bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3827: Caveolin-1 acts as a tumor suppressor promoting muscular differentiation in alveolar rhabdomyosarcomas.
Authors: Huertas-Martinez Juan , Barrau Ignasi , Sáinz-Jaspeado Miguel , Lagares-Tena Laura , Mateo-Lozano Silvia , Mora Jaume , Roma Josep , Gallego Soledad , Moran Sebastian , Esteller Manel , Garcia del Muro Xavier , Tirado Oscar M.
Published: August 2014

Rhabdomyosarcoma is the most common soft tissue sarcoma of childhood and adolescence. Despite advances in therapy, patients with a histological variant of rhabdomyosarcoma known as alveolar rhabdomyosarcoma (ARMS) have a 5-year survival of <30%. Caveolin-1 (CAV-1), encoding the structural component of cellular caveolae, is a suggested tumor suppressor gene involved in cell signaling. We report that CAV1 is expressed in embryonic rhabdomyosarcoma (ERMS) cell lines and tumor samples. However, in 3 out of 4 different ARMS cell lines and 8 out of 9 clinical samples CAV1 expression is either undetectable or very low. The epigenetic silencing of CAV1 by hypermethylation was tested in ARMS cells before and after azacytidine treatment. CAV1 methylation was evaluated by bisulfite genomic sequencing. Re-expression of CAV1 in two of these cell lines is cytostatic and promotes differentiation. In vivo, CAV1-expressing tumor cells showed growth delay and features of muscular differentiation in comparison with untransfected and vector transfected cells. Our results demonstrated that CAV1 could function as a potent tumor suppressor in ARMS tumors. Inhibition of CAV1 function could contribute to aberrant cell proliferation, leading to ARMS development. The use of peptides that mimic CAV1 function may be of therapeutic use for the treatment of ARMS.

Citation Format: Juan Huertas-Martinez, Ignasi Barrau, Miguel Sáinz-Jaspeado, Laura Lagares-Tena, Silvia Mateo-Lozano, Jaume Mora, Josep Roma, Soledad Gallego, Sebastian Moran, Manel Esteller, Xavier Garcia del Muro, Oscar M. Tirado. Caveolin-1 acts as a tumor suppressor promoting muscular differentiation in alveolar rhabdomyosarcomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3827. doi:10.1158/1538-7445.AM2013-3827
Abstract 3837: Effect of matrix and intercellular interactions on chemotherapeutic efficacy in a high-throughput format.
Authors: O'Day Christine L. , Ejiri Yoko , Hosoda Masaya
Published: August 2014

Many researchers believe that clinical results often do not coincide with two-dimensional in vitro assays, prompting the need for better correlative cellular models. 3D culture has gained much interest in recent years for the effects of matrix and inter-cellular contacts on tumor growth and progression. We have devised a high throughput assay to look at the effects of matrix and intracellular interactions using a standard 384 2D plate, matrigel in 384 well format and 3D Elplasia micro-space plates with a 200 micron grid. We standardized the media between all three platforms but decreased the cell number 3-4 fold in 2D plates to keep growth within the linear range for a 6 day assay. Media was changed once after 3 days. In all cell lines tested, there were at least one doubling of cell number in 6 days and the rate of growth was fairly consistent between 3D and 2D techniques, with 2D growth slightly faster. We examined differences in potency and efficacy of 6 different agents in 10 cell lines comparing 2D and 3D techniques. These agents are directed at PI3K, DR4/5, MEK, EGFR, and BCL2. Effects due to matrix interactions vs. suspension were noted along with potency and efficacy changes of several compounds when compared to 2D but not in all cells lines. Often EC-50 values for various agents were identical between platforms. In a few cell lines, the potency of a PI3K/mTorr inhibitor was increased three fold but the overall efficacy (defined by area under the curve) was decreased in 2D vs. 3D. DR4/5 agonists also had a mixed profile with some cell lines. TRAIL was dramatically more potent in 2D for some lines and conversely, more potent for others in 3D. Ranges of over 10 fold in EC-50 for Trail were observed between 2D and 3D in the same cell line. This interesting variation between 2D and 3D endpoints prompts the need to correlate 3D potency with clinical data and begs insight into the reasons behind the differences in receptor mediated signaling in 3D. If 3D is more correlative, which many studies suggest, there will be a clear benefit to screening multiple cell lines for efficacy and correlating them with gene expression data in 3D culture for biomarker identification.

Citation Format: Christine L. O'Day, Yoko Ejiri, Masaya Hosoda. Effect of matrix and intercellular interactions on chemotherapeutic efficacy in a high-throughput format. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3837. doi:10.1158/1538-7445.AM2013-3837
Abstract 384: NADH oxidase, Enox1 activity, is required for vasculogenesis in Zebrafish.
Authors: Venkateswaran Amudhan , Levic Daniel S. , Melville David B. , Sekhar Konjeti R. , Walsh Alexandra J. , Skala Melissa C. , Crooks Peter A. , Knapik Ela W. , Freeman Michael L.
Published: August 2014

Establishment of an adequate vascular network is a crucial event for tumor development. Emerging research has implicated the endothelial redox state as a significant regulator of vascular growth and remodeling. ENOX1 encodes a protein that is expressed in mammalian endothelial cells and exhibits NADH oxidase activity. Recent research suggests that ENOX1 functions as a positive regulator of angiogenesis. However, direct genetic evidence supporting this hypothesis is lacking. We used a zebrafish model to directly examine the role of Enox1 during embryonic vascular development. Whole mount in situ hybridization coupled with immunocytochemistry performed on zebrafish cryosections show that enox1 message and the translated protein are expressed in heart and blood vessels during embryonic development. Furthermore, use of immunocytochemistry demonstrates that Enox1 co-localizes with vascular expression of the ETS-domain transcription factor fli in heart and intersegmental vessels. Using in vivo multiphoton microscopy, we show that morpholino-mediated knockdown of Enox1 in Zebrafish embryos increases NADH fluorescence, an expected consequence of loss of Enox1 oxidase activity. Phenotypic manifestations resulting from loss of Enox1 function in embryos include vascular defects, most notably loss of fli-eGFP and flk-eGFP (receptor kdr, VEGFR2 in mammals) expression in intersegmental vessels and a loss of blood circulation in the intersegmental vessels. Pharmacological inhibition of Enox1 activity using a novel small molecule phenocopies the defects observed following morpholino-mediated functional knockdown. Together, these results show that Enox1-mediated redox regulation is essential for vasculogenesis in vertebrates in vivo. These results, in conjunction with previous work that demonstrated that inhibition of ENOX1 radiosensitized tumor vasculature, support the hypothesis that ENOX1 represents a novel target for impairing tumor angiogenesis.

Citation Format: Amudhan Venkateswaran, Daniel S. Levic, David B. Melville, Konjeti R. Sekhar, Alexandra J. Walsh, Melissa C. Skala, Peter A. Crooks, Ela W. Knapik, Michael L. Freeman. NADH oxidase, Enox1 activity, is required for vasculogenesis in Zebrafish. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 384. doi:10.1158/1538-7445.AM2013-384
Abstract 3859: Characterization of imaging-enhanced models of disseminated multiple myeloma.
Authors: Baugher Meridith , Baranski Jenifer , Balagurunathan Deepa , Bull Christopher , Winchell Noah , Shaw Darren , Trachet Erin , Lister Deanne , McConville Patrick , Leopold Wilbur R.
Published: August 2014

Multiple myeloma is a cancer of the plasma cells that is characterized by multiple localized lesions in the marrow, particularly of the spine, skull, and pelvis, although soft tissue lesions also occur. It is the second most common blood cancer, affecting approximately 45,000 people in the US. Most preclinical modeling of myeloma employs SC xenografts that mimic the less common plasmacytoma form of the disease. Systemic (IV) implants are also used, but studies typically are based on a single survival endpoint, limiting knowledge about the progression and response of the disease under treatment. In order to more quantitatively monitor disseminated disease progression and response to treatment, we have characterized two human (JJN3 and MM1S) and one murine (5TGM1) myeloma models that have been modified to express luciferase. All models were characterized by 100% tumor take rate and focal dissemination of the disease to the spine and skull that mimic clinical experience. These models showed individual and reproducible patterns of spread to other sites, and differed in their sensitivities to standards of care. Analysis of tumor doubling times, tumor titrations, luminescence-based growth delay, and survival all indicated that the bioluminescence signal was a reliable quantitative indicator of viable tumor burden, even under treatment with clinical standard of care agents. Luciferase labeling and the tight correlation between luminescence signal and viable tumor burden raises the possibility of differential real-time tracking of tumor progression and response at individual sites.

Citation Format: Meridith Baugher, Jenifer Baranski, Deepa Balagurunathan, Christopher Bull, Noah Winchell, Darren Shaw, Erin Trachet, Deanne Lister, Patrick McConville, Wilbur R. Leopold. Characterization of imaging-enhanced models of disseminated multiple myeloma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3859. doi:10.1158/1538-7445.AM2013-3859
Abstract 386: Inhibiting the CSF-1 receptor on tumor-infiltrating monocytes may enhance the efficacy of radiotherapy for glioblastoma multiforme.
Authors: Stafford Jason H. , Chernikova Sophia , West Brian , Brown J. Martin
Published: August 2014

Glioblastoma (GBM) is the most common brain tumor and is one of the most deadly forms of cancer with median survival only 15 months after diagnosis. Ionizing radiation (IR) is the most effective treatment for GBM, but the prognosis remains poor because the tumors invariably recur. We have previously shown that bone marrow-derived monocytes promote GBM recurrence after radiotherapy by homing to the irradiation site and stimulating the growth of new tumor blood vessels in a process referred to as vasculogenesis. We found that this infiltration of monocytes can be prevented and tumor recurrences delayed by inhibition of the SDF-1/CXCR4 chemokine axis.1 Here we present data that suggest Colony Stimulating Factor-1 (CSF-1) may also play an important role in this phenomenon. CSF-1 is a potent chemokine secreted by several different types of solid tumors and is also the main growth factor regulating the differentiation of monocytes into macrophages. RT-PCR analysis of U251 GBM tumors growing in mice revealed a more than 2.5-fold increase in expression of CSF-1 after treatment with a single dose of 12 Gy IR. Increased CSF-1 expression in these tumors correlated with increased recruitment of CD11b+ monocytes. Immunohistochemistry (IHC) and fluorescence activated cell sorting (FACS) showed that the number of CD11b+ monocytes in GBM tumors had approximately doubled within 2 weeks of IR treatment. F4/80 staining indicated that the majority of the CD11b+ cells were mature macrophages. Treatment with 40 mg/kg/day PLX3397, which inhibits the receptor for CSF-1 (CSF-1R), effectively blocked the recruitment of monocytes/macrophages to the irradiated tumors. Moreover, bioluminescence imaging (BLI) of intracranial U251-luciferase tumors growing in mice showed that combined treatment with IR and PLX3397 was more effective at inhibiting tumor growth than IR alone. Eighty percent of mice treated with IR and PLX3397 were alive at 50 days post-irradiation whereas there were no survivors among groups receiving monotherapy. These results suggest inhibition of CSF-1R may prevent post-irradiation recurrence of GBM by blocking the recruitment of tumor-infiltrating monocytes/macrophages.

Citation Format: Jason H. Stafford, Sophia Chernikova, Brian West, J. Martin Brown. Inhibiting the CSF-1 receptor on tumor-infiltrating monocytes may enhance the efficacy of radiotherapy for glioblastoma multiforme. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 386. doi:10.1158/1538-7445.AM2013-386
Abstract 3878: Bach1 promotes liver metastasis of colorectal cancer cells by regulating c-Myc and SOX4.
Authors: Yun Jieun , Jo Yeong Nang , Park Song-Kyu , Kim Jin-Ah , Yang Junguk , Lee Chang Woo , Yoon Won Kee , Lee Ok-Jun , Han Sang-Bae , Doebele Robert C. , Kang Jong Soon
Published: August 2014

Bach1 is a basic leucine zipper transcription factor that binds to Maf recognition elements (MAREs) and has recently been identified as the regulator of breast cancer metastasis. Bach1 promotes breast cancer metastasis to bone by directly upregulating MMP1 and CXCR4 and it is critical for the cell transformation and tumor growth induced by activated H-RasV12. Here, we demonstrate the oncogenic mechanisms of Bach1 in colorectal cancer. Ectopic expression of Bach1 increases invasion of colorectal cancer cells, whereas knockdown of Bach1 significantly reduces the invasive efficiency of the cancer cells. In a mouse model, suppression of Bach1 expression significantly inhibits intravasation and liver metastasis of colorectal cancer cells. The mechanism involves upregulation of c-Myc and SOX4 expression by Bach1 maintaining ERK signaling. Finally, we confirmed the expression of Bach1 is high in the patient samples of colorectal cancer and metastasis, unlike normal colon tissues. These results suggest Bach1 as a novel target for cancer therapeutic strategy.

Citation Format: Jieun Yun, Yeong Nang Jo, Song-Kyu Park, Jin-Ah Kim, Junguk Yang, Chang Woo Lee, Won Kee Yoon, Ok-Jun Lee, Sang-Bae Han, Robert C. Doebele, Jong Soon Kang. Bach1 promotes liver metastasis of colorectal cancer cells by regulating c-Myc and SOX4. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3878. doi:10.1158/1538-7445.AM2013-3878
Abstract 3884: Targeting ATX and LPA receptors melanoma invasion and metastasis.
Authors: Tigyi Gabor , Fujiwara Yuko , Lee SueChin , Liu Jianxiong , Patil Renukadevi , Gupte Renuka , Miller Duane D. , Oravecz Tamas , Balazs Louisa
Published: August 2014

ATX, a lysophospholipase D that generates LPA, is highly up-regulated in metastatic and chemotherapy-resistant carcinomas. Many cancers secrete ATX, and it contributes to their invasive properties. LPA produced by ATX activates LPA1-8 G protein-coupled receptors on the cell surface. Many cancers over-express multiple subtypes of LPAR and the overexpression of LPAR leads to malignant transformation, metastasis, and resistance to therapy. Therefore, ATX-LPA signaling axis is an emerging target for cancer therapy. We have previously shown that knockdown of ATX expression in B16F10 melanoma cells decreased tumor invasion in vitro and metastasis in vivo. In this study, we sought to understand the roles of the LPA GPCRs both in the tumor cells and tissues targeted by metastatic cancer cells. Using the in vivo B16 melanoma metastasis model syngeneic to the C57BL/6 mouse strain, we found that the incidence of lung metastasis was greatly reduced in the LPA5KO mice compared to WT, LPA1KO, and LPA2KO mice, respectively. In addition, we found that the selective inhibitor of LPA1&3, Ki16425 also reduced lung metastasis, suggesting that inhibition of LPA5 and LPA1 receptors in the target tissues may prevent cancer metastasis. B16 melanoma cells predominantly express LPA5 receptor. These cells do not invade across a matrigel layer in response to LPA. We found that siRNA knockdown of LPA5 receptors in B16 melanoma cells relieves this inhibition and resulted in an increase in invasion. These results suggest that the LPAR, in particular LPA5 receptors in tumor and stromal cells may play different roles in invasion and metastasis.

Citation Format: Gabor Tigyi, Yuko Fujiwara, SueChin Lee, Jianxiong Liu, Renukadevi Patil, Renuka Gupte, Duane D. Miller, Tamas Oravecz, Louisa Balazs. Targeting ATX and LPA receptors melanoma invasion and metastasis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3884. doi:10.1158/1538-7445.AM2013-3884
Abstract 3885: FXR is implicated in matrix metalloproteinases regulation, tumor invasion and metastasis in breast cancer.
Authors: Alasmael Noura , Meira Lisiane B. , Swales Karen , Plant Nick
Published: August 2014

Metastatic breast cancer is a principle cause of female mortality. In general, metastatic cancers show poor response to current treatments and therefore the development of new therapies is essential. Metastasis is a multi-step process where cancer cells migrate from the primary tumour to other distant organs. For this to occur, degradation of the extracellular matrix is required, allowing cancer cells to invade the vasculature and migrate to distant sites. Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that breakdown the extracellular matrix, with increased expression of MMP-2 and -9 associated with metastatic breast cancer. MMP activity is tightly regulated by the tissue inhibitors of metalloproteinases (TIMPs): TIMP-1 inhibits MMP-9 activity while TIMP-2 inhibits MMP-2 activity. Studies have shown that the nuclear receptor Farnesoid X-receptor (FXR) is involved in regulation of MMP and TIMP activity in hepatic and vascular tissues. As FXR is highly expressed in metastatic breast cancer, the aim of the current study was to investigate whether FXR is a novel regulator of MMP-2 and -9 in breast cancer cells. The viability of the breast cancer cell lines MCF7 (Estrogen receptor positive) and MDA-MB-468 (Estrogen receptor negative) was measured after exposure to the FXR agonists chenodeoxycholic acid (CDCA) and 3-[2-[2-Chloro-4-[[3-(2, 6-dichlorophenyl)-5-(1-methylethyl)-4 isoxazolyl]methoxy]phenyl]ethenyl]benzoic acid (GW4064). In both cases, these FXR ligands caused cell death in both cell lines, with reduction of cell viability being significantly higher in the triple negative MDA-MB-468 cells. mRNA and protein levels of MMP-2 and -9 within both cell lines did not change after FXR activation, as measured by real time PCR and Western blotting. However, GW4064 did increase secreted MMP activity, as measured by a fluorescent substrate, in a concentration dependent manner, whereas CDCA had no effect. Such data suggests that while FXR ligands may be cytotoxic to breast cancer cells, they may also increase the risk of invasion and metastasis.

Citation Format: Noura Alasmael, Lisiane B. Meira, Karen Swales, Nick Plant. FXR is implicated in matrix metalloproteinases regulation, tumor invasion and metastasis in breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3885. doi:10.1158/1538-7445.AM2013-3885
Abstract 389: The functional relevance of decreased ANGPTL4 in human urinary bladder cancer.
Authors: Cardenas Eduardo , Horton Tiffani , Ghosh Rita
Published: August 2014

Transitional cell carcinoma is the most common urothelial cancer in the US. Current therapeutic options have been unable to overcome issues with the highly recurrent nature of this tumor, which requires frequent follow-up making urinary bladder cancer, one of the most expensive solid tumors to manage. Microarray-based gene expression analysis in response to capsaicin treatment identified significant upregulation of angiopoietin-like 4 (ANGPTL4) compared with solvent-treated bladder cancer cells. The focus of this study was to determine the functional relevance of ANGPTL4 in normal urothelial and bladder cancer cells. The microarray data was validated in a panel of normal, immortalized and bladder cancer cells of increasing stage by real-time qPCR, immunoblotting, and ELISA. To determine the possible function of ANGPTL4, we knocked down and overexpressed ANGPTL4 in normal urothelial and bladder cancer cells respectively. The role of ANGPTL4 in growth, proliferation and angiogenesis was assessed. Our results show that normal urothelial cells have significantly higher levels of ANGPTL4 when compared to non-tumorigenic and bladder cancer cells. Upon capsaicin treatment ANGPTL4 levels increased in all cell lines tested. Overexpression of ANGPTL4 in the metastatic cell line TCCSUP, led to decrease in cell growth. Further, ANGPTL4 knockdown increased cell growth in the normal urothelial cell line. Capsaicin treatment decreased the expression of VEGF in bladder cancer cell lines UMUC-3 and T24 as well as reduced expression in normal urothelium, albeit initial results in ANGPLT4-knockdown in normal urothelium indicates no change in VEGF expression. Our results thus far suggest that the loss of ANGPTL4 expression may be involved in bladder cancer. The mechanism involved is under investigation. Capsaicin remains an attractive compound for further studies in bladder cancer. Supported by RO3 CA136058 (RG) and NIHT32 T32CA148724 (EC).

Citation Format: Eduardo Cardenas, Tiffani Horton, Rita Ghosh. The functional relevance of decreased ANGPTL4 in human urinary bladder cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 389. doi:10.1158/1538-7445.AM2013-389
Abstract 3891: Hornerin: a novel, non-VEGF mediated human tumor angiogenic protein.
Authors: Seaman Marc E. , Kelly Kimberly A.
Published: August 2014

Angiogenesis is vital to the growth, progression and ultimately metastasis of tumors. As such, many therapies have attempted to eradicate tumors through anti-angiogenic strategies. Most current and past preclinical and clinically approved therapies focus on the vascular endothelial growth factor (VEGF) pathway. Unfortunately, these VEGF therapies have not shown much success, leading oncologists and scientists to become pessimistic about the usefulness of targeting angiogenesis as an anti-tumor strategy. Compensatory pathways may explain the lack of efficacy. Therapies against novel and/or non-VEGF proteins could hold the key to effective anti-angiogenic cancer therapies. Therefore, we used an in vivo phage display screen to find new, non-VEGF mediated angiogenic proteins. After phage selection, non-VEGF binding phage clones were identified by screening on human umbilical vein endothelial cells (HUVEC) treated with tumor conditioned media, VEGF conditioned media or vehicle control. Those that bound tumor conditioned media, but not VEGF conditioned media were selected for binding partner identification via functional proteomics pioneered in our lab. The binding partner of one clone was identified as hornerin, a previously unknown player in tumor angiogenesis.

Subsequent validation of hornerin by mRNA expression, immunocytochemistry and immunohistochemistry showed that hornerin was indeed not induced by VEGF. Hornerin expression was 7-fold more in resected human PDAC tumor vasculature than adjacent normal pancreatic vasculature. Interestingly, hornerin expression was also detected in tumor cells of resected human PDAC and 20 other types of tumors, providing a potential new biomarker of pancreatic and other cancers. A functional role in angiogenesis was confirmed by intratumoral injection of short siRNA against endothelial hornerin; treated tumor growth was static and tumor vessels displayed a more physiologic phenotype. Combination therapy consisting of siRNA against hornerin and a VEGF inhibitor (AV-951) produced additive reduction of tumor growth, consistent with the existence of separate and/or compensatory pathways.

The discovery of novel, non-VEGF mediated angiogenic proteins provides the potential to develop therapies that synergize with current anti-VEGF therapies. One such protein, hornerin was identified by our in vivo phage screen and shown to be functionally important in angiogenesis. Future studies include identifying the growth factor responsible for endothelial expression, understanding hornerin's mechanism and role in pathologic angiogenesis, and designing new therapeutic strategies.

Citation Format: Marc E. Seaman, Kimberly A. Kelly. Hornerin: a novel, non-VEGF mediated human tumor angiogenic protein. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3891. doi:10.1158/1538-7445.AM2013-3891
Abstract 3894: A high phosphate environment induces angiogenesis, which is mediated by FOXC2.
Authors: Lin Yiming , Garneys Laura M. , Bennett Kelly E. , Beck George R.
Published: August 2014

Inorganic phosphate is a common dietary element that is critical to the human body on many levels however the effects of changes in serum phosphate on cell and tissue behavior are not well understood. Due, in part, to the increased consumption of processed foods, the amount of inorganic phosphate in the American diet continues to rise above levels already considered high by the FDA. There is a growing appreciation that changes in serum phosphate levels influence pathological states associated with aging such as cancer, bone metabolism, and cardiovascular function, even in individuals with normal renal function. The causes are only beginning to be elucidated but are likely a combination of endocrine, paracrine, autocrine, and cell autonomous effects. Recent studies in mice have demonstrated that a diet high in phosphate, relative to a reduced phosphate diet, increases tumorigenesis in the two-stage skin carcinogenesis model as well as the Kras lung cancer model. Using cell cultures models we have determined that high phosphate results in increased proliferation and transformation as measured by XTT, cell counts, soft agar and focus formation assays. Our in vitro studies have also identified specific signaling cascades induced by elevated extracellular phosphate which result in the temporal and dynamic regulation of hundreds of phosphate responsive genes. A subset of the Pi-responsive genes including the Forkhead box protein C2 (FOXC2), osteopontin (spp1), Cyr61, and Vegf among others, has been previously linked to stimulation of angiogenesis leading to the hypothesis that conditioned medium from phosphate treated cells would increase angiogenesis. We confirmed the phosphate-induced expression of these genes in a number of cancer cells including breast, colon, and lung. The increased expression of these genes was determined to be dependent on phosphate transport as well as FGF receptor and ERK1/2 signaling. Using human umbilical cord vascular endothelial cells (HUVECs) we determined that, in fact, conditioned medium from these phosphate treated cancer cells increased both migration and tube formation in the HUVEC model. Knockdown of FOXC2 inhibited phosphate induced gene expression in cancer cells as well as eliminating the stimulation of HUVEC migration in response to Pi-treated cancer cell conditioned medium. These results suggest that a cellular environment with elevated phosphate promotes multiple aspects of tumorigenesis including angiogenesis.

Citation Format: Yiming Lin, Laura M. Garneys, Kelly E. Bennett, George R. Beck. A high phosphate environment induces angiogenesis, which is mediated by FOXC2. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3894. doi:10.1158/1538-7445.AM2013-3894
Abstract 3898: Glioma stem cells promote brain endothelial cell motility: a new mechanism for the direct interaction of glioma stem cells with endothelial cells.
Authors: Burgett Monica E. , Huang Ping , Tipps Russell S. , Vasanji Amit , Lathia Justin D. , Li Meizhang , Bao Shideng , Rich Jeremy N. , Gladson Candece L.
Published: August 2014

Angiogenesis, the development of a neovasculature, is a key histologic feature of malignant glioma. It occurs through multiple mechanisms, including sprouting and migration of existing endothelial cells (ECs), and is driven by signals in the microenvironment. Glioma stem cells (GSC) inhabit a region near ECs (perivascular niche). Here we examined the hypothesis that GSCs interact directly with ECs and promote EC motility. In 4 of 5 glioblastoma biopsies we found that ∼40% of GSCs were localized within 25 μm of ECs, based on double-labeling for Sox2 and vWf. We then examined the effect of GSCs on EC motility when plated on a 2D laminin substrate, and found a significant increase in EC motility in the presence of GSCs as well as a direct interaction. In a 3D assay, EC migration was enhanced by GSC-conditioned media, but the physical presence and interaction between ECs and GSCs further increased EC migration through a laminin-coated, 3-μm pore filter, as compared to GSC-conditioned media. Integrin αvβ3 and L1CAM have been reported to be upregulated on ECs and GSCs, respectively, in GBM tumors, and integrin αvβ3 can bind the RGD-peptide in L1CAM. We examined the effect of blocking or downregulating integrin αvβ3 or L1CAM on EC interaction with GSCs. In a cell-cell binding assay, blocking or downregulating the integrin β3 subunit significantly inhibited EC-GSC direct contact, as did an RGD peptide and an antibody to L1CAM. Furthermore, downregulation of integrin β3 on ECs significantly reduced EC migration in the presence of GSCs in the transwell migration assay and reduced EC association with GSCs in mouse brain slices in organotypic culture, analyzed by 2-Photon-LSM. These data suggest GSCs promote angiogenesis in part by promoting EC motility, and that the EC-GSC direct interaction mediated by integrin αvβ3 on ECs and L1CAM on GSCs also stimulates EC motility.

Citation Format: Monica E. Burgett, Ping Huang, Russell S. Tipps, Amit Vasanji, Justin D. Lathia, Meizhang Li, Shideng Bao, Jeremy N. Rich, Candece L. Gladson. Glioma stem cells promote brain endothelial cell motility: a new mechanism for the direct interaction of glioma stem cells with endothelial cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3898. doi:10.1158/1538-7445.AM2013-3898
Abstract 3904: uPAR-regulated nuclear translocation of hand-1 mediates vascular radiosensitivity in medulloblastoma tumors.
Authors: Asuthkar Swapna , Velpula Kiran Kumar , Nalla Arun Kumar , Gogineni Venkateswara Rao , Dasari Venkata Ramesh , Gorantla Bharathi , Rao Jasti S.
Published: August 2014

Urokinase-type plasminogen activator (uPAR) is overexpressed in the tumor-stromal invasive microenvironment in many human cancers including medulloblastoma. The role of uPAR in tumor progression and angiogenesis has been well characterized. Most recently, in medulloblastoma cells, we showed that ionizing radiation (IR)-induced uPAR is a potent activator of cancer stem cell (CSC)-like properties and is associated with various transcription factors that are involved during embryonic development and cancer. In this study, we show that the uPAR protein is a cytoplasmic sequestration factor for a novel basic helix-loop-helix (bHLH) transcription factor, Hand-1. The Hand-1 protein plays an essential role in differentiation of trophoblast giant cells and cardiac morphogenesis, and yet its precise cellular function and its contributions to cancer remain mostly unknown. In the present study, we observed that Hand-1 protein is upregulated in uPAR shRNA-treated medulloblastoma cells and accompanies sustained cell growth and angiogenesis. Furthermore, IR-induced uPAR overexpression negatively regulates Hand-1 activity and results in the stabilization of angiogenesis promoting molecules, such as HIF-1 alpha. Finally, uPAR overexpression and its association with Hand-1 after IR treatment indicate that uPAR is capable of regulating Hand-1 and that uPAR has a role in the process of IR-induced tumor angiogenesis.

Citation Format: Swapna Asuthkar, Kiran Kumar Velpula, Arun Kumar Nalla, Venkateswara Rao Gogineni, Venkata Ramesh Dasari, Bharathi Gorantla, Jasti S. Rao. uPAR-regulated nuclear translocation of hand-1 mediates vascular radiosensitivity in medulloblastoma tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3904. doi:10.1158/1538-7445.AM2013-3904
Abstract 3925: Optical coherence tomography imaging in cancer research.
Authors: Vakoc Benjamin J. , Fukumura Dai , Padera Timothy P. , Jain Rakesh K. , Bouma Brett E.
Published: August 2014

Optical coherence tomography (OCT) is an established imaging technology with a greater than 10 year history in the clinic. Over the past 5 years, an increasing number of cancer investigators have employed OCT in preclinical studies and have found that its unique capabilities address some of the gaps left by existing intravital imaging approaches. However, because OCT is relatively new to the cancer research community, these capabilities and limitations are not broadly known. Therefore, to facilitate translation, we will summarize OCT to the cancer research community. This will include a description of how it works, what its capabilities are, and what its role in preclinical cancer research is now and what it is likely to become over the next decade.

The basic principles of OCT will be summarized to allow investigators to compare it with alternative imaging technologies and identify investigations where OCT is best suited. The imaging modes of OCT will be described and examples of use in cancer investigations will be given. The use of OCT for angiography at spatial and temporal scales not accessible to other technologies will be described. We will also present anatomical imaging of tumors using OCT and highlight its ability to assess tumor volume and to map tumor viability. Finally, the advantages of OCT-based label-free lymphangiography will be described and compared with existing approaches for lymphatic vessel imaging.

We will end our discussion by communicating briefly some of the innovations in OCT that are happening now in research and engineering laboratories and the new capabilities that such innovations are likely to enable. Through this presentation, we hope to establish dialogues to drive technology development toward answering critical problems in cancer research.

Citation Format: Benjamin J. Vakoc, Dai Fukumura, Timothy P. Padera, Rakesh K. Jain, Brett E. Bouma. Optical coherence tomography imaging in cancer research. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3925. doi:10.1158/1538-7445.AM2013-3925
Abstract 393: Clinical impact of pentraxin family expression on prognosis in pancreatic carcinoma.
Authors: Kondo Shunsuke , Ueno Hideki , Morizane Chigusa , Hosoi Hiroko , Koizumi Fumiaki , Tamura Kenji , Okusaka Takuji
Published: August 2014

Background: Inflammatory mediators may play decisive roles at different stages of tumor development. PTX3, a mediator that is a member of the pentraxin family, may be a biomarker for the early prognostic stratification of advanced pancreatic carcinoma patients.

Methods: Enzyme-linked immunosorbent assay and cellular migration assay were performed to determine PTX3 concentrations in pancreatic carcinoma cell lines (ASPC-1, BXPC-3, MIAPACA-2 and PANC-1) PTX3 levels and other inflammatory mediators (IL-1beta, IL-6, CCL2, CCL3, CCL4, CCL7, CXCL9, and MIF) were validated in a subgroup of pancreatic carcinoma patients. Univariate and multivariate Cox analysis were performed to investigate the extent of PTX3 production in the cell lines and the differences in outcomes between patients with high and low levels of inflammatory mediators, who had been divided into high and low groups based on mean values.

Results: In two cell lines (MIAPACA-2 and PANC-1), elevated PTX3 production was observed. A direct relationship was identified between migration activity and PTX3 level. In pancreatic carcinoma patients, median PFS was 76 days (95% confidence interval [CI], 43-109) in the PTX3high group and 150 days (95% CI, 131-197; P = 0.002) in the PTX3low group. Median OS was 117 days (95% CI, 82-152) in the PTX3high group and 357 days (95% CI, 239-475) in the PTX3low group (P < 0.001). Univariate and multivariate Cox model analysis of the pancreatic carcinoma clinical specimens revealed a strong correlation between pentraxin family member expression and PFS as well as OS. Examination of the relationship between PTX3 expression and the expression of other pro-inflammatory mediators indicated that PTX3 level is positively correlated with level of CRP, IL-6, and MIF.

Conclusion: Thus, pentraxin family member, especially PTX3, is a promising biomarker for the prognosis of pancreatic carcinoma, and its expression can promote pancreatic carcinoma progression.

Citation Format: Shunsuke Kondo, Hideki Ueno, Chigusa Morizane, Hiroko Hosoi, Fumiaki Koizumi, Kenji Tamura, Takuji Okusaka. Clinical impact of pentraxin family expression on prognosis in pancreatic carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 393. doi:10.1158/1538-7445.AM2013-393
Abstract 3935: AnnexinA2 regulates invasion and metastasis of pancreatic ductal adenocarcinoma through Semaphorin3d.
Authors: Foley Kelly , Zhou Donger , Xiao Qian , Rucki Agnieszka , Mo Guanglan , Huang Lanqing , Maitra Anirban , Jaffee Elizabeth , Zheng Lei
Published: August 2014

Pancreatic ductal adenocarcinoma (PDA), a devastating disease with a median survival of less than six months, is characterized by its high metastatic ability and a lack of effective therapies. The mechanisms involved in promoting metastasis of PDA have yet to be elucidated. We recently identified AnnexinA2 (AnxA2) as a PDA-associated antigen. Tyr23 phosphorylated AnxA2 promotes metastasis of PDA cells, while shRNA knockdown, knockout or therapeutic inhibition of AnxA2 prevents metastasis to the liver in a mouse model.

To identify downstream mediators of AnxA2 signaling, we evaluated gene expression differences in cell lines derived from the primary tumors of mice that express AnxA2 (KPC) and are metastatic or lack AnxA2 expression (KPCA) and are unable to metastasize. Using microarray analysis, we observed that expression of the ligand-receptor pair Semaphorin3d (Sema3d) and PlexinD1 (PlxnD1) was significantly down regulated in the non-metastatic KPCA cells. Sema3d and PlxnD1 were first identified as axon guidance genes. However, recently, axon guidance genes, including Sema3d and PlxnD1, were recently found by the International Cancer Genome Consortium to be among the cellular pathways that are most frequently altered by genetic mutations and amplification in human PDA (Biankin 2012). Therefore, we hypothesized that AnxA2 interacts with Sema3d and PlxnD1 in PDA to induce metastasis.

We confirmed by RT-PCR and immunofluorescence analysis in multiple independent tumors from KPC and KPCA mice that Sema3d and PlxnD1 expression is down regulated in the absence of AnxA2 expression. AnxA2, Sema3d and PlxnD1 colocalize at the cell surface during pancreatic tumor development when AnxA2 is expressed. Interestingly, co-immunoprecipitation analysis demonstrated a direct interaction between Sema3d and AnxA2 in PDA cells supporting our hypothesis that AnxA2 regulates the secretion of Sema3d from PDA cells to enable its paracrine interaction with PlxnD1 resulting in invasion of PDA cells.

Clinically, we have found that Sema3d expression correlates with increased lymphovascular invasion, increased perineural invasion, poor survival and increased metastatic potential in human PDA patients. Therefore, understanding the pathways that lead to metastasis of PDA will enable the development of more effective therapeutics and better clinical outcomes.

Citation Format: Kelly Foley, Donger Zhou, Qian Xiao, Agnieszka Rucki, Guanglan Mo, Lanqing Huang, Anirban Maitra, Elizabeth Jaffee, Lei Zheng. AnnexinA2 regulates invasion and metastasis of pancreatic ductal adenocarcinoma through Semaphorin3d. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3935. doi:10.1158/1538-7445.AM2013-3935
Abstract 3937: Collagen prolyl hydroxylases are essential for breast cancer metastasis.
Authors: Gilkes Daniele M. , Chaturvedi Pallavi , Bajpai Saumendra , Hubbi Maimon , Wirtz Denis , Semenza Gregg
Published: August 2014

Metastasis is the leading cause of death among patients who have breast cancer. Understanding the role of the extracellular matrix in the metastatic process may lead to the development of improved therapies to treat cancer patients. Intratumoral hypoxia is found in the majority of breast cancers and is associated with an increased risk of metastasis and patient mortality. Here we demonstrate that hypoxia-inducible factor 1 activates the transcription of genes encoding collagen prolyl hydroxylases that are critical for collagen deposition by breast cancer cells. We show that expression of collagen prolyl hydroxylases promotes cancer cell alignment along collagen fibers, resulting in enhanced cancer cell invasion and metastasis to lymph nodes and lungs. We establish the prognostic significance of collagen prolyl hydroxylase mRNA expression in human breast cancer biopsies and evaluate the novel use of ethyl 3,4-dihydroxybenzoate, a prolyl hydroxylase inhibitor, which decreases tumor fibrosis and metastasis in a mouse model of breast cancer.

Citation Format: Daniele M. Gilkes, Pallavi Chaturvedi, Saumendra Bajpai, Maimon Hubbi, Denis Wirtz, Gregg Semenza. Collagen prolyl hydroxylases are essential for breast cancer metastasis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3937. doi:10.1158/1538-7445.AM2013-3937
Abstract 3949: Hypoxia induces the conversion of tumor initiating cells from stationary to invasive by promoting cell surface trafficking of matrix metalloproteinase-14.
Authors: Cao Jian
Published: August 2014

Emerging experimental and clinical evidence has demonstrated that quiescent, stationary tumor initiating cells (TICs) can become invasive and metastatic. The mechanism underlying the conversion of TICs from stationary to invasive remains to be characterized. A better understanding of this conversion is crucial for the design of inhibitors aimed at preventing cancer metastasis. We report here for the first time that hypoxia, a low oxygen environment that often occurs in solid tumors, induces translocation of the membrane anchored protease, matrix metalloproteinase-14 (MMP-14) from the cytoplasmic storage pool to the cell surface resulting in conversion of stationary TICs to invasive TICs. Exposure of minimally invasive breast cancer TICs, SK-3rd, that displays CD44high/CD24low with high mammosphere-forming and tumorigenic capacities, to hypoxic conditions (1% O2) results in markedly enhanced cell invasive ability. This phenotypic change is also observed in SK-3rd TICs in the presence of cobalt chloride (CoCl2), an agent that mimics hypoxia. Given the role of MMP-14 in cancer invasion/metastasis, we explored a possible involvement of MMP-14 in hypoxia-induced TIC invasion. Silencing of MMP-14 by a shRNA approach resulted in diminution of hypoxia-induced cell invasion in vitro and metastasis in vivo. When SK-3rd cells were cultured under hypoxic conditions, MMP-14 redistributed from cytoplasmic storage pools to the cell surface of TICs, which coincides with the increased cell invasion. In addition, CD44, a cancer stem-like cell marker, inversely correlated with increased cell surface MMP-14. Interestingly, cell surface MMP-14 gradually disappeared when the hypoxia-treated cells were switched to normoxia, suggesting the plasticity of TICs in response to oxygen content. Furthermore, we dissected the pathways leading to upregulated MMP-14 in cytoplasmic storage pools under normoxic conditions. We demonstrated a cascade involving Twist1-miR10b-HoxD10 leading to enhanced MMP-14 expression in SK-3rd TICs. These observations suggest that MMP-14 is a key molecule capable of executing conversion of stationary TICs to invasive TICs under hypoxic conditions and thereby controlling metastasis.

Citation Format: Jian Cao. Hypoxia induces the conversion of tumor initiating cells from stationary to invasive by promoting cell surface trafficking of matrix metalloproteinase-14. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3949. doi:10.1158/1538-7445.AM2013-3949