bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 199: Increased killing potential of ex vivo differentiated human natural killer cells in the presence of HOXB4
Authors: Chouaib Salem , Nanbakhsh Arash , Pochon Cécile , Amsellem Sophie , Pittari Gianfranco , Bourhis Jean-Henri
Published: May 2015

We have previously shown that human umbilical cord blood CD34+ progenitor cells undergo in vitro differentiation into functional NK cells and that their co-culture in the presence of HOXB4 transduced stromal MS5 cells resulted in an increase in differentiated NK number. The present study was conducted to compare the stromal effect on NK lytic potential in the presence and absence of HOXB4. Our results provide evidence that HOXB4 transduced MS5 cells as compared to transduced, GFP (+) MS5 cells induced highly differentiated cytotoxic NK cells. Importantly, this difference was not due to the expression of activating NK receptors but was associated with an increased induction of granzyme B transcription and degranulation in response to stimulation with NK cell susceptible targets. DNA microarray-based global transcriptional profiling confirmed the up-regulation of Granzyme B. We provide evidence that HOXB4 is a crucial regulator of NK lytic function and that its use in generating functional NK cells with increased lytic potential may be significant for hematologic malignancies. Our findings provide further evidence that HOXB4 is a crucial regulator of NK function and that its use in generating functional NK cells with increased lytic potential may be significant for cancer immunotherapy.

Citation Format: Salem Chouaib, Arash Nanbakhsh, Cécile Pochon, Sophie Amsellem, Gianfranco Pittari, Jean-Henri Bourhis. Increased killing potential of ex vivo differentiated human natural killer cells in the presence of HOXB4. [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 199. doi:10.1158/1538-7445.AM2014-199
Abstract 1993: The role of metabotropic glutamate receptor 5 on the lymph node metastases in oral cancer
Authors: Kuribayashi Nobuyuki , Uchida Daisuke , Kinouchi Makoto , Tamatani Tetsuya , Nagai Hirokazu , Miyamoto Youji
Published: May 2015

We identified a metastasis-related gene, metabotropic glutamate receptor (mGluR)-5, as a target gene of stromal cell-derived factor (SDF)-1/CXCR4 system in oral cancer. In this study, we examined the role of mGluR5 on the lymph node metastasis of oral cancer. mGluR5-specific antagonists, 2-Methyl-6-(phenylethynyl)pyridine (MPEP) and 3-((2-Methyl-1,3-thiazol-4-yl)ethynyl)pyridine (MTEP) did not affect the growth of oral cancer cells, but did inhibit the SDF-1/CXCR4 dependent-migration of the cells. When the CXCR4 expressing-oral cancer cells were orthotopically transplanted into masseter muscle of nude mice, these antagonists significantly inhibited lymph node metastases of the cells. Immunohistochemical analysis revealed that there was a significant association between CXCR4 and mGluR5 protein expression. Furthermore, the mGluR5 protein expression in clinical specimen was significantly associated with the lymph node metastases and the modes of invasion. Moreover conditioned media derived from the oral cancer cells acquiring SDF-1/CXCR4 system enhanced the migration of lymphatic endothelial cells, which was significantly suppressed by the treatment with MPEP or MTEP. Although SDF-1/CXCR4 dependent-production of angiogenic factors, including interleukin-6, interleukin-8, and vascular endothelial growth factor (VEGF)-A were observed, mGluR5 antagonists did not affect the production of these factors. However, induction of a lymphangiogenic factor, VEGF-C was specifically impaired by the treatment with these mGluR5 antagonists. These results suggested that mGluR5 promotes the lymph node metastases of oral cancer cells by the SDF-1/CXCR4 system via enhancement of cancer cell migration and induction of lymphangiogenesis by VEGF-C.

Citation Format: Nobuyuki Kuribayashi, Daisuke Uchida, Makoto Kinouchi, Tetsuya Tamatani, Hirokazu Nagai, Youji Miyamoto. The role of metabotropic glutamate receptor 5 on the lymph node metastases in oral 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 1993. doi:10.1158/1538-7445.AM2014-1993
Abstract 2: Paxillin enhances angiogenesis through transcriptional regulation of Src in ovarian cancer
Authors: Choi Hyun Jin , Kim Seung-Wook , Pradeep Sunila , Dalton Heather J. , Rupaimoole Rajesha , Mangala Selanere , Lopez Gabriel , Anil Sood K.
Published: May 2015

Purpose: Paxillin (PXN) expression is increased in various cancer types, but its biological and clinical impact is not well understood. Here, we examined the biological effects of paxillin in ovarian cancer.

Methods: The in vivo (HeyA8 and HeyA8-MDR orthotopic mouse models of ovarian cancer) and in vitro (tube formation assays, reverse phase protein array (RPPA) analysis, RT-PCR chromatin-immunoprecipitation (ChIP) assay) effects of paxillin were examined in ovarian cancer.

Results: Paxillin silencing using siRNA incorporated in DOPC nanoliposomes resulted in 55% reduction in tumor growth in a HeyA8 orthotopic ovarian cancer mouse model (p<0.05). There was an unexpected effect of marked reduction in vascular density (67%, p<0.01). In addition, tumor proliferation was decreased by 60% (p<0.01) in PXN-siRNA treatment group. This anti-proliferative effect of siPXN was not observed in vitro. In contrast, a significant anti-angiogenic effect was observed in vitro using a tube formation assay, where condition media collected from ovarian cancer cells treated with siPXN compared to siControl resulted in a prominent (60%,p<0.01) reduction in tube formation ability of endothelial cells. RPPA analyses pointed to Src protein expression being significantly decreased following PXN silencing in HeyA8 cells. Using RT-PCR and western blotting techniques, we observed that Src mRNA and protein levels were significantly decreased in siPXN-treated cells. Moreover, p-Src protein level was also found to be decreased following PXN silencing. ChIP analysis showed direct PXN binding to the Src promoter.

Conclusion PXN exerts indirect effects on tumor angiogenesis, mediated through Src. These findings provide a new understanding of the role of PXN in malignant biology.

Citation Format: Hyun Jin Choi, Seung-Wook Kim, Sunila Pradeep, Heather J. Dalton, Rajesha Rupaimoole, Selanere Mangala, Gabriel Lopez, Sood K. Anil. Paxillin enhances angiogenesis through transcriptional regulation of Src in ovarian 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 2. doi:10.1158/1538-7445.AM2014-2
Abstract 2001: GATA2: Potential role as a prostate cancer metastasis-driving gene
Authors: Chiang Yan Ting , Wang Kendric , Crea Francesco , Collins Colin , Gout Peter , Wang Yuzhuo
Published: May 2015

Metastasis is thought to result from changes in the expression of specific metastasis-driving genes leading to a cascade of activated downstream genes that set the metastatic process in motion. The present study was aimed at identifying such metastasis-driving genes in prostate cancer for potential therapy and identification of primary prostate cancers that are likely to metastasize. To this end, a differential gene expression profile was established of metastatic LTL-313H and non-metastatic LTL-313B prostate cancer tissue xenografts, derived from one patient's specimen using sub-renal capsule grafting technology. The profile was then subjected to integrative analysis using the Ingenuity Upstream Regulator Analysis tool. Six candidate master regulatory genes were identified, including GATA2, a gene encoding a pioneer factor in prostate cancer. Elevated GATA2 expression in clinical metastatic prostate cancer tissues was found to correlate with poor patient prognosis. Furthermore, GATA2 gene silencing in human prostate cancer LNCaP cells led to a marked reduction in cell migration, tissue invasion, focal adhesion disassembly and to a drastic change in cell transcriptome. Furthermore, 582 genes were identified that are (i) differentially expressed after GATA2 gene silencing in LNCaP cells and (ii) whose changes in gene expression significantly correlated with changes in GATA2 expressions in a MSKCC prostate cancer patient cohort. Taken together, the data suggest that GATA2 could represent a prostate cancer metastasis-driving gene and that the expression patterns of GATA2 and its associated-genes could serve as signatures (biomarkers) for poor prognosis in prostate cancer.

Citation Format: Yan Ting Chiang, Kendric Wang, Francesco Crea, Colin Collins, Peter Gout, Yuzhuo Wang. GATA2: Potential role as a prostate cancer metastasis-driving gene. [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 2001. doi:10.1158/1538-7445.AM2014-2001
Abstract 2006: The role of neuronal predominant gene expression in breast cancer brain metastasis
Authors: Kanojia Deepak P. , Sarvaiya Purva , Qiao Jian , Zhang Lingjiao , Balyasnikova Irina , Lesniak Maciej S.
Published: May 2015

Breast cancer brain metastasis (BCBM) occur in about 10-30% of patients with breast cancer. It has been established that breast cancer is a heterogeneous disease with 3 predominant subtypes (1) ER/PR positive (3) HER2 positive (4) Triple negative breast cancer (TNBC)/Basal subtype. TNBC is associated with the highest incidence of metastasis to brain, mechanism of which is poorly understood. Therefore, in order to understand the proclivity of TNBC to the brain and identify potential targets for treatment BCMB, we employed two widely used paired cell model system, the MDA-MB-231/MDA-MB-231BrM2 and the CN-34/CN34BrM2. The levels of brain predominant markers [ST6GALNAC5 (ST6), Nestin, Tubulin β3 (TUBB3)] in brain-derived clones were assesed by qRT-PCR and found to be overexpressed in brain-derived MDA-MB-231BrM2 and CN34BrM2 in comparison with parental breast cancer cell lines. We also identified hyperactivated TGF-β signaling in brain derived clones in comparison with parental cells. Moreover, the treatment of parental cells with TGF-β increased the expression of ST6, Nestin and TUBB3 indicating that TGF-β plays role in the development of BCBM. To further delineate the role of these markers in BCBM, knockdown experiments using two different shRNA specific to TUBB3 (neuronal specific) were performed. TUBB3 knockdown resulted in a dramatic reduction in tumorspheres formation, migration, invasion and the ability to adhere to laminin in both MDA-MB-231BrM2. Furthermore, the downregulation of expression of β1 and β3 integrins in TUBB3 knockdown cells was discovered as assessed by qPCR and flow cytometry. These results implicate that overexpression of brain specific TUBB3 provides breast cancer cells with migration and invasion abilities and a possible survival advantage in the brain microenvironment.

Citation Format: Deepak P. Kanojia, Purva Sarvaiya, Jian Qiao, Lingjiao Zhang, Irina Balyasnikova, Maciej S. Lesniak. The role of neuronal predominant gene expression in breast cancer brain 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 2006. doi:10.1158/1538-7445.AM2014-2006
Abstract 2015: Thrombospondin-1 is a targetable marker of invasive, mesenchymal- like melanoma cells
Authors: Behren Andreas , Jayachandran Aparna , Anaka Matthew , Hudson Christopher , Cebon Jonathan
Published: May 2015

Resistance to anti-cancer therapies arising from genetic heterogeneity is well understood. Less well characterised is the resistance to therapy proposed to occur via the plastic changes in melanoma cells generated by the impact of the tumor microenvironment. In melanoma, phenotypic-switching as means of plasticity has been well described and resembles, at least partially, an epithelial-to-mesenchymal transition (EMT). The presence of mesenchymal-like, invasive and slow-proliferating cells in melanoma, a highly aggressive cancer with intrinsic resistance to chemotherapy, is controversial and their potential contribution to therapeutic resistance has yet to be determined.

Methods: We examined melanoma cell lines for the presence of mesenchymal-like cells using CM-Dil, a membrane dye evenly distributed to daughter cells with each cell division as a marker of reduced cell division. Functional assays included invasion and motility assays, in vivo mouse xenograft models as well as an in vivo chicken neural crest transplantation assay. Genome-wide gene-expression profiling was performed using Illumina HT12 arrays.

Results: Here we show that mesenchymal-like cells are present in cell lines and in xenograft tumors, survive exposure to cytotoxic drugs and are invasive. Importantly, resistance was associated with a gene expression signature characterized by the expression of thrombospondin, TGFBI, genes associated with the extracellular matrix, and acquisition of increased invasiveness. This gene-expression pattern is reminiscent of an epithelial-to-mesenchymal transition in other solid cancers and a similar phenotype was detected after acquired resistance to the BRAF inhibitors PLX4720 and dabrafenib. Moreover, targeting thrombospondin in an in vivo model of melanoma cell plasticity and invasion led to the complete abrogation of the plastic and invasive potential of melanoma cells.

Conclusion: The results link therapeutic resistance to the presence of mesenchymal-like and invasive cells, and identify potential targets for novel therapies directed at eradicating this key melanoma subpopulation.

Citation Format: Andreas Behren, Aparna Jayachandran, Matthew Anaka, Christopher Hudson, Jonathan Cebon. Thrombospondin-1 is a targetable marker of invasive, mesenchymal- like melanoma 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 2015. doi:10.1158/1538-7445.AM2014-2015
Abstract 2025: Multicellular tumor spheroid 3D models to decipher cancer cell biology and to evaluate anticancer drugs
Authors: Ducommun Bernard , Lobjois Valérie
Published: May 2015

MultiCellular Tumor Spheroid (MCTS) mimic the organization of a tumor and is considered as an invaluable model to study cancer cell biology and for the preclinical development of new antiproliferative drugs.

We will report how engineering of MCTS in association with new technological developments can be used to explore the regionalization and the effects of novel drugs. We will demonstrate the potential of these 3D models to study the kinetics aspects of the response on live spheroids expressing fluorescent reporters and markers using light sheet microscopy (SPIM).

We will also report the development of innovative microdevices dedicated to the study of the dynamics of MCTS formation and to the monitoring of mechanosensing and tumor growth-induced mechanical stress. We will present our latest advances on the characterization of these processes and on the use of these assays for the identification of new original targets and compounds.

Citation Format: Bernard Ducommun, Valérie Lobjois. Multicellular tumor spheroid 3D models to decipher cancer cell biology and to evaluate anticancer drugs. [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 2025. doi:10.1158/1538-7445.AM2014-2025
Abstract 2031: HMGB1 regulates pancreatic cancer initiation and pogression
Authors: Kang Rui , Zhang Qiuhong , Hou Wen , Chen Ruochan , Lotze Michael , Zeh Herbert , Tang Daolin
Published: May 2015

Most pancreatic ductal adenocarcinoma (PDAC), an almost uniformly lethal disease, is thought to arise from well-defined, non-invasive precursor lesions, termed pancreatic ductal intraepithelial neoplastic lesions (PanINs). Studies of human pancreatic carcinogenesis have been greatly facilitated by the development of a genetically engineered mouse model that expresses oncogenic K-Ras under a pancreatic promoter Pdx1-Cre:KrasG12D/+. A more detailed understanding of how this pathway accelerates pancreatic carcinogenesis may allow improved early detection, prevention, and therapeutic strategies. Our recent studies demonstrate that high mobility group box 1 (HMGB1) is a critical regulator of autophagy, a major pathway for degradation of effete proteins and damaged organelles. We found that conditional genetic ablation of HMGB1 limited to the pancreas (Pdx1-Cre;K-Ras G12D/+;HMGB1-/-; termed KCH mice) inhibits autophagy, promotes proliferation, activates normally quiescent pathways, and renders mice extraordinarily sensitive to K-RasG12D/+-driven pancreatic carcinogenesis. We found that the progression of PanINs from low grade PanIN1 to high grade PanIN3 was observed as early as three-seven days (normally three-nine months) after birth in KCH mice, suggesting a critical role of HMGB1 in regulation of the earliest events during pancreatic carcinogenesis. This extraordinarily rapid murine pancreatic cancer model that we have created will allow us to dissect the mechanism by which loss of HMGB1 contributes to the process.

Citation Format: Rui Kang, Qiuhong Zhang, Wen Hou, Ruochan Chen, Michael Lotze, Herbert Zeh, Daolin Tang. HMGB1 regulates pancreatic cancer initiation and pogression. [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 2031. doi:10.1158/1538-7445.AM2014-2031
Abstract 2032: TGF-β differentiates immortalized pancreatic epithelial cells to form duct architecture in 3D culture
Authors: Yamaguchi Takashi , Ikehara Sanae , Ikehara Yuzuru
Published: May 2015

A genetically engineered mouse (GEM) strain T26 having a transgene “CAG-loxP-βgeo-pA-loxP-SV40 tsA58 large T antigen (=tsTAg)-pA” was created by us, which expressed tsTAg in cells Cre/loxP recombination occurred. In a previous study, T26; Tie2-Cre mice were created, and their vascular and lymphatic endothelial cells expressing tsTAg could be cultured in vitro at 33°C (Yamaguchi et al., FEBS J., 2008). In this study, we established immortalized and cloned pancreatic epithelial cell lines from T26; Pdx1-Cre mice which expressed tsTAg in pancreatic epithelial cells in vivo by pancreas-specific Cre/loxP recombination. The cell lines were categorized into 1) ductal epithelial cell group which expressed CK19 and Sox9, and 2) acinar cell group which expressed Amylase2A of a digestive enzyme. In order to elucidate regulatory mechanisms of ductal cell differentiation and their plasticity, the immortalized ductal cell lines were cultured in collagen gel. A part of the cells formed spheroid or elongated and blanched architecture in the gel. Confocal or electron microscopic analysis indicated that their architectures had lumens with micro villi inside, and a part of them had tight junction at intercellular domain. It is noteworthy that the spheroid property changed into the elongated ductal (tubular) property by TGF-β stimulation in the 3D culture system, and the ductal architecture formation was suppressed by inhibitor of TGF-β signaling. 3D time-laps microscopic analysis demonstrated that a part of sphere-forming cells could change into duct-forming cells by TGF-β, which suggested existence of stem cells with susceptibility to TGF-β stimulation. The effect of TGF-β to change sphere-forming property into duct-forming property could be confirmed partially in some originally established murine pancreatic cancer cell lines. Taken together, these results suggested that TGF-β is an important factor for pancreatic duct and cancer differentiation and their architecture formation.

Citation Format: Takashi Yamaguchi, Sanae Ikehara, Yuzuru Ikehara. TGF-β differentiates immortalized pancreatic epithelial cells to form duct architecture in 3D culture. [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 2032. doi:10.1158/1538-7445.AM2014-2032
Abstract 2036: OncoPanel 3D: High-content characterization of 240 three-dimensional tumor spheroids for drug response profiling and biomarker discovery
Authors: Snead Katie , Hnilo Jim , Bernards Karen , Crane Jonathan M. , Nelson Brian , Angione Alison R. , McKinley Keith , Waikins Kate , Shah O. Jameel
Published: May 2015

While drug response profiling of cancer cells in two-dimensional culture has been a mainstay of predictive biomarker discovery and anti-cancer drug development, there are aspects of tumor biology that are not replicated in a two-dimensional cell culture environment. This is particularly true for solid tumor models where the three-dimensional organization of the tumor creates distinct, regional microenvironments that influence drug response. For example, as the radius of solid tumors approaches the diffusion limit of oxygen and other nutrients, the tumor core becomes hypoxic and accumulates metabolic waste products, resulting in cell death through mechanisms involving apoptosis and necrosis. Moreover, without proper vascularization, the tumor interior also exhibits increased interstitial pressure, which limits drug penetration into the tumor. We have characterized the three-dimensional growth of 240 cancer cell lines from various tumor origins by high-content imaging. This characterization involves the growth of tumor spheroids over 14 days and an assessment of tumor hypoxia in each model. The genomic traits associated with spheroid formation are also defined through genomic analysis. Finally, we provide validation and drug response data for several standard-of-care cancer therapeutics in pre-formed, three-dimensional tumor spheroid models. Drug profiling of this three-dimensional cell line panel offers new insights into drug response and can be used to prioritize drugs with improved tumor penetration or those with otherwise improved activity in three-dimensional models.

Citation Format: Katie Snead, Jim Hnilo, Karen Bernards, Jonathan M. Crane, Brian Nelson, Alison R. Angione, Keith McKinley, Kate Waikins, O. Jameel Shah. OncoPanel 3D: High-content characterization of 240 three-dimensional tumor spheroids for drug response profiling and biomarker discovery. [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 2036. doi:10.1158/1538-7445.AM2014-2036
Abstract 2039: Dynamic changes and regulation of circulating tumor cells that govern the metastasis of colorectal cancer
Authors: Tseng Ju-Yu , Yang Chih-Yung , Jiang Jeng-Kai , Lin Chi-Hung
Published: May 2015

Despite the growing public awareness and recent progress in multimodality therapy to the colorectal cancer (CRC), CRC still ranks high on the list of the most common and deadly cancers in the worldwide. The cause of CRC mortality is due mainly to the occurrence of distant metastasis, upon which the circulating tumor cells (CTCs), is thought to play an active role. CTCs could serve as a prognostic marker for mortality in clinical stage IV patients; however, the prognostic value of CTCs in early-stage patients remains uncertain. Here, we used EpCAM-postive/CD45-negative immunoselection and flow-cytometry to identify and quantify CTCs in CRC patients.

The number of mesenteric CTCs obtained from stage II patients was higher than those obtained for patients with stages I, III, or IV CRC. To explore the mechanisms underlying this clinical observation, we established an orthotopic (intra-cecal) tumor growth and metastasis mouse model. Cancer progression was monitored in real time with tumor cells expressing luciferase and GFP tag by using a bioluminescence imaging system and flow cytometery. We noticed that CTC dynamics was associated with a corresponding change in serum interleukin (IL)-17A and an opposing change in serum GM-CSF. Ablation of IL-17A by neutralizing antibodies and administration of GM-CSF effectively diminished the rise of CTCs and prevented metastasis in the mouse model. GM-CSF was through to help eliminating CTCs by boost host immunity, whereas IL-17A affected cancer cells intravasation by promoting cancer cell motility, matrix digestion and angiogenesis. We also noticed that IL-17 depletion could increase CD8+ to CD4+CD25+ ratio that might help CTCs clearance.

In CRC patients, serum levels of IL-17A positively correlated with the number of mesenteric CTCs and the patients' disease-free survival. Our results show that CTCs and IL-17A can serve not only as prognostic markers, but also therapeutic targets for CRC metastasis.

Citation Format: Ju-Yu Tseng, Chih-Yung Yang, Jeng-Kai Jiang, Chi-Hung Lin. Dynamic changes and regulation of circulating tumor cells that govern the metastasis of colorectal 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 2039. doi:10.1158/1538-7445.AM2014-2039
Abstract 2040: Adenomatous polyposis coli mediated signaling, self renewal and differentiation of breast epithelial cells
Authors: Lesko Alyssa , Nakshatri Harikrishna , Anjanappa Manjushree , Bhat-Nakshatri Poornima , Prosperi Jenifer
Published: May 2015

Breast cancer is the leading cause of cancer-related death in non-smoking women in the United States. Despite tremendous amounts of information about the etiology of breast cancer, many questions remain unanswered. It has been demonstrated that specific subtypes of breast cancer differ in their sensitivity to chemotherapeutic and targeted therapies. The oncogenic events and signaling pathways driving these tumor subtypes are distinct. This indicates that increased knowledge of the molecular events defining the unique subtypes will provide opportunities for novel therapeutic approaches tailored to fit the specific subtypes and will therefore lead to improvement in patient outcomes. The Adenomatous Polyposis Coli (APC) tumor suppressor is mutated or hypermethylated in 18-70% of sporadic breast cancers depending on subtype. Oncomine database analysis revealed reduced expression of APC in aggressive estrogen receptor (ER) negative breast cancers compared to ER-positive breast cancer or normal breast. Using the ApcMin/+ mouse model, we identified pre-neoplastic lesions in the breast and enhanced breast tumorigenesis in the presence of the Polyoma middle T antigen (PyMT) oncogene. Apc mutation changed the tumor histopathology from solid to squamous adenocarcinomas, resembling the highly aggressive human metaplastic breast cancer. In this model of mouse mammary tumorigenesis, these changes occurred independent of Wnt/b-catenin signaling and required activation of focal adhesion kinase (FAK)/Src signaling, with a subsequent increase in cyclooxygenase-2 (COX-2). The applicability of these signaling pathways to human breast cancer remains unclear. Using the epithelial cell reprogramming assay for growing normal breast epithelial cells, we have begun to characterize expression of cell surface markers to delineate subtypes of epithelial cells. We have also assessed APC expression and correlated APC expression with risk and well-established clinical markers, such as ErbB2. Future studies will modulate APC expression in specific compartments of breast epithelial hierarchy to accomplish our objectives of dissecting the molecular alterations specifically downstream of APC loss and its role in the self-renewal and differentiation of breast epithelial cells.

Citation Format: Alyssa Lesko, Harikrishna Nakshatri, Manjushree Anjanappa, Poornima Bhat-Nakshatri, Jenifer Prosperi. Adenomatous polyposis coli mediated signaling, self renewal and differentiation of breast epithelial 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 2040. doi:10.1158/1538-7445.AM2014-2040
Abstract 2043: Modeling the differential responses of cancer stem cells (CSCs) as heterogeneous versus homogenous populations in human cancers
Authors: Mancini Maria L.
Published: May 2015

Emerging data suggests that many human cancers including breast, brain, lung, colon, pancreatic and head and neck cancer are maintained by a subpopulation of self-renewing cells characterized as cancer stem cells that contribute to tumor growth and metastasis. Cancer stem cells have become an important target for new treatment paradigms as they may hold the key to solving treatment resistance and tumor recurrence. Paramount to the development of new therapies that preferentially target cancer stem cells is having predictable and clinically relevant models that accurately represent stem cell behavior and the tumor microenvironment. Examination of cancer stem cells both as a homogenous population and as a heterogeneous population interacting with other tumor cells is required for the development of effective treatments for stem cell driven tumor types. To address this we developed in vitro and in vivo cancer stem cell models of breast and pancreatic cancers to provide a comprehensive approach for preclinical research in these areas. Using the MDA-MB-231 and BxPC3 human cancer cell lines we are able to show the differential responses of cancer stem cells isolated from the parent lines to the standard chemotherapeutic agents docetaxel and gemcitabine with and without the small molecule inhibitor Reparixin specifically designed to inhibit cancer stem cells. Treatment response differences between homogeneous cancer stem cell versus heterogeneous populations were revealed in their growth, differentiation and invasiveness as determined by 2D and 3D tumorsphere assays. This response is also recapitulated in vivo using IVIS imaging to compare orthotopic tumor growth of isolated cancer stem cells and the parent lines treated with standard chemotherapeutics agents with or without combination Reparixin treatment. These results emphasize the need to employ a multifaceted approach to specifically target cancer stem cells in preclinical drug development to identify successful therapies for use as part of an overall treatment regimen in recurring or treatment resistant cancers.

Citation Format: Maria L. Mancini. Modeling the differential responses of cancer stem cells (CSCs) as heterogeneous versus homogenous populations in human cancers. [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 2043. doi:10.1158/1538-7445.AM2014-2043
Abstract 2063: Trojan horsing the blood-brain barrier with nanocarriers aided by adenosine receptor agonists
Authors: Awwad Nasir Al
Published: May 2015

Drug delivery to the central nervous system (CNS) is not easy compared to other body systems due to the presence of the blood brain barrier (BBB). It is essential that any delivery system to the CNS needs to be efficient and non-toxic. These features are fulfilled through tailored nanomicelles, non cross-linked and disulfide cross-linked micelles (NCM and DCM, respectively) and Polyvinyl alcohol-based polymeric nanoparticles. It has recently been reported that adenosine receptor agonists can modulate the permeability of the BBB. For example 5′-(N-Ethylcarboxamido)Adenosine (NECA) helps increase penetrability of the BBB membrane to macromolecules. We were able to demonstrate that similar strategy can be applied in mouse model for facilitating the penetration of NCM, DCM, and PVA-nanoparticles through the BBB. In the murine model we found that NECA stimulated the uptake of these nanoparticles to organs expressing adenosine receptors, especially the brain. DCM and NCM uptake was not seen in the brain without prior administration of NECA. NECA has been previously shown to alter tight junction adhesion molecules (Occludin) expressed on brain endothelial cells. NECA at 5.3 mg/kg best facilitated the penetration of both DCM and NCM at an optimal loading dosage of 33 mg/kg. NECA opened the BBB and facilitated the penetration of nanoparticles through the BBB at a constant rate. This allows potential for future drug-loading studies on a diverse scale for chemotherapeutics, kinase inhibitors, and other drug classes.

Note: This abstract was not presented at the meeting.

Citation Format: Nasir Al Awwad. Trojan horsing the blood-brain barrier with nanocarriers aided by adenosine receptor agonists. [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 2063. doi:10.1158/1538-7445.AM2014-2063
Abstract 207: Dual targeting of DLL4 and VEGF signaling by a novel bispecific antibody inhibits tumor growth and reduces cancer stem cell frequency
Authors: Yen Wan-Ching , Axelrod Fumiko , Bond Chris , Cain Jennifer , Chartier Cecile , Fischer Marcus , Ma Shirley , Meisner Rene , Raval Janak , Shah Jalpa , Gurney Austin , Lewicki John , Hoey Timothy
Published: May 2015

Both Notch/Delta-like ligand 4 (DLL4) and vascular endothelial growth factor (VEGF) pathways play a critical role in angiogenesis and tumor growth. Due to differential regulatory effects of VEGF and DLL4 on the vasculature, inhibition of DLL4 or VEGF inhibits tumor growth by distinct mechanisms: anti-DLL4 treatment induces an abnormal increase of poorly perfused blood vessels, which results in a nonproductive angiogenesis unable to support tumor growth, whereas the anti-VEGF therapy significantly decreases vasculature reducing the blood supply to tumors. We have recently developed a high binding affinity bispecific monoclonal antibody that targets both human DLL4 and human VEGF. In vitro, this antibody exhibited nanomolar affinity to hVEGF and hDLL4, and reduced HUVEC proliferation induced by VEGF. The bispecific antibody demonstrated significant in vivo anti-tumor efficacy in various solid tumors, delayed tumor recurrence following termination of chemotherapy, and decreased the frequency of tumor initiating cells. Analysis of tumor vasculature after treatment with anti-DLL4/VEGF revealed inhibition of vascular gene expression and endothelial cell proliferation, indicating that the anti-VEGF effect on the vasculature is dominant over the anti-DLL4 effect. Notably, at doses where both anti-DLL4 and anti-VEGF alone produced suboptimal anti-tumor effect, dual targeting resulted in additive tumor growth inhibition. These results indicate that our bispecific anti-DLL4/VEGF is broadly efficacious and may be useful for treatment of a variety of solid tumors.

Citation Format: Wan-Ching Yen, Fumiko Axelrod, Chris Bond, Jennifer Cain, Cecile Chartier, Marcus Fischer, Shirley Ma, Rene Meisner, Janak Raval, Jalpa Shah, Austin Gurney, John Lewicki, Timothy Hoey. Dual targeting of DLL4 and VEGF signaling by a novel bispecific antibody inhibits tumor growth and reduces cancer stem cell frequency. [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 207. doi:10.1158/1538-7445.AM2014-207
Abstract 2075: Resistin promotes MMP-2 production and migration in human chondrosarcoma cells through AMPK/p38/mir 519d pathway
Authors: Huang Ho-Ning , Tang Chih-Hish , Huang Yuan-Li
Published: May 2015

Chondrosarcoma is a primary malignant bone cancer, with a potent capacity to invade locally and cause distant metastasis; it has a poor prognosis and shows a predilection for metastasis to the lungs. Resistin the most potent pro-inflammatory and plays a crucial role in migration and metastasis of human cancer cells. However, the effects of resistin on human chondrosarcoma cells are largely unknown. In this study, we found that treatment of human chondrosarcoma cells (JJ012) with resistin increased migration and expression of matrix metalloproteinase (MMP)-2. Resistin-mediated cell migration and MMP-2 expression were reduced by pretreatment with inhibitors of AMP-activated protein kinase (AMPK), and p38 mitogen-activated protein kinases (p38). Resistin treatment induced phosphorylation of AMPK and p38. In addition, resistin reduced miRNA 519d expression. Transfect ion of 519d mimic also reduced resistin mediated migration and MMP2 expression. Furthermore, we analyzed samples from chondrosarcoma patients by immunohistochemical staining. The expression of resistin and MMP2 in chondrosarcoma patients was significantly higher than in normal cartilage. Taken together, these results suggest that resistin increased MMP-2 expression and migration in human chondrosarcoma cells.

Citation Format: Ho-Ning Huang, Chih-Hish Tang, Yuan-Li Huang. Resistin promotes MMP-2 production and migration in human chondrosarcoma cells through AMPK/p38/mir 519d pathway. [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 2075. doi:10.1158/1538-7445.AM2014-2075
Abstract 2076: βIII-tubulin is required for the tumorigenic phenotype and resistance to anoikis via the PTEN/AKT signaling axis in non-small cell lung cancer
Authors: McCarroll Joshua A. , Gan Pei Pei , Erlich Rafael B. , Liu Marjorie , Dwarte Tanya , Akerfeldt Mia C. , Chang Melissa , Shum Michael S. , Byrne Frances , Kavallaris Maria
Published: May 2015

BACKGROUND: Non-small cell lung cancer (NSCLC) has a dismal prognosis and remains the most common cause of cancer death worldwide. Expression of βIII-tubulin, encoded by the TUBB3 gene, is associated with clinical resistance and aggressive disease in NSCLC1. Herein, we interrogated the mechanistic role of βIII-tubulin in regulating the tumorigenic potential of NSCLC.

METHODS: Functional studies involved independent clones of NSCLC H460 cells stably expressing shRNA targeting βIII-tubulin, H460 controls stably expressing (non-functional) shRNA and βIII-tubulin rescue clones established in our laboratory2. Differential proteomics was conducted using fluorescence based 2D-DIGE and mass spectrometry. Gene and protein expression performed by qRT-PCR and western blotting respectively. To assess effects on tumor growth and incidence we used metastatic (tail vein) and subcutaneous models of NSCLC. Tumors monitored by CT or Xenogen imaging.

RESULTS: Functional and differential proteomics revealed that βIII-tubulin regulates expression of tumor growth- and metastases-associated proteins. In particular, the tumor suppressor maspin, associated with adhesion and metastasis, was differentially regulated by βIII-tubulin. Functionally, βIII-tubulin suppression led to altered cell morphology, increased cell adhesion and increased sensitivity to anoikis. Mechanistically, we identified PTEN and AKT kinase as a key signaling axis mediating anoikis and regulated by βIII-tubulin levels in NSCLC cells. Finally, βIII-tubulin suppression was shown to reduce NSCLC tumor growth and incidence in vivo. Collectively, these data identified βIII-tubulin as a regulator of tumor growth and metastasis through regulation of PTEN and AKT signaling. We conclude that suppressing βIII-tubulin may reduce tumor growth in NSCLC.

SIGNIFICANCE: This is the first study to show that silencing βIII-tubulin alters the expression of proteins involved in promoting tumorigenicity and increases sensitivity to anoikis, leading to reduced tumor incidence. Targeting βIII-tubulin could be a promising strategy for inhibiting tumor growth and metastasis in NSCLC.

1 Kavallaris, M. Nature Rev Cancer, 10:194-204, 2010

2 McCarroll et al. Cancer Res 70 :4995-5003, 2010

Citation Format: Joshua A. McCarroll, Pei Pei Gan, Rafael B. Erlich, Marjorie Liu, Tanya Dwarte, Mia C. Akerfeldt, Melissa Chang, Michael S. Shum, Frances Byrne, Maria Kavallaris. βIII-tubulin is required for the tumorigenic phenotype and resistance to anoikis via the PTEN/AKT signaling axis in non-small cell lung 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 2076. doi:10.1158/1538-7445.AM2014-2076
Abstract 2077: Snail modulates stemness properties in hypoxic Glioblastoma
Authors: srivastava Chitrangda , Irshad Khushboo , Sahu Arpit , Chattopadhyay Parthaprasad , Sarkar Chitra , Gupta Deepak kumar , Sinha Subrata , Chosdol Kunzang
Published: May 2015

Introduction: Glioblastoma (GBM) is the most aggressive form of glioma characterized by hypoxic regions with surrounding areas of necrosis. Hypoxia is responsible for inducing cancer invasiveness via epithelial-mesenchymal transition (EMT), thereby promoting tumor aggressiveness and therapeutic resistance. HIF-1α is known to upregulate Snail in various cancers. Snail over expression correlates with cancer stem cell-like properties in cancers. The association of EMT and stemness properties plays a vital role in tumors progression and might helps in therapeutic targeting. We have, therefore, investigated the correlation of EMT and Stem cell markers in hypoxic GBM.

Method: mRNA expression of 5 EMT, 3 stemness and 5 hypoxia markers were analyzed in the 31 GBM samples and U373MG glioma cell line maintained under normoxia (20% O2) and severe hypoxia (0.2% O2) conditions for 24, 48 and 72 hrs. Correlation and cluster analysis were applied using SPSS 11.5, Cluster 3.0 software etc and R-software.

Result: In tumors, spearman correlation was found to be significant (p≤0.05) between EMT markers and Hypoxia markers. Of all EMT markers, Snail correlated significantly with the maximum hypoxia markers, including HIF-1α. In addition, Snail was significantly correlated with Stem cell markers SOX2 (p<0.01) and Nestin (p<0.05). Gene Cluster analysis in 31 GBM arranged in decreasing order of Snail expression showed significant upregulation of SOX2 and Nestin in the high Snail expressing tertile. This correlation of gene expression in tumors was validated by in-vitro expression analysis of the molecules in U373MG cells, where we assessed significant increase in the expression of hypoxia markers (HIF-1α, VEGF and CA9), EMT (Snail and LOX) and SC markers (SOX2 and OCT4) were significantly increased under severe hypoxic conditions.

Interpretation & conclusions: Our finding for the first time suggests the role of Snail in provoking stemness of cancer cells in hypoxic GBM, thereby promoting aggressiveness of GBM.

Note: This abstract was not presented at the meeting.

Citation Format: Chitrangda srivastava, Khushboo Irshad, Arpit Sahu, Parthaprasad Chattopadhyay, Chitra Sarkar, Deepak kumar Gupta, Subrata Sinha, Kunzang Chosdol. Snail modulates stemness properties in hypoxic Glioblastoma. [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 2077. doi:10.1158/1538-7445.AM2014-2077
Abstract 208: Metformin represses stem cell properties and induces apoptosis in human breast cancer cells and mouse mammary epithelial cells
Authors: Kang Eun Joo , Seo Jae Hong , Kim Jun Suk , Kim Ji Young
Published: May 2015

Cancer stem cells (CSCs) are small portions of cancer cells that have important self-renewal and proliferation characteristics, and therefore, they play a significant role in cancer development and metastasis. Recently, several studies have revealed CSCs of several solid tumors. The identification and targeting of CSCs are expected to advance treatment outcomes, but until now, only a few drugs that specifically target CSCs have been developed.

Metformin, an antidiabetic drug used for the treatment of type 2 diabetes mellitus, suppresses gluconeogenesis in the liver and increases insulin sensitivity and glucose uptake in peripheral tissues such as muscles. Moreover, the drug has shown an anti-cancer effect by inhibiting cellular proliferation and protein synthesis in several cancer cell lines. Additionally, many clinical and epidemiological studies have suggested that metformin might be associated with a decreased risk of cancer development and an increased response to chemotherapy.

In addition to its anticarcinogenic effect, studies about the anti-cancer stem cell effect of metformin have been recently reported. Several reports revealed that metformin could preferentially kill cancer stem cells, including breast cancer stem cells. However, the mechanism of anti-cancer stem cell activity is not well known.

Therefore, this study was planned to confirm the cytotoxicity of metformin, especially for breast cancer stem cells as well as breast cancer cells. To this end, several human breast cancer cell lines and normal mouse mammary epithelial cell lines were used. In this study, metformin showed cell growth inhibition in human breast cancer cells and mouse mammary epithelial cells by inducing cell cycle arrest and apoptosis. Moreover, metformin specifically killed breast cancer stem cells. Metformin showed reduced mammosphere formation in all cell lines. In addition, decreasing breast cancer stem cell population by metformin treatment was identified through FACS analysis. For the mechanism of anti-cancer stem cell activity, western blot assay was performed. We identified decreased levels of cyclin D and cycline E and increased levels of Bax in secondary mammaospheres according to metformin treatment. In conclusion, this present study show that metformin kills breast cancer stem cells as well as breast cancer cells, and these phenomena occur by the induction of cell cycle arrest and apoptosis.

Citation Format: Eun Joo Kang, Jae Hong Seo, Jun Suk Kim, Ji Young Kim. Metformin represses stem cell properties and induces apoptosis in human breast cancer cells and mouse mammary epithelial 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 208. doi:10.1158/1538-7445.AM2014-208