bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2011: ETV4 promotes pancreatic cancer progression and metastasis
Authors: Tyagi Nikhil , Srivastava Sanjeev K. , Bhardwaj Arun , Arora Sumit , Grizzle William E. , Owen Laurie B. , Singh Ajay P. , Singh Seema
Published: May 2015

Pancreatic cancer is a highly metastatic disease with a 5 year survival rate of ∼6.0% after diagnosis. The poor outcome of pancreatic cancer is largely due to its highly aggressive nature and lack of effective therapies against metastatic disease. Therefore, it is imperative to identify novel molecular targets driving metastatic progression of pancreatic cancer. ETV4, proto-oncogenic transcription factor, has been shown to be expressed aberrantly in various cancers types and to play important roles in pathobiology; however, its roles in pancreatic cancer have not been defined. We investigated the pathological significance of ETV4 in pancreatic cancer by its silencing in two highly metastatic pancreatic cancer cell lines, Colo357 and ASPC1, and by forced overexpression in weakly tumorigenic BXPC3 cells. Silencing of ETV4 led to decreases in growth, clonogenicity, motility and invasiveness of pancreatic cancer cells, whereas its overexpression had opposite effects. Furthermore, our data demonstrate a correlation with changes in the expression of mesenchymal (N-cadherin, Vimentin, Slug and Snail) and epithelial (E-cadherin) markers upon ETV4 modulation, indicating its role in epithelial to mesenchymal transition (EMT). More importantly, our in vivo studies in an orthotopic mouse model of pancreatic cancer demonstrated a direct association of ETV4 overexpression with tumorigenicity and metastatic potential of pancreatic cancer cells. In summary, our studies establish a functional role of ETV4 in pancreatic cancer pathogenesis.

Citation Format: Nikhil Tyagi, Sanjeev K. Srivastava, Arun Bhardwaj, Sumit Arora, William E. Grizzle, Laurie B. Owen, Ajay P. Singh, Seema Singh. ETV4 promotes pancreatic cancer progression 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 2011. doi:10.1158/1538-7445.AM2014-2011
Abstract 2013: NEDD9 depletion leads to MMP14 inactivation by TIMP2 and prevents invasion and metastasis
Authors: Pugacheva Elena N. , McLaughlin Sarah , Ice Ryan , Rajulapati Anuradha , Kozyulina Polina , Livengood Ryan , Kozyreva Varvara , Loskutov Yuriy , Ivanov Alexey , Weed Scott
Published: May 2015

The scaffolding protein NEDD9 is an established pro-metastatic marker in several cancers. Nevertheless, the molecular mechanisms of NEDD9 driven metastasis in cancers remain ill defined. Here, using a comprehensive breast cancer (BCa) tissue microarray, we show that increased levels of NEDD9 protein significantly correlated with the transition from carcinoma in situ to invasive carcinoma. NEDD9 expression is crucial for the mesenchymal invasion of cancer cells at the primary site but not at the metastatic site. Depletion of NEDD9 is sufficient to suppress invasion, leading to decrease in circulating tumor cells (CTCs) and lung metastases in xenograft models. Mechanistically, NEDD9 localizes to invasive pseudopods and is required for local matrix degradation via regulation of MMP14 trafficking. Depletion of NEDD9 impaired invasion of cancer cells through inactivation of MMP14 by excess TIMP2 on the cell surface. Re-expression of NEDD9 is sufficient to restore the activity of MMP14 and the invasive properties of BCa cells. Collectively, these findings uncover critical steps in invasion of BCa cells with potential strategy to target metastasis through manipulation of NEDD9.

Citation Format: Elena N. Pugacheva, Sarah McLaughlin, Ryan Ice, Anuradha Rajulapati, Polina Kozyulina, Ryan Livengood, Varvara Kozyreva, Yuriy Loskutov, Alexey Ivanov, Scott Weed. NEDD9 depletion leads to MMP14 inactivation by TIMP2 and prevents invasion 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 2013. doi:10.1158/1538-7445.AM2014-2013
Abstract 2020: Spheroid culture of primary colorectal cancer cells from liver metastases as an in vitro model of patient tumors
Authors: Jeppesen Maria , Hagel Grith , Vainer Ben , Harling Henrik , Thastrup Ole , Jorgensen Lars Nannestad , Thastrup Jacob
Published: May 2015

More than 65 % of patients with colorectal cancer require chemotherapeutic treatment during the course of their disease. However, colorectal tumors demonstrate pronounced heterogeneity, leading to significant individual variation in sensitivity to standard chemotherapeutic drugs. Tools for assessing chemosensitivity and tailoring therapeutic regimens to the individual patient are therefore urgently needed. A promising approach is to isolate viable cancer cells from patient tissue and to test the chemosensitivity of the individual tumor in vitro.

The aim of the current study was to establish primary colorectal cancer cell cultures from patient liver metastases and investigate if the cultured cells retained important characteristics of the original tumor. Small cell clusters were isolated from metastatic tissue and allowed to form cell spheroids in suspension culture. The cell composition of patient spheroids was characterized by immunostaining with cell type specific antibodies. Cancer stem cells were identified by staining for the markers CD44, CD166 and Lgr5. Spheroid size was monitored over time and growth curves established for each patient. The distribution of proliferating and apoptotic cells in the spheroids was assessed by staining for specific markers. Overall, the 3-D culture system was verified as a tumor model by comparing the results obtained for the spheroids to paraffin-embedded sections of the original tumors.

Successful spheroid culture was obtained from more than 80 % of tissue samples. Immunostaining confirmed that the primary cultures were composed of epithelial cells with minimal contamination by other cell types. Cell morphology and expression of intestinal markers confirmed their malignant nature and colorectal origin. The dominating cell populations of the original tumor could generally be identified in the derived spheroids as well. The spheroids demonstrated significant growth in vitro with no decrease in proliferation or increase in apoptosis during short-term culture. In summary, spheroid cultures of colorectal cancer cells can be established from liver metastases of individual patients with a high success rate and the spheroids retain important cellular characteristics of the original tumor. Spheroid culture therefore represents a promising technique for establishing primary cultures for chemotherapeutic screening of the individual patient.

Citation Format: Maria Jeppesen, Grith Hagel, Ben Vainer, Henrik Harling, Ole Thastrup, Lars Nannestad Jorgensen, Jacob Thastrup. Spheroid culture of primary colorectal cancer cells from liver metastases as an in vitro model of patient tumors. [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 2020. doi:10.1158/1538-7445.AM2014-2020
Abstract 2026: Analysis of the impact of compounds derived from marine organisms on 3D co-spheroids generated from multiple myeloma cells and stromal cell types
Authors: Ganesan Jayanthi , Weber Holger , Umber Sarah , Siedentopf Oliver , Feger Daniel , García-Fernández Luis F. , Ávila Miguel Aracil , Kubbutat Michael H.G. , Ehlert Jan E.
Published: May 2015

Multiple myeloma (MM) is a severe leukemic disease of plasma B-cells in the bone marrow to which there is currently no cure. Stromal components such as mesenchymal stromal cells (MSC) or endothelial cells (EC) are suspected to play a central role in mediating resistance during treatment of patients. In the attempt to identify novel drugs for the treatment of MM in extracts of marine organisms, we established 3D co-spheroid systems which would allow to factor in micro-environmental signals already in the early phase of drug discovery. We report on the establishment of both an angiogenesis and a proliferation assay using 3D co-spheroids combining MM cells together with either endothelial or mesenchymal stromal cells, respectively. These co-spheroid systems were applied to characterize a variety of compounds for their anti-angiogenic activity on EC or anti-proliferative activity on MM as well as MSC.

Funded by European Union Seventh Framework Program (FP7/2007-13) under Grant Agreement Nr.278570

Citation Format: Jayanthi Ganesan, Holger Weber, Sarah Umber, Oliver Siedentopf, Daniel Feger, Luis F. García-Fernández, Miguel Aracil Ávila, Michael H.G. Kubbutat, Jan E. Ehlert. Analysis of the impact of compounds derived from marine organisms on 3D co-spheroids generated from multiple myeloma cells and stromal cell types. [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 2026. doi:10.1158/1538-7445.AM2014-2026
Abstract 2028: A novel human gastric primary cell culture system for modelling Helicobacter pylori infection in vitro
Authors: Schlaermann Philipp , Meyer Thomas F.
Published: May 2015

The significance of bacterial infections in the initiation of human cancers is increasingly being recognized. A well established example is Helicobacter pylori, a causative agent of gastric adenocarcinoma. Current research largely relies on the use of transformed human cell lines as in vitro models of these infections. Obviously, this concept is largely artificial and especially inappropriate when it comes to investigations on the mechanisms of cancerogenesis. A primary cell system would be preferable in order to approximate the in vivo situation, however, previous attempts to establish such models are hampered by the limited growth and reproducibility of normal human primary cell cultures. Here, we report on the establishment of specific long-term culture conditions that allow the formation of 3-dimensional human gastric organoids with epithelial domains that have the capacity to self-renew and can be maintained in culture indefinitely. Interestingly, these 3-dimensional structures can be transferred into 2-dimensional primary cell layers, which provide ideal conditions to study infections in vitro. We expect that our novel human gastric primary cell model will facilitate investigations on the mechanisms underlying infection and is suited to open up avenues of approach to investigations on the initial processes of cellular transformation from normal to a transformed cell stages including the tumor-promoting capacity of the gastric pathogen H. pylori.

Citation Format: Philipp Schlaermann, Thomas F. Meyer. A novel human gastric primary cell culture system for modelling Helicobacter pylori infection in vitro. [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 2028. doi:10.1158/1538-7445.AM2014-2028
Abstract 203: Inhibiting invasion and stemness in glioblastoma by thymosin beta 4 gene silencing: a new therapeutic target
Authors: Wirsching Hans-Georg , Krishnan Shanmugarajan , Florea Ana-Maria , Frei Karl , Krayenbühl Niklaus , Hasenbach Kathy , Reifenberger Guido , Weller Michael , Tabatabai Ghazaleh
Published: May 2015

Glioblastoma are incurable primary brain tumors with one of the worst 5-year survival rate among all cancers. Despite multimodality treatment involving neurosurgical intervention, radiation and temozolomide chemotherapy, the median overall survival is in the range of 15-16 months. Four large phase III trials investigating dose-dense temozolomide (RTOG0525) or the addition of the integrin inhitibor cilengitide (CENTRIC) or the addition of vascular endothelial growth factor (VEGF) antibody bevacizumab (RTOG 0828, AVAglio) to the current standard treatment failed to extend this range. Novel therapeutic targets are urgently needed. Thymosin beta 4 (TB4) is a pleiotropic actin-sequestering polypeptide that is involved in wound healing and developmental processes. TB4 gene silencing promotes differentiation of neural progenitor cells whereas TB4 overexpression initiates cortical folding of developing brain hemispheres. Moreover, TB4 is involved in migration invasion and epithelial to mesenchymal transition in other cancer entities, e.g. colon cancer. In the present study, we investigated the role of TB4 in malignant glioma.

TB4 expression increased with the grade of malignancy in gliomas and correlated with patient survival. In vitro, TB4 gene silencing decreased migration, invasion, growth and self-renewal, and promoted differentiation and the susceptibility to undergo apoptotic cell death upon nutrient depletion in LNT-229, U87MG and the glioma stem-cell line GS2, respectively. In vivo, survival of nude mice bearing tumors derived from TB4-depleted glioma cells was improved and the tumorigenicity of the GS2 glioma-initiating cell line was diminished. The gene expression pattern in TB4-silenced glioma cells was shifted from the mesenchymal towards the pro-neural gene signature. The clustering of differentially regulated genes involved TGF-beta and p53 signaling networks.

Taken together, our data indicate that TB4 may be a novel regulator of malignancy in glioblastoma and therefore a candidate therapeutic target.

Citation Format: Hans-Georg Wirsching, Shanmugarajan Krishnan, Ana-Maria Florea, Karl Frei, Niklaus Krayenbühl, Kathy Hasenbach, Guido Reifenberger, Michael Weller, Ghazaleh Tabatabai. Inhibiting invasion and stemness in glioblastoma by thymosin beta 4 gene silencing: a new therapeutic target. [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 203. doi:10.1158/1538-7445.AM2014-203
Abstract 2058: Molecular imaging of glutaminolysis as a tool for evaluating therapeutic response in preclinical models of colorectal cancer
Authors: Hight Matthew R. , Schulte Michael L. , Saleh Samir , Ayers Gregory D. , Revetta Frank L. , Washington M. Kay , Coffey Robert J. , Manning H. Charles
Published: May 2015

The metabolic repertoire of cancer cells diverge significantly from that of normal cells. Energy production in cancer cells tends to depend on aerobic glycolysis, a feature that is routinely monitored by positron emission tomography (PET) imaging using 2-[18F]fluoro-2-deoxy-D-glucose ([18F]FDG). In addition to predilection for glycolysis, cancer cells may possess other unique metabolic characteristics, such as increased consumption of glutamine. As with glucose, glutamine also serves as a key carbon source for ATP production and biosynthesis. Given this, quantitative measures of glutaminolysis may reflect critical processes in oncology. Accordingly, PET agents targeting glutamine uptake, such as 4-[18F]fluoro-glutamine ([18F]4F-GLN), have been reported and used in preclinical models of cancer.

The goal of this study was to elucidate the feasibility of using [18F]4F-GLN PET to predict response to targeted therapy within the context of colorectal cancer (CRC). Initially we validated expression of SLC1A5, the primary plasma membrane transporter of glutamine, in tumor and normal colon specimens for 58 patients with primary and advanced CRC. We found that SLC1A5 expression was elevated in over 80%of primary tumor specimens but did not correlate with grade or gender. Furthermore, elevated SLC1A5 expression correlated strongly with elevated Ki67, a molecular marker of proliferation.

Given this, we evaluated [18F]4F-GLN PET in preclinical models of V600EBRAF-expressing CRC as a means of detecting anti-proliferation responses to targeted therapeutics. Simulating a clinical study within the context of the Vanderbilt GI SPORE, the regimen included an inhibitor of mutant BRAF, a PI3K/mTOR inhibitor, and a combination there of. Strikingly, [18F]4F-GLN PET was found to correlate more closely to markers of anti-proliferative responses in vivo than analogously performed [18F]FDG PET imaging studies. We believe that these findings not only provide a greater understanding of the role that glutaminolysis plays in CRC but also illuminate the potential impact that glutaminolysis derived PET could have towards guiding drug development clinical trials as an imaging metric of early therapeutic response detection.

Citation Format: Matthew R. Hight, Michael L. Schulte, Samir Saleh, Gregory D. Ayers, Frank L. Revetta, M. Kay Washington, Robert J. Coffey, H. Charles Manning. Molecular imaging of glutaminolysis as a tool for evaluating therapeutic response in preclinical models 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 2058. doi:10.1158/1538-7445.AM2014-2058
Abstract 2071: The utility of FDG uptake as a surrogate biomarker to monitor tumor cell metabolism in response to anticancer therapy
Authors: Huser Nanni , Li Wenlin , Lira Maruja , Lappin Patrick , Kindt Erick , Fantin Valeria , Li Gary , Zhang Cathy C.
Published: May 2015

[18F]FDG-PET imaging has been emerging as a powerful clinical tool for tumor diagnosis, staging and monitoring anticancer therapy. Due to some physiologic variants and technical artifacts, one of the limitations for [18F]FDG-PET imaging is the potential to show negative scans despite the presence of malignant tumors. In this report, we aimed to gain insights into the FDG tracer biology in different tumor models and subsequently to validate the use of FDG tracer uptake for monitoring targeted therapy. To measure the FDG tracer uptake in tumor cells, a LC/MS method was established to monitor the FDG monophosphate; these results were compared with the outcome of [18F]FDG-PET imaging. In a subset tumor models including patient derived xenografts, not all growing tumors showed high FDG tracer avidity. RT-PCR and IHC assays were performed to understand the potential physiological factors that could impact the FDG tracer avidity including the intrinsic glucose level, the exposure and distribution of FDG, as well as the expressions of glucose transporters and hexokinase. The results indicated that LC/MS analysis of FDG monophosphate served as a cost-effective tool to provide a surrogate measure of [18F]FDG-PET imaging. By using FDG tracer avid tumor models, several anticancer agents including the PI3K/mTOR dual inhibitor PF-04691502, crizotinib and the γ-Secretase inhibitor PF-03084014 demonstrated suppression in FDG tracer uptake after treatment, which predicted their antitumor efficacies in the corresponding models. This work provides insights into the FDG tracer biology and the utility of [18F]FDG-PET imaging as a non-invasive biomarker for anticancer drug discovery.

Citation Format: Nanni Huser, Wenlin Li, Maruja Lira, Patrick Lappin, Erick Kindt, Valeria Fantin, Gary Li, Cathy C. Zhang. The utility of FDG uptake as a surrogate biomarker to monitor tumor cell metabolism in response to anticancer therapy. [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 2071. doi:10.1158/1538-7445.AM2014-2071
Abstract 2072: Chitinase-3-like-1 protein expression associated with pulmonary inflammation accelerates metastasis to the lung
Authors: Libreros Stephania , Garcia-Areas Ramon , Iragavarapu-Charyulu Vijaya
Published: May 2015

Disseminated metastasis accounts for majority of breast cancer deaths. Recently, elevated serum levels of a glycoprotein known as chitinase-3-like-1 protein (CHI3L1) has been correlated with poor prognosis and shorter survival of patients with metastatic breast cancer. We found that there are elevated levels of CHI3L1 in plasma of mammary tumor-bearing mice. However, the biological and physiological functions of CHI3L1 are still unclear. We previously found that CHI3L1 has an inhibitory role on the expression of interferon-gamma, and in up-regulation of pro-inflammatory mediators such MCP-1, IL-8 and MMP-9 all of which contribute towards tumor growth and metastasis. In determining the cause of high levels of CHI3L1 in plasma, we found that in addition to tumor cells, splenic macrophages and more interestingly pulmonary macrophages from mammary tumor bearers secrete high levels of CHI3L1. Additionally, allergic mice also expressed higher levels of CHI3L1 in circulation and lung tissue. More specifically, CD11b+Ly6C+ cells from the lungs of allergic mice produced higher amounts of CHI3L1 compared to control mice. We have previously established that pulmonary myeloid cells through their production of CHI3L1 set up the proper microenvironment for growth of incoming mammary tumor cells. Thus, we hyphothesize that CHI3L1 levels in the lungs of mice with allergic pulmonary inflammation could accelerate metatstasis. To understand the role of CHI3L1 in accelerating pulmonary metastasis, pulmonary inflammation was induced by sensitization with ragweed allergen. Allergic mice implanted with either 4T1 or 67NR mammary tumors had a 5-fold increase in tumor volume, and formation of metastatic foci in their lungs compared to control mammary tumor bearers. Further, allergic mice showed accelerated tumor growth and shorter survival. In contrast, CHI3L1 null mice showed decreased tumor volume and metastasis, and increased survival compared to control mice. In vivo administration of chitin (β-(1-4)-poly-N-acetyl D-glucosamine) microparticles, the ligand for CHI3L1, to control and allergic mammary tumor bearers resulted in a significant reduction in pulmonary metastasis and tumor growth. These studies suggest that CHI3L1 plays a role in tumor progression and that chitin microparticles can inhibit the pleiotropic effects of CHI3L1 giving support to the idea that CHI3L1 is a useful target for treatment of breast cancer.

Citation Format: Stephania Libreros, Ramon Garcia-Areas, Vijaya Iragavarapu-Charyulu. Chitinase-3-like-1 protein expression associated with pulmonary inflammation accelerates metastasis to the lung. [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 2072. doi:10.1158/1538-7445.AM2014-2072
Abstract 2074: Metastasis prevention using fibroblast activation protein-targeting prodrugs at the pre-metastatic niche
Authors: Murgai Meera , Giles Amber , Kasai Miki , Vicioso Yorleny , Mackall Crystal , Kaplan Rosandra
Published: May 2015

Metastasis is the major cause of cancer death, and therapies that effectively target this process are required in order to improve patient survival. Previous work by Kaplan et al demonstrated that activated resident stromal cells and recruited bone marrow derived cells (BMDCs) form an altered microenvironment at distant sites in response to the primary tumor, termed the pre-metastatic niche, that promotes metastatic progression. Current on-going studies in the laboratory focus on in-depth investigations of the role, composition and mechanism of action by which each cell population including the bone marrow-derived hematopoietic cells and the activated stromal cells within the pre-metastatic niche contribute to metastatic progression. Fibroblast activation protein (FAP) is a serine protease that is overexpressed by tumor associated stromal cells, including cancer-associated fibroblasts (CAFs) and is implicated in the suppression of tumor immunity. Using a mouse model that expresses FAP promoter-driven mCherry and luciferase, we demonstrate that FAP gene expression is up-regulated in both tumor-bearing mice in pre-metastatic tissue at pre-metastatic time-points and in non-tumor-bearing mice injected with tumor-conditioned media, suggesting that metastatic tumors secrete factors that increases the number of FAP-expressing cells at the pre-metastatic niche. The studies described herein investigate the hypothesis that FAP-expressing cells at the pre-metastatic niche promote metastasis, and FAP-targeted cytotoxic drug delivery will aid in targeting tumor colonization and inhibiting metastatic progression.

Citation Format: Meera Murgai, Amber Giles, Miki Kasai, Yorleny Vicioso, Crystal Mackall, Rosandra Kaplan. Metastasis prevention using fibroblast activation protein-targeting prodrugs at the pre-metastatic niche. [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 2074. doi:10.1158/1538-7445.AM2014-2074
Abstract 2093: LPP promotes ErbB2 overexpressing breast cancer cell migration, invasion and metastasis
Authors: Ngan Elaine , Siegel Peter M.
Published: May 2015

Metastatic breast cancer remains a major hurdle to the successful treatment of this disease. Overexpression of the HER2/ErbB2 receptor tyrosine kinase in breast cancer is associated with metastatic disease and correlates with poor patient prognosis. Growing evidence demonstrates that TGFβ can potentiate ErbB2 signaling to enhance the invasive and metastatic behavior of breast cancer cells. Nonetheless, the mechanisms underlying the synergy between TGFβ and ErbB2 signaling pathways remain to be elucidated.

We previously identified Lipoma Preferred Partner (LPP) as an indispensable mediator of TGFβ-induced migration, invasion and focal adhesion turnover of ErbB2-expressing breast cancer cells. Furthermore, we established domains within LPP that are required for its localization to focal adhesions and for its interaction with α-Actinin, an actin cross-linking protein, to promote cell migration and invasion.

Herein, we examine the requirement of LPP in tumorigenesis and metastasis formation. We observe that LPP is dispensable for primary tumor growth. Nonetheless, the loss of LPP leads to decrease in spontaneous lung metastasis formation of ErbB2-expressing breast cancer cells injected into the mammary fat pads of mice. Furthermore, we observe that LPP is a critical determinant for efficient dissemination of breast cancer cells. We show that shRNA-mediated reduction of LPP significantly impairs pulmonary metastasis formation in experimental metastasis tail vein assay.

Overall, we have identified LPP as a novel mediator that integrates TGFβ and ErbB2 signaling to promote the migration, invasion and metastasis of breast cancer cells. We identify LPP as a pro-metastatic mediator of ErbB2 overexpressing breast cancer cells and a potential new biomarker of metastatic breast cancer.

Citation Format: Elaine Ngan, Peter M. Siegel. LPP promotes ErbB2 overexpressing breast cancer cell migration, invasion 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 2093. doi:10.1158/1538-7445.AM2014-2093
Abstract 2479: Nuclear promyelocytic leukemia bodies tether to early endosomes during mitosis
Authors: Palibrk Vuk , Lång Emma , Lång Anna , Rowe Alexander , Bøe Stig Ove
Published: May 2015

During mitosis, the nuclear envelope breaks down, leading to potential interactions between cytoplasmic and nuclear components. PML bodies are nuclear structures with tumor suppressor functions. However, the protein responsible for generating these bodies, the promyelocytic leukemia protein (PML), has been shown to support both nuclear and cytoplasmic activities. Here we demonstrate that PML bodies form stable interactions with the cytoplasmic transport vesicles early endosomes immediately following entry into mitosis. The two compartments remain stably associated throughout the mitotic phase and dissociate in the cytoplasm of newly divided daughter cells.  We also show that PML promotes ligand-induced endosome-mediated degradation of the epidermal growth factor receptor, and that a minor subset of PML bodies become anchored to the mitotic spindle during cell division. The study demonstrates a stable mitosis-specific interaction between a cytoplasmic and a nuclear compartment. The implications of these findings will be discussed in relation to PMLs role in tumor suppression and as a target for cancer therapy.

Citation Format: Vuk Palibrk, Emma Lång, Anna Lång, Alexander Rowe, Stig Ove Bøe. Nuclear promyelocytic leukemia bodies tether to early endosomes during mitosis. [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 2479. doi:10.1158/1538-7445.AM2014-2479
Abstract 2488: Analysis of the salivary proteome of the prolactin-inducible-protein knockout mouse
Authors: Blanchard Anne , Ezzati Peyman , Ma Xiuli , Wilkins John , Myal Yvonne
Published: May 2015

The human Prolactin-Inducible-Protein (PIP)/Gross Cystic Disease Fluid Protein 15 (GCDFP-15) gene is abnormally expressed in human invasive breast cancer and gross cystic disease of the breast. In healthy individuals PIP is highly expressed in salivary, sweat and lacrimal glands, and the secreted protein abundantly found in saliva as well as other bodily fluids which bathe various ports of entry. PIP has been shown to bind numerous bacterial strains as well as a number of immunoregulatory molecules including the CD4-T cell receptor and IgG and is therefore implicated to play an immune role in host defense. In the present study we interrogated the salivary proteome of the PIP knockout (Pip KO) mouse by mass spectrometry to determine protein changes that occur in saliva in the absence of PIP. Proteomic analysis identified 165 proteins in the mouse saliva which had a minimum of 2 unique peptides at the 99% confidence level. Nineteen proteins, including 2 proposed markers of disease, S100A9 and adenosine deaminase, were significantly differentially expressed in the saliva of the Pip KO mouse, compared to the saliva of the wild type counterparts. Functional and biological determinations revealed that the differentially expressed proteins were associated with processes that include inflammatory responses, immunity and cell death. This study provides evidence that a number of proteins associated with host defense are affected by the loss of PIP, and lends support for an immunomodulatory role for PIP.

Citation Format: Anne Blanchard, Peyman Ezzati, Xiuli Ma, John Wilkins, Yvonne Myal. Analysis of the salivary proteome of the prolactin-inducible-protein knockout mouse. [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 2488. doi:10.1158/1538-7445.AM2014-2488
Abstract 2495: Quantitative screening of protein biomarkers of hepatocellular carcinoma by antibody array
Authors: Luo Shuhong , Huang Ruochun , Wang Zhongsheng , Chen Yun-Ru , Lin Jinfei , Huang Ruo-Pan
Published: May 2015

Protein biomarkers can be key indicators for evaluating the presence or progression of a human disease or therapeutic response to treatment. We recently carried out a biomarker screening in hepatocellular carcinoma (HCC) using our antibody array platform. Using 25 pairs of HCC and adjacent control tissue samples, the expression levels of 274 proteins including growth factors, inflammation factors, angiogenesis, apoptosis factors and adhesion molecules, were examined using an antibody arrays. A panel of proteins demonstrating significant differences in expression levels between HCC and adjacent control samples was identified by significance analysis of microarrays (SAM) and Wilcoxon signed-rank test, combined with fold-change determination. Additionally, proteomic analysis algorithms were used to capture GO annotations and to construct signaling pathways and protein-protein interaction networks for these proteins. Proteins selected through these comprehensive analyses were designated as potential biomarkers and were well able to distinguish hepatocellular carcinoma tissue samples from paired, adjacent controls with cluster tools. The expression levels of several proteins were further evaluated by enzyme-linked immunosorbent assay (ELISA). Finally, we performed follow-up verification studies on selected proteins (Nidogen-1 and CEACAM-1) for detection of HCC in 164 pairs of serum samples from HCC patients and normal subjects with reverse phase protein array (RPPA). Our work reveals promising protein biomarkers for detection of HCC.

Citation Format: Shuhong Luo, Ruochun Huang, Zhongsheng Wang, Yun-Ru Chen, Jinfei Lin, Ruo-Pan Huang. Quantitative screening of protein biomarkers of hepatocellular carcinoma by antibody array. [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 2495. doi:10.1158/1538-7445.AM2014-2495
Abstract 2496: Defining the cell surface landscape in cancer: Comprehensive profiling of surface antigens for biomarker discovery and immune monitoring
Authors: O'Donnell Erika A. , Alvendia Erica L. , Osetek Benjamin E. , Wehrman Tom S. , Krutzik Peter O.
Published: May 2015

Flow cytometric analysis of surface marker expression is a fundamental component of diagnosis, classification, and monitoring for many hematologic malignancies. However, most clinical implementations of surface immunophenotyping focus on only a handful of markers. A comprehensive survey of CD markers and cell-surface receptors could provide a higher resolution and more accurate characterization of cancer lineages, as well as a better understanding of the immune response to disease progression or therapeutic intervention. To this end, we collated and validated 400 antibodies to known markers and receptors. Antibodies were validated for their performance on live and formaldehyde-stabilized samples to ensure compatibility with standard processing procedures. To establish the baseline and variability of each marker in the normal human population, a protocol was established to co-stain each sample for CD3, CD19, CD16, CD56, and CD14 simultaneously with each of the 400 markers. Differences in antigen abundance and expression frequency were calculated to establish the variation inherent in each marker across the population. To further validate the application of this panel for cancer immunophenotyping, eight commonly studied leukemia and lymphoma cell lines were selected for comprehensive surface characterization. Prior to staining, each cell line was fixed and barcoded with a combination of fluorescent dyes to generate a unique fluorescent signature. This approach allowed multiple cell lines to be combined into a single well for staining and analysis, enabling the rapid evaluation of hundreds of surface antigens using less than five million cells from each line. These studies have resulted in the optimization of a protocol and antibody panel for use in profiling surface protein expression on fixed human blood cells and cell lines. Application of this procedure to clinical samples may provide information useful for biomarker discovery, improved stratification of hematologic malignancies, and the identification of new drug targets.

Citation Format: Erika A. O'Donnell, Erica L. Alvendia, Benjamin E. Osetek, Tom S. Wehrman, Peter O. Krutzik. Defining the cell surface landscape in cancer: Comprehensive profiling of surface antigens for biomarker discovery and immune monitoring. [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 2496. doi:10.1158/1538-7445.AM2014-2496
Abstract 2497: Extracellular vesicle-mediated reversal of the prostate cancer phenotype
Authors: Panagopoulos Kiriaki , Cross-Knorr Sam , Renzulli Joseph F. , Quesenberry Peter J. , Chatterjee Devasis
Published: May 2015

Extracellular vesicle (EV) trafficking is a fundamental cellular process that occurs in cells and is required for different aspects of pathophysiology. EV trafficking leads to changes in cellular function including apoptosis, angiogenesis and proliferation required for increased tumor formation.

In our current study we report several phenotypic changes mediated by EVs isolated from non-malignant and malignant prostate cells as well as patient biopsied prostate tumor samples. EVs can reverse the resistance of prostate cancer cells to camptothecin EVs isolated from non-malignant PrECs (Prostate Epithelial Cells) can reverse soft agar colony formation of malignant DU145 cells, with the reciprocal effect observed. Isolation of EVs from 2 Gleason grade 8 prostate cancer patients significantly induced soft agar colony formation of non-malignant PrECs. We have identified proteins via antibody and Mass spectrometry analysis that may be responsible for the phenotypic changes. Mass spectrometry analysis of protein lysates using ProteoIQ revealed protein candidates associated with gene ontology annotations that may be responsible for this phenotypic change. Ingenuity Pathway Analysis was used to identify statistically relevant canonical pathways and functions associated the protein IDs and expression values obtained using ProteoIQ. Western blot analysis confirmed the increase of 14-3-3 zeta, pRKIP and prohibitin protein levels in PrEC cells co-cultured with patient EVs. 14-3-3 proteins were also found as common proteins of 3 other Gleason grade 8 patients.

Our results provides a rational basis to further investigate putative proteins, such as 14-3-3 and prohibitin and genetic factors that may be responsible for phenotypic changes that are associated with prostate cancer progression.

Citation Format: Kiriaki Panagopoulos, Sam Cross-Knorr, Joseph F. Renzulli, Peter J. Quesenberry, Devasis Chatterjee. Extracellular vesicle-mediated reversal of the prostate cancer phenotype. [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 2497. doi:10.1158/1538-7445.AM2014-2497
Abstract 250: Sulindac delayed and suppressed the tumor progression, was not effective on inhibition of tumor initiation in a human colorectal cancer mouse model (CPC;Apc mouse)
Authors: Adachi Tomohiro , Hinoi Takao , Sasaki Yuu , Shimomura Manabu , Saito Yasufumi , Miguchi Masashi , Niitsu Hiroaki , Sotomaru Yuusuke , Oue Naohide , Yasui Wataru , Ohdan Hideki
Published: May 2015

Purpose

Non-steroidal anti-inflammatory drugs (NSAIDs) have shown potential as chemopreventive agents against cancer formation, especially colorectal cancers. However, the mechanisms by which these drugs act are not fully understood. In this study, CPC;Apc mice, a human colorectal cancer mouse model treated with sulindac, were evaluated according to tumor initiation and prognosis and gene expression profile of the tumors were analyzed.

Methods

CPC;Apc mice, originating from ApcFlox/wt mice harbor a Cdx2-Cre transgene in which colorectal tumorigenesis was driven by Apc allelic loss and represent a genetic model of human colorectal cancer (CRC) were treated with control (n=20) or sulindac (160ppm, n=22) for 12 weeks and their colorectal tumors were evaluated. Gene expression analysis was performed to identify genes differentially expressed in tumor cells from sulindac-treated mice. The gene expression was validated through quantitative real-time polymerase chain reaction in vivo.

Results

Body weight of mice with control or sulindac was no significantly difference ( P = 0.801). However, we observed that sulindac significantly suppressed the tumor progression relative to untreated controls (the colon tumors size per mouse: median of 4.2mm, range 2.3-5.9 versus 5.0mm, range 3.1-8.7, respectively ; P = 0.007 ), but not the initiation (the colon tumor numbers per mouse: median of 1.5, range 0-5 versus 1.0 tumors, range 0-8 respectively ; P = 0.979) in CPC;Apc mice. We identified some upregulated (n = 6, >1.4 fold ) and downregulated (n = 5, >1.4 fold) target genes by microarray assay. We validated expression of these candidate genes in human colorectal cancer.

Conclusion

We have identified for the first time that sulindac affected tumor progression by delayed and suppressing it, but had no effect on the initiation of tumor in CRC mice model.

Citation Format: Tomohiro Adachi, Takao Hinoi, Yuu Sasaki, Manabu Shimomura, Yasufumi Saito, Masashi Miguchi, Hiroaki Niitsu, Yuusuke Sotomaru, Naohide Oue, Wataru Yasui, Hideki Ohdan. Sulindac delayed and suppressed the tumor progression, was not effective on inhibition of tumor initiation in a human colorectal cancer mouse model (CPC;Apc mouse). [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 250. doi:10.1158/1538-7445.AM2014-250
Abstract 2501: Proteomics signature of lack of retinoic acid signal surveillance at early breast cancer stages
Authors: Rossetti Stefano , Bshara Wiam , Sacchi Nicoletta
Published: May 2015

Endogenous retinoic acid (RA), the bioactive derivative of Vitamin A, is necessary for breast acinar morphogenesis. One of the hallmarks of early stage breast cancer, such as ductal carcinoma in situ (DCIS), is the inability to respond to RA morphogenetic action, which results in the uncontrolled growth of the mammary duct luminal cells. By imaging endogenous RA signaling during normal and aberrant breast acinar development in three dimensional (3D) culture, we found that the inability to form the acinar lumen can be traced to lack of activation of the RA receptor alpha (RARA) genomic function by endogenous RA variation. We further found that aberrant acinar morphogenesis, regardless of the causing factors, is marked by a distinct protein signature, which includes the upregulation of Annexin A8 (ANXA8). Forced expression of ANXA8 in breast epithelial cells was sufficient to disrupt 3D acinar architecture. Moreover, by screening DCIS tissue microarrays (TMAs), we found that ANXA8 expression correlates with tumor grade and stage. Thus, further development of this proteomics signature indicative of lack RA signal surveillance is expected to assist in the early detection of initial breast cancer stages.

This work was funded by R01 CA127614, the Friends for an Earlier Breast Cancer Test Foundation, and the Terri Brodeur Breast Cancer Foundation.

Citation Format: Stefano Rossetti, Wiam Bshara, Nicoletta Sacchi. Proteomics signature of lack of retinoic acid signal surveillance at early breast cancer stages. [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 2501. doi:10.1158/1538-7445.AM2014-2501
Abstract 2505: Exposure to a therapeutically relevant dose of ionizing radiation reveals colorectal cancer biomarker and associated receptor alterations
Authors: Suman Shubhankar , Fornace Albert J. , Datta Kamal
Published: May 2015

Purpose: Diagnostic and therapeutic radiation exposure to humans is inevitable, and radiation exposure is known to enhance risks of colorectal cancer (CRC) and inflammatory bowel diseases. Radiation exposure has also been observed to promote obesity in humans as well as in rodents. While altered levels of insulin like growth factor 1 (IGF1), leptin, and adiponectin have been reported as potential markers of obesity-related CRC risk, there is a paucity of long-term radiation-risk-markers for intestinal pathologies. The goal of the current study was to evaluate in mice whether obesity-related CRC risk markers have the potential to serve as radiation-associated CRC risk markers after exposure to a clinically relevant γ radiation dose.

Methods and materials: Mice (C57BL/6J, 6-8 week old, female; n= 8 per group) were exposed to a γ radiation dose of 2 Gy, commonly used in fractionated radiotherapy. Serum and intestinal tissues were collected two and twelve months after radiation. Serum levels of IGF-1, IGF binding protein 3 (IGFBP3), leptin, and adiponectin were analyzed by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry was performed on small intestine and colon sections for IGF1 receptor (IGF1R), obesity receptor (OB-R), adiponectin receptor 1 (AdipoR1), adiponectin receptor 2 (AdipoR2) and proliferative marker Ki-67.

Results: The levels of free IGF1 and free leptin measured by calculating IGF1/IGFBP3 and leptin/adiponectin ratio showed a distinct pattern. While significantly higher free IGF1 was observed twelve months post-radiation, higher free leptin was observed two months after radiation exposure. Immunohistochemical analysis demonstrated increased IGF1R and OB-R, decreased AdipoR1 and R2, and increased Ki67 in intestinal as well as in colonic sections at both the time points.

Conclusions: Considering that one fraction of 2 Gy showed significant alterations in risk markers, we believe exposure to a higher total dose using more fractions will show more pronounced effects. Taken together, this study demonstrates that adipogenic risk markers of CRC could be effective tools to assess CRC risks after therapeutic γ radiation exposures.

Note: This abstract was not presented at the meeting.

Citation Format: Shubhankar Suman, Albert J. Fornace, Kamal Datta. Exposure to a therapeutically relevant dose of ionizing radiation reveals colorectal cancer biomarker and associated receptor alterations. [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 2505. doi:10.1158/1538-7445.AM2014-2505