bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract C17: Identifying novel genes critical to invasion in head and neck squamous cell carcinoma
Authors: Jayakar Sangeeta K. , Loudig Olivier D. , Brandwein-Gensler Margaret , Ryung Kim S. , Prystowsky Michael B. , Segall Jeffrey E. , Belbin Thomas J. 
Published: August 2014

Abstract
Oral cavity squamous cell carcinomas (OCSCC) have a poor patient prognosis, which is attributed to their invasive nature. Our goal was to identify specific genes that are important in OCSCC invasion, utilizing genome-wide expression data generated from HNSCC patient tumor samples of varying degrees of invasiveness. Invasion was assessed using the histological parameter pattern of invasion (POI), which describes the manner in which tumor cells infiltrate the normal tissue at the tumor-host interface. POI was used to categorize tumor resections into five types (1-5), type 5 being the most invasive with a “widely dispersed pattern of tumor infiltrate” and correlated significantly with the appearance of local recurrence and decreased overall survival of the patients after surgery. Global gene expression data was acquired from analysis of the surgical resections from 9 OCSCC patient samples collected at Montefiore Medical Center between 2001 and 2008. Tumor cell-enriched cores from formalin-fixed paraffin-embedded (FFPE) samples were used to generate global gene expression data using the Illumina® WG-DASL array to compare gene expression in four tumors that were WPOI type 5 and five tumors that were WPOI type 3. RNA extracted from flash-frozen tumor samples following surgical resection from the same patients was used to obtain global gene expression data using the Illumina® HumanHT-12 v4 Expression BeadChip. Candidate genes were assessed using in vitro invasion assays following siRNA knockdown of specific gene expression using siGENOME smart pools from Dharmacon. When both gene expression datasets were analyzed, we identified 104 genes that were overexpressed at least 1.5-fold in the WPOI type 5 tumors compared to the less invasive WPOI type 3 tumors. Fifty genes were initially tested based on functional criteria. In our initial screen, 16 of these genes showed a significant reduction in tumor cell invasion when expression was knocked down by siRNA in an OCSCC cell line (UMSCC1). These 16 genes are now being tested in other cell lines as well as in our in vivo mouse model to assess their invasive phenotype. This initial screen of global gene expression data in combination with pattern of invasion has revealed multiple novel genes which may play a critical role in OCSCC invasion and warrant further study.
Citation Format: Sangeeta K. Jayakar, Olivier D. Loudig, Margaret Brandwein-Gensler, Kim S. Ryung, Michael B. Prystowsky, Jeffrey E. Segall, Thomas J. Belbin. Identifying novel genes critical to invasion in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C17.
Abstract C2: High-content imaging facilities modeling of cancer epithelial-mesenchymal transition signaling network
Authors: Zhu Xiaoping , Zhen Yin , Zhao Hong , Wong Stephen 
Published: August 2014

Abstract
Background: The epithelial-mesenchymal transition (EMT) is critically involved in tumorigenesis and tumor progression except its fundamental role to development and tissue remodeling. Although the full spectrum of signaling agents that contribute to EMTs of carcinoma cells remains unclear, the interactions and coordination of individual known molecules or pathways that contribute to EMT phenotype as a network are not well studied. The EMT signaling network is complicate in nature, as such, a systems mathematical modeling is especially valuable to help understand and characterize genes that are mediated individually or in combination in the context of relevant signaling pathways, and such models would help to predict feedback loop in the network and provide candidate strategies to control the signaling transduction furthermore guide us make clinic research available after the further validation.
Experimental design and methods: siRNA-mediated knockdown of individual nine EMT-related genes in mesenchymal like cancer cell line (HMLE-Ras-Snail and U87), and epithelial cell lines (HMLE-Ras-vector and MCF10A) were imaged in a time-lapse manner at single cell resolution. Multiple cell phenotype features were extracted by image informatics analysis. Thirteen proteins including the 9 siRNA targeted genes as well as 4 epithelial and mesenchymal marker proteins were profiled by Western blot. Bioinformatics modeling was performed to create a gene EMT network and describe the genes or pathway interactions which are dominant for EMT or MET phenotypes.
Results: Each individual siRNA knockdown indicated by western blot, concomitantly incucyte captured clear cell morphology changes due to RNA silence: silencing TJP1, PXN, FLNA, CCNE1switched cells to mesenchymal-like, silencing Gli3, CTNNB1, PSEN1 induced cells more Epithelial-like. However not every maker protein showed as significant changes as images parameters (cell width/length ratio, protrusion ect) did. Therefore by using traditional biochemical and imaging two approaches, we will identify the confident genes for EMT regulation.
Conclusion: Our results demonstrate that high-content imaging is a reliable and convenient tool to help us to distinguish crucial metastasis genes and related pathway from other ambiguous, redundant and broad genes network that is valuable for us to interpret the intricate EMT mechanism.
Citation Format: Xiaoping Zhu, Yin Zhen, Hong Zhao, Stephen Wong. High-content imaging facilities modeling of cancer epithelial-mesenchymal transition signaling network. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C2.
Abstract C25: Hsp27 is required for EGF-induced epithelial to mesenchymal transition in prostate cancer
Authors: Cordonnier Thomas , Bishop Jennifer L. , Shiota Masaki , Takeuchi Ario , Nip Ka Mun , Gleave Martin , Zoubeidi Amina 
Published: August 2014

Abstract
Prostate cancer (PCa) is the most common cancer and the second leading cause cancer death in North American men. Understanding the molecular mechanisms underlying metastatic PCa development is essential to decrease morbidity and mortality. An important determinant for metastasis is epithelial to mesenchymal transition (EMT). We show that the molecular chaperone Hsp27 drives EMT in PCa cells by decreasing epithelial cell markers, upregulating mesenchymal markers and enhancing cell migration and MMP-9 activity. Mechanistically, Hsp27 is required for EMT induced by epidermal growh factor (EGF) via modulation of the beta-catenin/Slug signaling pathway. Hsp27 knockdown induces sequestration of beta-catenin in the cytoplasm by increasing its binding to GSK3beta and abrogating EGF induced beta-catenin nuclear translocation. Additionally, by decreasing the nuclear translocation of beta-catenin, Hsp27 knockdown decreases the binding of beta-catenin to the Slug promoter, thereby abrogating its transcriptional activity. As Slug is known to be a critical factor in EMT induction, we confirmed the role of Hsp27 in EGF mediated EMT using Hsp27 knockdown. To address a role for Hsp27 in promoting aggressive PCa in vivo, Hsp27 was suppressed in a murine model of PCa using the antisense drug OGX-427 (currently in phase II clinical trials). We found OGX-427 treatment significantly reduces tumor cell metastasis in mice, and moreover, decreases circulating tumor cell counts in patients with castration resistant prostate cancer. This study defines a role for Hsp27 as a critical regulator of EGF dependent EMT and highlights Hsp27 is an attractive anti-cancer target for metastatic PCa.
Citation Format: Thomas Cordonnier, Jennifer L. Bishop, Masaki Shiota, Ario Takeuchi, Ka Mun Nip, Martin Gleave, Amina Zoubeidi. Hsp27 is required for EGF-induced epithelial to mesenchymal transition in prostate cancer. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C25.
Abstract C26: Molecular determinants of the intravasation step in cancer metastasis
Authors: Deryugina Elena I. , Quigley James P. 
Published: August 2014

Abstract
Intravasation, the directional entry of primary tumor cells into host vasculature, remains the most understudied step in the metastatic cascade, likely due to deficiency in models recapitulating the intravasation process and allowing for molecular probing and intervention in vivo. By employing chick embryo models of spontaneous metastasis, where human tumor cells form microtumors and disseminate into secondary tissues strictly via an angiogenic vasculature, we have identified several rate-limiting processes and critical molecules of tumor and host origin, which functionally contribute to intravasation. We have demonstrated that the induction of an intravasation-sustaining tumor microenvironment involves tumor-produced cytokines, including IL-8 that triggers the influx of neutrophils into developing primary tumors. These inflammatory neutrophils deliver a potent pro-angiogenic protease, TIMP-1-free MMP-9, which releases stroma-bound angiogenic factors such as FGF-2 and VEGF. In addition, our recent findings indicate that M2-polarized macrophages also produce TIMP-1-deficient MMP-9, which explains the high angiogenic potential of this sub-type of inflammatory cells. Whereas host inflammatory cell MMP-9 promotes tumor angiogenesis, one of the tumor-produced MMPs, namely MMP-1, appears to regulate the microarchitecture of intratumoral vasculature. We have demonstrated that carcinoma cell MMP-1 facilitates tumor cell intravasation by specific induction of vascular permeability via activation of PAR1 expressed on the angiogenic endothelial cells. Finally, we show that intimate interactions between tumor cells and host endothelial cells result in de novo generation of plasmin that cleaves a transmembrane molecule, CDCP1, regulating tumor cell survival and motility signaling, which allows for successful intravasation. Together, our findings illuminate how different proteolytic enzymes, implicated mostly in path-clearing matrix proteolysis and tumor invasion, also orchestrate localized protein modifications and regulate cellular processes leading to, and following the intratumoral intravasation events.
Citation Format: Elena I. Deryugina, James P. Quigley. Molecular determinants of the intravasation step in cancer metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C26.
Abstract C37: Positive involvement of chemokine CXCL14 in osteolytic bone metastasis of lung cancer
Authors: Iguchi Haruo , Takiguchi Soichi 
Published: August 2014

Abstract
To investigate the molecular mechanisms for osteolytic bone metastasis, we established a human lung cancer cell line (HARA-B4) with elevated bone metastatic activity. All the experiments employed by HARA-B4 were performed with the parental HARA as a control cell line. Anchorage-independent growth ability was higher in HARA-B4 according to the results by soft agar colony formation assay. Murine macrophage-like and osteoblast-like cell lines (RAW264.7 and MC3T3-E1, respectively) migrated faster toward cultured HARA-B4 in transwell cell migration assay. The spans of bone metastatic emergence and survival were shortened in nude mice inoculated with HARA-B4. CXC chemokine ligand 14 (CXCL14) was overexpressed in HARA-B4 and responsible for the activities described above according to the results performed with the CXCL14 siRNA-treated HARA-B4. Furthermore, in the clinical specimens of the lung cancer-induced bone metastasis, expression of CXCL14 was observed in the infiltrated tumor cells of the bone marrow as well as bone stromal cells in all the specimens examined (n=7). These data indicates the positive involvement of CXCL14 in the bone metastasis from lung cancer.
Citation Format: Haruo Iguchi, Soichi Takiguchi. Positive involvement of chemokine CXCL14 in osteolytic bone metastasis of lung cancer. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C37.
Abstract C40: αvβ6 integrin promotes a TGFβ1-mediated cancer cell autonomous osteolytic program through upregulation of matrix metalloproteinase 2 (MMP2)
Authors: Dutta Anindita , Li Jing , Lu Huimin , Akech Jacqueline , Pratap Jitesh , Wang Tao , FitzGerald Thomas J. , Jiang Zhong , Violette Shelia M. , Stein Gary S. , Lian Jane B. , Languino Lucia R. 
Published: August 2014

Abstract
Metastatic prostate cancer to the bone is associated with considerable morbidity and mortality. Because the molecular basis of this process is not completely understood, current treatment is largely palliative, aiming at reducing severe bone pain, nerve compression and pathological fractures. The molecular circuitries controlling osseous prostate metastasis are known to depend on the activity of a pro-metastatic cytokine, TGFβ1, and of matrix metalloproteinases (MMPs). MMP2 is known to enhance prostate cancer-mediated bone degradation and has been reported to be induced by TGFβ1. Various studies show that the αvβ6 integrin activates and is induced by TGFβ1, thus potentiating TGFβ1 responses in the sites where it is expressed. However, a role for αvβ6 in regulating TGFβ1—induced MMP2 has not been described before. Here, we demonstrate that the αvβ6 integrin is upregulated in human prostate cancer bone metastasis and promotes osteolysis as observed by μ–computed tomography (μ–CT) in an in vivo model of metastatic bone disease. Using immunoblotting and real-time PCR analysis, we provide evidence that the αvβ6 integrin sustains TGFβ1—dependent upregulation of MMP2 mRNA and protein levels by associating with TGFβ receptor II. We also show that αvβ6 contributes to upregulation of MMP2 by TGFβ1 through a highly specific transcriptional program mediated by Smad3, and promotes osteolysis in vivo through upregulation of MMP2 in cancer cells. In conclusion, we have identified a novel role of the αvβ6 integrin in directing, in vivo, a cell-autonomous osteolytic program through activation of MMP2. These mechanistic studies will support the development of novel molecular strategies for prostate cancer bone metastasis through inhibition of the TGFβ1\αvβ6\MMP2 signaling pathway.
Acknowledgments: This work was supported by the following grants: NIH — RO1 CA89720 to Dr. Lucia R. Languino and NIH — PO1 CA140043 to Drs. Lucia R. Languino, Gary S. Stein, Jane B. Lian.
Drs. Dutta and Li share the first authorship.
Citation Format: Anindita Dutta, Jing Li, Huimin Lu, Jacqueline Akech, Jitesh Pratap, Tao Wang, Thomas J. FitzGerald, Zhong Jiang, Shelia M. Violette, Gary S. Stein, Jane B. Lian, Lucia R. Languino. αvβ6 integrin promotes a TGFβ1-mediated cancer cell autonomous osteolytic program through upregulation of matrix metalloproteinase 2 (MMP2). [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C40.
Abstract C50: CXCL12-triggered amoeboid cell motility is mediated through a RhoA-directed signaling hub
Authors: Wyse Meghan M. , Nestor-Kalinoski Andrea L. , Eisenmann Kathryn M. 
Published: August 2014

Abstract
Morphological plasticity is a common feature of cancer cells in response to environmental cues. Mesenchymal cancer cells, dependent on proteolysis for motility, can switch to a protease-independent amoeboid morphology and evade anti-invasion therapeutics that target matrix metalloproteases. The highly conserved formin family of proteins play a role in cell motility and play a critical role in mediating cancer cell invasion and metastasis. The cellular actin dynamics driving cell motility are highly regulated by mammalian diaphanous-related formin 2 (mDia2). mDia2 assembles non-branched actin filaments and maintains the cortical F-actin cytoskeleton. Our lab previously demonstrated a direct interaction between mDia2 and its inhibitor diaphanous-interacting protein (DIP), which subsequently acts as a trigger for mesenchymal/amoeboid interconvertability in breast cancer cells by disrupting F-actin polymerization and bundling. This interaction plays an essential role in the disruption of the cell cortex leading to a membrane blebbing phenotype, a hallmark of amoeboid cell motility (ACM). We recently reported that the chemokine CXCL12 stimulates DIP:mDia2 complex assembly. It is well established that CXCL12 and its receptor CXCR4 are involved in cancer metastasis, including breast cancer metastasis to lung and lymph node. Sites of distant metastasis are also often shown to express high levels of CXCL12. We, therefore, hypothesize that CXCL12 secreted from the tumor microenvironment may be a physiological trigger of amoeboid cell motility and invasion via the mDia2:DIP axis. Our current objective is to interrogate the requirement for CXCL12-driven, mDia2-dependent non-apoptotic membrane blebbing. Here we show in MDA-MB-231 human breast cancer cells that CXCL12 activates the GTPase RhoA, a known binding partner and activator of mDia2. Upon activation of mDia2, its inhibitor DIP can bind, disrupting F-actin polymerization within the cell cortex, leading to blebbing. Collectively, these data indicate a potential unexplored link between CXCL12/RhoA/mDia signaling and ACM in breast cancer cells. Understanding the molecular basis of ACM will lend novel insight into mechanisms controlling breast cancer progression and may translate into development of therapeutics controlling invasion/metastatic dissemination.
Citation Format: Meghan M. Wyse, Andrea L. Nestor-Kalinoski, Kathryn M. Eisenmann. CXCL12-triggered amoeboid cell motility is mediated through a RhoA-directed signaling hub. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C50.
Abstract C61: A macrophage autonomous α4β1integrin-Syk-Rac2 signaling axis controls macrophage differentiation, tumor growth, and metastasis
Authors: Joshi Shweta , Singh Alok Ranjan , Zulcic Muamera , Lei Bao , Messer Karen , Durden Donald L. 
Published: August 2014

Abstract
Macrophages, a major leukocyte population present in tumors, play an essential role in promoting tumor growth by affecting angiogenesis, immune suppression, invasion and metastasis. Despite considerable research efforts, the signal transduction events within macrophages which encode the complex cascade of events required for tumor growth and polarization of macrophages are poorly understood. Our previous findings demonstrate that α4β1 and αvβ3 integrin directed cell migration on specific extracellular matrices (ECM) requires a specific kinase-GTPase pair, Syk-Rac2. This pathway was shown to regulate the ECM dependent postnatal angiogenic response with no defect in vascular development. Herein, we have linked this ECM dependent signaling pathway in macrophages to the regulation of M2 macrophage differentiation and to the regulation of tumor growth, invasion and metastasis. Using mouse genetic models, we provide direct evidence that a macrophage specific, α4β1 integrin dependent Syk-Rac2 signaling axis acts in concert with the p110γ isoform (PTEN-PI-3 kinase pathway) to control HIF1α levels, tumor growth and metastasis. We report a novel proteasome/E3 ligase dependent mechanism by which PI-3 kinase regulates HIF1α levels under hypoxic conditions in the stromal compartment. Moreover, treatment with Syk or PI-3 kinase inhibitors demonstrate potent activity in multiple metastatic models. The results define a novel molecular mechanism for the regulation of HIF1α and a macrophage autonomous signaling pathway that is required for alterations in the ECM and the provisional integrin to regulate tumor metastasis and suggest treatment for metastatic disease targeting this pathway in the M2 macrophage compartment.
Citation Format: Shweta Joshi, Alok Ranjan Singh, Muamera Zulcic, Bao Lei, Karen Messer, Donald L. Durden. A macrophage autonomous α4β1integrin-Syk-Rac2 signaling axis controls macrophage differentiation, tumor growth, and metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C61.
Abstract C84: Application of sensitive EMT reporter to identification of breast cancer stem cell-related genes
Authors: Yoshikawa Kiyotsugu , Matsumoto Yoshiaki , Ito Junji , Shiojiri Yoshie , Reinherz Ellis , Toi Masakazu 
Published: August 2014

Abstract
Epithelial-mesenchymal transition (EMT) is a key feature in breast cancer stem cell. In order to identify genes responsible for EMT/stemness, we have established a novel reporter system, in which mammary epithelial cells undergoing EMT can be visualized and positively selected by expression of GFP and blasticidine-resistant (BsR) genes, respectively. By incorporating tetracycline-responsive transactivator (rTA) and tetracycline-responsive element (TRE), vimentin promoter activity could be successfully amplified, which make it possible to induce sufficient Bsr/GFP fusion protein. Snail introduction can induce GFP+ CD44hi population by 30%. Using this reporter cell line, cDNA libraries derived from aggressive breast cancer cell lines MDA-MB-231 and BT549 were screened. Snail cDNA was isolated from several clones with GFP+ CD44hi. An shRNA library screening identified several candidate genes, which might be responsible for maintenance of epithelial phenotype. GFP-positive epithelial cells were identified in the original cells, which enable us to identify early EMT markers. By replacing the vimentin promoter with E-cadherin promoter, we could successfully establish a reporter system for mesenchymal-epithelial transition (MET). Through these attempts, we are aiming to identify molecular targets in breast cancer stem cell and triple-negative breast cancer.
Citation Format: Kiyotsugu Yoshikawa, Yoshiaki Matsumoto, Junji Ito, Yoshie Shiojiri, Ellis Reinherz, Masakazu Toi. Application of sensitive EMT reporter to identification of breast cancer stem cell-related genes. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C84.