bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 397: Brain natriuretic peptide (BNP) as molecular marker for bladder, ovarian, and glioblastoma cancers
Authors: Lee Cynthia , Yeh William , Lee Dongwoon , Dixon Samuel , Trieu Vuong
Published: July 2016

Background: BNP has been reported as molecular marker for ovarian cancer. This study evaluated BNP as a molecular marker for bladder, ovarian, glioblastoma, and pancreatic cancers. Methods: Serum samples collected at the time of diagnosis were tested using rapid and quantitative point-of-care (POC) devices for BNP blood biomarkers and the data was evaluated using JMP® 11.2.1 statistical analysis software. Samples were collected from bladder cancer patients (48 transitional cell carcinoma of the bladder), glioblastoma patients (51), pancreatic cancer patients (39 adenocarcinoma, 1 mucinous adenocarcinoma), ovarian cancer patients (24 serous adenocarcinoma, 20 serous papillary adenocarcinoma, 7 endometrioid adenocarcinoma, 3 clear cell carcinoma, 1 endometrial adenocarcinoma, 1 endometrioid papillary carcinoma, 1 serous papillary carcinoma), and normal subjects (n = 60). Results: Compared to normal subjects, bladder patients are more likely to be smokers (p = 0.0064). All four patient groups were more likely to be hypertensive (bladder, p = 0.0165; glioblastoma, p<0.0001, ovarian, p = 0.0002; pancreatic, p < 0.0001). No significant differences among the groups versus normal for weight were observed. Significant differences between the groups versus normal for BMI (ovarian, p = 0.0061), age (bladder, p = 0.0006; pancreatic = 0.0085), BNP (ovarian, p = 0.0012; glioblastoma, p = 0.0016; bladder, p = 0.0112) were noted. There was positive correlation between age and BNP level among all four cancer groups (bladder, p = 0.0133; glioblastoma, p = 0.0629; ovarian, p = 0.0304; pancreatic, p = 0.0172); whereas there was a negative correlation between age and BNP level among normal controls (p = 0.0109). Age-adjusted BNP level was highly significant different versus normal (bladder, p <0.0001; ovarian, p = 0.0013; glioblastoma, p = 0.0047). Neither BNP nor age-adjusted BNP level was significantly different from normal for pancreatic cancer patients. Hypertensive condition had no impact on BNP or age-adjusted BNP. Conclusions: Bladder, ovarian, and glioblastoma cancers are associated with significant reduction in BNP and age-adjusted BNP blood levels. Pancreatic cancer was an exception. BNP level could be useful in the monitoring of treatment efficacy in which the BNP POC device described here would be highly appropriate due to its ease in deployment at the patient's home or doctor's office.

Citation Format: Cynthia Lee, William Yeh, Dongwoon Lee, Samuel Dixon, Vuong Trieu. Brain natriuretic peptide (BNP) as molecular marker for bladder, ovarian, and glioblastoma cancers. [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 397.
Abstract 3970: Impact of endoscopic stent insertion on detection of viable circulating tumor cells for obstructing colon cancer
Authors: Yamashita Shinya
Published: July 2016

The obstructive colorectal cancer(CRC) was reported to be 7-20% of CRC. For the patients (pts) with obstructive colorectal cancer, self-expanding metallic stents (SEMS) have been useful as a palliative treatment. Insertion of SEMS for obstructive colorectal cancer is used as a bridge to surgery approach. But suspicion have been raised about whether shear forces that act on the tumor during expansion of the stent may result in a release of cancer cells into the circulation (i.e., CTCs). This study was conducted to clarify whether insertion of colonic stents results in increased levels of viable CTCs (v-CTCs). For detecting viable CTCs(v-CTCs) of CRC, we employed a novel method, TelomeScan F35 detection system. The system was constructed with a GFP-expressing attenuated adenovirus, in which the telomerase promoter regulates viral replication. 7.5 ml of peripheral blood samples were obtained before and after stent insertion and after operation. GFP-positive and CD45-negative cells were counted as v- CTCs. This study was a single institution trial, approved by the Kure Medical Center IRB. Surgical operation was scheduled within 1 month after stenting. Between October 2013 and June 2015,10 pts aged 58-89 years (5 males and 5 females) were enrolled. The surgical operation (R0) was performed in 9 cases. CEA was reduced to <50% of baseline value in 6 of 9 measurable pts after resection. 6 pts had no v-CTCs before stenting. Among them, 3 pts had increasing number of 1-3 v-CTCs after stenting and 1 of them developed multiple liver metastasis 12 month after resection. While, 4 pts had v-CTCs (1-19 cells) before stenting and 3 of these 4 patients had significant increase of number of v-CTCs (1-21 cells) after stent insertion. 2 of them had 12 and 14 v-CTCs, respectively after operation, despite the normalized value of CEA. The present study suggested that endoscopic stent insertion may result in tumor cell dissemination into the peripheral circulation and may induce early distant metastasis. In conclusion, v-CTCs may become useful for prognosis assessment, especially when SEMS insertion is considered.

Citation Format: Shinya Yamashita. Impact of endoscopic stent insertion on detection of viable circulating tumor cells for obstructing colon 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 3970.
Abstract 3980: Combined detection of serum Dickkopf-1 and its autoantibodies to diagnose esophageal squamous cell carcinoma
Authors: Peng Yu-Hui , Xu Yi-Wei , Li En-Min , Xu Li-Yan
Published: July 2016

Esophageal squamous cell carcinoma (ESCC) can be treated effectively if diagnosed at an early stage. We evaluated whether measurement of Dickkopf-1 (DKK1) in combination of DKK1 autoantibodies in serum may benefit early diagnosis of ESCC. Serum DKK1 and DKK1 autoantibodies were measured by enzyme-linked immunosorbent assay in a training cohort (185 ESCC samples vs. 97 normal controls) and validated in a validation cohort (104 ESCC samples vs. 53 normal controls). Receiver operating characteristics (ROC) was applied to calculate diagnostic accuracy. The serum levels of DKK1 and DKK1 autoantibody were significantly higher in ESCC patients than in controls, respectively (P<0.0001). Measurement of serum DKK1 demonstrated an area under curve (AUC) of 0.709 [95% confidence interval (CI), 0.647-0.771], 37.3% sensitivity, and 90.7% specificity in the training set and an AUC of 0.697 (95% CI, 0.613-0.780), 41.3% sensitivity, and 84.9% specificity in the validation set. Testing of DKK1 and DKK1 autoantibodies together effectively improved the diagnostic accuracy for ESCC versus normal controls compared with either test alone (AUC 0.769, 95% CI 0.715-0.823, 50.3% sensitivity, and 90.7% specificity in the training cohort; AUC 0.752, 95% CI 0.675-0.829, 50.0% sensitivity, and 84.9% specificity in the validation cohort). Importantly, the diagnostic performance of the combination of DKK1 and DKK1 autoantibodies persisted in early ESCC patients (AUC 0.780, 95% CI, 0.699-0.862, 50.0% sensitivity, and 90.7% specificity in the training cohort; AUC 0.745, 95% CI, 0.626-0.865, 53.8% sensitivity, and 84.9% specificity in the validation cohort). Furthermore, the levels of serum DKK1 or DKK1 autoantibody after surgical resection were lower, respectively, compared with the corresponding preoperative samples (P<0.05). Our results suggest that measurement of DKK1 combined with DKK1 autoantibodies is a potentially valuable tool for the early detection of ESCC.

Citation Format: Yu-Hui Peng, Yi-Wei Xu, En-Min Li, Li-Yan Xu. Combined detection of serum Dickkopf-1 and its autoantibodies to diagnose esophageal 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 3980.
Abstract 3985: First case of B LAL with MLL - MAML2 rearrangement
Authors: Usseglio Fabrice , Beaufils Nathalie , Costello Régis , Gabert Jean
Published: July 2016

Over 70 MLL partner genes have been reported so far. MLL gene (or 11q23) translocation is occurring in 7-10% of B acute lymphoblastic leukemias and AF4 being the most frequent partner gene. The prognostic is usually bad, specially for AF4 partner. A rare partner gene, mastermind like 2 (MAML2) gene has been reported in 4 cases of myeloid neoplasms after chemotherapy so far: 2 AML and 2 MDS. Here we report a case with MLL - MAML2 discovered by NGS analysis in front of an inv11 (q21q23) present in a 47 years old female previously treated for a sarcoma in 2014. It is, to our knowledge, the first case of B acute lymphoblastic leukemia (CD79a, CD22, CD19, CD45, CD10-) with this fusion gene. At the molecular level, the rearrangement consists in the fusion of exons 7 and 2 of the MLL and MAML2 genes respectively. We will discuss the biological, clinical, physiological and therapeutic consequences of this rare event. Its prognostic impact is unknown as the reported cases are too rare.

Citation Format: Fabrice Usseglio, Nathalie Beaufils, Régis Costello, Jean Gabert. First case of B LAL with MLL - MAML2 rearrangement. [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 3985.
Abstract 3998: Sequencing the B-cell and T-cell repertoire
Authors: Stewart Fiona J. , Karaca Mehmet , Adams Kris , Clouser Chris , Patel Bonny , Timberlake Sonia , Donahue William , Apone Lynne , Russello Salvatore , Dimalanta Eileen T. , Davis Theodore B. , Vigneault Francois
Published: July 2016

Immune sequencing allows for the study of complex immunological diseases by sequencing millions of V(D)J combinations from B-cell antibody and T-cell receptors. The popularity of this technique has increased due to recent throughput and read length improvements in next-generation sequencing technologies. However, structural and sequence complexities of antibody genes have made reliable targeting approaches challenging.

We have developed and optimized a method for accurate sequencing of full-length immune gene repertoires of B-cells and T-cells. The method uses a unique barcoding scheme specifically designed to tag every mRNA molecule with a unique identifier (UID) so that all PCR copies of each mRNA fragment can be collapsed into a single consensus sequence. This makes the assay extremely accurate, by resolving PCR bias and sequencing errors as well as allowing quantitative digital molecule counting.

Immune sequencing libraries were generated from total RNA extracted from Peripheral Blood Mononuclear Cells in duplicate from a single patient. The use of UIDs enabled absolute quantification of starting RNA molecules present in the original sample and therefore accurate ranking of the antibody clone abundance, by avoiding the bias incorporated by PCR or sequencing when total reads only were measured. Using the same sequencing method, tumor samples were analyzed for abundance of expanded clones via grouping clones by V gene, J gene and CDR3 similarity and ranking by mRNA abundance. Additionally, the use of isotype-specific primers (IgM, IgD, IgG, IgA and IgE) enabled measurement of the heavy chain isotype proportions within the samples. Further, alignment of full-length heavy chain antibody sequences generated using this method to germline genes from reference databases enabled quantitation of the mutation level of each antibody sequence, thereby providing information on the overall maturity and mutational profile of the sample repertoire.

Citation Format: Fiona J. Stewart, Mehmet Karaca, Kris Adams, Chris Clouser, Bonny Patel, Sonia Timberlake, William Donahue, Lynne Apone, Salvatore Russello, Eileen T. Dimalanta, Theodore B. Davis, Francois Vigneault. Sequencing the B-cell and T-cell repertoire. [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 3998.
Abstract 3999: Altering the balance between immune activation versus regulation in the skin to promote CD8 T-cell activity within epithelial cancers
Authors: Bridge Jennifer A. , Overgaard Nana H. , Steptoe Raymond J. , Frazer Ian H. , Wells James W.
Published: July 2016

The Human Papilloma Virus (HPV) 16 is a high-risk HPV known to be a causative agent in numerous cancers including cervical cancer. While prophylactic vaccines exist to combat the spread of HPV16, successful therapeutic vaccines to combat established HPV16-associcated disease remain elusive. The expression, in a mouse model (“E7”), of the HPV16 E7 gene in keratinocytes under the control of the K14 promoter, leads to a local immune suppressive environment, as evidenced by the lack of graft rejection when E7 skin grafts are placed on WT recipient mice. Furthermore, well healed (>30 days) E7 skin grafts are not rejected when mice are immunised with E7 peptide in combination with Quil A- or CASAC-based adjuvants. This is despite a substantial increase in E7 peptide/H-2Db pentamer staining in the blood, and marked killing of E7-peptide expressing TC-1 cells when injected i.v., confirming that CD8 T-cells respond to vaccination and differentiate into CTL capable of killing E7-expressing target cells. We hypothesised that the removal of regulatory T-cells (T-reg) might lead to E7 graft rejection in immunised mice. The co-administration of an anti-CD4-depeting antibody at the time of immunisation led to rejection of ∼50% of grafts. To confirm a role for T-reg, E7-grafted T-reg-deficient Rag1-/- mice received purified donor CD8 T-cells from E7-vaccinated WT mice. FACS staining of Rag1-/- lymph nodes 30 days post CD8+ T-cell transfer confirmed the absence of classical CD4+FoxP3+ Treg, however the E7 grafts did not reject. As in the WT mice however, rejection could be induced through the co-administration of an anti-CD4 antibody. The data suggest that the removal of a CD4+, non T-reg cell, leads to CD8+ T-cell activity in the skin as evidenced by E7 skin graft destruction.

Citation Format: Jennifer A. Bridge, Nana H. Overgaard, Raymond J. Steptoe, Ian H. Frazer, James W. Wells. Altering the balance between immune activation versus regulation in the skin to promote CD8 T-cell activity within epithelial cancers. [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 3999.
Abstract 40: Metabolic rewiring in Rb deficient cells during cancer progression
Authors: Kohno Susumu , Okahashi Nobuyuki , Kitajima Shunsuke , Suzuki Sawako , Tanaka Tomoaki , Matsuda Fumio , Shimizu Hiroshi , Takahashi Chiaki
Published: July 2016

Tumor suppressors control critical steps of cellular metabolism and its inactivation induces cancer specific-metabolic features including aerobic glycolysis. Glycolysis contributes to energy generation and supplying precursors of biomass, e.g., nucleotide, amino sugar, amino acid and lipids. These macromolecules were required for proliferation and cancer survival. Although aerobic glycolysis is now widely accepted as a metabolic feature of cancer, its causal relationship with cancer progression is still unclear.

In common type cancers, inactivation of RB tumor suppressor is prevalently observed during tumor progression. Previous studies demonstrated RB inactivation induces inflammation, angiogenesis, drug resistance, etc., as well as cell cycle progression. We currently focus on the impact of RB deficiency on the central carbon metabolism, e.g., glycolysis, TCA cycle and glutamine metabolism. To address this point, we established a mouse model in which RB depletion in p53-null soft tissue sarcoma cells induced undifferentiated phenotype associated with “glycolytic to glutaminolytic” switch (metabolic rewiring) most probably due to down-regulation of Pgam2 activity. Up-regulation of Pgam2 in these cells antagonized the metabolic rewiring, and suppressed spherogenic and tumorigenic activities of RB deficient cells, however did not affect cell proliferation in 2D culture conditions. Moreover, Pgam2 depletion suppressed RB-dependent myogenic differentiation. These findings suggest that RB controls tumorigenicity and differentiation by influencing central carbon metabolism via Pgam2.

Citation Format: Susumu Kohno, Nobuyuki Okahashi, Shunsuke Kitajima, Sawako Suzuki, Tomoaki Tanaka, Fumio Matsuda, Hiroshi Shimizu, Chiaki Takahashi. Metabolic rewiring in Rb deficient cells during cancer 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 40.
Abstract 4031: Different expression of programmed death 1 (PD1) and its ligand (PD-L1) in esophageal and gastric cancer
Authors: Däster Silvio , Eppenberger-Castori Serenella , Droeser Raoul A. , Schaefer Hannah M. , Spagnoli Giulio C. , Terracciano Luigi , Tornillo Luigi , von Holzen Urs
Published: July 2016

Background: Prognosis of gastric and esophageal cancer remains poor. Improvements in gastric and esophageal cancer treatments are urgently needed. Programmed cell death 1 receptor (PD1) and its ligand 1 (PD-L1) are known to interact with T cells promoting epithelial cancer tolerance. Several therapeutic monoclonal antibodies blocking this interaction are reaching the clinical praxis. Therefore it is relevant to investigate the role of these biomarkers in different types of malignancy.

Methods: Four different copies of tissue microarrays (TMA), each including healthy mucosa (n = 74) and a total of 241 clinically annotated malignancies of the esophagus (n = 80) and stomach (n = 161) were constructed and stained with PD1, PD-L1, and CD8 specific reagents.

Results: Interestingly, only two cancer samples out of the 241 specimens investigated weakly expressed PD-L1. None of the normal mucosa epithelial cells expressed PD-L1. Stromal PD1 expression correlated with infiltration by CD8+ lymphocytes (rho = 0.4; P<0.0001). Elevated intraepithelial presence of CD8+ and/or PD1+ cells in gastric cancer was significantly correlated with longer patients’ survival. No correlation could be found in esophageal cancer.

Conclusion: In contrast to other epithelial cancers, PD-L1 expression was virtually absent in the investigated malignancies of the esophagus and stomach. PD1 expression in the stroma surrounding such malignancies correlated with intraepithelial presence of T-cells CD8+ expression, and with better prognosis in gastric cancer patients.

Citation Format: Silvio Däster, Serenella Eppenberger-Castori, Raoul A. Droeser, Hannah M. Schaefer, Giulio C. Spagnoli, Luigi Terracciano, Luigi Tornillo, Urs von Holzen. Different expression of programmed death 1 (PD1) and its ligand (PD-L1) in esophageal and gastric 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 4031.
Abstract 4037: Enhancement of efferocytosis by rhoA kinase inhibitor following doxorubicin treatment synergistically reduces tumor growth
Authors: Nam Gihoon , Kim In-San
Published: July 2016

Massive dying of tumor cells after chemotherapy may promote chronic inflammation which favors tumor growth by uncleared secondary necrotic cells. Given that defects in apoptotic cell clearance have been linked with various inflammatory diseases and autoimmunity, the prompt removal of apoptotic cells by phagocytes is important for anti-tumor immunity. To improve the anti-tumor effect of doxorubicin, we investigated the role of RhoA-kinase (ROCK) inhibitors on efferocytosis by macrophages, which is important for anti-tumor immunity and tissue regeneration. Treatment of bone marrow-derived macrophages (BMDMs) with ROCK inhibitor increased efferocytosis of dying tumor cells by doxorubicin in vitro. ROCK-blockade after doxorubicin treatment group showed effective inhibition of tumor growth and also T cell-mediated elimination of immunogenic tumors more than the doxorubicin monotherapy group in established tumor models. Furthermore, we also confirmed that macrophage depletion abrogated the therapeutic effect of ROCK inhibitor after doxorubicin treatment on tumor growth. Taken together, our findings indicate that ROCK inhibitor following doxorubicin treatment reduced tumor growth via efferocytosis enhancement.

Citation Format: Gihoon Nam, In-San Kim. Enhancement of efferocytosis by rhoA kinase inhibitor following doxorubicin treatment synergistically reduces tumor growth. [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 4037.
Abstract 4052: The role of the phosphorylated RXRα on diethylnitrosamine-induced liver tumorigenesis in mice
Authors: Sakai Hiroyasu , Shirakami Yohei , Shimizu Masahito
Published: July 2016

We have previously reported that a malfunction of RXRα due to aberrant phosphorylation at serine 260 by MAPK is associated with the development of hepatocellular carcinoma (HCC). The phosphorylated RXRα impairs normal receptor functions of either RXR or RAR, which are respective homodimeric or heterodimeric partner of RXRα, in a dominant-negative manner. Besides, these impaired receptor functions result in the downregulation of their target gene expression and inducing hepatocarcinogenesis by stimulating cell proliferation and inhibiting apoptosis, suggesting that phosphorylated RXRα plays a crucial role in the development of HCC. However, these findings were revealed primarily using HCC cell lines that express the phosphorylated-RXRα (p-RXRα) proteins and it still remains unclear whether p-RXRα may impact in the process of hepatocarcinogenesis in vivo. Therefore, in order to investigate the biological functions of p-RXRα on hepatocarcinogenesis in vivo, we generated doxycycline-inducible ES cell line and transgenic mouse, both of which overexpress phosphomimic mutant of RXRα (T82D/S260D). Interestingly, the transcriptional activities of Retinoid X Receptor Response Element (RXRE) were significantly reduced in the Mouse Embryonic Fibroblasts (MEFs) derived from the transgenic mice, suggesting that the retinoid signaling via RXRα were impaired in the mice. Moreover, the development of liver tumors induced by liver carcinogen diethylnitrosamine (DEN) was enhanced in the transgenic mice that overexpress T82D/S260D protein in the liver. Notably, the increased liver tumors in the transgenic mice resulted from the promotion of cell cycle progression, not the inhibition of apoptosis. Interestingly, expression of β-catenin and its target genes, cyclin D1 and c-myc, were increased in transgenic liver tumors, although the expression of retinoid-related genes, such as RARβ, p21 and p27, were not altered compared with control mice. These results suggest that the phospho-modification of RXRα may promote the development of DEN-induced liver tumors through the activation of β-catenin signaling pathways in mice.

Citation Format: Hiroyasu Sakai, Yohei Shirakami, Masahito Shimizu. The role of the phosphorylated RXRα on diethylnitrosamine-induced liver tumorigenesis in mice. [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 4052.
Abstract 4059: Molecular role of cytochrome P450 (CYP)1A2 in the regulation of hepatic and pulmonary CYP1A1 expression in mice exposed to 3-Methycholanthrene
Authors: Moorthy Bhagavatula , Maturu Paramahamsa , Wang Lihua , Jiang Weiwu
Published: July 2016

3-Methylcholanthrene (MC) is one of the most potent polycyclic aromatic hydrocarbons (PAHs) carcinogens known to date. PAHs are present in cigarette smoke, diesel exhausts, and charcoal broiled meats, etc. Cytochrome P450 (CYP) 1A enzymes play key roles in the activation of PAHs to carcinogenic metabolites. In this investigation, we tested the hypothesis that CYP1A2 plays a mechanistic role in the regulation of hepatic and pulmonary CYP1A1 by MC. Wild type (WT) (C57B6) and Cyp1a2_null mice were divided into two groups. Group I was treated with vehicle corn oil (CO) (8 ml/kg) and group II was treated with four doses of MC (100 μmol/kg), i.p. once daily for 4 days. Four animals from each group were sacrificed at 6, 12, 24, and 48 h, 8 and 15 days after MC withdrawal. The mRNA levels, protein content and enzyme activities of CYP1A1 and 1A2 were determined at different time points. In addition, the binding of MC-AHR-AHR nuclear translocator (ARNT) to the AHREs on the CYP1A1 promoter region was determined by chromatin immunoprecipitation (ChIP) assay in liver and lung tissues. In WT mice, ChIP experiments indicated that transcriptional activation of CYP1A1 was most pronounced at 6 h, followed by 12 h, but declined at later time points. In Cyp1a2_null mice, on the other hand, transcriptional activation of CYP1A1 persisted till 48 h. In lungs, AHR complex binding peak was observed at 12 h and declined later, but Cyp1a2 null mice showed more persistent binding till 48 h with a peak at 24 h. On the other hand, the expression of CYP1A1/1A2 at the mRNA, protein, and enzyme levels persisted for up to 48 h both in lung and liver tissues. These results suggest a mechanistic role for CYP1A2 in the molecular regulation of CYP1A1, and that transcriptional activation of CYP1A1 at 6-12 h is sufficient to result in sustained induction of CYP1A mRNA and protein expression for up to 48 h. These studies have important implications for carcinogenesis mediated by PAHs.

Citation Format: Bhagavatula Moorthy, Paramahamsa Maturu, Lihua Wang, Weiwu Jiang. Molecular role of cytochrome P450 (CYP)1A2 in the regulation of hepatic and pulmonary CYP1A1 expression in mice exposed to 3-Methycholanthrene. [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 4059.
Abstract 4066: The chemokine CXCL13 in lung cancers associated with environmental polycyclic aromatic hydrocarbons pollution
Authors: Wang Gui-Zhen , Cheng Xin , Zhou Guang-Biao
Published: July 2016

More than 90% of lung cancers are caused by cigarette smoke and air pollution, with polycyclic aromatic hydrocarbons (PAHs) as key carcinogens. In Xuanwei City of Yunnan Province the lung cancer incidence is among the highest in China attributed to smoky coal combustion-generated PAH pollution. Here we screened for abnormal inflammatory factors in non-small cell lung cancers (NSCLCs) from Xuanwei and control regions (CR) where smoky coal was not used, and found that a chemokine CXCL13 was overexpressed in 63/70 (90%) of Xuanwei NSCLCs and 44/71 (62%) of smoker and 27/60 (45%) of non-smoker CR patients. CXCL13 overexpression was associated with the region Xuanwei and cigarette smoke. The key carcinogen bezo(a)pyrene (BaP) induced CXCL13 production in lung epithelial cells and in mice prior to development of detectable lung cancer. Deficiency in Cxcl13 or its receptor, Cxcr5, attenuated BaP-induced lung cancer in mice, demonstrating CXCL13's critical role in PAH-induced lung carcinogenesis.

Citation Format: Gui-Zhen Wang, Xin Cheng, Guang-Biao Zhou. The chemokine CXCL13 in lung cancers associated with environmental polycyclic aromatic hydrocarbons pollution. [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 4066.
Abstract 4068: Role of cytochrome P450 (CYP)1A2 in the molecular regulation of CYP1A1 by the carcinogen, 3-methylcholanthrene (MC) in mouse heptoma cells (hepa-1)
Authors: Kondraganti Sudha , Chu Chun , Moorthy Bhagavatula
Published: July 2016

Cytochrome P4501A (CYP1A) enzymes play important roles in the activation of PAHs such as 3-methylcholanthrene (MC) to carcinogenic DNA-binding metabolites. We reported earlier that MC causes persistent induction of hepatic and pulmonary CYP1A1 in mice for several weeks after MC withdrawal, and that the phenomenon of sustained hepatic CYP1A1 induction is lost in Cyp1a2-null mice. In this study, we tested the hypothesis that MC elicits induction of persistent CYP1A1 induction in hepa-1 cells, and that CYP1A2 contributes mechanistically to this phenomenon. Hepa-1 cells were treated with the MC (2.5 μM), or dimethylsulfoxide (DMSO) as control, and at selected time points, CYP1A1 promoter activity, CYP1A1 enzyme activities, contents, and CYP1A1 mRNA levels were studied. A 24 h treatment induced the CYP1A1 promoter activity, CYP1A1 mRNA, and ethoxyresorufin O-deethylase (EROD) activity by 2-, 1000-, and 15-fold, respectively. The induction was sustained for 4-5 days. MC persistently induced CYP1A1 apoprotein level as well. Electrophoretic mobility shift assay indicated that MC induced a nuclear protein that bound to aryl hydrocarbon response elements in the CYP1A1 promoter region. Transfection of CYP1A2 siRNA resulted in knockdown of CYP1A2 mRNA by 70%, but a statistically significant increase of basal CYP1A1 mRNA by 35-40%. The induction of CYP1A1 promoter activity, CYP1A1 mRNA, CYP1A1 protein, and EROD activity by MC were not affected by CYP1A2 siRNA at the 24 h time point, but was significantly attenuated by CYP1A2 siRNA on day 5. These results suggest that CYP1A2, possibly via a metabolite, contributes to the sustained induction of CYP1A1 by MC in hepa-I cells. Further investigations into the mechanisms of persistent induction of CYP1A1 by MC could lead to novel preventative/therapeutic strategies against PAH-mediated carcinogenesis in humans.

Citation Format: Sudha Kondraganti, Chun Chu, Bhagavatula Moorthy. Role of cytochrome P450 (CYP)1A2 in the molecular regulation of CYP1A1 by the carcinogen, 3-methylcholanthrene (MC) in mouse heptoma cells (hepa-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 4068.
Abstract 4071: Targeted disruption of T-cell protein tyrosine phosphatase in mouse epidermis reveals its critical role in chemically-induced skin carcinogenesis
Authors: Lee Hyunseung , Kim Mihwa , Morales Liza D. , Slaga Thomas J. , DiGiovanni John , Kim Dae J.
Published: July 2016

Environmental factors such as chemical toxicants contribute to the development of skin cancer by creating mutations in housekeeping genes and proto-oncogenes which disrupt intracellular signaling mechanisms. One vital signaling mechanism is tyrosine phosphorylation signaling. Phosphotyrosine signaling is regulated by the counter-activities of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Recently we reported TC-PTP (PTPN2) is critical regulator of STAT3 signaling in mouse keratinocytes. To assess the role of TC-PTP in skin carcinogenesis, we generated epidermal-specific TC-PTP-deficient (K14Cre.PTPN2fl/fl) transgenic mice. Loss of TC-PTP led to a desensitization to tumor initiator 7,12-dimethylbenz[a]anthracene (DMBA)-induced apoptosis both in vivo epidermis and in vitro keratinocytes. TC-PTP deficiency in epidermis resulted in a significant increase in epidermal thickness and hyperproliferation (assessed by bromodeoxyuridine (BrdU) labeling index) compared to control mice following treatment with the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). Consistently, primary keratinocytes derived from TC-PTP-deficient mice showed a faster growth rate. To further investigate the role of TC-PTP in skin carcinogenesis, TC-PTP-deficient mice were subjected to two-stage skin carcinogenesis analysis. TC-PTP-deficient mice showed a shortened latency of tumor development and significantly increased numbers of tumors compared to control mice, demonstrating TC-PTP deficiency predisposes mice to skin carcinogenesis. The data reveals TC-PTP may be a novel potential target for the prevention of skin cancer due to its role in the regulation of proliferation and apoptosis in epidermis.

Citation Format: Hyunseung Lee, Mihwa Kim, Liza D. Morales, Thomas J. Slaga, John DiGiovanni, Dae J. Kim. Targeted disruption of T-cell protein tyrosine phosphatase in mouse epidermis reveals its critical role in chemically-induced skin carcinogenesis. [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 4071.
Abstract 4083: Tumor associated macrophages mediates the oncogenic effect of FGF18 in ovarian cancer
Authors: Wei Wei , Birrer Michael
Published: July 2016

Epithelial ovarian cancer is the fifth leading cause of cancer-related death among women and has the highest case-fatality rate among gynecologic cancers. Effective management of this deadly disease still remains scanty due to the inadequate understanding of the molecular mechanisms associated with the tumor progression and prognosis. We have previously reported the tumor-promoting role of Fibroblast growth factor 18 (FGF18), which have been identified through genomic analysis of ovarian tumors. Initial studies suggested FGF18 promotes ovarian tumor progression through NF-κB pathway mediated elevation of proinflammatory cytokines, which mediate the increased tumor burden, angiogenesis and infiltration of macrophages in FGF18 expressing xenografts. Based on these findings, the present study aims to delineate the mechanism by which FGF18 modulates the phenotype of tumor associated macrophages (TAMs) as well as the reciprocal tumor-promoting feedback from the TAMs which potentially contributes FGF18 mediated ovarian tumorigenesis.

Immunohistochemical staining of a tissue array comprising 216 archived high-grade advanced stage serous ovarian tumor specimens revealed a significant correlation between the FGF18 expression and infiltration of M2-polarized macrophages (by CD163). In addition, the density of intratumoral M2 TAMs inversely associated with patients’ overall survival as a negative prognostic factor independent to age, tumor grade and debulking status. These findings suggest intratumoral TAMs may play an important role in ovarian tumorigenesis. In vitro co-culture studies further demonstrated that human monocyte cell line THP-1 or murine peritoneal macrophage cell line IC-21 could significantly activate the NF-κB pathway and increase the production of various proinflammatory cytokines in A224 ovarian cancer cells. Co-culture also increased the migratory potential of A224 cells. Addition of an IKKβ inhibitor into the co-culture system nullified the cytokine production and migration of tumor cells promoted by monocytes/macrophages, implicating the involvement of canonical NF-κB signaling in the crosstalk. Conversely, liposome clodronate mediated in vivo macrophage depletion significantly shrunk the tumor burden of the intraperitoneal xenograft derived from FGF18 overexpressing SKOV3 cells, while minimal effect was observed over the control RFP overexpressing xenograft. Besides, macrophage depletion also markedly decreased the intratumoral microvessel density induced by FGF18 overexpression. Taken together, our data potentially suggest TAMs may mediate the FGF18 related ovarian tumorigenesis by augmenting the proinflammatory status of the tumors.

Citation Format: Wei Wei, Michael Birrer. Tumor associated macrophages mediates the oncogenic effect of FGF18 in ovarian 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 4083.
Abstract 4086: Critical roles of FRS2beta, a feedback inhibitor of ErbB-ERK signaling, for communication between cancer stem cells and their niche during mammary tumorigenesis
Authors: Machida Yukino , Kimura Natsuko , Tojo Arinobu , Akashi Ko-ichi , Saya Hideyuki , Kitabayashi Issay , Gotoh Noriko
Published: July 2016

Inflammatory microenvironment contributes to tumorigenesis. Although it is thought that breast cancer stem cells (CSCs) appear and grow in the inflammatory microenvironment that is called CSC niche, it remains largely unclear how it occurs. Here, we uncovered FRS2beta, a feed-back inhibitor of ERK activity in progenitor cells, plays critical roles for various cytokine production to increase progenitor cell- and CSC- state, and create the CSC niche. Expression levels of FRS2beta were increased in mammary tumors of MMTV-ErbB2 mice than in normal mammary tissues. Deficiency of FRS2beta greatly delayed mammary tumorigenesis, decreased mammosphere-forming ability and tumor stroma, a component of the CSC niche. Expression levels of various cytokines, including IGF1 and CXCL12, stemness- and stroma-inducing cytokines, respectively, were reduced in FRS2beta-mutant mammospheres, pre-cancerous mammary and tumor tissues. Furthermore, expression levels of FRS2beta were higher in CD44highCD24low CSCs-enriched population in patient-derived breast cancer tissues. Thus the mechanisms to maintain progenitor cell-state by prevention of excess ERK activity may permit production of various cytokines to create CSC niche, and allow appearance and growth of CSCs. These results suggest that treatment with MEK inhibitors may allow survival of CSCs, raising the risk of recurrence. We provide a rationale of combination therapy targeting both FRS2beta and MEK inhibitors to eradicate tumors.

Citation Format: Yukino Machida, Natsuko Kimura, Arinobu Tojo, Ko-ichi Akashi, Hideyuki Saya, Issay Kitabayashi, Noriko Gotoh. Critical roles of FRS2beta, a feedback inhibitor of ErbB-ERK signaling, for communication between cancer stem cells and their niche during mammary tumorigenesis. [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 4086.
Abstract 4088: Adipocytes-mediated autophagy activation and metabolic reprogramming promotes colon cancer survival
Authors: Wen Yang-an , Xiong Xiaopeng , Harris Jennifer , Zaytseva1 Yekaterina , Gao Tianyan
Published: July 2016

Obesity has been associated with increased incidence and mortality of a wide variety of human cancers including colorectal cancer. However, the molecular mechanism underlying the functional interaction between obesity and cancer remains elusive. Here, we demonstrated that adipocytes isolated from adipose tissues of colon cancer patients play an important role in promoting tumor cell survival and progression by altering cellular metabolism. In stage IV colon cancer cases, abundant adipocytes were found in close association with invasive colon cancer cells. Co-culture of adipocytes with colon cancer cells led to direct transfer of free fatty acids (FAs) released by the adipocytes to colon cancer cells. Uptake of FAs allowed the cancer cells to survive nutrient deprivation conditions by upregulating mitochondrial fatty acid β-oxidation, suggesting that FAs from adipocytes were used as an energy source by the cancer cells. In addition, we found that co-culture of adipocytes or treating cells with fatty acids induced autophagy in colon cancer cells as a result of AMPK activation. Inhibition of autophagy attenuated the ability of cancer cells to utilize FAs and blocked the growth promoting effect of adipocytes. Furthermore, we found that adipocytes induced dedifferentiation of tumor cells in primary colon cancer cells and mouse tumor organoids. Taken together, these results identify adipocytes as active contributors to the tumor microenvironment that promote tumor growth and survival by serving as an energy provider and a metabolic regulator for the embedded colon cancer cells.

Citation Format: Yang-an Wen, Xiaopeng Xiong, Jennifer Harris, Yekaterina Zaytseva1, Tianyan Gao. Adipocytes-mediated autophagy activation and metabolic reprogramming promotes colon cancer survival. [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 4088.
Abstract 4096: Fibroblast growth factor-2-derived from cancer-associated fibroblasts stimulates proliferation and migration of human breast cancer cells
Authors: Suh Jinyoung , Kim Do-Hee , Surh Young-Joon
Published: July 2016

Cancer-associated fibroblasts (CAFs) constitute a major compartment of the tumor microenvironment. CAFs produce a variety of cytokines, growth factors and extracellular matrix proteins, thereby stimulating tumor progression. CAFs are distinct from normal fibroblasts for their overexpression of α-smooth muscle actin and fibroblast specific protein-1. Recent studies suggest that CAFs play an important role in proliferation and migration of cancer cells through crosstalk with them.In the present study, we investigated the role for CAFs in proliferation and migration of breast cancer cells and underlying molecular mechanisms.When triple-negative human mammary MDA-MB-231 cells were treated with the conditioned medium (CM) collected from cultured CAFs, the cell viability and migration were significantly elevated. Furthermore, these cells manifest a more proliferative phenotype, exhibiting enhanced mRNA expression of cycinD1, c-Myc and PCNA as well as increased phosphorylation of Akt and STAT3.In addition, MDA-MB-231 cells exhibited elevated expression of proliferative and invasive genes including MMP2 and MMP9 when incubated with CAFs in the indirect co-culture system. Notably, mRNA levels of fibroblast growth factor-2 (FGF2), stromal-derived factor-1, interleukin-6 and interleukin-8 detected in CAFs were higher than those in normal fibroblasts of the same patients. In contrast, FGF2 was expressed at a relatively low level in breast cancer cells. FGF2 exerts its biological effects by binding to and activating FGF receptor (FGFR), a subfamily of cell surface receptor tyrosine kinases. Notably, the expression of FGFR1 was up-regulated in triple-negative breast cancer cells including MDA-MB-231 cells while another major FGF2-corresponding receptor FGFR2 was rarely expressed. Therefore, we focused on FGF2-FGFR1 signaling in the context of paracrine communication between breast cancer cells and CAFs. CAF-stimulated MDA-MB-231 cell migration as well as FGFR1 expression was abolished when FGF2-neutralizing antibody was added to the CAF-CM. In addition, treatment of MDA-MB-231 cells with FGF2 induced the phosphorylation of FRS2 and Akt. FGF2-induced cell migration and up-regulation of cyclinD1 expression were abrogated by siRNA-mediated FGFR1 silencing. Furthermore, FGF2 promotes nuclear localization of FGFR1. Taken together, above findings suggest that secretion of FGF2 by CAFs stimulates proliferation and migration of breast cancer cells through interaction with FGFR1 which may contribute to human breast cancer progression.

Citation Format: Jinyoung Suh, Do-Hee Kim, Young-Joon Surh. Fibroblast growth factor-2-derived from cancer-associated fibroblasts stimulates proliferation and migration of human breast cancer 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 4096.
Abstract 4114: Blocking Ezrin can inhibit HGF/Met signaling-induced metastasis
Authors: Yu Yanlin , Huang Liping , Merlino Glenn
Published: July 2016

Hepatocyte Growth Factor (HGF) binds its receptor Met and induces autophosphorylation of MET, which in turn activate appropriate signaling pathways. Aberrant HGF/Met signaling promotes tumor progression by facilitating cell proliferation, migration, invasion, and metastasis. Previously, we found that constitutive HGF/Met signaling promotes melanoma metastasis through a nonautocrine mechanism in engineered mouse models (Cancer Res, 2002); however, the molecular mechanisms by downstream of HGF/Met signaling that induce metastasis are not known. We recently identified the cytoskeletal organizer Ezrin as a key metastatic regulator using an HGF transgenic mouse model (Nature Medicine, 2004), implicating Ezrin is in HGF/Met signaling-induced metastasis. We hypothesize that HGF/Met signaling promotes metastasis by regulating expression of Ezrin. To test hypothesis, we are: 1) evaluating the relationship between ezrin expression and HGF/Met signaling; 2) validating the role of Ezrin in HGF/Met signaling -induced metastasis; 3) elaborating mechanisms underlying this HGF/Met signaling-mediated regulation of Ezrin expression; 4) evaluating the clinical therapeutic utility of blockage of Ezrin function in preclinical mouse models. We found that HGF is able to stimulate the expression of Ezrin. Moreover, analysis of the established model systems with HGF and Met alone or HGF with Met demonstrated that Ezrin expression correlated with activated HGF/Met signaling, suggesting that HGF/Met signaling regulates the expression of Ezrin. Interestingly, we showed that HGF/Met signaling could activate transcription factor Sp1 through the MAPK pathway. We further found that activated Sp1 could directly bind to the promoter of the Ezrin gene and regulate the transcription of Ezrin. Notably, knockdown of Ezrin expression by shRNAs inhibited the metastasis induced by both HGF/Met autocrine and notautocrine signaling in syngeneic host and HGF GEM hosts. We also used small molecule drugs in preclinical mouse models to confirm that Ezrin is at least one of downstream molecules mediating HGF/Met signaling-induced metastasis in melanoma. We conclude that Ezrin is a key downstream substrate involved in the regulation of HGF/Met signaling-induced metastasis. We demonstrate a link between Ezrin and HGF/Met/MAPK/Sp1 activation in the metastatic process. Our data indicate that Ezrin represents a promising therapeutic target for patients with activated HGF/Met signaling.

Citation Format: Yanlin Yu, Liping Huang, Glenn Merlino. Blocking Ezrin can inhibit HGF/Met signaling-induced metastasis. [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 4114.