1158/1538-7445.AM2017-4903
Abstract 4906: Sphingolipid regulates pro-inflammatory cytokines to subdue the neoplastic lesions in hepatocellular carcinoma
Authors: Chatterjee Nabanita , Saha Krishna Das
Published: July 2017

Hepatocellular carcinoma (HCC), is the most leading cause of cancer-related death in all over the world (seems to more than 80% of death cases). It is frequently linked with continuous hepatocytes death, inflammatory cell infiltration, and compensatory liver regeneration with higher angiogenic rate. Considering the different signalling process during this tumorogenesis with the involvement of new ‘lead’ identification of different sources, desire to be less toxic and higher bioavailability. The use of live, attenuated or genetically compensated microbes or its cellular component(s) or metabolites has begun to emerge as a potential new approach in medicinal research to deliver bioactive entities. Thus, advancing our knowledge of such microbes-mediated therapy may suggest new avenues for therapeutic intervention in many fatal diseases. Lipid (LSPL-1) isolated from Leishmania donovani could induce apoptosis in human HepG2 (Hepatocellular carcinoma cell, human) cells, as characterized by phosphatydylserine serine exposure, DNA fragmentation, alteration of BaX/Bcl2 ratio with loss of mitochondrial membrane potential and cytochrome C release, activation of series of caspases and cleaved PARP. LSPL-1 also modulates the various transcription factors like NF-kb65, p53 through regulation of cell proliferation exerted the death cause. Moreover this study also focused that the LSPL-1 effectively suppressed diethylnitrosamine (DEN) induced liver inflammation and hyperplasia in murine also decline in lipid peroxidation and increase in enzymic antioxidants (SOD, CAT, GPx, GR and GST) status with improvement of their survival rate. It reduced the elevated level of cytokines including tumor necrosis factors, interleukins, interferon (alpha and gamma), and transforming growth factors in hepatocytes from liver fibrosis bearing mice. As evident from immunochemical analysis, inflamed hepatocytes and liver tissue may up regulated the various growth factors including EGF, HGF, endothline-1 regulation with promoting angiogenic activity via modulation of CD34, VEGF, HIF-1α expression and alteration of these changes were balanced by LSPL-1. So taken together, these finding indicated that, LSPL-1 may induce apoptosis and in hyper proliferating cancer cells via altering cytokines level with tumour growth regulation in carcinogen exposed rodent system.

Note: This abstract was not presented at the meeting.

Citation Format: Nabanita Chatterjee, Krishna Das Saha. Sphingolipid regulates pro-inflammatory cytokines to subdue the neoplastic lesions in 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 4906. doi:10.1158/1538-7445.AM2017-4906
Abstract 4915: Alcohol-induced reprogramming of tumor plasticity is mediated via suppression of CCN5 signaling in breast cancer cells
Authors: Haque Inamul , Subramanian Arvind , Gupta Vijayalaxmi , Sarkar Sandipto , Banerjee Snigdha , Banerjee Sushanta K.
Published: July 2017

Background and Objective: The epidemiological and experimental studies suggest that alcohol consumption is associated with increased risk for breast cancer development and metastasis. However, the mechanisms of alcohol-induced breast cancer progression and metastasis remain unknown. In this study, we examined the roles of CCN5, a tumor suppressor gene in alcohol induced breast cancer cell migration/invasion, epithelial to mesenchymal transition (EMT) and stemness.

Methods: Estrogen receptor-positive (ER+) human breast cancer cell line MCF-7 and immortalized normal human epithelial breast cell line MCF-12A were treated with ethanol at various concentrations in the presence and absence of human recombinant human CCN5 protein (hrCCN5) . Invasive/migratory ability of treated and untreated cells was measured by Boyden chamber assays. The mRNA and protein expression level of CCN5 was determined by real-time qRT-PCR and Western blot. EMT and stemness markers were evaluated by Western blot. Single-cell suspensions from pre-treated cells were re-suspended at a density of 500 cells/ml mammocult media in ultralow attachment dishes. Number as well as the size of the mammosphere in specified experimental set-up was monitored and recorded alternate day for 8-10 days.

Results: The studies demonstrated that alcohol promotes significantly the invasive/migratory ability and EMT phenotypes of breast cancer cells through the suppression of CCN5. Interestingly, hrCCN5 protein treatment suppresses the effect of alcohol and reprograms MET (mesenchymal to epithelial transition). In addition, alcohol-induced mammosphere formation efficiency is also suppressed by hrCCN5 protein-treatment.

Conclusions: Our findings suggest that the CCN5 signaling plays a preventive role in alcohol-induced reprogramming of tumor plasticity in breast cancer cells.

Citation Format: Inamul Haque, Arvind Subramanian, Vijayalaxmi Gupta, Sandipto Sarkar, Snigdha Banerjee, Sushanta K. Banerjee. Alcohol-induced reprogramming of tumor plasticity is mediated via suppression of CCN5 signaling 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 4915. doi:10.1158/1538-7445.AM2017-4915
Abstract 4916: E3 ubiquitin ligase Cbl-b prevents tumor metastasis by maintaining the epithelial phenotype in multiple drug-resistant gastric and breast cancer cells
Authors: Xu Ling , Qu Xiujuan , Zhang Ye , Che Xiaofang , Guo Tianshu , Cai Ying , Li Aodi , Li Danni , Li Ce , Wen Ti , Fan Yibo , Hou Kezuo , Ma Yanju , Hu Xuejun , Liu Yunpeng
Published: July 2017

Background: Multiple drug resistance (MDR) and metastasis are two major unfavorable factors causing failure of cancer treatment. However, the relationship between MDR and metastasis is not characterized. Additionally, the role of the E3 ubiquitin ligase Cbl-b in metastasis of MDR gastric and breast cancer is little known.

Methods: The effect of Cbl-b on tumor invasion and metastasis was analyzed in P-gp-positive gastric adenocarcinoma tissues. Transfections of Cbl-b WT and epidermal growth factor receptor (EGFR) WT plasmid were used to assess Cbl-b and EGFR involvement in epithelial-mesenchymal transition (EMT) and tumor metastasis.

Results: MDR gastric and breast cancer cells possess a typical mesenchymal phenotype and an enhanced cell migration capacity. Additionally, Cbl-b is poorly expressed in MDR gastric and breast cancer cells. We performed immunohistochemical analysis on P-gp-positive gastric adenocarcinoma tissues and identified associations between Cbl-b expression and clinical pathological parameters. Gastric cancer patients with low Cbl-b expression were more likely to have tumor invasion (P=0.016) and lymph node metastasis (P=0.007). Moreover, over-expression of Cbl-b reduced cell migration in MDR cell cultures both in vitro and in vivo. Cbl-b over-expression also prevented EMT by inducing ubiquitination and degradation of EGFR, leading to inhibition of the EGFR- ERK/Akt-miR-200c-ZEB1 axis. However, further over-expression of EGFR on a background of Cbl-b over-expression restored both the mesenchymal phenotype and the cell migration capacity of MDR gastric and breast cancer cells.

Conclusions: These results suggest that Cbl-b is an important factor for maintenance of the epithelial phenotype and inhibition of cell migration in MDR gastric and breast cancer cells.

Note: This abstract was not presented at the meeting.

Citation Format: Ling Xu, Xiujuan Qu, Ye Zhang, Xiaofang Che, Tianshu Guo, Ying Cai, Aodi Li, Danni Li, Ce Li, Ti Wen, Yibo Fan, Kezuo Hou, Yanju Ma, Xuejun Hu, Yunpeng Liu. E3 ubiquitin ligase Cbl-b prevents tumor metastasis by maintaining the epithelial phenotype in multiple drug-resistant gastric and 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 4916. doi:10.1158/1538-7445.AM2017-4916
Abstract 4920: Inflammation and estrogen receptor beta modulate bacterial diversity of the gut
Authors: Ibrahim Ahmed F. , Thomas Quentin , Saxena Ashish , Williams Cecilia
Published: July 2017

Background: Chronic inflammation is a risk factor of colorectal cancer (CRC), and the microbiota of the gut is important in both colitis and carcinogenesis. Estrogen receptor beta (ERβ) is the predominant nuclear receptor in the colon, and is downregulated in CRC. Here, we explore the role of colitis and ERβ signaling with respect to the microbiota diversity.

Methods: Intestinal ERβ-/- mice were generated using the Cre-loxP system. WT and knockout mice were treated with AOM/DSS inducing colitis (at 9 weeks) and CRC (at 16 weeks). After 9 and 16 weeks, stool pellets were collected from treated and untreated mice (n=39), and DNA was extracted. Library preparation and 16S rRNA-sequencing was performed using the illumina Miseq platform with specific primers targeting the V3-V4 regions, and QIIME software was used to analyze microbiota diversity. Sequencing results were validated by qPCR

Results: High-throughput 16S rRNA gene sequencing demonstrated that treatment induced significant differentiation of both alpha (p=0.01) and beta (p=0.01) microbiota diversity. Alpha diversity indicators implied a significant reduction of microbiota richness and evenness after treatment. qPCR confirmed that 9-weeks treatment increased OTUs affiliated with Bifidobacteria (p=0.03), Bacteroidetes (p=0.04), Lactobacilli (p=0.0014), Cyanobacteria (p=0.0023) and Gammaproteobacteria (p=0.05) in WT mice. After 16 weeks treatment, 16s rRNA sequencing show enrichment of Tenercutes, Proteobacteria and Cyanobacteria in all tumor-bearing mice (treated WT and ERβ-/-), while Verrucomicrobia increased in untreated mice (No tumors). Further treated ERβ-/- but not WT mice displayed significant reduction of Gammaproteobaceria (p=0.0083).

Conclusion: Our data demonstrate a clear effect of colitis-induced treatment on the microbiota, and that ERβ attenuates this response. Lactobacilli and Bifidobacteria has, for example, been shown to attenuate inflammatory response. We note an increase in Lactobacilli and Bifidobacteia after treatment in WT mice, but not in ERβ-/- mice. This can result in increased colitis in ERβ-/- mice and help explain the increased expression of pro-inflammatory cytokines we note in treated ERβ-/- mice.

Citation Format: Ahmed F. Ibrahim, Quentin Thomas, Ashish Saxena, Cecilia Williams. Inflammation and estrogen receptor beta modulate bacterial diversity of the gut [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 4920. doi:10.1158/1538-7445.AM2017-4920
Abstract 4923: MyD88 and IL-17 have essential local and systemic functions in promoting tumors on mouse skin
Authors: Klosterman Mary , Cataisson Christophe , Salcedo Rosalba , Shibuya Kelly , Waters Jennifer , Trinchieri Giorgio , Yuspa Stuart H.
Published: July 2017

NF-kB has been causally implicated with cancer-associated inflammation. In the skin, ligands of the Toll-like receptor/IL-1 receptor superfamily induce the recruitment of MyD88 (Myeloid differentiation primary response gene 88), ultimately leading to NF-kB activation. We previously reported that MyD88 is required for two stage chemical carcinogenesis on mouse skin, and that MyD88 exerts a cell-intrinsic function in RAS-mediated transformation of keratinocytes (Ras-keratinocytes). Now, we explore the role of MyD88 during tumor promotion with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Epidermal hyperplasia is reduced in MyD88 deficient mice relative to controls after multiple applications of TPA. Furthermore, the upregulation of the chemokines Cxcl1 and S100a8 in response to TPA was deficient in MyD88 null cultured keratinocytes revealing a necessary role for MyD88 in the intrinsic response of keratinocytes to phorbol ester. We also uncovered a systemic requirement for MyD88 in carcinogenesis since lineage specific ablation of MyD88 in T cells conferred significant resistance to skin tumorigenesis and reduced infiltration of IL-17 positive T cells in TPA treated MyD88 null mouse skin. IL-17 treatment of normal primary keratinocytes increased their proliferation while reducing their response to calcium-induced differentiation but had no effect on the proliferation of RAS-keratinocytes. The atypical nuclear IkBzeta protein can act as an activator of so-called secondary NF-kB response genes. IkBzeta deficiency blocked the keratinocyte proliferative response to IL-17 but had no effect on altered differentiation. Finally, both IL-17 and oncogenic RAS required IkBzeta to upregulate the expression of S100a8, lipocalin2 and Steap4 mRNAs. Collectively, these results demonstrate that MyD88 plays a pro-tumorigenic function during tumor promotion in both epithelial and T-cell compartments and that IL-17/IkBzeta could play a facilitating role on keratinocytes during tumor promotion beyond its pro-inflammatory activity.

Citation Format: Mary Klosterman, Christophe Cataisson, Rosalba Salcedo, Kelly Shibuya, Jennifer Waters, Giorgio Trinchieri, Stuart H. Yuspa. MyD88 and IL-17 have essential local and systemic functions in promoting tumors on mouse skin [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 4923. doi:10.1158/1538-7445.AM2017-4923
Abstract 4925: Microbiome-TP53 gene interaction in human lung cancer
Authors: Greathouse Leigh , White James , Bliskovsky Valery , Vargas Ashley , Polley Eric , Bowman Elise , Khan Mohammed , Robles Ana , Ryan Brid , Dzutsev Amiran , Trinchieri Giorgio , Pineda Marbin , Bilke Sven , Meltzer Paul , Walther-Antonio Marina , Ehrlich Garth , Mell Joshua , Earl Joshua , Balashov Sergey , Bhat Archana , Valm Alex , Deming Clayton , Conlan Sean , Oh Julia , Segre Julie , Harris Curtis
Published: July 2017

Lung cancer is the leading cancer diagnosis worldwide and the number one cause of cancer deaths. Exposure to cigarette smoke, the primary risk factor in lung cancer, reduces epithelial barrier function and increases susceptibility to infections. Herein, we hypothesized that somatic mutations together with cigarette smoke create a dysbiotic microbiota that is associated with lung carcinogenesis. Using lung tissue from controls (n=33) or cancer cases (n=143), we conducted 16S rRNA gene sequencing (MiSeq), with RNA-seq data from lung cancer cases in The Cancer Genome Atlas (n=1112) serving as the validation cohort. We demonstrate a lower alpha diversity in normal lung as compared to non-tumor adjacent or tumor tissue, indicating a shift in the overall microbial community in lung cancer patients as compared to those without cancer. Lung cancer cases were classified by the relative abundance of two taxa, Variovorax and Streptococcus, with an increase in Variovorax abundance in tumors as compared to non-tumor adjacent paired lung tissue (FDR corrected P=0.013). The species of Variovorax was identified histologically, and also by two additional 16S rRNA strategies (Resphera Insight analysis and PacBio sequencing). A group of taxa were associated with squamous cell carcinoma (SCC), of which Acidovorax were enriched in smokers (P =0.0013). Further, these taxa, including Acidovorax, exhibited higher abundance among the subset of SCC cases with TP53 mutations, an association not seen in adenocarcinomas (AD). Therefore, we observed a microbiome-gene and a microbiome-exposure interaction in SCC lung cancer tissue. Together, these results open the door for future biomarker research that could be used to improve screening and direct mechanistic studies of lung cancer therapy.

Citation Format: Leigh Greathouse, James White, Valery Bliskovsky, Ashley Vargas, Eric Polley, Elise Bowman, Mohammed Khan, Ana Robles, Brid Ryan, Amiran Dzutsev, Giorgio Trinchieri, Marbin Pineda, Sven Bilke, Paul Meltzer, Marina Walther-Antonio, Garth Ehrlich, Joshua Mell, Joshua Earl, Sergey Balashov, Archana Bhat, Alex Valm, Clayton Deming, Sean Conlan, Julia Oh, Julie Segre, Curtis Harris. Microbiome-TP53 gene interaction in human 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 4925. doi:10.1158/1538-7445.AM2017-4925
Abstract 4928: Impact of E. hirae on the tumoricidal activity of cyclophosphamide
Authors: Daillere Romain , Vetizou Marie , Zitvogel Laurence
Published: July 2017

The tumor microenvironment is influenced by anticancer therapies, and even more so by those affecting the gut homeostasis. We previously reported that a deviated repertoire of the intestinal microbiome called « dysbiosis », caused by broad spectrum antibiotics compromised the efficacy of cyclophosphamide (CTX), an immunomodulatory alkylating agent exerting cytotoxic effects against cancer, highlighting the importance of the gut microbiota in this mechanism (Viaud, Science, 2013). CTX is responsible for disrupting the gut barrier integrity as well as intestinal homeostasis, allowing a NOD2-dependent translocation of several Gram-positive bacteria into secondary lymphoid organs. CTX breaks the intestinal tolerance towards the intestinal microbiota and leads to immunization of the host against some bacterial strains. Here, we identified a Gram positive bacteria, Enterococcus hirae, which markedly modulates the intestinal and systemic immunity through the elicitation of bacterial-specific Th1 and pathogenic Th17 cells. Moreover, we have shown that E. hirae is capable of enhancing tumor-specific CD4+ and CD8+ T cell responses against candidate tumor antigens. In parallel, we have demonstrated that intestinal epithelial cells NOD2 receptors represent ‘‘gut immune checkpoints’’ restricting the immunogenicity of E. hirae. Finally, E. hirae specific-memory Th1 immune responses selectively predicted longer progression-free survival in advanced lung or ovarian cancer patients treated with immunotherapy and chemotherapy (platinum salts- or CTX-based chemotherapy) (Daillère, Immunity, 2016). Altogether, E. hirae represents a valuable probiotic against cancer, an oncomicrobiotic ameliorating the efficacy of the most common alkylating immunomodulatory compound.

Citation Format: Romain Daillere, Marie Vetizou, Laurence Zitvogel. Impact of E. hirae on the tumoricidal activity of cyclophosphamide [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 4928. doi:10.1158/1538-7445.AM2017-4928
Abstract 4930: Genetic and epigenetic characteristics of esophageal cancer tissues with microbiome fusobacterium nucleatum
Authors: Baba Yoshifumi , Yamamura Kensuke , Nakagawa Shigeki , Mima Kosuke , Ishimoto Takatsugu , Iwatsuki Masaaki , Sakamoto Yasuo , Yoshida Naoya , Baba Hideo
Published: July 2017

Background: Research into the microbiome is a rapidly advancing field in human cancers. The gut microbiome has recently been shown to play an important role in health, as well as in diseases such as obesity, inflammatory bowel disease, diabetes, non-alcoholic fatty liver disease, and several types of cancers. Fusobacterium nucleatum is a component of the human microbiome that primarily inhabits the oral cavity. It causes periodontal disease and has also been implicated in the development of human cancers. We have recently reported that Fusobacterium nucleatum in esophageal cancer tissues was associated with shorter survival, suggesting a potential role as a prognostic biomarker. In addition, our KEGG enrichment analysis using microarray data showed that Fusobacterium nucleatum might also contribute to aggressive tumor behavior through activation of chemokines such as CCL20. Nonetheless, molecular features of Fusobacterium nucleatum-positive esophageal cancers have been still unknown. Methods: Using 325 esophageal cancers, we detected Fusobacterium nucleatum DNA in 74 tumors (23%). We assessed other genetic and epigenetic events including mutations in KRAS, BRAF, PIK3CA gene, p53 expression, phosphorylated-AKT expression, LINE-1 methylation level (global DNA methylation level), IGF2 DMR0 methylation, MGMT methylation, and MLH1 methylation. Results: The LINE-1 methylation levels of Fusobacterium nucleatum-positive tumors was significantly lower compared with that of Fusobacterium nucleatum-negative tumors (P<0.0001). In addition, Fusobacterium nucleatum was significantly associated with PIK3CA wild type (P=0.010). There was no relationship of Fusobacterium nucleatum with KRAS, BRAF mutation, p53 expression, p-AKT expression, IGF2 DMR0 methylation, MGMT methylation, or MLH1 methylation. Conclusion: Fusobacterium nucleatum was related with LINE-1 hypomethylation and PIK3CA wild type. Our findings may shed lights on understanding the mechanism by which Fusobacterium nucleatum affects aggressive tumor behavior. .

Citation Format: Yoshifumi Baba, Kensuke Yamamura, Shigeki Nakagawa, Kosuke Mima, Takatsugu Ishimoto, Masaaki Iwatsuki, Yasuo Sakamoto, Naoya Yoshida, Hideo Baba. Genetic and epigenetic characteristics of esophageal cancer tissues with microbiome fusobacterium nucleatum [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 4930. doi:10.1158/1538-7445.AM2017-4930
Abstract 4932: Oxygen sensing by T cells establishes an immunologically favorable metastatic niche
Authors: Clever David , Restifo Nicholas
Published: July 2017

Cancer cells must evade immune responses at distant sites to establish metastases. The lung is a frequent site for metastasis. We hypothesized that lung-specific immunoregulatory mechanisms create an immunologically permissive environment for tumor colonization. We found that T-cell-intrinsic expression of the oxygen-sensing prolyl-hydroxylase (PHD) proteins is required to maintain local tolerance against innocuous antigens in the lung but powerfully licenses colonization by circulating tumor cells. PHD proteins limit pulmonary type helper (Th)-1 responses, promote CD4(+)-regulatory T (Treg) cell induction, and restrain CD8(+) T cell effector function. Tumor colonization is accompanied by PHD-protein-dependent induction of pulmonary Treg cells and suppression of IFN-γ-dependent tumor clearance. T-cell-intrinsic deletion or pharmacological inhibition of PHD proteins limits tumor colonization of the lung and improves the efficacy of adoptive cell transfer immunotherapy. Collectively, PHD proteins function in T cells to coordinate distinct immunoregulatory programs within the lung that are permissive to cancer metastasis.

Note: This abstract was not presented at the meeting.

Citation Format: David Clever, Nicholas Restifo. Oxygen sensing by T cells establishes an immunologically favorable metastatic niche [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 4932. doi:10.1158/1538-7445.AM2017-4932
Abstract 4938: A vicious cycle between osteoblasts-derived WISP-1 and mesenchymal-like cancer cells is essential to prostate cancer metastasis
Authors: Chang An-Chen , Chuang Show-Mei , Tang Chih-Hsin
Published: July 2017

Prostate cancer (PCa) is the most commonly diagnosed malignancy in the United States and other Western countries. It is well established that tumor microenvironment plays an important role in cancer progression and metastasis. Wnt-induced secreted protein-1 (WISP-1) belongs to the CCN family (CTGF/CYR61/NOV) that plays a key role in osteogenesis and tumor growth. We found that osteoblast-conditioned media (OBCM) stimulated migration and invasion in human PCa cells. Furthermore, the medium was collected 2 days after treatment of osteoblasts with PCa conditioned media which were designed as PCa-OBCM. The results showed that treatment of PCa cells with PCa-OBCM further increased the migration and invasion activity, as well as epithelial-mesenchymal transition (EMT), including the epithelial marker (E-cadherin) down-regulation and mesenchymal marker (N-cadherin and twist) up-regulation. Moreover, human cytokine array was used to measure the secretion of 32 Cytokines into PCa-OBCM. The data showed that WISP-1 is highly up-regulated with PCa-OBCM treatment compared to OBCM. Stimulation of osteoblasts with PCa conditioned media induced WISP-1 expression. In contrast, knockdown WISP-1 in osteoblasts impacted PCa-OBCM-mediated cancer migration, invasion, and EMT. In addition, conditioned media from PCa cells was incubated with another cytokine array membranes. The data showed that ADAM-9 and cathepsinL were overexpressed in PCa conditioned media but only ADAM-9 could regulate WISP-1 expression in osteoblasts. Taken together, these results demonstrated that primary PCa-secreted ADAM-9 promotes osteoblasts-derived WISP-1 up-regulation, which in turn enhances primary PCa cells migration, invasion, and EMT, resulting in cancer metastasis. Our study suggests that inhibition of osteoblasts-derived WISP-1 is a new therapeutic target for treating metastasis in human PCa patients.

Citation Format: An-Chen Chang, Show-Mei Chuang, Chih-Hsin Tang. A vicious cycle between osteoblasts-derived WISP-1 and mesenchymal-like cancer cells is essential to prostate cancer metastasis [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 4938. doi:10.1158/1538-7445.AM2017-4938
Abstract 4939: Activation of GSK3β/β-catenin pathway promotes organ-specific metastasis in prostate cancer
Authors: Qin Siyuan , Song Qiong , Dai Jinlu , Tong Haibo , Keller Evan , Zhang Jian , Lu Yi
Published: July 2017

Metastasis is the major cause of treatment failure in various malignancies, including prostate cancer (PCa). Accumulating data has suggested that primary cancer cells prefer to spread to certain distant organs. To explore the mechanisms of different cancer cells preferentially metastasizing to certain organs, we established a prostate cancer lung metastasis model. Murine prostate cancer cells (RM1), which labeled with luciferase, were used in this study. RM1 cells were injected into left tibia of mouse (C57BL/6J), and monitored the tumor growth and metastasis every week by Bioluminescence Imaging. The mice would be sacrificed when metastatic tumor appeared in multi-organs. We isolated the lung metastatic tumor, made single cell suspension and amplified. The lung metastatic tumor cells were specifically metastasized to lung when the tumor cells were injected intratibially again. After several cycles we obtained highly lung metastatic tumor cells, named RM1-LM cells. Further, we investigated property and function changes in RM1-LM cells compared to parental cells (RM1) using in vitro assays. We firstly found the highly lung metastatic tumor cells showed more mesenchymal properties. Western blot results showed that hallmarks of EMT including E-cadherin, vimentin, snail, and ZO-1 were altered significantly in RM1-LM cells. Then, we observed that the highly lung metastatic tumor cells exhibited increased proliferative, colony formation, migratory and invasive abilities. Mechanistic analyses indicated that prostate cancer organ-specific metastasis mediated by the activation of GSK-3β/β-catenin pathway. The current study may provide potential therapeutic targets and diagnostic biomarkers of PCa metastasis. This work was supported by Natural Science Foundation of China (NSFC) Key Project (81130046); NSFC (81171993; 81272415); Guangxi Projects of China (2013GXNSFEA053004; 2012GXNSFCB053004).

Note: This abstract was not presented at the meeting.

Citation Format: Siyuan Qin, Qiong Song, Jinlu Dai, Haibo Tong, Evan Keller, Jian Zhang, Yi Lu. Activation of GSK3β/β-catenin pathway promotes organ-specific metastasis in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4939. doi:10.1158/1538-7445.AM2017-4939