bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3927: The presence of COX-2 and tumor-associated macrophages as a prognostic marker for invasive breast carcinoma patients
Authors: Esbona Karla , Saha Sandeep , Yi Yanyao , Yu Menggang , Wisinski Kari , Wilke Lee G. , Keely Patricia J.
Published: July 2016

Inflammation is an important mediator of tumor progression. The objective of this study was to assess whether the tissue localization of COX-2 and tumor-associated macrophages were associated with clinicopathological features of invasive carcinoma, including collagen deposition and patient survival outcome. A tumor microarray (TMA) of 371 biopsy specimens from patients with invasive breast carcinoma was analyzed for expression of high levels of COX-2, and the macrophage markers CD68 and CD163 in either the tumor nest (TN) or the tumor-associated stroma (TS). The study population for this TMA was female patients, 18 to 80 years of age with a median follow up of 8.4 years. Survival tables were calculated according to the Kaplan-Meier method. We found that elevated collagen deposition was associated with high stromal expression of COX-2 (P < 0.0001); however, the amount of collagen deposition was not a predictor for survival outcome. In order to better analyze patient data, samples were divided into quartiles based on their levels of COX-2 expression and/or macrophage infiltration. Tumor nests with high COX-2 expression had worse patient outcome (P = 0.011). One possible mechanism for this is COX-2 dependent recruitment of macrophages to the tumor microenvironment, supported by our finding of high infiltration of CD68+ macrophages in the TS, and CD163+ macrophages in both TN and TS. This recruitment of macrophages was associated with worse overall survival (OS). Furthermore, patient survival was worsened even more if macrophages expressed COX-2, suggesting positive amplification of COX-2 signaling via macrophage recruitment. This notion is further established by the finding of high presence of CD163+ macrophages in the TN as an independent prognostic factor as revealed by multivariate analysis (P < 0.0001, HR = 4.67). These results suggest that in invasive carcinoma the localization of inflammatory markers within the tumor are biomarkers for patient survival outcome. Therefore, we propose that these patients may benefit from therapy with a selective COX-2 inhibitor such as celecoxib.

Citation Format: Karla Esbona, Sandeep Saha, Yanyao Yi, Menggang Yu, Kari Wisinski, Lee G. Wilke, Patricia J. Keely. The presence of COX-2 and tumor-associated macrophages as a prognostic marker for invasive breast carcinoma patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3927.
Abstract 3957: Optimized plasma collection procedures for liquid biopsy analyses in cancer
Authors: Parpart-Li Sonya T. , Bartlett Bjarne , Popoli Maria , Adleff Vilmos , Brahmer Julie , Azad Nilo , Bonerigo Sarah , Browner Ilene , Ryan Amy , Velculescu Victor , Sausen Mark , Diaz Luis A.
Published: July 2016

Analyses of genomic alterations in cell-free DNA (cfDNA) shed from tumors into the blood stream is rapidly advancing as a method to detect and genotype cancer. Liquid biopsy analyses are a favorable alternative to invasive tissue approaches, particularly for cancers in organs where tissue is difficult to obtain. As these methods become a clinical standard, the method for blood collection and plasma isolation will be an important consideration to ensure optimal assay performance with limited degradation of circulating tumor DNA (ctDNA). We compared blood collection and plasma isolation using both K2EDTA and cell-free DNA Streck™ BCT tubes to determine the stability of cfDNA and ctDNA in blood at room temperature or at 4°C. Blood was collected from nine cancer patients with KRAS mutations using K2EDTA and Streck™ BCT tubes. Over the course of seven days, cell lysis occurred in K2EDTA tubes at room temperature, releasing germline DNA into the blood (1300% increase over seven days), which significantly increased the number of total genomic equivalents. Additionally, we found that the volume of plasma collected from both tube types during blood fractionation decreased over time when tubes were at room temperature (an average of 3.5 mL vs. 1.0 mL, respectively). To evaluate the effect of storage conditions on performance of each tube type, blood was also collected from six cancer patients with KRAS or EGFR mutations and either maintained at room temperature or stored at 4°C for up to three days. Taken together, K2EDTA tubes stored at 4°C prevented cell lysis and preserved the ctDNA in a manner equivalent to Streck™ BCT tubes. These data demonstrate that liquid biopsies can be collected using either tube type with similar performance and appropriate consideration of storage conditions.

Citation Format: Sonya T. Parpart-Li, Bjarne Bartlett, Maria Popoli, Vilmos Adleff, Julie Brahmer, Nilo Azad, Sarah Bonerigo, Ilene Browner, Amy Ryan, Victor Velculescu, Mark Sausen, Luis A. Diaz. Optimized plasma collection procedures for liquid biopsy analyses in cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3957.
Abstract 3979: Metabolic pathways associated with urinary metabolite biomarkers differentiate bladder cancer patients from healthy controls
Authors: Kim Won Tae , Yun Seok Joong , Yan Chunri , Jeong Pildu , Kim Ye Hwan , Lee Il-Seok , Park Sunghyouk , Moon Sung-Kwon , Choi Yung-Hyun , Choi Young Deuk , Kim Jayoung , Kim Wun-Jae
Published: July 2016

Our high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry study identified bladder cancer (BCA)-specific urine metabolites, including carnitine, acylcarnitines, and melatonin. The objective of the current study was to determine which metabolic pathways are perturbed in BCA based on our previously identified urinary metabolome. Significant alterations in the carnitine-acylcarnitine and tryptophan metabolic pathways were detected in urine specimens from BCA patients compared to those of healthy controls. The expression of eight genes involved in the carnitine-acylcarnitine metabolic pathway (CPT1A, CPT1B, CPT1C, CPT2, SLC25A20, and CRAT) or tryptophan metabolism (TPH1 and IDO1) was assessed by RT-PCR in our BCA cohort (n = 135). CPT1B, CPT1C, SLC25A20, CRAT, TPH1, and IOD1 were significantly downregulated in tumor tissues compared to adjacent normal bladder tissues (p < 0.05 all) of patients with non-muscle invasive bladder cancer, whereas CPT1B, CPT1C, CRAT, and TPH1 were downregulated in those with muscle invasive bladder cancer (p < 0.05), with no changes in IDO1 expression. In this study, alterations in the expression of genes associated with the carnitine-acylcarnitine and tryptophan metabolic pathways, the most perturbed pathways in BCA, were determined. Our findings provide insight into the mechanisms underlying BCA-associated metabolic flux perturbation.

Citation Format: Won Tae Kim, Seok Joong Yun, Chunri Yan, Pildu Jeong, Ye Hwan Kim, Il-Seok Lee, Sunghyouk Park, Sung-Kwon Moon, Yung-Hyun Choi, Young Deuk Choi, Jayoung Kim, Wun-Jae Kim. Metabolic pathways associated with urinary metabolite biomarkers differentiate bladder cancer patients from healthy controls. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3979.
Abstract 3990: FGFR3 mutations as novel oncogenic targets
Authors: Martinez Cardona Gabriela , Gaffney Dana , Bell Katherine , Portale Joseph , Dunworth Matthew , Lorenzi Matthew , Platero Suso , Karkera Jayaprakash
Published: July 2016

Fibroblast growth factors (FGFs) are a family of homologous secreted glycoproteins involved in signaling pathways responsible for embryonic development, cell proliferation, survival, and migration. FGF activity is mediated by four transmembrane fibroblast growth factor receptors (FGFRs) which are receptor tyrosine kinases. Typically, FGF binding induces FGFR dimerization, leading to phosphorylation of the intracellular tyrosine kinase domain. This leads to downstream activation of multiple signaling pathways, including the mitogen-activated protein kinase (MAPK), PI3K/AKT, signal transducer and activator of transcription (STAT), and phospholipase-C-γ cascades. Deregulated FGFR activity, through mutations or translocations, is often associated with oncogenic events. Aberrations in FGFR genes have been observed in several tumor types including bladder, gastric, colorectal, ovarian, and hematologic cancers. To date, several FGFR3 mutations have been identified in bladder cancer. In this study, we developed a TaqMan qRT-PCR-based approach to detect four FGFR3 mutations in formalin-fixed paraffin embedded tissue (FFPET) samples. Cell lines overexpressing FGFR3 mutations were generated to determine impact on cell signaling, and sensitivity to the small-molecule pan-FGFR inhibitor JNJ-42756493. To determine the role and significance of these FGFR3 mutations in cancer, mutation expression constructs were designed and individually transfected into normal rat kidney epithelial cells. Cells harboring the FGFR3 mutations exhibited anchorage-independent growth, increased proliferation, and showed increased sensitivity to the FGFR inhibitor JNJ-42756493 in vitro compared to parental lines. These findings underline the oncogenic potential of the FGFR3 mutation genes and highlight their unique potential as predictive biomarkers in the selection of patients for FGFR-targeted therapy.

Citation Format: Gabriela Martinez Cardona, Dana Gaffney, Katherine Bell, Joseph Portale, Matthew Dunworth, Matthew Lorenzi, Suso Platero, Jayaprakash Karkera. FGFR3 mutations as novel oncogenic targets. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3990.
Abstract 4004: Development of a successful combination therapy for hepatocellular cancer by targeting Treg and PD-1
Authors: Liu Dai , Li Guangfu , Cooper Timothy , Kimchi Eric , Qi Xiaoqiang , Li Ningfei , Rockey Don , Schell Todd , Staveley-O’Carroll Kevin
Published: July 2016

We have established a clinically relevant murine model to reveal tumor-induced immunotoerance in hepatocellular carcinoma (HCC). Critical factors are targeted to develop immune-based therapies for HCC control.

Intrasplenic (ISPL) inoculation of oncogenic hepatocytes and intraperitoneal (IP) injection of carbon tetrachloride (CCl4) are combined to induce progressive HCCs in fibrotic livers of immunocompetent mice. We characterize the features of this model, examine tumor-antigen-specific (TAS) immunity during tumor initiation and progression, and identify the critical factors in tumor-induced immune tolerance. The established murine model recapitulates human HCC and reflects its typical features. TAS CD8+ T cells initially maintain a naive phenotype and function in early-stage tumor-bearing mice, then become profound exhaustion with the tumor progression to the advanced stage. The deep immunosuppression is associated with the significant upregulation of Programmed cell death protein 1(PD-1), Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and the increase of Tregs. While the changes in Tregs and PD-1 are systemic, they are particularly pronounced in the tumor microenvironment. Sunitinib-mediated reduction of Treg together with antibody-mediated blockade of PD-1 synergistically suppress tumor growth and activate anti-tumor immunity.

Our data provide evidence that oncogenic hepatocytes escape immune surveillance during tumor initiation via immune ignorance, while later-stage established HCCs evade immune destruction via tumor-induced immunotolerance. Synergy of sunitinib and anti-PD-1 Ab generates favorable effect on suppressing tumor growth and activating anti-tumor immunity. This combinational strategy has high translational potential in HCC treatment.

Citation Format: Dai Liu, Guangfu Li, Timothy Cooper, Eric Kimchi, Xiaoqiang Qi, Ningfei Li, Don Rockey, Todd Schell, Kevin Staveley-O’Carroll. Development of a successful combination therapy for hepatocellular cancer by targeting Treg and PD-1. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4004.
Abstract 401: Preservation of cells in blood and culture
Authors: Browne Cecille , Lu Dan , Roth Daniela , Liberal Vasco
Published: July 2016

Introduction: Liquid biopsies and cell-based therapies are emerging approaches for the treatment of cancer. Accuracy of diagnosis using liquid biopsies and effectiveness of treatment using cell-based therapies will rely upon the preservation cells for both viability and the information contained in them.

Experimental procedures: Biomatrica's proprietary chemical library was used to develop formulations that could preserve cells in multiple formats. Cell preservation was assessed by a variety of cell-based and molecular techniques such as microscopy, flow cytometry and real-time PCR.

Results: Blood samples spiked with circulating tumor cells (CTCs) were preserved for several days at room temperature to allow for downstream morphological and transcriptomic analyses. CTC recoveries in preserved samples were as much as 4 times those in non-protected samples over 4 days. Changes in RNA expression of CTC-specific genes in preserved samples ranged from 0 to +/- 2 delta Ct values, while in non-protected samples, changes ranged from 0 to +/-5 delta Ct values over 4 days. In some formulations, CTC viabilities were as much as twice the viabilities of CTCs in non-protected samples. Supporting these viability results was a demonstrated culturability of isolated CTCs. Furthermore, cell-free DNA in plasma of preserved blood samples changed by less than 2 Ct values over 4 days, whereas in control samples, plasma DNA changed by as much as 10 Ct values. Hemolysis in preserved blood samples was also lower by as much as 69% compared to non-protected samples.

In a separate set of experiments, activated T-cells were stored in the presence of common buffers (RPMI, PBS) or formulations at room temperature. T-cells cultured with formulations displayed greater 40 to 60% greater cell viability after 3 days compared to those stored in common buffers.

Conclusions: Biomatrica has developed a library of cell preservatives that can be used to preserve cells in blood samples and in cell culture. The ability to recover cells in blood samples and determine their genetic information would enable monitoring of a patient's cancer status and personalizing drug treatments. In addition, cell preservation may allow further advances in cell-based therapies as cells could be more reliably recovered from patients, modified (e.g. CAR-T, stem cells), and administered to patients.

Citation Format: Cecille Browne, Dan Lu, Daniela Roth, Vasco Liberal. Preservation of cells in blood and culture. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 401.
Abstract 4015: Exposure of lymphatic endothelial cells to ionizing radiation increases the surface expression levels of integrin ligands
Authors: Rodriguez-Ruiz Maria E. , Garasa Saray , Rodriguez Inmaculada , Yanguas Alba , Teijeira Álvaro , Melero Ignacio , Rouzaut Ana
Published: July 2016

Radiotherapy has been proven an efficacious tool to combat cancer by directly destroying tumor cells. However, effects of radiotherapy have been also described that occur through the re-education of the immune tumor microenvironment, being leukocyte traffic regulation across the tumor vasculature of key importance.

Here, we address the effects that radiotherapy may be causing on the adhesive properties of the tumor lymphatic vasculature. To study this, we irradiated primary and immortalized human and murine lymphatic endothelial cells grown in vitro, and fresh tumor explants obtained from colon cancer patients. As a result, we observed dose- and time- dependent increases in the expression of the integrin counter-receptors ICAM-1 and VCAM on the surface of the tumor lymphatic endothelial cells. These effects are also observed in vivo on colon (MC38) and melanoma (B16) tumors transplanted into syngeneic mice and irradiated with a single dose of 20 Gy. These findings are consistent with increased ICAM-1 and VCAM expression levels on Lymphatic endothelial cells in tumor biopsies from cancer patients which were taken before and four weeks after fractionated radiotherapy, including head and neck, colon and oropharyngeal human carcinomas. While the TGFβ pathway was found to be involved in these effects, NF-κB inhibitors did not hamper ICAM-1 or VCAM induction.

In summary, these results show ICAM-1 and VCAM surface expression was enhanced on lymphatic vessels after radiotherapy and may explain differential leukocyte traffic following irradiation.

Citation Format: Maria E. Rodriguez-Ruiz, Saray Garasa, Inmaculada Rodriguez, Alba Yanguas, Álvaro Teijeira, Ignacio Melero, Ana Rouzaut. Exposure of lymphatic endothelial cells to ionizing radiation increases the surface expression levels of integrin ligands. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4015.
Abstract 402: Multiplex TaqMan assays for rare mutation analysis using digital PCR
Authors: Laig Marion -. , Chan Frances , Lac Le , Straub Ted , Varma Kamini , Keys David
Published: July 2016

Introduction

Detection of rare mutations for research purposes in tumor tissue and cell free DNA (cfDNA) allows for monitoring of tumor progression and regression. cfDNA isolated from plasma combined with a sensitive detection method like digital PCR is non-invasive and enables earlier detection compared to conventional imaging techniques.

Building on the TaqMan based Rare Mutation assay set for detection of rare mutations using digital PCR on the QuantStudio 3D Digital PCR System, we are now developing multiplex assays for simultaneous detection of several mutations. We selected relevant mutations in the EGFR and KRAS genes for our initial multiplex application: EGFR G719, EGFR exon 19 deletions, and KRAS G12/G13. These mutations may have implications for potential future targeted therapy.

Methods

Primers and probes of singleplex Rare Mutation Assays were reformulated to generate multiplex assays detecting the EGFR and KRAS mutations. All multiplex assays were tested on template composed of wild-type genomic DNA background mixed with mutant plasmid reflecting each of the mutations detected by the multiplex assays.

Summary

Initial experimental results were successful and showed excellent signal intensity and clear cluster separation when analyzed with the QuantStudio 3D AnalysisSuite™ Cloud Software. The EGFR G719 mutations (COSM6239, COSM6253, COSM6252) were detected using a 3plex assay, EGFR exon 19 deletions (COSM12383, COSM12422, COSM12678, COSM6223, COSM6254, COSM6255) were detected using a 6plex assay, and KRAS G12/G13 mutations (COSM516, COSM517, COSM518, COSM520, COSM521, COSM522, COSM527, COSM532) were detected using an 8plex.

Conclusion

Multiplexing assays for three relevant mutation loci proved feasible and presents an efficient way to assess the presence and the percentage of mutations at these loci.

Citation Format: Marion -. Laig, Frances Chan, Le Lac, Ted Straub, Kamini Varma, David Keys. Multiplex TaqMan assays for rare mutation analysis using digital PCR. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 402.
Abstract 4029: MEK pathway inhibition upregulates PD-L1 and MHC class I expression levels in head and neck squamous cell carcinoma
Authors: Kang Seong-Ho , Keam Bhumsuk , Kim Soyeon , Kim Miso , Ahn Yong-Oon , Kim Tae Min , Kim Dong-Wan , Heo Dae Seog
Published: July 2016

Background: Expression of programmed death-ligand 1 (PD-L1) and down-regulation of MHC class I are well known tumor immune evasion mechanisms. The purpose of this study is to investigate the regulation of PD-L1 and MHC class I expression in head and neck squamous cell carcinoma (HNSCC) cell lines by MEK inhibitor trametinib.

Methods: Six HNSCC cell lines (SNU-1041, SNU-1066, SNU-1076, PCI-13, Detroit 562, and FaDu) were used. Trametinib was purchased from Selleckchem. Growth inhibition of trametinib was measured using MTT assay. Trametinib was dosed at 10 - 640 nM for investigating their effect on PD-L1 and MHC class I molecule expression. Changes of PD-L1 and MHC class I (including HLA-A2) expression levels were analyzed by flow cytometry after treatment with trametinib (20 nM) and/or IFN-γ (1 - 10 ng/ml). Total and phosphorylated STAT1 and STAT3 were analyzed by western blot.

Results: Overall IC50 values of trametinib were over 10 μM in cell lines except SNU-1066 and Detroit 562 with IC50 values still higher than 100 nM. All the six cell lines have no upstream RAS or RAF mutation. Although trametinib treatment did not inhibit growth of these cells, it up-regulated MHC class I expression level in all cells. Furthermore, trametinib enhanced exogenous IFN-γ-induced MHC class I up-regulation in four out of six cell lines. Mechanistically, trametinib treatment increased phosphorylated STAT1 levels by IFN-γ in all cells, and subsequent total STAT1 levels in two out of six cell lines. Interestingly, trametinib treatment directly phosphorylated STAT3 although total STAT3 level was not changed. Either trametinib alone or with IFN-γ induce PD-L1 expression, but the effect was various in these cells. These results imply that blockade of MAPK pathway may alternatively activate JAK/STAT pathway in HNSCC cells, and MHC class I and PD-L1 expression levels were changed as the results

Conclusion: Our results suggest that MEK pathway is closely related with regulation of PD-L1 and MHC class I expression. Up-regulated levels of PD-L1 and MHC class I by inhibiting MEK pathway suggest a possible synergistic role of MEK inhibitor with anti-PD-1/PD-L1 immunotherapy in HNSCC.

Citation Format: Seong-Ho Kang, Bhumsuk Keam, Soyeon Kim, Miso Kim, Yong-Oon Ahn, Tae Min Kim, Dong-Wan Kim, Dae Seog Heo. MEK pathway inhibition upregulates PD-L1 and MHC class I expression levels in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4029.
Abstract 4041: Glioma cells display a prognostically significant hematopoietic phenotype
Authors: Moncayo Gerald
Published: July 2016

Glioblastoma multiforme (GBM) is one of the most aggressive human cancers. Glioma cells expressed immunoreceptor tyrosine-based activation motif (ITAM) bearing molecules, such as Fc receptors, T cell receptor components and C-type lectins and expression was up regulated in two distinct tumor-derived prognostic signatures identified in GBM and low-grade gliomas. Targeting SYK, downstream of ITAM molecules, attenuated GBM tumor growth and invasiveness, and reduced B and CD11b+ cell mobility and infiltration resulting in improved survival. In addition, GBM tumor cells were found to display phagocytic characteristics in vitro and in vivo by flow cytometry and two photon imaging. Our data demonstrate that gliomas are hematopoietic-like tumors that express an immune signaling network important in glioma progression, and identify targets for therapeutic intervention.

Citation Format: Gerald Moncayo. Glioma cells display a prognostically significant hematopoietic phenotype. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4041.
Abstract 4045: Urban-rural disparity of overweight/obesity distribution and its potential association with breast cancer among Chinese females
Authors: Gao Ying , Huang Yubei , Dai Hongji , Wang Peishan , Li Haixin , Song Fengju , Zheng Hong , Dong Henglei , Han Jiali , Wang Yaogang , Chen Kexin
Published: July 2016

Background: Disparity of overweight/obesity distribution had been investigated in developed countries, but rarely been investigated in developing countries, especially for China with rapid socio-economic development in the past decades.

Methods: A total of 1 210 762 participants were recruited from the Chinese National Breast Cancer Screening Program. Overweight and obesity were defined as body mass index (BMI) ranged 24.0-27.9 kg/m2 and BMI ≥ 28.0kg/m2, respectively.

Results: The prevalence of overweight/obesity for Chinese rural women (35.2%, 29.2% for overweight and 6.0% for obesity) was significantly higher than that for Chinese urban women (33.4%, 27.7% for overweight and 5.7% for obesity) (P <0.001). For either rural or urban women, the prevalence of overweight/obesity was highest in north region, followed by east region for rural women and north-east region for urban women. For rural women, higher prevalence of overweight/obesity was significantly positively associated with elder age, Han nationality, low level of education, no occupation, high family income, less number of family residents, insurance, and elder age at marriage. Similar positive associations were also found for urban women, except negative associations for high family income, less number of family residents, and elder age at marriage. A non-significant positive association between overweight/obesity and breast cancer was found for rural women [odds ratio (OR) = 1.06; 95% confidence interval (CI): 0.87-1.29], but a significant positive association for urban women (OR = 1.55; 95%CI: 1.19-2.02).

Conclusions: There was an obvious urban-rural disparity of overweight/obesity distribution among Chinese females, which could also lead to an obvious disparity of breast cancer distribution.

Citation Format: Ying Gao, Yubei Huang, Hongji Dai, Peishan Wang, Haixin Li, Fengju Song, Hong Zheng, Henglei Dong, Jiali Han, Yaogang Wang, Kexin Chen. Urban-rural disparity of overweight/obesity distribution and its potential association with breast cancer among Chinese females. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4045.
Abstract 4046: The role of TFPI2 and FGF2 in asbestos-induced mesothelial to fibroblastic transition
Authors: Thompson Joyce K. , Miller Jill , MacPherson Maximilian B. , Shukla Arti
Published: July 2016

Mechanisms involved in the tumorigenesis of the devastating cancer, malignant mesothelioma (MM) are poorly understood. We have recently shown that interleukin-1β (IL-1β), an inflammatory cytokine is upregulated by asbestos via the activation of the inflammasome (a molecular platform that assembles for the activation of caspase-1) in mesothelial cells. Furthermore we have demonstrated that IL-1β secretion may lead to the activation of downstream signaling cascades involved in malignant transformation of mesothelial cells.

Preliminary data from our lab indicate that in addition to IL-1β, asbestos exposure upregulated the secretion of basic fibroblast growth factor (bFGF/FGF2) and tissue factor pathway inhibitor 2 ((TFPI2) a kunitz type protease inhibitor). These factors were also regulated by the inflammasome and have never before been implicated in asbestos-induced mesothelial to fibroblastic transition (MFT). Based on our preliminary data, we hypothesized that upregulation of IL-1β by asbestos-induced inflammasome activation increases FGF2 secretion and signaling. Furthermore, we hypothesize that FGF2 together with increased TFPI2 secretion induces transition of mesothelial cells to a fibroblastic phenotype that facilitates MM carcinogenesis. In the proposed study we will delineate the role of TFPI2 (siRNA) and FGF2 (pan FGFR inhibitor, BGJ398) in the process of asbestos-induced MFT.

Data from this study will provide added insight into the mechanisms involved in the initiation of MM and indicate whether TFPI2 and FGF2 can serve as drugable targets for combination therapy against MM. This work is supported by NIH grant, RO1 ES021110.

Citation Format: Joyce K. Thompson, Jill Miller, Maximilian B. MacPherson, Arti Shukla. The role of TFPI2 and FGF2 in asbestos-induced mesothelial to fibroblastic transition. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4046.
Abstract 4058: DNA damage and tumour burden in mouse colon is increased in response to carcinogen exposure after induction of chronic inflammation - a more disease relevant model of colitis-associated colorectal cancer
Authors: Lannagan Tamsin RM , Mano Mark T. , Head Richard J. , Lockett Trevor , Ernst Matthias , Cosgrove Leah J.
Published: July 2016

Chronic inflammation (CI) such as inflammatory bowel disease increases the risk of developing colorectal cancer (CRC). Tumorigenesis is supported by inflammatory cytokines through increased cell proliferation and inhibiting apoptosis of neoplastic cells. The classic mouse model of colitis-associated CRC (CA-CRC) requires treatment with a mutagen (azoxymethane, AOM) followed by induction of inflammation through ingestion of a luminal irritant (dextran sulfate sodium, DSS). AOM is metabolized by colonic epithelium causing DNA damage through formation of mutagenic DNA adducts. Although the AOM + DSS model of CA-CRC is a valuable research model, it “reverses” events that take place in humans where CI typically precedes CRC. We hypothesise that CI sensitises colonic epithelium to DNA damage and predisposes the epithelium to neoplastic transformation.

In order to investigate this we reversed the AOM + DSS model to more accurately represent the disease process in humans. Using two experimental approaches we treated wildtype (WT) mice with AOM alone or with DSS then AOM and collected colonic tissue after 6 and 48 hours (short-term) or colonic tumours (long-term). Tumour number and size were quantified. Immunohistochemistry was used to analyse DNA damage, apoptosis, and proliferation. Formation of the DNA adduct O6MeG was measured by a modified method of HPLC, and qPCR was used to analyse expression of DNA damage signalling pathway components.

Compared with an AOM challenge alone, prior CI induction by DSS treatment increased DNA damage, reduced apoptosis, and increased tumour number and tumour size. This data reveals how prior CI modulates the colonic epithelial response to mutagen exposure in the short-term and how it promotes tumour formation in the long-term. In addition, this data demonstrates that the “reverse” model of CA-CRC can be successfully used as a more appropriate experimental framework to better understand the molecular mechanisms underpinning CA-CRC and will allow us to further investigate the relationship between prior CI, DNA damage, and tumour development.

Citation Format: Tamsin RM Lannagan, Mark T. Mano, Richard J. Head, Trevor Lockett, Matthias Ernst, Leah J. Cosgrove. DNA damage and tumour burden in mouse colon is increased in response to carcinogen exposure after induction of chronic inflammation - a more disease relevant model of colitis-associated colorectal cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4058.
Abstract 406: Enhancer templated RNAs as predictors of therapeutic response to epigenetic therapy
Authors: Firestein Ron , McCleland Mark , Mesh Kathryn , Gnad Florian
Published: July 2016

Colon tumors arise in a stepwise fashion from either discrete genetic perturbations or epigenetic dysregulation. To uncover the key epigenetic regulators that drive colon cancer growth, we utilized a CRISPR loss of function screen and identified a number of essential genes, including the Bromodomain and Extraterminal (BET) protein, BRD4. We find BRD4 is critical for colon cancer proliferation and its loss leads to differentiation effects in vivo. JQ1, a BET inhibitor, preferentially reduced growth in a subset of epigenetically dysregulated colon cancers, characterized by the CpG island methylator phenotype (CIMP). Integrated transcriptomic and genomic analyses defined a distinct super-enhancer in CIMP(+) colon cancers that regulates cMYC transcription. We find that the CCAT1 long non-coding RNA (lncRNA) is transcribed from this super-enhancer and is exquisitely sensitive to BET inhibition. Concordantly, cMYC transcription and cell growth were tightly correlated with the presence of CCAT1 RNA in a variety of tumor types. Taken together, we propose CCAT1 as a clinically tractable biomarker for identifying patients likely to benefit from BET inhibitors.

Citation Format: Ron Firestein, Mark McCleland, Kathryn Mesh, Florian Gnad. Enhancer templated RNAs as predictors of therapeutic response to epigenetic therapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 406.
Abstract 4060: The role of p62-dependent regulation of COX-2 in UVA response and skin tumor progression
Authors: Sample Ashley , Qiang Lei , Zhao Baozhong , He Yu-Ying
Published: July 2016

Skin cancer is the most common type of cancer in the US, with an estimated 3.5 million cases diagnosed each year. Exposure to ultraviolet (UV) radiation, namely UVA (320-400 nm) and UVB (290-320 nm), is the major risk factor for the development of skin cancer. UVA is 20-fold more abundant in sunlight than UVB and is the major component of tanning beds. However, the mechanism of UVA's contribution to skin cancer remains unclear. One putative effector of UVA in skin cancer is p62, a selective autophagy cargo protein and substrate. We have found that UVA upregulates p62 expression, independent of autophagy, by inducing transcription of p62. p62 is upregulated in many types of cancer, including squamous cell carcinoma (SCC) and melanoma, and can facilitate the activation of a number of pathways to promote cell proliferation, invasion, inflammation, and survival. We have previously shown that p62 stabilizes Twist1 to promote invasion and proliferation, indicating p62 can function independent of autophagy to promote tumor progression. However, the function of p62 in UVA-induced tumor progression is unknown. Using a candidate gene approach, we identified a novel interaction between p62 and cyclooxygenase-2 (COX-2), which suggests a putative function for p62 in UVA-induced skin cancer. COX-2 is a prostaglandin synthase often overexpressed in cancer, where it correlates with poor prognosis. COX-2 promotes cell proliferation and survival, and inhibition of COX-2 prevents skin cancer development. COX-2 protein expression is induced concomitantly with p62 in response to UVA and knockdown of p62 prevents induction of COX-2 by UVA. As COX-2 transcription is not induced by UVA in skin cancer cells along with p62, we assessed a possible protein-level regulatory mechanism. Co-immunoprecipitation of endogenous p62 and COX-2 in skin cancer cells reveals a novel physical interaction between these proteins. Therefore, we hypothesize that p62 transcription is induced by UVA to promote stability of cyclooxygenase-2 (COX-2) and consequently, tumor progression. This project aims to further understand how UVA regulates p62 and thus COX-2 availability, and the functional significance for skin tumor progression. As both p62 and COX-2 are critical for skin tumor progression, understanding the link between these proteins will unravel a signaling axis central to skin cancer progression.

Citation Format: Ashley Sample, Lei Qiang, Baozhong Zhao, Yu-Ying He. The role of p62-dependent regulation of COX-2 in UVA response and skin tumor progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4060.
Abstract 4082: Epithelial-stromal interactions are altered at the earliest stages of colon cancer development
Authors: Mo Allen , Jackson Stephen , Varma Kamini , Carpino Alan , Giardina Charles , Devers Thomas J. , Rosenberg Daniel W.
Published: July 2016

While progression of nonmalignant colonic cells carrying somatic mutations to early malignancy is dependent upon interactions between mutated epithelium and surrounding stroma, the nature of these interactions is poorly understood. Here we report the development of an ultra-sensitive laser capture microdissection/RNA-seq approach for profiling the epithelial and stromal compartments of mutated aberrant crypt foci (ACF). ACF are the earliest detectable pre-neoplastic lesion found in human colon and were identified using high-definition endoscopy with contrast dye-spray. We focused on epithelial and stromal cells of ACF carrying somatic mutations to either KRAS, BRAF or APC, and compared these to control samples from each patient. By comparing the gene expression from each group, we identified an increase in a number of pro-inflammatory NF-κB target genes that are specific to ACF epithelium (including TIMP1, RELA and RELB). We confirmed distinct transcriptional changes associated with each somatic mutation and demonstrate that a subset of ACF display a BRAFV600E-mediated senescence-associated transcriptome, characterized by increased expression of CDKN2A (p16). Furthermore, ACF-associated stroma is transcriptionally distinct from adjacent normal stroma and genes related to immune cell infiltration and activation of fibroblasts are up-regulated. Immunofluorescence analysis confirms the abundance of activated fibroblasts in ACF stroma regardless of mutational status. These results provide new insight into the cellular interplay that occurs at the very early stages of colon cancer development, highlighting the role of activated stromal fibroblasts and inflammation.

Citation Format: Allen Mo, Stephen Jackson, Kamini Varma, Alan Carpino, Charles Giardina, Thomas J. Devers, Daniel W. Rosenberg. Epithelial-stromal interactions are altered at the earliest stages of colon cancer development. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4082.
Abstract 4085: Notch1 signaling determines melanoma-regulating role of tumor stromal fibroblasts
Authors: Shao Hongwei , Kong Ranran , Moller Mecker , Ferrari Massimiliano L. , Zhang Leiming , Radtke Freddy , Capobianco Anthony J. , Liu Zhao-Jun
Published: July 2016

Objectives. Tumor stromal fibroblasts are crucial in regulating tumor growth, invasion and metastasis, yet the molecular mechanisms that determine the tumor regulatory role of stromal fibroblasts remains unknown. Here, we uncover the Notch1 signaling pathway as a molecular determinant that controls the tumor regulatory role of tumor stromal fibroblasts in melanoma growth and invasion. Methods. Murine melanoma cells B16-F10, stably transduced with Luciferase 2 (Luc2)/lentivirus, were xenografted on the skin of two new mouse lines: the Gain-Of-Function of Notch1 (GOFNotch1: Fsp1.Cre+/-;ROSALSL-N1IC+/+) and Loss-Of-Function of Notch1 (LOFNotch1: Fsp1.Cre+/-;Notch1LoxP/LoxP+/+), respectively. GOFctrl (FSP1.Cre-/-;ROSALSL-N1IC+/+) and LOFctrl (FSP1.Cre-/-; Notch1LoxP/LoxP+/+) mice were used as control. Tumor growth was measured based on tumor weight and melanoma local invasion were examined by H&E staining of resected tumor tissue.

Results. Using a mouse melanoma model in which exogenous melanoma cells were grafted on the skin of two lines of mice where the genetic activation or inactivation of Notch1 signaling specifically occurs in natural host stromal fibroblasts, we demonstrated that Notch1 pathway activity could determine the tumor-promoting or tumor-suppressing phenotype in tumor stromal fibroblasts. Tumor stromal fibroblasts carrying elevated Notch1 activity significantly inhibited melanoma growth and invasion, while those with a null Notch1 promoted melanoma invasion.

Conclusions. These findings identify the Notch1 pathway as a molecular determinant that controls the regulatory role of tumor stromal fibroblasts in melanoma skin growth and invasion, unveiling Notch1 signaling as a potential therapeutic target for melanoma and potentially other solid tumors.

Citation Format: Hongwei Shao, Ranran Kong, Mecker Moller, Massimiliano L. Ferrari, Leiming Zhang, Freddy Radtke, Anthony J. Capobianco, Zhao-Jun Liu. Notch1 signaling determines melanoma-regulating role of tumor stromal fibroblasts. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4085.
Abstract 4092: High mast cell density is associated with improved prognosis in colon cancer patients
Authors: Sjölander. Anita
Published: July 2016

Background. Inflammatory cells and inflammatory mediators play an important role in colon cancer. However, the role of mast cells (MCs) in colon cancer progression remains unclear. Pro-inflammatory cysteinyl leukotrienes (CysLTs) have been associated with colorectal cancer (CRC) development. In particular, previous studies have shown that CRC patients with increased expression of cysteinyl leukotriene receptor 1 (CysLTR1) have poorer prognoses, and Cysltr1−/− mice display fewer intestinal polyps. The aim of the present study was to investigate the clinical and prognostic significance of mast cell density (MCD) in patients with CRC and to determine the significance of CysLTR1 on MCD in a mouse model of colitis-associated colon cancer (CAC).

Materials and methods. A tissue microarray (TMA) of primary colon cancers from 72 patients was stained with MC anti-tryptase and anti-chymase antibodies. Mouse colon tissue was stained with MC anti-tryptase antibody. Immunohistochemistry was used to identify MCD in cancerous and normal areas of patient tissues and in polyp/tumour, adjacent and normal areas of mouse tissues.

Results. In patient samples, the mean MCD was significantly higher in normal tissue than in cancer tissue (p = 0.0009). Patients with a high MCD in cancer tissue showed better overall survival compared with those with a low MCD (hazard ratio (HR) 0.575; 95% confidence interval (CI), 0.293 - 1.130). The same result was observed in patients aged 55-75 years (HR 0.332; 95% CI 0.121 - 0.911) and among patients without distant metastasis at diagnosis (HR 0.311; 95% CI 0.118 - 0.821). In the CAC mouse model, Cysltr1−/− mice had fewer intestinal polyps and significantly higher MCD in polyp/tumour areas compared with controls.

Conclusion. A high MCD in cancer tissue correlates with longer patient survival. In Cysltr1−/− CAC mice, a high polyp/tumour MCD also appears beneficial. Thus, these results indicate that a high MCD is a good prognostic factor.

Citation Format: Anita Sjölander. High mast cell density is associated with improved prognosis in colon cancer patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4092.
Abstract 4099: Oligodendrocyte stimulates the invasion ability of glioblastoma cells
Authors: Kawashima Toshiyuki
Published: July 2016

Background & Aims: Glioblastoma (GBM, WHO Grade IV) is characterized by rapid cancer cell infiltration, and is one of the most lethal neoplasms among solid cancers. GBM might arise de novo, or following progression from lower grade gliomas. Recently, solid tumor progression has been recognized as the product of an evolving crosstalk between the cancer cells and its surrounding glial cells. The intensive invasion activity of GBM cells might be regulated by normal cells such as oligodendrocytes or fibroblasts. In this study, we evaluated the interaction between GBM cells and glial cells.

Methods: Two GBM cell lines, T98G and U251, were used. Two oligodendrocyte cell lines, ODC1 and ODC2, were derived from surgical tissues of each patient with low-grade glioma (WHO Grade II). Two fibroblasts cell lines, GF1 and GF2, were derived from surgical tissues of each patient with GBM. The culture medium was composed of DMEM with 10% fetal calf serum. We examined the effects of glial cells on the invasion ability of GBM cells by wound-healing assay and invasion assay. Using IncuCyte invasion assay and matrigel invasion assay, the invasiveness of T98G and U251 cells was analyzed in the presence or absence of conditioned medium from oligodendrocyte cells or fibroblasts. The effect of the proliferation of T98G and U251 cells was examined by MTT assay or cell count.

Results: ODC1 and ODC2 cells were positive for Olig2 staining but not for GFAP staining, suggesting that ODC1 and ODC2 were oligodendrocyte cells. GF1 and GF2 cells were spindle-shape and were negative for both Olig2 staining and GFAP staining, suggesting that GF1 and GF2 cells were fibroblasts. Oligodendrocyte cells, ODC1 and ODC2 cells, from grade II glioma significantly (p<0.01) increased the migration and invasion ability of GBM cells, T98G and U251 cells. In contrast, GF1 and GF2 fibroblasts from GBM did not affect on the migration and invasion ability of GBM cells. Both oligodendrocyte cells and fibroblasts did not stimulate the proliferation of T98G and U251 cells.

Conclusion: Oligodendrocytes might up-regulate the invasiveness of malignant glioma cells. An invasion-stimulating factor(s) from oligodendrocyte cells might be partially associated with the progression from lowe-grade gliomas to GBM.

Citation Format: Toshiyuki Kawashima. Oligodendrocyte stimulates the invasion ability of glioblastoma cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4099.