bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4009: Comparative microRNA profiling of prostate carcinomas with increasing tumor stage by deep-sequencing
Authors: Nolte Elke , Hart Martin , Wach Sven , Szczyrba Jaroslaw , Taubert Helge , Rau Tilman , Hartmann Arndt , Grässer Friedrich A. , Wullich Bernd
Published: August 2015

Background: MicroRNAs (miRNAs) are short, non-coding RNAs that post-transcriptionally regulate gene expression and play an important role in tumorigenesis. Previous deep-sequencing identified the microRNA profile of prostate carcinoma (PCa) vs. non-malignant prostate tissue. As an extension of our recent analyses, we now compared organ-confined prostate tumors (pT2), tumors with extra capsular growth (pT3, pN0), and lymph node metastasizing (pN1) PCa relative to corresponding non-malignant prostate tissue and healthy prostate tissue.

Material and Methods: We performed comparative miRNA expression profiling using deep sequencing of cDNA libraries. Potential target genes of deregulated miRNAs were identified by database analysis and subsequently validated in vitro using reporter gene constructs. miRNA and mRNA expression was determined using quantitative real-time PCR.

Results: The miRNA expression from the healthy and non-malignant tissue is consistent with our previous findings, indicating a high fidelity of the method employed. Some miRNAs (miR-375, miR-148a, miR-141 and miR-21) are expressed at increasing levels with increasing tumor stage, while other miRNAs (miR-200b/c, miR-25, miR-17 or miR-93) are up-regulated only from organ-confined tumors (pT2) to tumors with extra-capsular growth (pT3) but then expressed at lower levels after the transition to lymph node metastasizing tumors (pN0 vs. pN1). The same trend was seen in qRT-PCR analysis but due to the limited sample number, no significance was reached. Interestingly, miR-375, found to be up-regulated in tumor vs. normal tissue, and miR-15a, found induced by sequencing in T2 and T3 tumors but not in lymph node metastasizing tumors, target the phosphatases PHLPP1 and PHLPP2, respectively. PHLPP1 and PHLPP2 dephosphorylate members of the AKT-family of signal transducers thereby inhibiting cell growth. Co-expression of miR-15a and miR-375 resulted in down-regulation of PHLPP1/2 and a strongly increased PCa cell growth.

Conclusion: miRNA profiling of pT2 (pN0), pT3 (pN0), and lymph node metastasizing prostate carcinoma (pN1) by deep sequencing revealed clearly changed miRNA levels.

Citation Format: Elke Nolte, Martin Hart, Sven Wach, Jaroslaw Szczyrba, Helge Taubert, Tilman Rau, Arndt Hartmann, Friedrich A. Grässer, Bernd Wullich. Comparative microRNA profiling of prostate carcinomas with increasing tumor stage by deep-sequencing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4009. doi:10.1158/1538-7445.AM2015-4009
Abstract 4032: Functional analysis of exosomes derived from human esophageal squamous cell carcinoma
Authors: Matsumoto Yasunori , Kano Masayuki , Akutsu Yasunori , Nishimori Takanori , Hanari Naoyuki , Hoshino Isamu , Murakami Kentaro , Toyozumi Takeshi , Suito Hiroshi , Takahashi Masahiko , Sekino Nobufumi , Nabeya Yoshihiro , Matsubara Hisahiro
Published: August 2015

Exosomes are small membrane vesicles (30-100 nm) released by many types of cells including cancer cells, and are thought to play an important role in cell-to-cell communications. Exosomes contain lipids, proteins, and nucleic acids such as mRNAs or microRNAs, they are tought to have relationship with tumor growth or metastasis. Our laboratory was previously reported the level of microRNA-1246, expressed mainly in tumor tissues of esophageal squamous cell carcinoma (ESCC), was significantly high in exosomes extracted from the serum of ESCC patients. However, the exosome dynamics and transportation between cancer cells and the other cells are still unclear. In this study, we assessed the cell-to-cell communication via exosomes with fluorescent imaging and analyzed the effect of cancer cell derived exosome to tumor progression.

In order to image and analyze the movement of cancer cell-derived exosomes, green fluorescent protein (GFP)-tagged CD63, which is a general marker of exosomes, was expressed in human esophageal squamous cancer cell line TE2. Exosomes were extracted from supernatant with ultracentrifuge method. Extracted GFP-tagged exosomes were added to the other cells to obtain the images of transportation. The effects of exosomes to tumor progression were also assessed with cell proliferation assay, invasion oassay, and migration assay. Our data indicates that cancer cell derived exosomes affect tumor progression.

Citation Format: Yasunori Matsumoto, Masayuki Kano, Yasunori Akutsu, Takanori Nishimori, Naoyuki Hanari, Isamu Hoshino, Kentaro Murakami, Takeshi Toyozumi, Hiroshi Suito, Masahiko Takahashi, Nobufumi Sekino, Yoshihiro Nabeya, Hisahiro Matsubara. Functional analysis of exosomes derived from human esophageal squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4032. doi:10.1158/1538-7445.AM2015-4032
Abstract 4034: Macrophage-dependent activation of a non-canonical NOTCH-RhoA signaling pathway regulates tumor cell intravasation
Authors: Bravo Cordero Jose Javier , Roh-Johnson Minna , Pignatelli Jeanine , Oktay Maja , Hodgson Louis , Condeelis John
Published: August 2015

Intravital imaging of rodent mammary tumors has shown that direct contacts between a Mena expressing tumor cell, a perivascular macrophage and an endothelial cell forms a microanatomical structure named TMEM (Tumor Microenvironment of Metastasis). Clinical studies have shown that the number of TMEM is correlated with an increased risk of developing distant metastasis in breast cancer patients. While TMEM is an excellent prognostic marker for predicting metastasis the mechanisms of TMEM assembly and function are not understood. Recently, we showed that heterotypic cell contact between tumor cells and macrophages induces the formation of invadopodia in tumor cells, invasive structures necessary for matrix degradation and required for tumor cell intravasation. Using high resolution FRET imaging, we further found that macrophage-induced invadopodium formation is dependent on RhoA activation in the tumor cell. However, what remained to be determined was the signaling pathway that regulated this heterotypic cell contact-mediated phenomenon. We explored the role of NOTCH, a known receptor involved in heterotypic cell contact signaling, in stimulating macrophage-dependent tumor cell invadopodium formation and transendothelial migration. We found that, in the absence of Notch signaling, macrophage-induced invadopodium formation and tumor cell intravasation are abolished. Moreover, RhoA is no longer activated in tumor cells upon macrophage contact when Notch signaling is perturbed. These results show that Notch signaling regulates heterotypic cell contact mediated invadopodium formation through RhoA activation, and reveals a novel non-canonical NOTCH/RhoA pathway as a molecular target to prevent TMEM function and therefore metastasis.

Citation Format: Jose Javier Bravo Cordero, Minna Roh-Johnson, Jeanine Pignatelli, Maja Oktay, Louis Hodgson, John Condeelis. Macrophage-dependent activation of a non-canonical NOTCH-RhoA signaling pathway regulates tumor cell intravasation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4034. doi:10.1158/1538-7445.AM2015-4034
Abstract 4035: Extracellular secreted Ref-1 triggers triple negative breast cancer cells apoptosis
Authors: Lee Yu Ran , Kim Ki Mo , Joo Hee Kyoung , Jeon Byeong Hwa , Choi Sunga
Published: August 2015

Triple-negative breast cancers (TNBCs) are most aggressive type and resistant to standard chemotherapy. There is a high demand for developing approaches to treat TNBCs. Recently, some agents inducing post-translational modification (PTM) were suggested to treat therapeutic-resistant cancer cells. We previously discovered that trichostatin A (TSA), histone deacetylase inhibitor (HDACi), - mediated acetylation induced extracellular secretion of the acetylated apurinic apyrimidinic endonuclease 1/redox factor-1 (Ac-APE1/Ref-1). This is the first report showing that the Ac-APE1/Ref-1 is not merely a secreted protein but also a signal molecule that plays a critical role in the apoptotic cell death pathway in TNBC. To study the functional significance of the secreted Ac-APE1/Ref-1, we induced PTM, hyperacetylation of intracellular proteins with co-treatment of acetylsalicylic acid (ASA) and TSA in MDA-MB-231 cells. In response to hyperacetylation, the secretion of Ac-APE1/Ref-1 in a form of exosome-like vesicles was observed in both electron microscopy and confocal image analysis. Accordingly, it was caused a statistically significant inhibition of cell viability and induction of apoptotic DNA fragmentation, leading activation of p38 mitogen-activated protein kinase and apoptotic cell death. Blockade of Ac-APE1/Ref-1 thoroughly recovered from the cell death effects. The hyperacetylation-mediated apoptotic cell death was comparably efficient in treatment of another TNBC cell lines, BT549, and MDA-MB-468. Keeping in view secreted APE1/Ref-1 following acetylation - causing apoptotic cell death we suggest that PTM qualifies as an excellent candidate to be exploited for therapeutic agents to treat TNBCs. For future clinical investigations of TNBCs, this study offers a novel paradigm, which stimulation of apoptotic pathway by action of PTM-induced auto- or paracrine signaling molecules is prior to treatment of standard chemotherapeutic resistant cancer cells.

Citation Format: Yu Ran Lee, Ki Mo Kim, Hee Kyoung Joo, Byeong Hwa Jeon, Sunga Choi. Extracellular secreted Ref-1 triggers triple negative breast cancer cells apoptosis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4035. doi:10.1158/1538-7445.AM2015-4035
Abstract 404: MK2461 suppress progression of pancreatic cancer disrupting interaction between pancreatic cancer cells and stellate cells
Authors: Inoue Koetsu , Ohtsuka Hideo , Motoi Fuyuhiko , Douchi Daisuke , Kawasaki Shuhei , Kawaguchi Kei , Masuda Kunihiro , Fukase Koji , Ohnuma Shinobu , Naitoh Takeshi , Katayose Yu , Egawa Shinichi , Unno Michiaki
Published: August 2015

Pancreatic cancer is characterized excessive desmoplasia, which occupies 80% of pancreatic cancer tissue and mainly consists of pancreatic stellate cells (PSCs). Desmoplasia has been shown to play an important role in the progression, invasion, and metastasis of pancreatic cancer and has been implicated in the development of resistance to chemotherapy and radiotherapy. Growth factors are one of the components modulating the interactions between pancreatic cancer cells (PCCs) and PSCs. Therefore, blocking growth factor signaling could suppress cancer progression. In this study, we profiled the expression of receptor tyrosine kinases (RTKs) with quantitative targeted absolute proteomics using liquid chromatography tandem mass spectrometry (LC-MS/MS) in PCCs and PSCs. In PCCs, epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor (MET) levels were elevated compared with those in PSCs. In PSCs, platelet-derived growth factor receptor beta (PDGFRβ) and MET were upregulated compared with other RTKs. Conditioned medium from PSCs promoted the proliferation of PCCs, and vice versa. In addition, MK2461, a multikinase inhibitor targeting MET and PDGFRβ, suppressed the effects of conditioned medium on PCCs and PSCs. In addition, MK2461 significantly inhibited tumor growth in mice xenograft model. In conclusion, PDGFRβ and MET may play a critical role in the interaction between PCCs and PSCs. Moreover, MK2461 may have therapeutic potential in the treatment of pancreatic cancer by targeting the interaction between PCCs and PSCs.

Citation Format: Koetsu Inoue, Hideo Ohtsuka, Fuyuhiko Motoi, Daisuke Douchi, Shuhei Kawasaki, Kei Kawaguchi, Kunihiro Masuda, Koji Fukase, Shinobu Ohnuma, Takeshi Naitoh, Yu Katayose, Shinichi Egawa, Michiaki Unno. MK2461 suppress progression of pancreatic cancer disrupting interaction between pancreatic cancer cells and stellate cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 404. doi:10.1158/1538-7445.AM2015-404
Abstract 4084: Transient SNAIL1 expression is necessary for metastatic competence in breast cancer
Authors: Tran Hung D. , Kim Michael , Luitel Krishna , Zhang Kun , Longmore Gregory , Tran David D.
Published: August 2015

SNAIL1 has been suggested to regulate breast cancer metastasis based on analyses of human breast tumor transcriptomes and experiments using cancer cell lines and xenografts. However, in vivo genetic experimental support for a role for SNAIL1 in breast cancer metastasis that develops in an immunocompetent tumor microenvironment has not been determined. To address this question, we created a genetic SNAIL1 model by coupling an endogenous SNAIL1 reporter with an inducible SNAIL1 transgene. Using multiple genetic models of breast cancer, we demonstrated that endogenous SNAIL1 expression was restricted to primary tumors that ultimately disseminate. SNAIL1 gene deletion either during the premalignant phase or after primary tumors have reached a palpable size blunted metastasis, indicating that late metastasis was the main driver of metastasis and that this was dependent on SNAIL1. Importantly, SNAIL1 expression during breast cancer metastasis was transient and forced transient, but not continuous, SNAIL1 expression in breast tumors was sufficient to increase metastasis.

Citation Format: Hung D. Tran, Michael Kim, Krishna Luitel, Kun Zhang, Gregory Longmore, David D. Tran. Transient SNAIL1 expression is necessary for metastatic competence in breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4084. doi:10.1158/1538-7445.AM2015-4084
Abstract 4085: Depletion of Runx1 promotes epithelial to mesenchymal transition in breast cancer
Authors: Hong Deli , Lian Jane , Stein Janet , Stein Gary
Published: August 2015

Runx1 is an essential transcription factor for definitive hematopoiesis and Runx1 translocations are well established as the causative factor of various leukemias. Recent studies have identified Runx1 as one of the most frequently mutated genes in breast cancer patients, and also that Runx1 expression levels are decreased in high-grade primary breast tumors compared to low/mid-grade tumors. The recognition of Runx1 expression in epithelial lining cells of glands suggests that Runx1 functions in maintaining the integrity of that cell layer. Therefore we examined the role of Runx1 during cancer progression. We found that Runx1 is highly expressed in immortalized human mammary epithelial cells (MCF10A) compared with breast cancer epithelial cells (MCF7) and invasive breast cancer cells (MDA-MB-231). To test the hypothesis that Runx1 contributes to the epithelial to mesenchymal transition (EMT) in breast cancer, the EMT was induced in MCF10A cells which resulted in a 50% decrease of Runx1 mRNA, but complete loss of Runx1 protein. This finding suggests a role for Runx1 as an EMT suppressor. To further address the relationship between Runx1 and EMT, Runx1 depleted stable cell lines were generated in both MCF10A and MCF7 cells. We observed that Runx1 knockdown (90%) in MCF10A cells leads to a dramatic change in their morphology towards a mesenchymal state, with a concomitant decrease in expression of the epithelial marker E-cadherin. In addition Runx1 depletion in MCF7 cells directly induced EMT state by repression of E-cadherin, as well as increased expression of mesenchymal genes, including vimentin, N-cadherin and fibronectin. Our results highlight a crucial role for Runx1 in preventing epithelial to mesenchymal transition and tumor progression in breast cancer. This Runx1 mediated mechanism points to novel intervention strategies for early stage breast cancer.

Citation Format: Deli Hong, Jane Lian, Janet Stein, Gary Stein. Depletion of Runx1 promotes epithelial to mesenchymal transition in breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4085. doi:10.1158/1538-7445.AM2015-4085
Abstract 4128: Extensive morphological heterogeneity of circulating and selected high metastatic variants and differential chemosensitivity of human prostate cancer in mouse models
Authors: Zhang Lei , Wu Chengyu , Hoffman Robert. M
Published: August 2015

Circulating PC-3-GFP human prostate cancer cells were collected from the blood from nude mice orthotopically implanted with PC-3-GFP. In parallel experiments, a highly metastatic PC-3-GFP cell line was developed by injecting parental PC-3-GFP to the footpad of nude mice which metastasized to inguinal lymph nodes from which the cancer cells were collected and were re-injected to the footpad. After 6 cycles, the PC-3-GFP cells collected from inguinal lymph nodes (PC-3-GFP-LN) and were injected to the footpad. PC-3-GFP-LN showed 100% metastasis to major lymph nodes (popliteal, inguinal, axillary, and cervical), and 100% metastasis to bone and lung. PC-3-GFP-CTC and PC-3-GFP-LN were cultured in vitro, and their morphology and chemo-sensitivity were comparing to the parental PC-3-GFP cells. Highly pleomorphic subpopulations of prostate cancer cells were identified in PC-3-GFP-CTC and PC-3-GFP-LN. The different cell subtypes included characteristic high malignancy features such as high nucleus to cytoplasmic (N/C) ratios and giant cell formation as a result of abnormal mitoses. Variations in growth rate, plating efficiency, size, shape, and number of the nuclei and nucleoli were observed. The results suggest extensive tumor heterogeneity in both sublines. The giant cells, contain multiple nuclei, cannot survive by themselves and need small cells around them in order to proliferate. However, giant cells also can produce smaller cells, which divide further. The ratio of giant cells/total cells were significantly higher in PC-3-GFP-LN compared to parental cells. However, the morphology of cells from lung metastasis and bone metastasis showed lung and bone epithelial features such as squamous on shape and tightly packed together. PC-3-GFP-CTC had similar sensitivity to both cisplatinum and docetaxel when compared to PC-3-parental cells. PC-3-GFP-LN was resistant. In contrast, PC-3-GFP-LN was sensitive to the traditional Chinese medicine (TCM) herbal mixture LQ similar to the parental cells. These results demonstrate that circulating tumor cells (CTCs) and selected high metastatic variants can be extensively heterogeneous and have different chemosensitivity profiles, suggesting new approaches to treatment such as with TCM.

Citation Format: Lei Zhang, Chengyu Wu, Robert. M Hoffman. Extensive morphological heterogeneity of circulating and selected high metastatic variants and differential chemosensitivity of human prostate cancer in mouse models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4128. doi:10.1158/1538-7445.AM2015-4128
Abstract 4131: MEK inhibitor diminishes nasopharyngeal carcinoma (NPC) cell growth and NPC-induced osteoclastogenesis via modulating CCL2 and CXCL16 expression
Authors: Zhu Yu , Song Qiong , Lu Yi
Published: August 2015

Nasopharyngeal carcinoma (NPC) is a common malignant cancer in Guangxi, as well as in southern China and Southeast Asia. NPC preferentially metastasizes to bone in advanced patients, major factor resulting in high mortality. The molecular basis of NPC development and cancer-induced bone lesions are still unclear. We and others have reported that chemokine, like CCL2, CXCL16, CXCL12, and their corresponding receptors played important roles in cancer bone metastasis. In this study, we firstly investigated that CCR2 and CXCR6 expressions in clinical specimens. Then, we measured CCL2, CCR2, CXCL16 and CXCR6 expression levels by RT-PCR, ELISA and western blot in NPC cell lines. Further, we used MEK inhibitor, U0126, to treat the NPC cell lines and found CCL2 and CXCL16 expressions significantly decreased. The NPC cell lines proliferation were inhibited by U0126 in dose- and time-dependently manner. Finally, we collected conditioned medium (CM) from the NPC cell lines absent or present U0126 treatments, to treat mouse bone marrow non-adherent cells in order to detect NPC CM-induced multinucleated osteoclast-like formation. The CMs collected from U0126 treated NPC cells were dramatically diminished the numbers of multinucleated osteoclasts. These results indicate that U0126 inhibits NPC cell growth and diminishes cancer-induced osteoclast formation partially through modulating the expression of CCL2 and CXCL16. Thus this study may provide a novel therapeutic strategy for advanced NPC patients, especially for treatment of NPC bone metastasis.

Citation Format: Yu Zhu, Qiong Song, Yi Lu. MEK inhibitor diminishes nasopharyngeal carcinoma (NPC) cell growth and NPC-induced osteoclastogenesis via modulating CCL2 and CXCL16 expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4131. doi:10.1158/1538-7445.AM2015-4131
Abstract 4133: ODM-203, a novel, selective and balanced FGFR and VEGFR inhibitor with strong activity on primary and metastatic tumor growth in FGFR-dependent and angiogenic cancer models
Authors: Holmström Tim , Moilanen Anu , Björkman Mari , Linnanen Tero , Wohlfahrt Gerd , Karlsson Stefan , Oksala Riikka , Korjamo Timo , Samajdar Susanta , Rajagopalan Srinivasan , Chelur Shekar , Narayan Kishore , Ramachandra Raghuveer , Anthony Thomas , DS Samiulla , Ramachandra Murali , Kallio Pekka
Published: August 2015

Genomic alterations in fibroblast growth factor receptors (FGFR) and upregulation of vascular endothelial growth factor receptors (VEGFR) are often found in the same cancer types such as gastric, lung and breast and these alterations correlate with patient survival and disease progression. In addition, both FGFR and VEGFR signaling promote tumor angiogenesis. Activation of FGFR signaling has also been described to function as a compensatory angiogenic signal following development of resistance to VEGF inhibition.

ODM-203 is a novel and selective inhibitor of the FGFR and VEGFR families exhibiting equal potency to both receptors both in vitro and in vivo. In recombinant kinase assays, ODM-203 inhibits both the FGFR and VEGFR family kinases in the low nanomolar range (IC50 6-35 nM). In cells, ODM-203 suppresses FGFR and VEGFR signaling with similar potency. It also inhibits VEGF-induced endothelial cell (HUVEC) proliferation (IC50 33 nM) with similar potency as it inhibits proliferation in FGFR-dependent cell lines (IC50 50-150 nM). In vivo, ODM-203 shows strong anti-tumor activity in FGFR-dependent xenograft models and in angiogenic xenograft models with similar well tolerated doses. In addition ODM-203 inhibits metastatic tumor growth in angiogenic xenograft models. Lack of activity in a FGFR and VEGFR independent tumor models further supports the selective mode of action of ODM-203. Oral administration of ODM-203 was well tolerated with only FGFR or VEGFR inhibition related physiological events observed in vivo in rat and dog.

Based on our findings, ODM-203 has a unique profile with equal potency for both FGFR and VEGFR kinase families which correlates with with strong activity on primary and metastatic tumor growth in FGFR-dependent and angiogenic tumor models. Therefore, a clinical trial with ODM-203 is ongoing in patients with solid tumors (NCT 02264418).

Citation Format: Tim Holmström, Anu Moilanen, Mari Björkman, Tero Linnanen, Gerd Wohlfahrt, Stefan Karlsson, Riikka Oksala, Timo Korjamo, Susanta Samajdar, Srinivasan Rajagopalan, Shekar Chelur, Kishore Narayan, Raghuveer Ramachandra, Thomas Anthony, Samiulla DS, Murali Ramachandra, Pekka Kallio. ODM-203, a novel, selective and balanced FGFR and VEGFR inhibitor with strong activity on primary and metastatic tumor growth in FGFR-dependent and angiogenic cancer models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4133. doi:10.1158/1538-7445.AM2015-4133
Abstract 4146: Overexpression of C5a receptor becomes risk factor in gastric cancer
Authors: Kaida Takayoshi , Nitta Hidetoshi , Arima Kota , Takeyama Hideaki , Higashi Takaaki , Taki Katunobu , Hayashi Hiromitsu , Hashimoto Daisuke , Chikamoto Akira , Ishiko Takatoshi , Beppu Toru , Baba Hideo
Published: August 2015

Background: C5a is one of the important factors produced on complement pathway and a strong chemokine. There is emerging evidence that C5a is generated in the cancer microenvironment and C5a receptor (C5aR) is expressed in various cancer cells and associated with invasive ability in several cancer cells. However, the role of C5aR in gastric cancer (GC) is still mostly unknown.

Aim: To investigate the clinical significance of C5aR expression in human GC.

Method: 168 patients underwent gastrectomy after the diagnosis of gastric cancer from January 1st 2001 to December 31th 2009. C5aR-expression of GC cells was investigated in all 168 patients and was analyzed in regard to an association with clinicopathological factors and outcomes in the patients. Additionally, in vitro experiments were used to examine the proliferation and invasion of GC cells by using the GC cell lines that had high-expression of C5aR.

Result: High-expression of C5aR was found in 55 patients (32.7%). High-expression of C5aR was significantly related to depth of invasion, stage, lymphatic and vascular invasion. Among the 146 patients with a tumor stage lower than stageIV, the 5-years recurrence-free and overall survival rates of patients in the high-expression of C5aR group were significantly lower than those in the low-expression of C5aR group (91.0% vs 52.4%, p = 0.007 and 87.2% vs 59.1%, p = 0.003, respectively). The high-expression of C5aR group had more frequently distant metastasis than the low-expression of C5aR group during the follow-up time, presenting itself mainly in liver metastasis and peritoneal dissemination (p = 0.020 and p = 0.026). We found that the gastric cancer cell lines MKN1 and MKN7 had the high-expression of C5aR with western blotting in vitro. Moreover, matrigel chamber assay revealed that 10 nM of recombinant C5a enhances cell invasiveness of MKN1 by 2.55-fold and the cell invasiveness of MKN7 by 2.90-fold, but such an enhancement by C5a was not observed in control cells.

Conclusions: The gastric cancer patients with a high expression of C5aR were associated with poor prognosis, and the invasive ability of gastric cancer cell lines highly expressing of C5aR were promoted by C5a. Thus, targeting this C5a-C5aR axis might provide a useful therapeutic option for gastric cancer treatment.

Citation Format: Takayoshi Kaida, Hidetoshi Nitta, Kota Arima, Hideaki Takeyama, Takaaki Higashi, Katunobu Taki, Hiromitsu Hayashi, Daisuke Hashimoto, Akira Chikamoto, Takatoshi Ishiko, Toru Beppu, Hideo Baba. Overexpression of C5a receptor becomes risk factor in gastric cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4146. doi:10.1158/1538-7445.AM2015-4146
Abstract 4147: Synergistic effect of combined EGFR and Src inhibitor in colon cancer cell migration
Authors: Wu Yi-Che , Wei Wan-Chen , Chao Wei-Ting
Published: August 2015

EGFR signaling plays the important role in cancer cell proliferation and cell migration. Cetuximab which is the antibody against EGFR was approved for colon cancer treatment. Recently, Src inhibitor, dasatinib has been demonstrated to improve drug sensitivity to cetuximab in KRAS mutant colorectal cancer cell growth. Src is a non-receptor tyrosine kinase which is necessary for full activation of EGFR and also functions in focal adhesion kinase activity in cell migration, therefore, the combination of cetuximab and dasatinib may has effect on colon cell migration. In this study, cetuximab and dasatinib were applied to SW480 colon cell which is KRAS mutant cell line. Cell viability and cell migration were analyzed, localization of Src and focal adhesion kinase (FAK) were observed by confocal microscopy. In the result, Combination of 5μM cetuximab and 40 nM dasatinib treatment resulted in synergistic effect on cell viability. However, when cetuximab combined with 25nM dasatinib was applied in trans-well migration assay, the synergistic inhibition on cell migration was observed. The immunofluorescence confocal microscopic image demonstrated cetuximab treatment induced phospho-Src accumulation on the cell membrane and colocalized with phospho-FAK. When the combination of 5μM cetuximab and 25 uM dasatinib were applied in the cell, the membrane accumulation of phospho-Src and Phospho-FAK were decreased. Taken together, cetuximab combined with dasatinib has synergistic efficacy in colon cancer cell proliferation, however, cetuximab combined with lower concentration of dasatinib (25nM) suppressed cell migration by inhibited FAK membrane localization through Src inactivation.

Citation Format: Yi-Che Wu, Wan-Chen Wei, Wei-Ting Chao. Synergistic effect of combined EGFR and Src inhibitor in colon cancer cell migration. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4147. doi:10.1158/1538-7445.AM2015-4147
Abstract 416: Carbon monoxide targets Notch1 and MAPK-ERK1/2 signaling pathways to block growth of lung carcinoma
Authors: Nemeth Zsuzsanna , Csizmadia Eva , Vikstrom Lisa , Li Mailin , Bisht Kavita , Feizi Alborz , Gallo David , Otterbein Leo , Fillinger Janos , Dome Balazs , B. Costa Daniel , Wegiel Barbara
Published: August 2015

We have recently shown that inhalation of low, non-toxic doses of carbon monoxide (CO) is a potential therapeutic adjuvant treatment for lung cancer and prostate cancer (Cancer Research, 2013, Wegiel B et al). We found that CO doses of 100 ppm daily or twice per week were the most effective in suppressing A549 xenograft tumors growth, Ki67 expression and increased cleaved caspase-3 staining. Additionally, we found that inhibition of xenografts and KRAS-driven lung tumor growth in response to CO was associated with increased expression of M1 macrophage markers (CD86, CD197, RANTES) and decreased M2 markers (MMR) and HO-1 blockage. We demonstrated a direct interaction between HO-1 and Notch1. Further, that CO promotes Notch 1 cleavage as well as phosphorylation of ERK1/2 in the stroma cells, which are the markers of M1 polarization. Moreover, we demonstrated a significant correlation between low HO-1 expression and high p-ERK ½ expression in primary human lung cancers (n = 30 patients). In summary, we describe a mechanism by which CO affects cancer growth through regulation of macrophage polarization via Notch1 and Erk1/2 pathways.

Citation Format: Zsuzsanna Nemeth, Eva Csizmadia, Lisa Vikstrom, Mailin Li, Kavita Bisht, Alborz Feizi, David Gallo, Leo Otterbein, Janos Fillinger, Balazs Dome, Daniel B. Costa, Barbara Wegiel. Carbon monoxide targets Notch1 and MAPK-ERK1/2 signaling pathways to block growth of lung carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 416. doi:10.1158/1538-7445.AM2015-416
Abstract 4180: TGF-beta promotes angiogenesis in an RB-deficient, Kras-driven mouse model of pancreatic cancer
Authors: Gore Jesse , Craven Kelly E. , Wilson Julie L. , Korc Murray
Published: August 2015

Pancreatic ductal adenocarcinoma (PDAC) is a deadly disease that is projected to become the 2nd leading cause of cancer deaths by 2030. PDACs are associated with a high frequency of KRAS mutations (95%) and overexpression of many pro-angiogenic cytokines and growth factors. Using a genetically engineered mouse model (GEMM) that we established in which oncogenic Kras is combined with loss of RB (KRC), we have determined that PDACs arising in these mice (mPDACs) harbor many endothelial cells (ECs) and sinusoidal-like vessels that have blood flow, as determined by intravital confocal microscopy. Array analysis of pancreatic cancer cells (PCCs) derived from KRC tumors revealed gene expression profiles that correlate with active TGF-β signaling pathways, as determined by gene set enrichment analysis (GSEA), and include increased expression of multiple pro-angiogenic genes. In silico analysis indicated that many of these pro-angiogenic genes were TGF-β targets, and inhibition of the type I TGF-β receptor (TβRI) with SB505124 confirmed that TGF-βs drive pro-angiogenic gene expression in KRC PCCs. Moreover, TGF-β increased the levels of pro-angiogenic cytokines in conditioned media (CM) prepared from KRC PCCs, which when added to ECs, activated canonical TGF-β signaling pathways and stimulated EC proliferation and migration. Although SB505124 blocked TGF-β pathway activation in ECs, it failed to suppress CM-enhanced EC proliferation and migration. By contrast, SB505124 attenuated tumor angiogenesis, growth and metastasis in a syngeneic orthotopic mouse model using KRC PCCs. Taken together, these results suggest that TGF-β promotes angiogenesis in an indirect manner, by up-regulating pro-angiogenic factors in PCCs that act on ECs in a paracrine manner. Therefore, targeting TGF-β in PDAC could be a beneficial anti-angiogenic strategy.

Citation Format: Jesse Gore, Kelly E. Craven, Julie L. Wilson, Murray Korc. TGF-beta promotes angiogenesis in an RB-deficient, Kras-driven mouse model of pancreatic cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4180. doi:10.1158/1538-7445.AM2015-4180
Abstract 4181: Chronic stress and beta adrenergic signaling promote angiogenesis and prostate cancer progression through suppressing the expression of Thrombospondin 1
Authors: Hulsurkar Mohit M. , Sang Meixiang , Song Haiping , Li Wenliang
Published: August 2015

Several recent studies have revealed that the activation of beta adrenergic signaling induced by chronic biobehavioral stress promotes the progression of ovarian, breast and prostate cancers. Conversely, the use of hypertension medicine, beta blockers (beta adrenergic antagonists), has been found to be associated with better prognosis in several human cancers. Although the beta adrenergic signaling has been shown to be activated by chronic stress and responsible for promoting cancers, the mechanisms through which beta adrenergic signaling promotes tumor angiogenesis and cancer progression are still unclear. We found that chronic stress and activation of beta adrenergic signaling suppressed the expression of Thrombospondin 1 (TSP1) in prostate cancer cells in vitro, as well as in mouse xenograft tumors in vivo, which in turn promoted tumor angiogenesis and cancer progression. TSP1 was the first endogenous angiogenesis inhibitor to be discovered and its down-regulation is associated with increased angiogenesis. We recently demonstrated that G-protein coupled receptor kinase 3 (GRK3, ADRBK2) promotes angiogenesis and prostate cancer progression through suppressing TSP1. Interestingly, GRK3 expression is up-regulated by chronic stress and activation of beta adrenergic signaling, suggesting that GRK3 acts as a mediator for chronic stress and beta adrenergic signaling in repressing TSP1 expression. Taken together, these results suggest that beta adrenergic signaling pathway stimulates angiogenesis through GRK3 mediated suppression of TSP1.

Citation Format: Mohit M. Hulsurkar, Meixiang Sang, Haiping Song, Wenliang Li. Chronic stress and beta adrenergic signaling promote angiogenesis and prostate cancer progression through suppressing the expression of Thrombospondin 1. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4181. doi:10.1158/1538-7445.AM2015-4181
Abstract 4182: Gemcitabine triggers angiogenesis-promoting molecular signals in pancreatic cancer cells: Therapeutic implications
Authors: Khan Mohammad Aslam , Srivastava Sanjeev K. , Bhardwaj Arun , Singh Seema , Carter James E. , Singh Ajay P.
Published: August 2015

Pancreatic tumor microenvironment (TME) is characterized by poor tumor vasculature and extensive desmoplasia that together contribute to poor response to chemotherapy. It was recently shown that targeting of TME to inhibit desmoplastic reaction in a mouse model of pancreatic cancer resulted in increased microvessel density and intratumoral drug concentration, leading to improved therapeutic response. This approach; however, failed to generate a favorable response in a clinical trial. In that regard, we have previously demonstrated that gemcitabine-treatment of pancreatic cancer cells induces CXCR4 expression in a counter-defense mechanism and promotes pancreatic tumor cell invasiveness. The goal of present study was to examine the effect of chemotherapy on endothelial cell growth and migration. Pancreatic cancer cells were treated with gemcitabine or vehicle for 8 h and subsequently incubated in fresh low serum-containing media for next 48 h. Conditioned media was collected from vehicle- (V-CM) and gemcitabine-treated (Gem-CM) pancreatic cancer cells and used for the treatment of human umbilical vein endothelial cells (HUVEC). The data demonstrated stimulatory effects of Gem-CM on endothelial cell proliferation, migration, invasion and capillary-like structure (CLS) formation. Expression analysis of vehicle- and gemcitabine-treated pancreatic cancer cells and their conditioned-media revealed an upregulation of CXCL8/IL8, which is a known regulator of angiogenesis. Currently, we are investigating whether IL8 mediates the effect of Gem-CM on endothelial cells. Together, our findings suggest that gemcitabine triggers angiogenesis-promoting molecular signals in pancreatic cancer cells, which, in light of our previous observations, may hold important clinical and translational significance.

Citation Format: Mohammad Aslam Khan, Sanjeev K. Srivastava, Arun Bhardwaj, Seema Singh, James E. Carter, Ajay P. Singh. Gemcitabine triggers angiogenesis-promoting molecular signals in pancreatic cancer cells: Therapeutic implications. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4182. doi:10.1158/1538-7445.AM2015-4182
Abstract 4183: Differential expression of VEGF in breast cancer cells induced by gp130 cytokines
Authors: Hedeen Danielle S. , Tawara Ken , Nandakumar Madhuri , Fox Ryan , Chang David , Ide Alex , Oler Andrew , LaJoie Dollie , Jorcyk Cheryl L.
Published: August 2015

Glycoprotein 130 (gp130) is a glycosylated type 1 membrane protein and functions as a cytokine receptor modulating activity of the interleukin-6 (IL-6) family cytokines, which include IL-6, IL-11, IL-27, oncostatin M (OSM), leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF), cardiotrophin 1 (CT-1), cardiotrophin-like cytokine (CLC), and neuropoietin (NP). This study was designed to determine differential expression of VEGF in breast cancer cells by IL-6 family cytokines, and to elucidate the molecular mechanisms and signaling pathways involved in cytokine-mediated VEGF induction. Triple negative breast cancer (TNBC) MDA-MB-231 and estrogen receptor positive/progesterone receptor positive/Her2 negative (ER+/PR+/Her2-) T47D cells were treated with three cytokines (25 ng/mL), OSM, IL-6, and LIF, and analyzed. Our results showed that all three cytokines increased levels of HIF1α by Western blot analysis. OSM lead to a greater production of VEGF compared to IL-6 and LIF, as measured by enzyme-linked immunosorbent assay (ELISA) and as seen by luciferase reporter assay. Of the three cytokines, OSM alone induced VEGF in TNBC MDA-MB-231 cells, and this effect was mediated by signal transducer activator of transcription 3 (STAT3) signaling. However, both OSM and IL-6 increased VEGF secretion levels in ER+/PR+/Her2- T47D breast cancer cells, and this signaling was mediated by both HIF1α and pSTAT3, utilizing ELISA and Western blot analysis. These results implicate OSM as the key gp130 cytokine inducing VEGF secretion, and also suggest that this induction is not always HIF1α dependent and is cell-line specific.

Citation Format: Danielle S. Hedeen, Ken Tawara, Madhuri Nandakumar, Ryan Fox, David Chang, Alex Ide, Andrew Oler, Dollie LaJoie, Cheryl L. Jorcyk. Differential expression of VEGF in breast cancer cells induced by gp130 cytokines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4183. doi:10.1158/1538-7445.AM2015-4183
Abstract 4207: Secreted proteins from breast cancer cell lines as a source of cancer biomarkers
Authors: Birkeland Even , Mannelqvist Monica , Akslen Lars Andreas
Published: August 2015

Introduction:

Breast cancer is a heterogeneous disease whose molecular diversity is not well reflected in clinical and microscopic markers used for prognostic information and treatment decisions. Traditionally, most biomarkers are based on tissue samples. To increase efficiency, precise liquid biopsies are needed to detect breast cancer at an early stage including its molecular phenotype, to monitor disease progression, and to predict treatment response. Cell line secretomes are enriched with proteins already linked to tumorigenesis, many of which will be present in biological fluids.

Methods:

Here, we studied the secreted proteins in conditioned media (CM) from two basal-like (HS-578, MB-231) and two luminal-like breast cancer cell lines (MCF-7, BT-474), using liquid chromatography-tandem mass spectrometry. The resulting data were analyzed using SearchGUI, PeptideShaker, Progenesis software and DAVID.

Results:

In total, 954 proteins were identified, of which 93 significantly differentially abundant (P-value ≤ 0.05 Mann-Whitney U, two or more peptides quantified) in the CM between basal-like and luminal-like cell lines. 76 proteins were more abundant in the CM from basal-like cell lines, including several proteins related to angiogenesis and extra-cellular matrix remodeling.

Conclusion:

This study shows the potential for using the secretome of breast cancer cell lines combined with mass-spectrometry for biomarker discovery.

Citation Format: Even Birkeland, Monica Mannelqvist, Lars Andreas Akslen. Secreted proteins from breast cancer cell lines as a source of cancer biomarkers. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4207. doi:10.1158/1538-7445.AM2015-4207
Abstract 4211: The proteomic characterization of HNSCC patient-derived xenografts
Authors: Li Hua , Wheeler Sarah E. , Park Yongseok , Ju Zhenlin , Egloff Ann Marie , Fichera Michele , Thomas Sufi , Mills Gordon B. , Grandis Jennifer R.
Published: August 2015

Head and neck squamous cell carcinoma (HNSCC) is the fifth most common cancer in the US and a leading cancer worldwide. There has been little change in the prognosis for these patients despite extensive research and FDA approval of the targeted therapy cetuximab. Cetuximab has been effective in immortalized HNSCC cell lines but demonstrates only a modest benefit for a subset of patients. This gap between pre-clinical discovery and testing and final clinical efficacy can be narrowed improving the translational relevance of preclinical model systems. Patient derived xenografts (PDXs) have emerged as a promising approach in several cancers. We sought to define and compare the protein expression patterns between primary HNSCC and PDXs derived from 13 patient tumors and test predictive biomarkers to cetuximab therapy.

We compared 13 PDXs propagated at the University of Pittsburgh and analyzed for the expression levels of 190 proteins by reverse-phase protein array (RPPA) to 212 HNSCC specimens and 4566 cancer specimens from TCGA also analyzed by RPPA. We found that 12/13 PDXs were more like HNSCC than any of 14 other cancer types analyzed. Unsupervised clustering of the primary HNSCC and the PDXs demonstrated that PDXs clustered together, indicating some differences in protein expression between HNSCC and PDXs. Further analysis demonstrated that a subset of the proteins were different between the mouse derived PDXs and the primary HNSCC while a second subset were similar between mouse derived PDXs and primary HNSCC. Proteins that were different between these two groups may be poor candidates for translational models, but proteins that were similar may prove useful. We tested the different responses to cetuximab using the PDXs. We found that a PDX with high EGFR/pEGFR expression was more sensitive to cetuximab treatment than a PDX with low EGFR/pEGFR expression.

These results suggest that improved model systems used for translational may predict clinical responses. RPPA measurement of protein expression in PDXs and primary cancers demonstrates that PDXs do represent HNSCC overall. Comparison of the PDXs specifically with primary HNSCC reveals that PDXs are not an appropriate model system for all targeted therapies/studies but provide an effective model for many studies of interest in the field.

Citation Format: Hua Li, Sarah E. Wheeler, Yongseok Park, Zhenlin Ju, Ann Marie Egloff, Michele Fichera, Sufi Thomas, Gordon B. Mills, Jennifer R. Grandis. The proteomic characterization of HNSCC patient-derived xenografts. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4211. doi:10.1158/1538-7445.AM2015-4211