bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5146: Administration of Parabacteroides distasonis in the attenuation of colorectal tumorigenesis in a carcinogenic murine model of colorectal cancer
Authors: Gacasan Camilo Anthony G.
Published: August 2018

Colorectal cancer (CRC) is the second leading cause of related deaths in the United States. Interindividual differences in genetic background result in a complex and dynamic system between the gut microbiome and variable phenotypic response to diet as it relates to CRC risk. Previous research from our laboratory investigated CRC susceptibility in an azoxymethane (AOM) murine model using FVB/NJ (FVB) and C57Bl/6J (B6) mice fed a ketogenic (high fat, no carbohydrate) American (high fat, high carbohydrate) or standard (moderate fat, moderate carbohydrate) mouse chow. Tumor multiplicity was dependent on both strain and diet with a four-fold increase reported in FVB mice fed ketogenic diet relative to those fed standard chow. This response was not observed in FVB mice fed the American diet or in B6 mice on any of the three diets. A subsequent study that investigated the physiological response to diet in multiple genetic backgrounds revealed a decrease in the abundance of the bacterial species Parabacteroides distasonis in FVB mice fed a ketogenic diet. This established an emphasis on the interaction between variable response to diet, gut microbiome and further implications to gut health. To further investigate this relationship in FVB mice we are using an AOM-induced carcinogenic model on FVB mice fed a ketogenic diet supplemented with a biweekly gavage of P. distasonis. Upon necropsy, tumor load of mice receiving the experimental gavage will be compared to the control mice. Droplet digital PCR will be used to quantify the absolute abundance of P. distasonis in fecal samples at several time points before and after the initiation of diet and administration of AOM. Ultimately, we aim to determine the relationship between P. distasonis and CRC tumorigenesis, which will lead to a further understanding of the etiology of colorectal cancer.

Citation Format: Camilo Anthony G. Gacasan. Administration of Parabacteroides distasonis in the attenuation of colorectal tumorigenesis in a carcinogenic murine model of colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5146.
Abstract 515: Integrative analysis of LncRNA-drug interactions in cancer identified EPIC1 as an oncogenic lncRNA that regulates breast cancer tumorigenesis and iBET resistance through regulating MYC transcriptional activity
Authors: Wang Yue , Wang Zehua , Yang Bo , Zhang Min , Yang Da
Published: August 2018

Emerging evidence suggests that lncRNAs can serve as promising biomarkers and therapeutic targets in cancer. Here, by integrating multiple dimensional pharmacogenomic data of 11,950 lncRNA in 5,605 tumors and 1,005 cancer cell lines, we build lncRNA drug response models for 265 anti-cancer compounds across 27 cancer types. Our study reveals the cancer cell lines can realistically recapitulate patient tumors in IncRNA genomic and epigenetic alterations. Using Elastic Net (EN) regression model combined with bootstrap cross validation, our analysis identified 162,327 unique lncRNA-drug interactions. The robustness of lncRNA based EN-models is validated by independent cancer cell line drug response data. By applying lncRNA EN-models of 49 FDA approved drugs to TCGA pan-cancer patient samples across 21 caner types, we have shown that the cancer cell line based EN-models could readily predict chemotherapy responses in patients with breast, stomach, thyroid, and colon rectal cancer. Further lncRNA-protein coding gene co-expression analysis reveals that drug-predictive lncRNAs regulated drug-metabolism and drug-target pathways, which provide underlying mechanism for lncRNAs' regulation of drug response. This analysis has identified a novel intergenic lncRNA, EPIC1, regulates breast cancer tumorigenesis and iBET (Bromodomain Inhibitors) resistance through directly interacting with MYC and regulating MYC transcriptional activity. Knockdown of EPIC1 in breast cancer cells leads to inhibition of colony formation, cell cycle arrest, and suppression of tumor growth in vitro and in vivo. Overexpression of EPIC1 increased MYC target expression, breast tumorigenesis in vitro and in vivo, and promote iBET resistance in breast cancer. Mechanistically, EPIC1 directly interacts with MYC through EPIC1's 129-283 nt region. EPIC1 knockdown reduces the occupancy of MYC protein to the promoters of its target genes (e.g., p21, CCNA2, CDC20, and CDC45) without influencing MYC expression. The oncogenic effect of EPIC1 can be abolished by MYC knockdown. To our best knowledge, this is the first study to link noncoding genotypes with drug response phenotypes in both cancer cell lines and patient tumors.

Citation Format: Yue Wang, Zehua Wang, Bo Yang, Min Zhang, Da Yang. Integrative analysis of LncRNA-drug interactions in cancer identified EPIC1 as an oncogenic lncRNA that regulates breast cancer tumorigenesis and iBET resistance through regulating MYC transcriptional activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 515.
Abstract 5150: Loss of CYLD promotes cell invasion via ALK5 stabilization in oral squamous cell carcinoma
Authors: Shinriki Satoru , Jono Hirofumi , Maeshiro Manabu , Nakamura Takuya , Guo Jianying , Li Jian-Dong , Yoshida Ryoji , Shinohara Masanori , Nakayama Hideki , Matsui Hirotaka , Ando Yukio
Published: August 2018

Oral squamous cell carcinoma (OSCC) has a very poor prognosis because of its highly invasive nature, and the 5-year survival rate has not changed appreciably for the past 30 years. Although cylindromatosis (CYLD), a deubiquitinating enzyme, is thought to be a potent tumor suppressor, its biological and clinical significance in OSCC is largely unknown. This study aimed to clarify the roles of CYLD in OSCC progression. Our immunohistochemical analyses revealed significantly reduced CYLD expression in invasive areas in OSCC tissues, whereas CYLD expression was conserved in normal epithelium and carcinoma in situ. Furthermore, downregulation of CYLD by siRNA led to acquisition of mesenchymal features and increased migratory and invasive properties in OSCC cells and HaCaT keratinocytes. It is interesting that CYLD knockdown promoted TGF-β signaling by inducing stabilization of TGF-β receptor I (ALK5) in a cell autonomous fashion. In addition, the response to exogenous TGF-β stimulation was enhanced by CYLD downregulation. The invasive phenotypes induced by CYLD knockdown were completely blocked by an ALK5 inhibitor. Also, lower CYLD expression was significantly associated with the clinical features of deep invasion, poor overall survival, and increased Smad3 phosphorylation, which is an indicator of activation of TGF-β signaling in invasive OSCC. These findings suggest that downregulation of CYLD promotes invasion with mesenchymal transition via ALK5 stabilization in OSCC cells.

Citation Format: Satoru Shinriki, Hirofumi Jono, Manabu Maeshiro, Takuya Nakamura, Jianying Guo, Jian-Dong Li, Ryoji Yoshida, Masanori Shinohara, Hideki Nakayama, Hirotaka Matsui, Yukio Ando. Loss of CYLD promotes cell invasion via ALK5 stabilization in oral squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5150.
Abstract 516: Long noncoding RNA NEAT1 promotes lung metastasis of soft tissue sarcoma
Authors: Huang Jianguo , Xu Eric , Sachdeva Mohit , Robinson Timothy , Qin Xiaodi , Zhang Dadong , Owzar Kouros , Gokgoz Nalan , Seto Andrew , Andrulis Irene , Wunder Jay , Okada Tomoyo , Singer Samuel , Lazar Alexander , Rubin Brian , Kirsch David G.
Published: August 2018

Soft tissue sarcomas (STS) are malignant tumors from diverse mesenchymal tissues. About 40% STS patients develop fatal lung metastasis with a median survival of 15 months. The mechanisms driving the development of lung metastasis in sarcoma patients are poorly understood. Therefore, our lab has developed a genetically engineered mouse model (GEMM) of high-grade primary STS with conditional mutations in Kras and Trp53 (KP) where 40% of mice tumors develop lung metastasis. This KP model recapitulates human patients with Undifferentiated Pleomorphic Sarcoma (UPS), one of the most common subtypes of STS diagnosed in adults. RNA sequencing (RNA-Seq) was performed on paired primary and lung metastases in KP mouse sarcomas and determined that the expression of the long non-coding RNA (lncRNA) Neat1 is increased in lung metastases compared to paired primary tumors. Real time PCR (qPCR) in 22 paired KP mouse primary sarcomas and lung metastases further confirmed that Neat1 is significantly upregulated in lung metastases. In addition, RNA-Seq data from 5 pairs of human primary sarcomas and matched lung metastases also showed that NEAT1 levels are increased in lung metastases. Furthermore, NEAT1 RNA in situ hybridization (ISH) on tissue microarrays (TMAs) of human primary UPS and lung metastases determined that the expression of NEAT1 is upregulated in lung metastases. Next, CRISPR/Cas9 technology was applied to delete Neat1 in primary mouse sarcoma cells and loss of expression of Neat1 was confirmed by qPCR and northern blot in knockout (KO) clones. In addition, loss of Neat1 significantly reduced lung metastasis in vivo following tail vein injection of these modified cells into nude mice. To further test that loss of Neat1 reduces lung metastasis in primary sarcoma mouse model, primary tumors with Neat1 deletion are generated in GEMMs using in vivo CRISPR/Cas9 technology and KrasLSL-G12D/+; Trp53Flox/Flox; Neat1-/- (KPN) mouse model. Finally, RNA-Seq and capture hybridization analysis of RNA targets (CHART) are performed to determine mechanisms by which Neat1 regulates lung metastasis. Overall, these results suggest that upregulation of Neat1 promotes lung metastasis of soft tissue sarcoma and NEAT1 is a potential target to prevent or treat lung metastasis in sarcoma patients.

Citation Format: Jianguo Huang, Eric Xu, Mohit Sachdeva, Timothy Robinson, Xiaodi Qin, Dadong Zhang, Kouros Owzar, Nalan Gokgoz, Andrew Seto, Irene Andrulis, Jay Wunder, Tomoyo Okada, Samuel Singer, Alexander Lazar, Brian Rubin, David G. Kirsch. Long noncoding RNA NEAT1 promotes lung metastasis of soft tissue sarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 516.
Abstract 5166: Lysyl oxidase regulates cell surface EGFR, and directionality of cancer cell invasion
Authors: Tang Haoran , Springer Caroline , Marais Richard
Published: August 2018

Lysyl oxidase (LOX) remodels the tumour microenvironment by cross-linking the extracellular matrix. LOX overexpression is associated with poor cancer outcomes. We find that LOX regulates the epidermal growth factor receptor (EGFR) to drive tumour progression. LOX regulates EGFR by suppressing TGFβ1 signalling through the secreted protease HTRA1. This increases the expression of Matrilin2 (MATN2), an EGF-like domain containing protein that traps EGFR at the cell surface to facilitate its activation by EGF. A novel pharmacological inhibitor of LOX, CCT365623, disrupts EGFR cell surface retention in vitro and delays the growth of primary tumour cells in vivo. In addition to its role in controlling primary tumour growth, LOX expression is also implicated in the processes of cancer cell invasion, and thereby the process of metastasis. Notably, we find that when LOX is depleted or inhibited, cancer cells are no longer able to sence a chemo-attractive gradient, and invade in a directional manner. Importantly, LOX produced by both cancer cells and fibroblasts can mediate this behaviour. Together, we show that LOX regulates EGFR cell surface retention to drive tumour progression. We also find that LOX does not regulate cancer cell invasion per se, but rather that it regulates the directionality of cancer cell migration, and both the cancer and stromal cells appear to contribute to this behaviour.

Citation Format: Haoran Tang, Caroline Springer, Richard Marais. Lysyl oxidase regulates cell surface EGFR, and directionality of cancer cell invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5166.
Abstract 5168: Understanding the role of integrins in breast cancer
Authors: Adorno-Cruz Valery , Liu Xia , Liu Huiping
Published: August 2018

Breast cancer is a highly prevalent disease and the second-leading cancer related death in women. Mortality rates have declined a 38% in recent years thanks to early detection and development of treatments. However, the five year survival rate of patients with a distant metastasis is only 26%. Hence, it is important to identify key players that mediate or promote the migration of cells to other organs. Integrins are cell surface receptors that mediate cell adhesion to the extracellular matrix. They are involved in roles such as cell migration and proliferation. In this project we seek to understand the role of integrin α2 in breast cancer metastasis. It has been previously reported that integrin α2 is a metastasis suppressor of a Her2+ model of breast cancer. However, its role in triple negative breast cancer is unknown. We found the integrin α2 gene (ITGA2) a target of has-miR-206 which controls metastasis and stemness of breast cancer. We hypothesize integrin α2 has a metastasis regulating role in triple negative breast cancer. The triple negative breast cancer cell line, MDA-MB-231 has been used to characterize the role of integrin α2 in migration, cell viability and lung colonization of mice injected though tail vein injection. Lung colonization of breast cancer cells in NSG mice is significantly altered by the knockdown expression of integrin α2. Consistent with the phenotypes in migration and invasion. Taken together, these results suggest integrin α2 regulates breast cancer metastasis through modified cell migration and lung colonization.

Citation Format: Valery Adorno-Cruz, Xia Liu, Huiping Liu. Understanding the role of integrins in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5168.
Abstract 5183: Using transposon elements to elucidate the genetic mechanisms of HCC-associated lung metastases
Authors: Lo Lilian H. , Chiu Amy P. , Li Xiao-Xiao , Tschida Barbara R. , Rowlands Dewi K. , Largaespada David A. , Keng Vincent W.
Published: August 2018

Metastases account for most cancer-related deaths as a result of primary cancer spreading to distant sites. Amongst human hepatocellular carcinoma (HCC), there is a drastic decrease in the survival rate observed in patients with early stage localized HCC to late stage metastasis-associated HCC. Importantly, the genetic mechanisms associated with metastasis-associated HCC remains elusive. Therefore, a forward genetic screen in mice using Sleeping Beauty (SB) insertional mutagenesis system was developed to identify genes associated with metastasis-associated HCC.

Truncated version of the epidermal growth factor receptor (Egfr) gene was frequently detected in both mouse primary liver tumors and lung metastases, which was confirmed by DNA sequencing and RNA sequencing. This indicates the importance of Egfr mutations for liver tumorigenesis and the metastasis process. More importantly, additional candidate genes involved in the metastasis process were also identified by our forward genetic screen and showed to be relevant in human HCC patients at the M1 cancer metastasis stage. Taken together, our study shows that the SB screen yields a high fraction of relevant events in human liver cancer and can provide valuable information on the evolution of metastasis-associated HCC.

The authors declare that they have no competing interests.

Citation Format: Lilian H. Lo, Amy P. Chiu, Xiao-Xiao Li, Barbara R. Tschida, Dewi K. Rowlands, David A. Largaespada, Vincent W. Keng. Using transposon elements to elucidate the genetic mechanisms of HCC-associated lung metastases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5183.
Abstract 5184: Expression and role of regulator of G-protein signing 5 in squamous cell carcinoma of the tongue
Authors: Abe Yushi , Yano Hirohisa , Akiba Jun , kondou Reiichirou , Ogasawara Sachiko , Nakamura Ken , Kusukawa Zingo
Published: August 2018

Objective: Regulator of G-protein signing (RGS) 5 is a member of the RGS family and acts as GTPase-activating proteins for heterotorimeric G protein αsubunits, negatively regulating G-protein signaling. RGS5 was reportedly expressed in the heart, lung, skeletal muscle and small intestine and relates with tumor angiogenesis and gestational hypertension. It was reported that RGS5 was related with the invasion and metastasis of cancers, such as non-small lung cancer and hepatocellular carcinoma. In the present study, we examined RGS5 expression and its relation with invasion in squamous cell carcinoma (SCC) of the tongue. Materials and Methods: For immunohistochemical analysis for RGS5, we used SCC tissues of the tongue obtained from 43 patients who underwent resection at Kurume University Hospital from 2011 to 2015. The staining intensity was graded as 0, negative; 1, weak; 2, moderate; 3, strong. The staining intensity in the normal epithelium was used as an internal control. We examined relationship between RGS5 expression in invasive portions and clinicopathological features (e.g., lymph node metastasis, depth of tumor, and mode of invasion, and so forth). Mode of invasion was classified into 3 levels, i.e., IFNa (expansive), IFNc (invasive), IFNb (between INFa and INFc). Depth of tumor was measured and classified into 2 levels: < 5 mm and ≥ 5mm. Comparison and estimation of cumulative survival rates were performed using the Kaplan-Meier curves and log rank test. Result: Non-cancerous squamous cells expressed RGS5 solely in nucleus. SCC cells in superficial portion of tumor nodules often expressed weak nuclear RGS5 expression. In invasive portions of the tumor nodules, SCC cells expressed significantly lower nuclear and higher cytoplasmic RGS5 expression. A significant correlation was found between the expression score of RGS5 at invasive portions and mode of invasion (P=0.009). When the 43 cases were subdivided into two groups, i.e., expression group (score 1-3) and non-expression group (score 0), significant differences were observed in mode of invasion (P=0.0042), lymph node metastasis (p=0.0314), depth of tumor (p=0.0239) between the two groups. The log-rank test revealed that postoperative early lymph node metastasis was significantly higher in the expression group (p= 0.0415). Conclusion: This study suggests that RGS5 expression is related with tumor invasion in SCC of the tongue and that RGS5 can be a predictor of postoperative early lymph node metastasis.

Citation Format: Yushi Abe, Hirohisa Yano, Jun Akiba, Reiichirou kondou, Sachiko Ogasawara, Ken Nakamura, Zingo Kusukawa. Expression and role of regulator of G-protein signing 5 in squamous cell carcinoma of the tongue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5184.
Abstract 5185: Identification of a new candidate therapeutic target for gastric cancer by in silico analysis
Authors: Kohmoto Tomohiro , Masuda Kiyoshi , Shoda Katsutoshi , Fujita Yuji , Tange Shoichiro , Ichikawa Daisuke , Otsuji Eigo , Imoto Issei
Published: August 2018

Gastric Cancer (GC) is a leading cause of global cancer mortality, with high incidence rates in Asia including Japan. Although recent studies have provided valuable insights into identification of major driver genes, there are still little therapeutic targets and prognostic markers of GC. Here, we report that a novel driver gene, OverExpressed in Gastric Cancer 1 (OEGC1), would induce cell proliferation and migration of GC.Analyzing gene expression profiles by RNA-Seq and relevant clinical data in TCGA dataset, we identified candidate genes whose mRNA expression was higher in tumor tissues than in non-tumorous and associated with poor prognosis. Through further expression analysis and Cox proportional hazards regression analysis of those candidates, we identified OEGC1 as the most prominent candidate for driver gene. OEGC1 mRNA was expressed highly especially in intestinal type GC. The prognosis of the OEGC1 high-expressed intestinal type GC patients was as poor as that of diffuse type GC patients. Immunohistochemical analysis showed that overexpression of OEGC1 was ectopically found in the tumor cell junctions located in the infiltrated region of tumor. By silencing endogenous OEGC1 using siRNAs, cell proliferation was significantly suppressed and G0/G1 arrest was induced in NUGC3, AGS and MKN7 cells with relatively higher OEGC1 expression. Transwell assay showed that the number of migrated cells was significantly lower for siRNA-OEGC1-transfected cells than for control cells. Our findings suggested that aberrant expression of OEGC1 might play an important role in tumorigenesis of GC, and that OEGC1 might be a possible therapeutic target in GC.

Citation Format: Tomohiro Kohmoto, Kiyoshi Masuda, Katsutoshi Shoda, Yuji Fujita, Shoichiro Tange, Daisuke Ichikawa, Eigo Otsuji, Issei Imoto. Identification of a new candidate therapeutic target for gastric cancer by in silico analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5185.
Abstract 5193: A subpopulation of circulating epithelial tumor cells positive for fatty acid receptor CD36 is linked to aggressiveness of breast cancer disease
Authors: Pizon Monika , Schott Dorothea , Pachmann Ulrich , Pachmann Katharina
Published: August 2018

Background: Breast cancer is one of the leading causes of cancer death for women worldwide. Major hurdles for a successful treatment are cancer metastases, resistance to therapy and disease recurrence. The presence of circulating epithelial tumor cells (CETCs) is closely related to metastasis formation, but the mechanisms through which CETCs promote recurrence of disease are still unclear. CD36 is found in the cell membrane of tumor cells and responsible for taking up fatty acid. It is known that CD36 is a general marker of metastatic cells and its amplification specifically correlated with metastasis in a large number of human tumors. Therefore, the aim of this study was to determine the metastatic marker CD36 on CETCs and to correlate it's frequency with clinicopathological features in breast cancer patients.

Methods: CETCs were determined from blood of 42 patients suffering from breast cancer. The number of vital CETCs and their expression of CD 36 was investigated using the maintrac® method.

Results: CD36 expressing CETCs were detected in 95 % of breast cancer patients. Breast cancer patients with metastatic disease had significantly more CD36 positive CETCs as compared to patients without metastases (median 72% vs. 48%; p<0.05). The fraction of CD36 positive CETCs was significantly higher in patients with HER2 positive primary tumor (69% vs. 54%; p<0.05). Furthermore, triple negative breast cancer patients had significantly more CD36 positive CETCs as compared to hormone receptor positive tissue (median 62% vs. 52%; p<0.05). Interestingly, frequency of CD36 positive CETCs correlated significantly with a blinded determination of Ki-67 proliferation index in tumor tissue (r=0.75; p<0.01). We observed a significant heterogeneity in CD36 immunostaining intensity across CETCs from the same patients.

Conclusion: We showed for the first time that breast cancer patients have detectable CETCs with a high frequency of CD36 which highly correlated with progression of cancer disease. Our findings suggest that using a CD36 antibody treatment may be an effective strategy to fight cancer.

Citation Format: Monika Pizon, Dorothea Schott, Ulrich Pachmann, Katharina Pachmann. A subpopulation of circulating epithelial tumor cells positive for fatty acid receptor CD36 is linked to aggressiveness of breast cancer disease [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5193.
Abstract 5194: In vivo capture of early metastatic cells through a CXCL12 loaded-dermal filler
Authors: Ieranò Caterina , D'Alterio Crescenzo , Giarra Simona , Napolitano Maria , Santagata Assunta , Barbieri Antonio , Campani Virginia , Luciano Antonio , Arra Claudio , Anniciello Annamaria , Botti Gerardo , Mayol Laura , Rosa Giuseppe De , Pacelli Roberto , Scala Stefania
Published: August 2018

The chemokine CXCL12 and its receptor CXCR4 are critical for orienting and seeding cells in metastatic sites. To distract CXCR4 expressing circulating tumor cells (CTCs) a “fake” pre-metastatic niche was built with a hyaluronic acid based dermal filler hydrogel-CXCL12-loaded (CLG). CXCR4 expressing human T leukemia cells, CCRF-CEM, and murine melanoma cells human CXCR4 transduced, B16-hCXCR4-GFP, were evaluated for migration in vitro toward CLG. CLGs and EGs (empty gels) were subcutaneously injected in C57BL/6 mice and five days later B16-hCXCR4-GFP (5x105 cells) were intravenously injected. Twenty two days later mice were euthanized and hydrogels recovered. Gel trapped cells were recovered through ialuronidase digestion of the gels and lungs were analyzed to detect metastases. Circulating tumor cells (CD45neg/hCXCR4 plus/GFP plus) were assessed in the blood at day 10 and 22 from cells injection. In vitro, CEM and B16-hCXCR4-GFP cells more efficiently migrated toward CLG compared with EG. In vivo, a significantly higher number of B16-hCXCR4-GFP tumor cells was detected within the CLGs compared with EGs, indicating that the CXCL12 in hydrogel enabled recruitment of tumor cells. Concomitantly, lung metastases were significantly reduced in mice carrying CLG, but not EG, as assessed by reduced B16-hCXCR4-GFP positive tumor cells in the lung (75% reduction in lung metastases in mice carrying CLG; p<0.05). CLG recovered cells by hydrogel enzymatic digestion recapitulated the features of melanoma cell line B16-hCXCR4-GFP (epithelial, melanin rich, MELAN A/ S100/ c-kit/CXCR4 +; α-SMA -). CTCs, evaluated at 10 and at 22 days post cells injection, significantly increased in CLG carrying mice compared to EG carrying mice. Our study suggests that a hyaluronic acid (HA) hydrogel, commercially used as dermal filler, loaded with CXCL12 can divert CXCR4 positive circulating tumor cells B16-hCXCR4-GFP. The cells trapped in the CLG can be recovered and characterized. As a corollary, a reduction of CXCR4 dependent lung metastasis was detected. It suggests that hydrogel-CXCL12 trap may be a therapeutic and diagnostic tool to recover and analyze the biology of circulating tumor cells.

Citation Format: Caterina Ieranò, Crescenzo D'Alterio, Simona Giarra, Maria Napolitano, Assunta Santagata, Antonio Barbieri, Virginia Campani, Antonio Luciano, Claudio Arra, Annamaria Anniciello, Gerardo Botti, Laura Mayol, Giuseppe De Rosa, Roberto Pacelli, Stefania Scala. In vivo capture of early metastatic cells through a CXCL12 loaded-dermal filler [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5194.
Abstract 5197: Single-cell analysis of the brain metastatic microenvironment in breast cancer
Authors: Evans Katrina , Nguyen Quy , Blake Kerrigan , Lawson Devon
Published: August 2018

Breast cancer brain metastasis affects 10-20% of patients, and there are no effective treatments. Activated infiltrating microglia and astrocytes are a major component of metastatic breast tumors in the brain, but it is not known whether these cells are involved in the promotion or rejection of breast cancer cells. To characterize the brain metastatic microenvironment, we used an FACS-based approach to capture pure and viable microglia and astrocytes from normal and metastatic mouse brains for single-cell mRNA sequencing analysis. Using this approach, we have captured more than 10,000 microglia and astrocytes from normal and metastatic brains. Applying our analysis pipelines to this data revealed distinct pathways and markers associated with microglia and astrocytes in metastatic brains. Future work will aim to identify spatial locations and function of specific microglia and astrocyte subpopulations in the metastatic brain.

Citation Format: Katrina Evans, Quy Nguyen, Kerrigan Blake, Devon Lawson. Single-cell analysis of the brain metastatic microenvironment in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5197.
Abstract 5198: Androgen receptor signaling affects macrophage differentiation in the human prostate cancer microenvironment
Authors: Cioni Bianca , Nevedomskaya Ekatarina , Stelloo Suzan , Melis Monique , Burgsteden Johan van , Jong Jeroen de , Poel Henk van der , Boer Jan Paul de , Zwart Wilbert , Bergman Andries M.
Published: August 2018

Increasing evidence from epidemiologic and pathologic studies suggests a role of macrophages in the initiation and progression of prostate cancer (PCa). During PCa development, macrophages (CD68+) are recruited into the human prostate cancer microenvironment, including inflammation-associated M1 and cancer-promoting M2 macrophages. Like other prostate cancer-associated stromal cells, macrophages express the androgen receptor (AR); however, its functionality is not known. CD68+ cells generated in vitro from peripheral white blood cells and PMA activated and IFN and LPS polarized human acute monocytic leukemia THP-1 cells also expressed AR. In these cells AR is an 80KD splice variant, lacking parts of exon 1 and exon 2. AR translocated to the nucleus upon testosterone stimulation, which was counteracted by the AR signaling inhibitor RD162. In vitro generated M1 macrophages expressed the M2 markers CD163 and CD206 upon testosterone stimulation, which was reversed by RD162. AR-ChIP sequencing suggested AR regulated Macrophage Triggering Receptor 1 (TREM1) signaling. Expression of key cytokines (e.g., CCL2, CXCL8, CCL7, IL10, IL1B) involved in TREM1 signaling was upregulated upon testosterone stimulation. Actions of these cytokines is generally associated with a protumor phenotype of macrophages. Conditioned medium from macrophages stimulated with testosterone for 8 hrs resulted in enhanced proliferation and migration of human prostate LNCap and CWRR1 cells. In conclusion, AR signaling might affect differentiation of prostate cancer-associated macrophages into cancer-promoting M2, which affects prostate cancer cell proliferation and migration. Maintaining macrophages in M1 might be a novel mechanism of action of androgen receptor inhibitors.

Citation Format: Bianca Cioni, Ekatarina Nevedomskaya, Suzan Stelloo, Monique Melis, Johan van Burgsteden, Jeroen de Jong, Henk van der Poel, Jan Paul de Boer, Wilbert Zwart, Andries M. Bergman. Androgen receptor signaling affects macrophage differentiation in the human prostate cancer microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5198.
Abstract 5200: Extracellular vesicles in the ascites from gastric cancer patients play critical roles to promote peritoneal dissemination in the intraperitoneal cancer microenvironment
Authors: Ito Atene , kagawa shunsuke , Sakamoto Shuichi , Kuwada Kazuya , Kikuchi Satoru , Kuroda Shinji , Yoshida Ryuichi , Tazawa Hiroshi , Fujiwara Toshiyoshi
Published: August 2018

Although gastric cancer often develops peritoneal dissemination, the underlying mechanism of peritoneal metastasis is still unclear. Extracellular vesicles (EVs) are known to play roles in cell-cell communication. Here, we investigated the existence of EVs in peritoneal ascites of gastric cancer patients and explored their roles in the intraperitoneal cancer microenvironment. We found that EVs actually existed in peritoneal ascites of advanced gastric cancer patients. On the other hand, immune-fluorescent assay of the ascites by using antibodies against CD14 (Monocytes) and CD45 (Leukocytes) revealed that many macrophages existed along with the cancer cells in the peritoneal cavity. EVs purified from peritoneal ascites with cancer dissemination made peripheral blood mononuclear cells-derived macrophages M2-like phenotype which was confirmed by their morphology and expression of CD163/206 (M2 marker). Next, we purified EVs from the culture medium of gastric cancer cell lines. Gastric cancer-derived EVs induced the M2- differentiation in macrophages and the differentiated macrophages promoted migration ability of gastric cancer cells. Finally, we observed that gastric cancer-derived EVs induced apoptosis of cultured mesothelial cells in vitro. These results suggested that EVs in the ascites of gastric cancer patients are possibly derived from gastric cancer and play critical roles in the intraperitoneal cancer microenvironment by affecting the macrophages phenotype and inducing apoptosis in the mesothelial cells, which would be in part the underlying mechanism of peritoneal metastasis.

Citation Format: Atene Ito, shunsuke kagawa, Shuichi Sakamoto, Kazuya Kuwada, Satoru Kikuchi, Shinji Kuroda, Ryuichi Yoshida, Hiroshi Tazawa, Toshiyoshi Fujiwara. Extracellular vesicles in the ascites from gastric cancer patients play critical roles to promote peritoneal dissemination in the intraperitoneal cancer microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5200.
Abstract 5207: HIF-1α regulates the Tie2 receptor on Tie2-expressing monocytes in PyMT breast tumors and augments angiogenic function and metastatic potential
Authors: Steinberger Kayla J. , Forget Mary , Mo Xiaokui , Evans Randall , Gross Amy , Moldovan Leni , Bobko Andrey , Khramstov Valery , Marsh Clay B. , Eubank Timothy D.
Published: August 2018

Tissue oxygenation in the tumor microenvironment serves as a significant parameter in tumor pathophysiology. In fact, normalization of tumor oxygen is associated with inhibition of tumor growth and metastatic capacity. Irregular tumor vessels produced during tumor angiogenesis have low blood perfusion; thus, oxygen transport is decreased in tumors. Tie2-expressing monocytes (TEMs) are a distinct subset of proangiogenic monocytes selectively recruited to tumors in breast cancer patients. Due to the hypoxic nature of the tumor microenvironment, we investigated if oxygen regulates the trafficking of these cells into tumors or if oxygen regulates monocyte differentiation into TEMs once inside the tumor proper. To understand the differentiation of F4/80+/Tie2- cells to Tie2-positivity, we orthotopically implanted PyMT breast tumor cells containing particulate lithium octa-n-butoxy-naphthalocyanine (LiNcBuO) into the mammary fat pads of LysM-Cre control and HIF-1αfl/fl/LysM-Cre mice and evaluated tissue oxygenation by electron paramagnetic resonance (EPR), TEM infiltration, tumor angiogenesis, and metastatic potential. Longitudinal monitoring of physiologic oxygen in tumors using EPR demonstrates the significant role of oxygen-driven TEM regulation in tumor angiogenesis and progression.

Citation Format: Kayla J. Steinberger, Mary Forget, Xiaokui Mo, Randall Evans, Amy Gross, Leni Moldovan, Andrey Bobko, Valery Khramstov, Clay B. Marsh, Timothy D. Eubank. HIF-1α regulates the Tie2 receptor on Tie2-expressing monocytes in PyMT breast tumors and augments angiogenic function and metastatic potential [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5207.
Abstract 5210: Liver prometastatic genes in patients with and without colorectal cancer
Authors: Vidal-Vanaclocha Fernando
Published: August 2018

Metastasis-related hepatic genes were investigated in the tumor-unaffected hepatic tissue from patients with hepatic colorectal cancer (CRC) metastases. In addition, the contribution of CRC and patient's clinical parameters and comorbidities to hepatic expression of CRC prometastatic genes was also studied in new hepatic biopsies from metastasis-free CRC patients and from CRC-free patients used as controls. The expression level of 58% metastasis-associated hepatic genes significantly changed between patients with and without CRC. Principal component and Linear discriminant statistical analyses, and a hierarchical cluster analysis based on Pearson's correlation indices uncovered a subset of 37 CRC-dependent liver prometastatic genes able to segregate patients with and without CRC. Functionally, they belonged to immune regulation, inflammation, metabolic bioprotection, hepatokines and regeneration factors. More importantly, a multivariate canonical correlation analysis showed the additional contribution of patient's pathophysiology and comorbidities to hepatic gene expression and metastasis risk. The liver prometastatic gene expression pattern represented a patient-specific and heterogeneous hepatic reaction to CRC-specific factors, which developed prior to metastatic recurrence and varied according to location of such CRC in the left and right-sided colonic area and rectum.

Citation Format: Fernando Vidal-Vanaclocha. Liver prometastatic genes in patients with and without colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5210.
Abstract 5213: HRH1 inhibition enhances antitumor therapeutic responses through ERK activation in breast cancer
Authors: Fernandez-Nogueira Patricia , Castells Aleix Noguera , Orellana Gemma Fuster , Percaz Leire Recalde , Garcia Nuria Moragas , Plana Anna Lopez , Jimenez Patricia Jauregui , Carbó Maria Neus Carbó , Villaplana Pedro Gascon , Domingo Paloma Bragado , Mancino Mario
Published: August 2018

Histamine receptor 1 (HRH1) belongs to the rhodopsin-like G-protein-coupled receptor family. Its activation by histamine, a ubiquitous messenger molecule released mostly from mast cells, has been demonstrated to be involved in cell proliferation, embryonic development, and tumor growth. In particular, high levels of HRH1 have been associated with dual behavior in the regulation of several tumors. Most of the physiological and pathological actions of HRH1 have been shown to depend strictly on its level of expression and the tumor cell type under study. We recently established that HRH1 is upregulated in patients with basal and HER2-enriched breast tumors and that its expression correlates with a worse prognosis. Despite this, the functional role of HRH1 basal and HER2 targeted therapies for resistant breast cancer (BC) progression has not so far been addressed. We assessed HRH1 expression in a wide panel of breast cancer cell lines and found HRH1 overexpressed in basal and HER2-targeted therapy resistant to BC cell lines at the protein and mRNA levels, corroborating data previously published using patient databases. Using terfenadine, a selective chemical inhibitor of HRH1, we showed that inhibition of HRH1 activity in basal BC cells leads to sub-G0 accumulation, suppresses proliferation, promotes cell motility and triggers the activation of ERK signaling, initiating the mitochondrial pathway of caspase cascade activation, inducing apoptosis. Moreover, in vivo experiments showed that terfenadine therapy reduced basal and HER2-targeted therapies for resistant BC cell tumor growth but does not affect luminal and parental tumor growth. In conclusion, our results suggest that targeting HRH1 might be an effective therapeutic treatment in patients with basal and HER2-targeted resistant BC tumors.

Citation Format: Patricia Fernandez-Nogueira, Aleix Noguera Castells, Gemma Fuster Orellana, Leire Recalde Percaz, Nuria Moragas Garcia, Anna Lopez Plana, Patricia Jauregui Jimenez, Maria Neus Carbó Carbó, Pedro Gascon Villaplana, Paloma Bragado Domingo, Mario Mancino. HRH1 inhibition enhances antitumor therapeutic responses through ERK activation in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5213.
Abstract 5223: IL-6 regulates the OCT4 expression by epigenetic modulation in hepatocellular carcinoma
Authors: Lai Ssu-Chuan , Chang Te-Sheng , Su Yu-Ting , Wu Yu-Chih , Kuo Yung-Che , Lan Pei-Chi , Huang Yen-Hua
Published: August 2018

Purpose: Chronic hepatic inflammatory cytokine interleukin 6 (IL-6) appears important role in pluripotent transcription factor OCT4 expression particularly in HBV-related hepatocellular carcinomas (HBV-HCC). However, the underlying mechanism still remains largely unknown. Experimental Design: Serum levels of IL-6 were detected using ELISA assays (n = 120). The effects of IL-6 on DNMT/OCT4 expression in HCC were examined using real-time QPCR analysis, RNA interference, and xenograft animal models. The DMNT/OCT4 protein and phospho-STAT (p-STAT) were detected by western blotting. OCT4, and DNMT expression levels in tissues (n = 144) were analyzed by real-time Q-PCR and was correlated with early tumor recurrence using Kaplan-Meier survival analysis. Results: A high positive correlation between the expression levels of OCT4 and epigenetic DNMT family in human HCC tissues was observed. High serum levels of IL-6 were significantly correlated with high OCT4 and DNMT expression. IL-6 stimulated the DNMT expression STAT3-dependently, which stimulated the OCT4 expression in both the cell lines and the xenografted mouse tumors. The inhibition STAT phosphorylation/DNMT expression by dominant negative STAT-Y705F plasmid transfection or DNMT RNA interference significantly suppressed the IL-6-induced OCT4 expression in vitro. Conclusions: The expression of pluripotency-related OCT4 is associated with early tumor recurrence and is regulated by IL-6-induced epigenetic DNMTs in HCC.

Citation Format: Ssu-Chuan Lai, Te-Sheng Chang, Yu-Ting Su, Yu-Chih Wu, Yung-Che Kuo, Pei-Chi Lan, Yen-Hua Huang. IL-6 regulates the OCT4 expression by epigenetic modulation in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5223.
Abstract 5225: PAR1 signaling on tumor cells limits tumor growth by maintaining a mesenchymal phenotype in pancreatic cancer
Authors: Tekin Cansu , Shi Kun , Spek Arnold , Bijlsma Maarten F.
Published: August 2018

Background: Pancreatic cancer is a dismal disease with limited treatment options. Importantly, in recent years an oncogenic role of protease-activated receptor 1 (PAR1) emerged in various cancer types, and PAR1 is generally accepted to promote tumor progression. PAR1 depletion in stroma limits cancer growth in an orthotopic pancreatic cancer model, suggesting that PAR1 would be a prognostic marker for PDAC.

Aim: To determine PAR1 expression in tumor compartment of PDAC microenvironment and its contribution to tumor progression and invasion.

Methods: Kaplan-Meier overall survival analysis for PAR1 expression on PDAC tumor expression sets was performed. In an orthotopic tumor model, short hairpin RNA mediated PAR1 (shPAR1) knockdown cells derived from p48-CRE/LSL-KRAS/p53flox/flox mice (KP cells) and Panc02 cells were used. Additionally, gene set enrichment analysis was done for EMT signatures on PDAC tumor and pancreatic cancer cell lines. Furthermore, in vitro migratory capacity and mRNA levels for emerging mesenchymal signatures were analyzed in PAR1 knockdown PANC-1 cell line.

Results: PAR1 expression does not associate with the overall survival of PDAC patients. However, tumor cell-specific PAR1 silencing potentiates pancreatic tumor growth but decreases metastatic potential in subsequent murine orthotopic pancreatic cancer model. Further analysis of PAR1 expression in PDAC tumors and pancreatic cancer cell lines show positive correlation with mesenchymal signatures and epithelial to mesenchymal transition genes. Indeed, PAR1 knockdown results in well-differentiated pancreatic tumors in vivo, with enhanced epithelial characteristics both in vitro and in vivo. Furthermore, decreased PAR1 activity decreases migratory capacity and in vitro metastatic potential in vivo.

Conclusion: Our results suggest that growth promoting effects of PAR1 in stromal cells are counterbalanced by the growth inhibitory contributions of PAR1 in tumor cells. Overall, tumor cell PAR1 is linked to the maintenance of a mesenchymal-like state and silencing PAR1 in this setup results in an increased epithelial phenotype of cancer cells with an increased proliferative index but a decreased metastatic potential.

Citation Format: Cansu Tekin, Kun Shi, Arnold Spek, Maarten F. Bijlsma. PAR1 signaling on tumor cells limits tumor growth by maintaining a mesenchymal phenotype in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5225.