bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3920: Understanding the molecular nature of cancer cell lines
Authors: Volpe Lysa A. , Bal Anupreet , Foulke John , Jackson Michael , Chen Luping , Tian Fang
Published: May 2015

Large-scale cancer genome programs have generated rich data of genetic abnormalities observed in thousands of clinical patient tumors, which provides a major opportunity to develop a landscape of molecular aberrations across tumor types and propose new therapeutic targets. Although human tumor cell lines have been important tools for the cancer researcher for decades, there is a lack of bio-functional validation of genetic alterations and systematic molecular characterization across commonly used cell lines in this genomic age. To meet the need of applying cancer genome knowledge to facilitate basic and translational cancer research, an updated and accurate knowledge of the cell lines in use, particularly from a molecular viewpoint, is critical for in vitro studies. Here, we show systematic molecular characterization and clustering of 90 authenticated human tumor cell lines, which represent the most common human cancer types found in the clinic, such as lung, breast, colon, pancreatic, skin cancer and so on. By next generation sequencing and molecular profiling, those tumor cell lines were fully analyzed to capture the driver gene mutations, DNA copy number variations, gene expressions, protein expressions and relevant cell signaling pathway activations. In addition to driver mutations such as BRAF V600, KRAS G12, PI3K E545 and EGFR T790, the gene copy number amplifications of AKT, FGFR, MET, ERBB2 and deletion of PTEN are presented in this work. Moreover, correlating with the verified genetic alternations, the endogenous basal levels of protein expression and signaling pathway activations within the cells were analyzed by western blot and immunofluorescence staining. A set of 6 wild type control cell lines that were derived from either normal tissues or tumors were characterized in parallel. Furthermore, live cell growth kinetics was continually monitored by label-free cell based assay. Finally, the cell lines are clustered into 10 genetic alteration panels according to driver genes, critical protein kinases, or key components of signaling pathways. Cell lines are essential models for progressing our understanding of the cancer genome, as they allow for the functional and biological validation of the genetic alterations proposed by NGS data. The genetic alteration tumor cell panels and their molecular signature profiles provide powerful tools to accelerate the discoveries in basic cancer research, compound screening, biomarker selection, pathway analysis, and targeted therapeutic development.

Citation Format: Lysa A. Volpe, Anupreet Bal, John Foulke, Michael Jackson, Luping Chen, Fang Tian. Understanding the molecular nature of cancer cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3920. doi:10.1158/1538-7445.AM2014-3920
Abstract 3921: Generation of a T-cell lymphoma mouse model based on chronic LPS challenge and TGF-β signaling disruption
Authors: Muñoz Nina M. , Katz Lior H. , Gagea Mihai I. , Rashid Asif , Mishra Lopa
Published: May 2015

Background: It has been demonstrated that elevated alcohol intake affects the permeability of the gut allowing endotoxins such as LPS to enter into the portal circulation and to promote liver ethanol toxicity. It has also been shown that increased ethanol intake and LPS challenge impairs liver function and affects liver tumor formation. Hence, with the aim of generating a mouse model of alcoholic liver disease that would rapidly develop steatosis, inflammation and fibrosis, we formulated a regimen that combined chronic injections of low dose LPS with ethanol feeding to mice with impaired TGF-β signaling through constitutive disruption of β2-spectrin (Sptbn1+/-) , Smad3 (Smad3+/-) or both. Methods: Wild type, Sptbn1+/-, Smad3+/- and Sptbn1+/-;Smad3+/- female mice between 8-10 weeks of age were fed orally with a Lieber-DeCarli-based diet containing 6.7% ethanol, and 2-4 weeks later they were injected with 2mg/kg LPS via i.p. 3 times/week for up to 20 weeks or death, whichever occurred first. Survival analysis was done using the Gehan-Breslow-Wilcoxon test. Results: Unexpectedly, the mice treated with chronic low dose LPS and fed the alcohol-containing diet only developed a mild liver phenotype, i.e. steatosis but not hepatitis or fibrosis. Instead, these mice developed very aggressive T-cell lymphomas with massive abdominal and thoracic hemorrhages to which the TGF-β mutant mice succumbed more rapidly than the wild type mice. The median survival of the mutants oscillated between 39 to 42 weeks vs. undefined of the wild type mice. We found statistically significant decreased survival of the mutant mice vs. the wild type (p-values between 0.0015 and 0.028). Conclusions: To our knowledge, this is the first report of a mouse model of aggressive T-cell lymphoma based on chronic challenge with low dose LPS and alcohol feeding. We are currently assessing the cross-talk of the LPS/TLR4 and TGF-β pathways in T-cells from these mice to understand the molecular mechanism underlying the accelerated rate of lymphoma development in these mice.

Citation Format: Nina M. Muñoz, Lior H. Katz, Mihai I. Gagea, Asif Rashid, Lopa Mishra. Generation of a T-cell lymphoma mouse model based on chronic LPS challenge and TGF-β signaling disruption. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3921. doi:10.1158/1538-7445.AM2014-3921
Abstract 3923: Molecular characterization of an animal model for neuroendocrine thyroid cancer
Authors: Pozo Karine , Hillmann Antje , Augustyn Alexander , Singh Tanvir , Plattner Florian , Minna John , Chen Herbert , Cote Gilbert J. , Bibb James A. , Nwariaku Fiemu E.
Published: May 2015

Neuroendocrine tumors are rare forms of cancers originating from hormone-producing cells that are scattered throughout the body. Medullary thyroid carcinoma (MTC) develops from calcitonin-secreting, C cells, in the thyroid gland. Surgical removal of the thyroid is the best treatment currently available for early stage cancer patients but advanced forms of MTC are often incurable and fatal. Thus, better therapies are needed. A lack of relevant animal models for this disease has slowed the development of new treatments. We have recently reported a novel conditional mouse model for MTC. In these mice, overexpression of the cyclin-dependent kinase 5 (Cdk 5), activator, p25 in C cells induces MTC tumors. Furthermore, interruption of p25 overexpression results in growth arrest of these tumors. Here we take advantage of our conditional mouse model to investigate the molecular pathways underlying MTC tumorigenesis. We used Illumina Mouse WG-6 v2.0 Expression BeadChip to compare the gene expression profile of proliferating versus arrested mouse MTC tumors. We found that a total of 920 genes encoding transcription factors, cell cycle proteins, motor proteins and angiogenesis factors were differentially regulated in growing versus arrested tumors. Particularly the genes encoding cell cycle proteins that are involved in the regulation of Rb pathway were significantly up-regulated in the proliferating tumors. Selected genes with higher degrees of change were evaluated by quantitative PCR and immunoblotting analyses. Taken together these findings suggests that the Rb signaling pathways is involved in MTC tumorigenesis and that Cdk5 may act as a critical regulator of the Rb signaling pathway in this cancer. Thus, targeting Cdk5 and/or the Rb pathway may be a valid strategy to stop the progression of neuroendocrine thyroid cancer.

Citation Format: Karine Pozo, Antje Hillmann, Alexander Augustyn, Tanvir Singh, Florian Plattner, John Minna, Herbert Chen, Gilbert J. Cote, James A. Bibb, Fiemu E. Nwariaku. Molecular characterization of an animal model for neuroendocrine thyroid cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3923. doi:10.1158/1538-7445.AM2014-3923
Abstract 394: TET1, which act as an tumor suppressor gene, is suppressed by DNA hypermethylation in gastric cancer
Authors: Park Jong-Lyul , Kwon Oh-Hyung , Song Kyu Sang , Kim Seon-Young , Kim Yong Sung
Published: May 2015

5-hydromethylcytosine (5hmC) is a hydroxmethylated form of DNA cytosine catalyzed by TET proteins. Recently, it was reported that aberrant regulation of 5hmC is associated with many cancers such as breast cancer, glioma, leukemia and melanoma. In this study, we examined the status of 5hmC and the role of TET1, which converts 5mC to 5hmC, in gastric cancer. Dot blot and ELISA assay results revealed global loss of 5hmC level in gastric cancer, and that decreased TET1 expression was associated with decreased 5hmC levels. Importantly, in gastric cancer patients, TET1-low group had advanced metastasis and poor survival compared with TET1-high group. To understand the mechanisms of TET1 suppression in gastric cancer, we performed pyrosequencing in 80 paired normal and tumors samples from gastric cancer patients, and found that the promoter region of TET1 was hypermethylated in tumors than adjacent normal tissues. Treating AGS cells with 5-Aza-2′deoxycytidine (Aza-dC) restored the expression of TET1. Transient transfection of in vitro transcribed TET1 mRNA into AGS cells significantly reduced the growth of AGS cells. In summary, our results showed that DNA methylation is responsible for the suppression of TET1 which has a tumor suppressor role in gastric cancer

Citation Format: Jong-Lyul Park, Oh-Hyung Kwon, Kyu Sang Song, Seon-Young Kim, Yong Sung Kim. TET1, which act as an tumor suppressor gene, is suppressed by DNA hypermethylation in gastric cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 394. doi:10.1158/1538-7445.AM2014-394
Abstract 3942: Cell death resistance promoted by radiotherapy in a colorectal cell line
Authors: Baptistella Antuani R. , Rodrigues Bruna , Dias Marcos , Giudice Fernanda , Lacerda Tonielli , Santos Tiago , Silva Felipe , Silva Petrus Paulo , Aguiar Samuel , Martins Vilma
Published: May 2015

Colorectal adenocarcinoma is the fourth more frequent cause of death by cancer in the world. Despite some similarities, the rectal cancer has characteristics that differentiate it from the colon cancer, resulting in different treatments and outcomes. The rectal cancer in stages II and III is treated with neoadjuvant chemoradiation followed by surgery, and the response to neoadjuvant treatment in patients is variable and unpredictable. Up to now, surgery intervention is required to assess the pathologic response to therapy. The identification of predictive biomarkers for response to therapy is of great importance, in order to identify the patients that will benefit from such treatment. To better understand the molecular alterations in cells that resist to the radiotherapy, we started our research using WiDr, a colon tumor cell line. Cell cycle synchronized WiDr cells were submitted to two cycles of irradiation (4Gy in each cycle and 2 week of interval between the cycles) and resistant cells were isolated. Irradiated WiDr cells (RxTWiDr) shown a better capacity to form colonies, in colony growth assays, when compared to parental WiDr cells. This characteristic was maintained when they were submitted to a new round of irradiation. In addition, WiDrRxT cells treated with Staurosporine presented a higher viability in MTT assays than WiDr cells. We also collected the conditioned medium (CM) from RxTWiDr and WiDr cells, and treated the parental WiDr cells. The parental cells treated with CM from RxTWiDr cells for 15 minutes shown an increase in the ERK1/2 activation when compared to cells treated CM from WiDr cells. These results demonstrated that radio resistant cells are more resilient than parental cells and that their secreted factors activated specific signaling pathways via ERK1/2 phosphorylation. The nature of these factors is under investigation using proteomics.

Supported by São Paulo State Foundation (FAPESP)

Citation Format: Antuani R. Baptistella, Bruna Rodrigues, Marcos Dias, Fernanda Giudice, Tonielli Lacerda, Tiago Santos, Felipe Silva, Petrus Paulo Silva, Samuel Aguiar, Vilma Martins. Cell death resistance promoted by radiotherapy in a colorectal cell line. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3942. doi:10.1158/1538-7445.AM2014-3942
Abstract 3945: Inhibition of MMP2 expression enhances the efficacy of radiation therapy for a murine astrocytoma
Authors: Yu Ching-Fang , Yang Ying-Chieh , Hong Ji-Hong , Chiang Chi-Shiun
Published: May 2015

Radiation therapy (RT) remains the front-line treatment for high-grade gliomas; however, tumor recurrence remains the main obstacle for the clinical success of RT. Using a murine astrocytoma tumor cell line, ALTS1C1, we have previously shown that RT can effectively reduce primary tumor volume and increase mice surviving time, but the mice eventually died associated with increased tumor infiltration. In this study, we explore the role of MMP2 (Matrix metalloproteinase-2), a reported molecule associated with brain tumor invasion, on ALTS1C1 tumor response to RT. In this study, the MMP2 expression was inhibited by specific lentiviral-shRNA construct. Eight Gray of whole brain irradiation could only prolong the median survival days of brain tumor-bearing mice from 25 days to 28 days, but the inhibition of MMP2 expression by siRNA prolonged the median survival days of tumor bearing mice from 25 days to 42 days. The median survival days were further prolonged to 53 days when 8 Gy of radiation and MMP2 inhibition were concurrently applied. Immunohistochemical (IHC) results showed that the synergistic effect as a result of RT-caused vessel density decrease (inadequate vasculature) and the retardation of vessel maturation as a result of MMP2 inhibition-caused the decrease of pericyte-covered vessels (insufficient vessel function). This study provides a new treatment option for treating invasive brain tumors. (This study was supported by NHRI-EX103-10132BI and NSC 102-2314-B-007-003-MY3 grants)

Citation Format: Ching-Fang Yu, Ying-Chieh Yang, Ji-Hong Hong, Chi-Shiun Chiang. Inhibition of MMP2 expression enhances the efficacy of radiation therapy for a murine astrocytoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3945. doi:10.1158/1538-7445.AM2014-3945
Abstract 3958: Investigation of HMGB1 as a potential marker for radiation response in muscle invasive bladder cancer
Authors: Mansure Jose J. , Shrivastava Sanhita , Nassim Roland , Ferbeyre Gerardo , Kassouf Wassim
Published: May 2015

Purpose: Radical cystectomy remains the standard treatment for invasive Bladder Cancer (BC). Radiotherapy is an attractive alternative; however lack of local control of the disease and toxicity associated with it remain problematic. As HMGB1 plays a major role in the cancer associated radio-resistance pathways, our objective is to evaluate its role as a predictive marker for radiation response in BC.

Methods: Expression of HMGB1 was characterized in a panel of eight BC cell lines via RT-PCR and Western Blotting. Knockdown of HMGB1 was carried out via shRNA lentiviral system. Correlation of HMGB1 expression and radiation response was done using clonogenic assays in control and shRNA transduced cell lines. Alkaline Comet Assay was used to measure the extent of DNA damage and repair in different cell lines 24hr post 5Gy radiation. Olive Tail moment was used as to quantify the DNA Damage.

Results: All cell lines expressed different levels of HMGB1, with BV and UC3 having the lowest and the highest expression of HMGB1 simultaneously. Clonogenic assay results showed that UC3 was resistant to radiation while BV was sensitive. Knockdown of HMGB1 caused the radio-resistant UC3 cells to become highly radio-sensitive, with more than 60% cells dying at 2Gy. Comet assay suggest the involvement of HMGB1 in DNA repair, as a significant DNA damage post radiation was observed in the HMGB1 knockdown cell line.

Conclusion: Our results demonstrate that high levels of HMGB1 protein are linked to a more radio-resistant outcome and HMGB1 knockdown leads to an increase in radio-sensitivity. Hence HMGB1 could potentially be used as a radiation response marker in invasive BC.

Citation Format: Jose J. Mansure, Sanhita Shrivastava, Roland Nassim, Gerardo Ferbeyre, Wassim Kassouf. Investigation of HMGB1 as a potential marker for radiation response in muscle invasive bladder cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3958. doi:10.1158/1538-7445.AM2014-3958
Abstract 3967: BET protein inhibitor OTX015 has selective anti-tumoral activity in preclinical models of MYCN- amplified neuroblastoma
Authors: Schulte Johannes H. , Althoff Kristina , Bell Emma , Odersky Andrea , Beckers Anneleen , Speleman Frank , Schäfers Simon , Schramm Alexander , Eggert Angelika , Westermann Frank , Riveiro Eugenia , Cvitkovic Esteban , Henssen Anton
Published: May 2015

Neuroblastomas harboring MYCN amplifications are highly lethal tumors. They are often resistant to standard chemotherapy, yet the development of targeted therapies has been hampered by a lack of compounds targeting MYCN. We and others have recently discovered that targeting BET bromodomain proteins, especially BRD4, disrupts epigenetic regulation of MYCN and its targets in neuroblastoma. OTX015, a new BET protein inhibitor, is the first lead into clinical phase I/II trials and has shown promising pharmacological properties in adults. Here, we investigate the preclinical efficacy of OTX015 in MYCN-amplified neuroblastoma.

We tested in vitro OTX015 efficacy in 6 established neuroblastoma (NB) cell lines. We performed cell cycle profiling and analyzed markers for apoptosis and proliferation after 72h-treatment at 500 nM OTX015. The effect of OTX015 on MYCN expression and global MYCN-associated transcriptional activity was assessed by quantitative real time PCR and gene expression microarray profiling, respectively. In vivo efficacy of orally OTX015 was assessed in IMR5 xenografts, a N-MYC driven NB model, using diffent treatment schedules (50mg/kg/day, 100mg/kg/day and 50mg/kg/bidaily).

Treatment of MYCN-amplified neuroblastoma cells with OTX015 resulted in decreased cell viability, induction of apoptosis and reduced proliferation. Concentrations of 50% inhibition (IC50) ranged between 50nM and 500nM. OTX015 treatment also resulted in an increase in the percentage of cells in G1 phase. This corresponded with the downregulation of MYCN mRNA and protein levels and MYCN-associated transcriptional activity. Interestigly, MYCN amplified cell lines were most sensitive to OTX015 treatment. In contrast, no effect was observed with OTX15 on normal cells. In vivo treatment with OTX015, significantly decreased tumor burden after 4 weeks and prolonged survival as compared to vehicle-treated mice. These preclinical findings highlight the promise of BET bromodomain inhibitors as novel agents for MYCN-driven neuroblastomas and serve as rationale to move forward with early phase clinical trials for children with these highly lethal tumors.

Citation Format: Johannes H. Schulte, Kristina Althoff, Emma Bell, Andrea Odersky, Anneleen Beckers, Frank Speleman, Simon Schäfers, Alexander Schramm, Angelika Eggert, Frank Westermann, Eugenia Riveiro, Esteban Cvitkovic, Anton Henssen. BET protein inhibitor OTX015 has selective anti-tumoral activity in preclinical models of MYCN- amplified neuroblastoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3967. doi:10.1158/1538-7445.AM2014-3967
Abstract 3976: Expression of formyl peptide receptor 1 (FPR1) in neuroblastoma: Implications in tumorigenesis
Authors: Snapkov Igor , Kogner Per , Johnsen John-Inge , Sveinbjørnsson Baldur
Published: May 2015

The formyl peptide receptor 1 (FPR1) is a G protein-coupled receptor with pattern recognition properties. It is mainly expressed by myeloid cells and is involved in a broad range of host defense mechanisms. However, a variety of host-derived agonists of FPR1 have been identified, including formyl peptides released from disrupted mitochondria of necrotic cells. In the present study, we demonstrate that FPR1 is expressed in neuroblastoma cell lines and primary tumors and high expression is correlated with poor overall survival. Addition of the FPR1 agonist N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) to neuroblastoma cells in vitro causes enhanced proliferative activity, increase of intracellular calcium response and activation of STAT3 and MAPK/Erk signaling pathways. Currently, clonal neuroblastoma cell line with inducible activity of FPR1 gene is under testing in a mouse model to assess the significance of this receptor in vivo. Our findings so far suggest that FPR1 may play a significant role in neuroblastoma tumorigenesis and that therapeutic intervention of the FPR1 pathway may become an important clinical strategy in neuroblastoma therapy.

Citation Format: Igor Snapkov, Per Kogner, John-Inge Johnsen, Baldur Sveinbjørnsson. Expression of formyl peptide receptor 1 (FPR1) in neuroblastoma: Implications in tumorigenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3976. doi:10.1158/1538-7445.AM2014-3976
Abstract 4000: Biological basis of aggressive phenotype in African-American prostate cancer cell line
Authors: Nickkholgh Bita , Fang Xiaolan , Gyabaah Kenneth , Balaji K.C.
Published: May 2015

Introduction: It is well established that African American men (AAM) with prostate cancer (PC) present more frequently with higher grade tumors compared to Caucasians. While the disparity is likely multifactorial, the biological basis is perhaps the strongest contender. Protein kinase D1 (PKD1) is a tumor and metastasis suppressor in PC. Down regulation of PKD1 was shown to increased cell proliferation, migration and metastasis in PC. Aim and experimental procedure: In order to address the fundamental question whether PC in AAM is inherently aggressive, we compared the proliferation, invasion and the expression of PKD1-centered signaling based 13-biomarker panel in primary and metastatic AAM cell lines (E006AA and MDApca2b) to benign (RWPE) and metastatic Caucasian cell lines (LNCaP and C4-2) using Western blot within different cell compartments. The results were further validated by RT-PCR and immunofluoroscent staining. Results: The proliferation, viability and invasion assays showed that E006AA cells proliferate and invade matrigel significantly more rapidly than metastatic LNCaP and MDAPca2b cells. The 13-biomarker panel analysis showed PKD1-centered dysregulaiton in AAM prostate cancer cell lines. Although E006AA is a primary PC cell line, PKD1 was down regulated similar to more aggressive C4-2 cells. Nuclear androgen receptor (AR), which shows active AR, was higher in LNCaP and MD2Pca2b; Vimentin showed higher expression in C4-2 and E006AA. Localization of E-Cadherin, N-Cadherin, T120 phosphorylated ß- Catenin, MMP-2 and S11 phosphorylated snail in MD2Pca2b was similar to LNCaP cells but different to C4-2 and E006AA. Conclusion: Our Data demonstrates that the dysregulaiton of PKD1-centered signaling occurs in primary AAM prostate cancer cell line E006AA is akin to other metastatic PC cell lines suggesting an inherently aggressive nature of PC in AAM.

Acknowledgment: Authors would like to thank Dr. Shahriar Koochekpour (Roswell park institute, University at Buffalo,State University of NewYork) for his kind gift E006AA cell line.

Citation Format: Bita Nickkholgh, Xiaolan Fang, Kenneth Gyabaah, K.C. Balaji. Biological basis of aggressive phenotype in African-American prostate cancer cell line. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4000. doi:10.1158/1538-7445.AM2014-4000
Abstract 4027: Traditional Chinese Medicine herbal mixture LQ prevents cyclophosphamide-induced metastasis and toxicity
Authors: Zhang Lei , Wu Chengyu , Li Lingna , Zhang Yong , Hoffman Robert M.
Published: May 2015

Cyclophosphamide (CTX) can enhance metastasis (Cancer Res. 66, 303-306, 2006; Cancer Res. 68, 516-520, 2008) and has toxic side effects such as immune-suppression, weight loss, and hair loss. In the present study, the traditional Chinese medicine (TCM) herbal mixture LQ (J. Cell. Biochem. 114, 2131-2137, 2013) was tested to determine if it could enhance CTX efficacy and reduce side effects. HT1080 human fibrosarcoma cells, labeled with GFP in the nucleus and RFP in the cytoplasm, were injected into the epigastric cranialis vein of nude mice that were pre-treated with CTX and LQ. Lewis lung carcinoma (LLC)-RFP cells were injected into the tail vein of mice that were pre-treated with CTX and LQ. Efficacy was monitored with dynamic subcellular imaging of trafficking of cancer cells in blood vessels (Cancer Res. 68, 516-520, 2008) and imaging metastasis to the lung. LQ significantly reduced CTX-induced lung metastasis and cancer-cell adhesion to the inner wall of blood vessels. LQ also reduced kidney toxicity induced by CTX. Treatment with LQ induced hair growth after CTX administration. This result demonstrates that LQ provides safe and effective prevention of metastasis and toxicity in combination with chemotherapeutic agents such as CTX.

Citation Format: Lei Zhang, Chengyu Wu, Lingna Li, Yong Zhang, Robert M. Hoffman. Traditional Chinese Medicine herbal mixture LQ prevents cyclophosphamide-induced metastasis and toxicity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4027. doi:10.1158/1538-7445.AM2014-4027
Abstract 405: Mbd3 localizes at promoters, gene bodies and enhancers of active genes
Authors: Shimbo Takashi , Du Ying , Grimm Sara A. , Dhasarathy Archana , Mav Deepak , Shah Ruchir R. , Shi Huidong , Wade Paul A.
Published: May 2015

The structure of chromatin is tightly regulated; its disregulation is causal for developmental abnormalities and for cancer. The nucleosome remodeling and deacetylase (NuRD) complex regulates chromatin structure and gene expression via its nucleosome remodeling and histone deacetylase activities. MBD3, a component of NuRD, has pleiotropic roles in embryonic stem (ES) cells and tumor biology. However, the results of recent studies are conflicting, resulting in an unclear model for NuRD function. Here, we investigated the localization of MBD3 in two human breast cancer cell lines (MCF-7 and MDA-MB-231) with two independent genomic technologies (DamID and ChIP-seq). DamID and ChIP-seq data were in an excellent agreement and led consistent conclusions in both cell lines. MBD3 localized not only at promoters and enhancers of active genes but also accumulated at gene bodies; this previously undescribed localization suggests an active role of MBD3 in transcription. MBD3 preferred hypomethylated, CpG rich promoters marked by H3K4me3. We also noted a subset of MBD3 that localized at silent promoters marked by H3K9me3 or H3K27me3, consistent with biochemical studies. Another subset of MBD3 co-localized with H3K27ac and was in physiological proximity with promoters, suggesting a role of MBD3 in the regulation of high-order structure of the genome. Functional analysis of chromatin demonstrated that MBD3 regulates nucleosome occupancy near promoters and in gene bodies. These data suggest multiple roles of MBD3/NuRD in the regulation of chromatin structure and gene expression for both active and silent genes. The findings serve as an important initial step in defining regulatory mechanism of MBD3/NuRD in ES cells and in tumor biology.

Citation Format: Takashi Shimbo, Ying Du, Sara A. Grimm, Archana Dhasarathy, Deepak Mav, Ruchir R. Shah, Huidong Shi, Paul A. Wade. Mbd3 localizes at promoters, gene bodies and enhancers of active genes. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 405. doi:10.1158/1538-7445.AM2014-405
Abstract 4050: A central role for mTORC1 in CXCR4-mediated directional migration and metastasis
Authors: Dillenburg-Pilla Patricia , Patel Vyomesh , Mikelis Constantinos M. , Zarate-Blades Carlos Rodrigo , Amornphimoltham Panomwat , Wang Zhiyong , Martin Daniel , Leelahavanichkul Kantima , Doçi Colleen L. , Dorsam Robert T. , Masedunskas Andrius , Nohata Nijiro , Weigert Roberto , Molinolo Alfredo A. , Gutkind J. Silvio
Published: May 2015

Tumor cells can co-opt the promigratory activity of chemokines and their cognate G protein-coupled receptors to disseminate to regional lymph nodes and distant organs. Indeed, the migration to SDF-1 of tumor cells expressing its receptor, CXCR4, is implicated in lymphatic and organ-specific metastasis in multiple human malignancies. However, the critical role of CXCR4 in bone marrow stem cell retention has hampered the development of CXCR4 inhibitors for metastasis prevention. Therefore the study of CXCR4 downstream signaling circuitry may provide new opportunities for the treatment of many human malignancies that depend on CXCR4 for their metastatic spread. The PI3K/mTOR pathway represents a major player in normal and aberrant cell growth. Consistently, mTOR inhibitors targeting its complex 1 (mTORC1) alone or mTORC1/mTORC2 have already shown promising therapeutic responses in many tumor types. Our data show that mTOR is activated upon SDF-1 stimulation of CXCR4 on epithelial-derived cancer cells, and that the disruption of mTORC1 or mTORC2 abrogates CXCR4-mediated directional migration. To study the CXCR4/mTOR axis in vivo we developed a simple and robust model for CXCR4-mediated spontaneous metastasis. Surprisingly, disruption of mTORC1 alone was sufficient to decrease tumor cell proliferation, angiogenesis, lymphangiogenesis, and CXCR4-mediated metastasis, whereas mTORC2 impairment had no effect on tumor dissemination or growth in vivo. Taken together, our data suggest that mTORC1 blockade could inhibit the migration of CXCR4 expressing cancer cells to their secondary sites, while disrupting the establishment of a permissive tumor microenvironment thereby halting the spread of highly aggressive tumors that require CXCR4 to metastasize.

Citation Format: Patricia Dillenburg-Pilla, Vyomesh Patel, Constantinos M. Mikelis, Carlos Rodrigo Zarate-Blades, Panomwat Amornphimoltham, Zhiyong Wang, Daniel Martin, Kantima Leelahavanichkul, Colleen L. Doçi, Robert T. Dorsam, Andrius Masedunskas, Nijiro Nohata , Roberto Weigert, Alfredo A. Molinolo, J. Silvio Gutkind . A central role for mTORC1 in CXCR4-mediated directional migration and metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4050. doi:10.1158/1538-7445.AM2014-4050
Abstract 4053: Formin-binding protein-17 (FBP17) is a target of p53 and promotes invadopodia formation in breast cancer cells
Authors: Chander Harish , Watts Kathleen , Pogmore Justin , Truesdell Peter , Brien Colin , Craig Andrew W B
Published: May 2015

Invadopodia are extracellular matrix (ECM)-degrading structures that promote tissue invasion and metastasis of tumor cells. A family of adaptor proteins with F-BAR and SH3 domains, including FBP17 and Toca-1, have been identified as key scaffolds for recruiting actin regulatory proteins (eg. Cdc42, formins, N-WASP, dynamin) to promote formation of invadopodia in bladder and breast cancers, respectively. In this study, we investigated the potential role of FBP17 in breast cancer models. We observed FBP17 expression in normal breast epithelial cells (MCF10A), and at higher levels in basal breast cancer cell lines (MDA-MB-231) compared to luminal cell lines. Since basal breast cancers frequently harbor mutations in tumor suppressor p53, we tested whether wild-type (WT) p53 regulates expression of FBP17, which may explain higher expression in basal breast cancers. Camptothecin (CPT) treatment of WT p53-expressing breast cancer cells (MCF-7, DU4475) led to a dose dependent reduction in FBP17 mRNA and protein levels, and this was inversely correlated with p53 levels or its target p21WAF1. The downregulation of FBP17 was also observed using Nutlin-3, an Mdm2 inhibitor, indicating that WT p53 activation by multiple methods causes a rapid downregulation of FBP17. Currently we are testing whether this is a direct or indirect effect of p53 on FBP17 gene expression. As expected, FBP17 formed complexes with actin regulatory proteins (dynamin, cortactin, Arp2/3) that accumulate and promote invadopodia formation in MDA-MB-231 cells. Stable silencing of FBP17 in these cells led to a significant reduction in ECM degradation. This is consistent with FBP17 promoting invadopodia formation. Tumor xenograft models are in progress to determine whether FBP17 also promotes tumor metastasis. FBP17 expression levels in human breast tumors are currently being tested to determine whether altered expression occurs in molecular subtypes with frequent mutations in p53 (HER2, basal) and in their metastases. Overall, these results are consistent with FBP17 participating in cancer cell invasion, and supports further studies of its role in cancer metastasis.

Citation Format: Harish Chander, Kathleen Watts, Justin Pogmore, Peter Truesdell, Colin Brien, Andrew W B Craig. Formin-binding protein-17 (FBP17) is a target of p53 and promotes invadopodia formation in breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4053. doi:10.1158/1538-7445.AM2014-4053
Abstract 4080: Smad and NFAT pathways cooperate to regulate CD103 expression in human CD8 T lymphocytes and TCR-mediated epithelial tumor cell killing
Authors: Mami-Chouaib Fathia , Mokrani M'barka , Bismuth Georges
Published: May 2015

The interaction of integrin αE(CD103)β7, often expressed on tumor-infiltrating lymphocytes (TIL), with epithelial cell marker E-cadherin on tumor cell surface plays a major role in antitumor cytotoxic T lymphocyte (CTL) responses. CD103 is encoded by ITGAE gene and is induced in CD8 T cells upon T-cell receptor (TCR) engagement and exposure to transforming growth factor (TGF)-β1, abundant within the tumor microenvironment. However, the transcriptional mechanisms underlying the cooperative role of these two signaling pathways in inducing CD103 expression in CD8 T cells remain unknown. Using a model system based on a CD8+/CD103- T-cell clone, we demonstrated that the transcription factors Smad2/3 and NFAT-1 are required for CD103 induction on T lymphocytes activated with a combination of TGF-β1 and anti-CD3 monoclonal antibodies. We also identified the promotor and enhancer elements of human ITGAE gene, and showed that Smad2/3 and NFAT-1 are necessary for the activity of the ITGAE enhancer, and thus for CD103 expression. Overall, our results explain how TGF-β1 induces CD103 expression on locally activated CD8 T cells, thus contributing to antitumor CTL responses and malignant cell killing.

Citation Format: Fathia Mami-Chouaib, M'barka Mokrani, Georges Bismuth. Smad and NFAT pathways cooperate to regulate CD103 expression in human CD8 T lymphocytes and TCR-mediated epithelial tumor cell killing. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4080. doi:10.1158/1538-7445.AM2014-4080
Abstract 4082: CD47 blockade to enhance adaptive anti-tumor immune responses
Authors: Anderson Katie L. , Ito Daisuke , Lins Debra C. , Curtsinger Julie M. , Mescher Matthew F. , Modiano Jaime F.
Published: May 2015

The success of immunotherapy has been hindered by the ability of tumors to evade or inhibit the host immune system. One strategy tumors use to achieve this is upregulation of CD47, a cell surface protein that delivers a “don't eat me” signal. CD47 binds to SIRP-alpha on the surface of macrophages and disables their phagocytic function. Previous data showed CD47 blockade enhanced phagocytosis of tumor cells in vitro and improved survival of mice transplanted with human tumors. However, CD47 blockade has yet to be examined in a fully syngeneic system where the potential for tumor phagocytosis by macrophages and/or dendritic cells to promote presentation of tumor antigens to T cells can be defined. It has been reported that CD47 blockade can hinder T cell activation; however, this also has not been examined in a syngeneic system. Our hypothesis was that inducing tumor cell phagocytosis by CD47 blockade would enhance T-cell-mediated anti-tumor responses. Our data show that CD47 was expressed on the EL4 thymoma and B16-F10 melanoma cell lines that are syngeneic to H-2b, as well as on their ovalbumin-expressing derivatives EG7 and B16-OVA. CD47 also was expressed on CD8+ T cells and was further upregulated upon stimulation, suggesting it might play a role in immune effector function. However, in our hands CD47 blockade using antibody MIAP-301 did not inhibit activation of CD8+ T cells in vitro, and in preliminary experiments, CD47 blockade appeared to enhance activation of antigen-specific CD8+ T cells in vivo. Ongoing work is evaluating the effects of CD47 blockade and phagocytosis on antigen presentation in vitro and the effects of CD47 blockade on the generation of tumor-specific CD4 and CD8 T cells in vivo. We also plan to confirm that T cells activated under conditions of CD47 blockade can recognize endogenous tumor antigens to which there may be self-tolerance.

Supported by an HHMI Medical Student Fellowship (to KA), by the GREYlong Foundation, and by grant P01 AI035296 (to MFM).

Citation Format: Katie L. Anderson, Daisuke Ito, Debra C. Lins, Julie M. Curtsinger, Matthew F. Mescher, Jaime F. Modiano. CD47 blockade to enhance adaptive anti-tumor immune responses. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4082. doi:10.1158/1538-7445.AM2014-4082
In VivoAntineoplastic Effects of the NSAID Sulindac in an Oral Carcinogenesis Model
Authors: Katoumas Konstantinos , Nikitakis Nikolaos , Perrea Despina , Dontas Ismene , Sklavounou Alexandra
Published: May 2015

The antineoplastic properties of the NSAID sulindac have long been studied. The purpose of this study was to explore sulindac's in vivo effects on oral squamous cell carcinoma (SCC) oncogenesis using the hamster cheek pouch oral carcinogenesis model (HOCM). Thirty Syrian golden hamsters were divided into three experimental and two control groups (n = 6 each). The animals' right buccal pouches were treated with carcinogen for 9 weeks in one experimental and one control group and for 14 weeks in all other three groups. The animals of two experimental groups received sulindac from the 1st week and those of the third experimental group from the 10th week. After the end of carcinogenesis, treated buccal pouches were removed and examined. In animals treated with carcinogen for 14 weeks, development of oral SCC and tumor volume were significantly lower in animals that received sulindac from the first week of the experiment. Oral SCC developing in animals that received sulindac were more frequently well differentiated compared with the control group. In animals treated with carcinogen for 9 weeks, the animals that received sulindac developed lower grade of epithelial dysplasia. Proliferation index Ki-67 and positivity for the antiapoptotic molecule survivin were lower in the animals that received sulindac. Treatment with sulindac appears to delays the progression of oral premalignant lesions to oral SCC in the HOCM, also resulting in smaller and better differentiated tumors. These in vivo antineoplastic effects may be related to sulindac's ability to decrease cell proliferation and to prevent survivin expression. Cancer Prev Res; 8(7); 642–9. ©2015 AACR.
Determining Pre-Conception Risk Profiles Using a National Online Self-Reported Risk Assessment: A Cross-Sectional Study
Authors: Vink-van Os Lena C.A. , Birnie Erwin , van Vliet-Lachotzki Elsbeth H. , Bonsel Gouke J. , Steegers Eric A.P.
Published: May 2015

Aim: To measure the prevalence of health risk factors in women who are preparing for pregnancy, using an online publicly available questionnaire aimed at identifying personal and pre-conception risks and at providing tailored information. Methods: A nation-wide available, free, web-based, self-reported questionnaire for pre-conception use (in Dutch). Between May 2006 and August 2009, 89,946 questionnaires were completed (78,732 were from unique respondents) and available for research purposes, from which those of non-pregnant women (n = 66,617) were selected. Socio-demographic subgroups were distinguished by age, ethnicity, urban living area and living in a deprived neighbourhood. The four pre-conception risk domains were lifestyle, medical, reproductive and family history; together they were defining the risk profile. χ2 tests were used to compare the risk profiles among the subgroups. Results: The prevalences of the reported risk factors are given. The risk factor profiles revealed that the average, responding, non-pregnant, Dutch woman is exposed to a substantial number of risk factors. Different risk profiles were observed in the different socio-demographic subgroups. Women older than 36 years, of non-Western origin, living in urban areas and those in deprived neighbourhoods showed higher risk profiles, based on a larger number of risks, with significantly higher prevalences. Conclusion: Self-reported data from a large, self-selected, non-pregnant population who actively visited a web-site for reproductive information suggest the need for active general pre-conception care as risk factors were abundant. A considerable increase in attention for pre-conception care is justified; different subpopulations most likely require adapted approaches.
Independent Proteolytic Activities Control the Stability and Size of Drosophila Inhibitor of Apoptosis 2 Protein
Authors: Guntermann Silvia , Fraser Brittany , Hazes Bart , Foley Edan
Published: May 2015

The Drosophila immune deficiency pathway defends many bacterial pathogens and bears striking molecular similarities to the mammalian tumor necrosis factor signal transduction pathway. Orthologous inhibitors of apoptosis ubiquitin ligases act at a proximal stage of both responses to coordinate the assembly of signal transduction platforms that shape host immune responses. Despite the importance of inhibitor of apoptosis proteins within evolutionarily conserved innate immune responses, we know relatively little about the cellular machinery that controls inhibitor of apoptosis activity. In this study, we examined the molecular basis for inhibitor of apoptosis 2 protein regulation in the immune deficiency pathway. Our studies identified two distinct proteolytic events that determine the stability and composition of cellular inhibitor of apoptosis 2 protein pools. We found that apoptotic caspase activity cleaves inhibitor of apoptosis 2 at an N-terminal aspartate to generate a truncated protein that retains the ability to interact with immune deficiency pathway members. We also showed that a C-terminal ubiquitin ligase activity within inhibitor of apoptosis 2 directs the proteasomal destruction of full-length and truncated inhibitor of apoptosis 2 isoforms. These studies add to our appreciation of the regulation of innate immunity and suggest potential links between apoptotic caspases and innate defenses.
Outcomes of Male Sling Mesh Kit Placement in Patients with Neuropathic Stress Urinary Incontinence: A Single Institution Experience
Authors: Vainrib Michael , Reyblat Polina , Ginsberg David
Published: May 2015

Background and Objectives: The mainstay of therapy in patients with neuropathic stress urinary incontinence (nSUI) has been through the use of artificial urinary sphincter (AUS). AUS infection/erosion rates are higher in nSUI patients and these patients tend to be younger, increasing the likelihood of multiple AUS revisions in the future. We review our experience with mesh male slings for patients with nSUI. Methods: A retrospective review of patients who had mesh sling placement. Results: Twenty patients were identified between 2003 and 2011. 14/20 (70%) (5 = AdVance, 8 = InVance, 1 = Virtue) were available for long-term evaluation; in 6/20 (30%) the sling was removed for either infection or perineal wound breakdown. The mean time from injury to male sling was 148.2 (29-449) months. Pre-/post-op fluorourodynamic study was performed in 13 and 7 patients, respectively. There were no significant differences in ALPP (46.4 vs. 55.7 cm H2O, p = 0.106) and MCC (456.6 vs. 608 ml, p = 0.21) in the 7 patients who had a post-op study: five patients had new onset low bladder compliance and two had new onset detrusor overactivity post-sling. With a mean follow-up of 24.7 (1-66) months, 4/14 (28.6%) had no UI. Conclusions: With short-term follow-up, mesh male slings are a feasible option to treat nSUI. There appears to be a lower success rate for UI resolution, which may be attributable to new onset detrusor failure or wound infection requiring sling removal.