bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5079: Parasympathetic signaling suppresses pancreatic cancer development
Authors: Renz Bernhard W. , Macchini Marina , Hayakawa Yoku , Westphalen C. Benedikt , Churchill Michael , Kar Suchetak , Chen Xiaowei , Nagar Karan , Tailor Yagnesh , Worthley Daniel L. , Iuga Alina C. , Olive Ken P. , Wang Timothy C.
Published: August 2015

Background/Aims: The parasympathetic nervous system plays an important role in the regulation of epithelial homeostasis and has also been postulated to play a role in tumorigenesis. However, the exact role of the vagus nerve in pancreatic carcinogenesis is not well understood. Here we study the effects of muscarinic signaling on pancreatic tumorigenesis in a genetically engineered mouse model of pancreatic cancer (PDx1-Cre/KRasG12D (KC).

Methods: Mice were either vagotomized at 8 weeks or treated with a cholinergic agonist. Pancreatic tissue was collected and analyzed by immunohistochemistry and RT-PCR at 20 weeks of age; cells were isolated and assayed for colony and sphere forming assays. Different human (AsPC-1, BxPC-3, Mia PaCa-2, Panc-1) and murine (K-2548 and K-8282) pancreatic cancer cell lines were subjected to cholinergic and anti-cholinergic drugs and assayed by RT-PCR, Western blot and flow cytometry.

Results: In pancreatic organoid cultures derived from pancreata harboring an oncogenic KRas mutation, cholinergic agonists suppressed sphere formation significantly. Furthermore, pharmacological inhibition or genetic knockout of the muscarinic M3 receptor abolished this effect in vitro. In human and murine pancreatic cancer cells, anchorage independent growth and tumor sphere forming capacity were reduced by pretreatment with cholinergic agonists. Further evaluation revealed that parasympathetic agonists decrease the CD44+CD24+EpCAM+ proved CSC's population. Vagotomy, when performed in KC mice at 8 weeks resulted in pancreatic cancer development in 20% of the animals at 20 week of age. Treatment with the direct muscarinic agonist bethanechol caused a significant delay of PanIN progression in KC mice.

Conclusions: Taken together, our findings suggest that vagal innervation has a regulatory role in pancreatic tumorigenesis via M3 receptor-mediated suppression of CSCs. Since current surgical approaches to resection of PDAC from the head of the pancreas are necessarily associated with a parasympathetic denervation of the pancreas, and thereby a loss of its suppressive effect, this fact may might partly explain the high local recurrence rate of this dismal disease. Additional treatment with cholinergic agonists should be considered in future regimen.

Citation Format: Bernhard W. Renz, Marina Macchini, Yoku Hayakawa, C. Benedikt Westphalen, Michael Churchill, Suchetak Kar, Xiaowei Chen, Karan Nagar, Yagnesh Tailor, Daniel L. Worthley, Alina C. Iuga, Ken P. Olive, Timothy C. Wang. Parasympathetic signaling suppresses pancreatic cancer development. [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 5079. doi:10.1158/1538-7445.AM2015-5079
Abstract 5094: Capsaicin inhibits invasion of human non-small cell lung cancer cells in a TRPV1 receptor independent manner
Authors: Eskew Matthew N. , Brown Kathleen C. , Dasgupta Piyali
Published: August 2015

Non-small cell lung cancer (NSCLC) comprises 85% of lung cancer cases and is characterized by aggressive clinical course and rapid metastasis. One of the challenges of NSCLCs is that the tumor has frequently metastasized to distant organs at the time of diagnosis. The invasion of tumor cells is a key step in the metastatic cascade. Dietary compounds like capsaicin have been known to cause apoptosis in human NSCLCs. However, the effect of capsaicin on the pro-invasive ability of NSCLC cells has not yet been studied. The present study investigates the impact of capsaicin on the invasion of human NSCLCs. Boyden chamber assays show that capsaicin inhibits the invasion of NSCLC in a concentration-dependent and time-dependent manner. The pro-invasive activity of capsaicin was verified by a second invasion assay, the spherical invasion assay, and similar results were obtained. The anti-invasive activity of capsaicin was independent of its growth-inhibitory activity and manifested at a lower concentration. The heat-sensation activity of capsaicin is mediated by the transient receptor potential vanilloid (TRPV) family of proteins. Capsaicin functions as an agonist of the TRPV1 receptor. However, the anti-invasive activity of capsaicin was not affected by the generalized TRPV receptor antagonist ruthenium red. Specific antagonists to TRPV1, namely capsazepine and SB36671, did not reverse the anti-invasive activity of capsaicin. Taken together, capsaicin potently suppresses invasion of human NSCLCs independent of the TRPV receptor family.

Citation Format: Matthew N. Eskew, Kathleen C. Brown, Piyali Dasgupta. Capsaicin inhibits invasion of human non-small cell lung cancer cells in a TRPV1 receptor independent manner. [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 5094. doi:10.1158/1538-7445.AM2015-5094
Abstract 5097: APE1/Ref-1 promotes cell adhesion and migration in cervical cancer cells
Authors: Kim Mihwa , You Ho Jin , Kim Dae Joon
Published: August 2015

Apurinic and apyrimidinic endonuclease 1 (APE1)/redox effector factor-1 (Ref-1) is known to be involved in DNA base excision repair pathway and redox-dependent regulation of a number of transcription factors, such as AP1, p53, and Pax-8. It is expressed in numerous human cancers such as pancreatic cancer, prostate cancer, breast cancer, cervical cancer, and colon cancer. Our previous studies suggested APE1/Ref-1 can promote cancer progression through its involvement in cell proliferation, metastasis and angiogenesis; however, the molecular mechanism by which APE1/Ref-1 functions in these processes has not been characterized. In this study, we are the first to report that APE1/Ref-1 activates cell adhesion-related proteins to trigger cell migration. To investigate whether APE1/Ref-1 is involved in cell adhesion in normal fibroblasts, an adhesion assay was performed using GM00637 human fibroblast cell lines stably overexpressing APE1/Ref-1. Cell adhesion to vitronectin, a known ligand of the adhesion molecules integrin ανβ3 and integrin ανβ5, was significantly increased in APE1/Ref-1-overexpressing GM00637 cells compared to control fibroblast cells. FACS analysis showed APE1/Ref-1 induces the expression and activation of adhesion molecules including integrin αν and β3 in APE/Ref-1-overexpressing GM00637 cells. To evaluate whether APE1/Ref-1-mediated regulation of integrin ανβ3 is involved in cell migration in cervical cancer cells, a migration assay was performed using APE1/Ref-1-specific siRNA and an integrin ανβ3-specific antibody. Cell migration was significantly decreased in the absence of APE1/Ref-1 expression in cervical cancer cells, which was comparable with inhibition of integrin ανβ3. Consistent with these results, APE1/Ref-1 increased the activation and expression of integrin ανβ3 in both cervical cancer cells and cervical cancer patient tissue. Combined, these studies suggest that APE/Ref-1 can promote cell adhesion and migration through integrin ανβ3 signaling and thereby this signaling mechanism could contribute to cervical cancer progression.

Citation Format: Mihwa Kim, Ho Jin You, Dae Joon Kim. APE1/Ref-1 promotes cell adhesion and migration in cervical cancer 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 5097. doi:10.1158/1538-7445.AM2015-5097
Abstract 510: A novel role for Olig2 in the regulation of glioma invasion
Authors: Singh Shiv K. , Giannonatti Nicole , Kupp Robert , Cascio Costanza Lo , Mehta Shwetal
Published: August 2015

Glioblastoma multiforme (GBM) are highly infiltrative, aggressive and lethal primary brain tumors in adults that are resistant to conventional treatments. A major clinical obstacle in the treatment of GBMs is the ability of glioma cells to invade the surrounding brain parenchyma, preventing their complete surgical resection. Several studies have shown that invasive glioma stem cells (GSCs) extend beyond the surgically resected regions. These invasive cells are the cause for tumor recurrence and poor prognosis in GBM patients. The molecular and cellular mechanism involved in the regulation of the switch from proliferation to invasion/migration in GBM is poorly understood. We investigated the role of Olig2, a Central Nervous System (CNS) specific transcription factor, in promoting the switch from proliferating to invading glioma cell. Genetically relevant murine glioma model as well as patient-derived glioma stem cells (GSCs) were utilized to identify the effect of Olig2 on glioma invasion. Functionally, Olig2-mediated invasion was assessed by 3D-transwell invasion assays, organotypic slice cultures as well as PDX tumors in immunocompromised mice. In addition, murine and patient-derived GSCs were utilized to identify direct genetic targets of Olig2 involved in glioma invasion by ChIP-seq and RNA-seq.

Our data demonstrate that Olig2-expressing murine glioma stem/progenitor-like cells are highly invasive as compared to Olig2 knockout cells. We further corroborated the requirement of Olig2 for glioma invasion in patient-derived GSCs by utilizing shRNA-mediated knockdown of Olig2. Olig2 depletion resulted in reduced glioma cell invasion in at least four independent human GSCs. Recent studies have shown that activation of transcription factors such as Zeb1 and Twist1 occurs in invasive high grade gliomas. We show that knockdown of Olig2 in murine as well as human GSCs leads to a significant decrease in the expression of invasion genes such as Zeb1, Vimentin, Twist1, and Stat3 as compared to control cells. In conclusion, our studies highlight a novel role for the Olig2 signalling network in promoting the switch from proliferation to invasion in GBM, and provide a plausible target for inhibiting glioma invasion.

Citation Format: Shiv K. Singh, Nicole Giannonatti, Robert Kupp, Costanza Lo Cascio, Shwetal Mehta. A novel role for Olig2 in the regulation of glioma invasion. [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 510. doi:10.1158/1538-7445.AM2015-510
Abstract 5108: Oscillatory calcium signal drives melanoma invasion and metastasis
Authors: Sun Jianwei , Lu Fujian , He Huifang , Cheng Heping , Yang Shengyu
Published: August 2015

Ca2+ signaling has been increasingly implicated in cancer invasion and metastasis, and yet the underlying mechanisms remained largely unknown. Here, we report that STIM1- and Orai1- mediated Ca2+ oscillations promote melanoma invasion by orchestrating invadopodium assembly and extracellular matrix (ECM) degradation. Ca2+ oscillation signals facilitate invadopodial precursor assembly by activating Pyk2 phosphorylation at tyrosine 402, which directly binds to and activates Src to promote invadopoium assembly. Disruption of Ca2+ oscillations inhibited invadopodium assembly. Futhermore, STIM1 and Orai1 regulate the proteolysis activity of individual invadopodium. Mechanistically, Orai1 blockade inhibits the recycling of MT1-MMP to the plasma membrane and entraps MT1-MMP in the endocytic compartment to inhibit ECM degradation. STIM1 knockdown significantly inhibited melanoma lung metastasis in a xenograft mouse model, implicating the importance of this pathway in metastatic dissemination. Our findings provide a novel mechanism for Ca2+ -mediated cancer cell invasion and shed new light on the spatiotemporal organization of store-operated Ca2+ signals during melanoma invasion and metastasis.

Note: This abstract was not presented at the meeting.

Citation Format: Jianwei Sun, Fujian Lu, Huifang He, Heping Cheng, Shengyu Yang. Oscillatory calcium signal drives melanoma invasion and metastasis. [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 5108. doi:10.1158/1538-7445.AM2015-5108
Abstract 5132: Evaluation of multi-marker in tissue-derived cancer cells from patients with epithelial ovarian carcinoma
Authors: Lee Shin-Wha , Lee Sang-Eun , Lee Ha-Young , Kim Dae-Yeon , Kim Jong-Hyeok , Kim Yong-Man , Kim Young-Tak , Nam Joo-Hyun
Published: August 2015

Objective: The objective of this study is to refine a simple, rapid and reproducible method for the isolation and characterization of epithelial ovarian cancer (EOC) cells from ovarian tumor tissues and to evaluate the intracellular expression of CK-7, CA-125, HE4 and FOLR-1 in primary ovarian cancer cells.

Methods: Fourteen patients diagnosed with papillary serous adenocarcinoma of ovary were selected in ASAN Medical Center. Isolation of tumor EOC cells involves the mechanical disruption of the tumor tissue and placed directly into culture with very little manipulation. Primary ovarian cancer cells were monitored after initial plating and every day for following 15 days. Confocal laser scanning microscopy was used for the imaging of intracellular immunofluorescence of CK-7, CA-125, HE4 and FOLR-1.

Results: Two days after plating primary ovarian cancer cells began to adhere to the plate by the presence of small adherent clusters. The primary ovarian cancer cells became a confluent monolayer and displayed the typical epithelial cobblestone morphology. The mean duration of passage was 8.5 days, it was revealed variously in the primary ovarian cancer cells. Immunofluorescent detection of CK-7 expression in EOC cells demonstrates the epithelial origin, however their expression was not constant. CA-125, HE-4 and FOLR-1 were expressed by all tissue-derived EOC cells. The expression pattern of CA-125, HE-4 and FOLR-1 was different in each patient and that was not correlated with the serum level of CA-125.

Conclusions: The isolation and characterization of tissue-derived cancer cells from patients with EOC using the expression of CK-7, CA-125, HE4 and FOLR-1 were reasonable method. This result could provide clinically relevant models suitable for investigation of the ex vivo biological characteristics of ovarian cancers.

Citation Format: Shin-Wha Lee, Sang-Eun Lee, Ha-Young Lee, Dae-Yeon Kim, Jong-Hyeok Kim, Yong-Man Kim, Young-Tak Kim, Joo-Hyun Nam. Evaluation of multi-marker in tissue-derived cancer cells from patients with epithelial ovarian 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 5132. doi:10.1158/1538-7445.AM2015-5132
Abstract 5141: A comprehensive system of congenic mouse melanoma models for evaluation of immune therapies
Authors: Meeth Katrina M. , Damsky William E. , Bosenberg Marcus
Published: August 2015

There has been longstanding interest in the relationship between the immune system and malignant melanoma, the deadliest form of skin cancer. Melanoma has been the tumor of choice for the evaluation of a wide variety of therapeutic approaches that rely on stimulation of the immune system to combat cancer. The striking responses to immune checkpoint inhibitors noted in some melanoma patients have now been observed in several other malignancies, including renal cell carcinoma, lung cancer, and prostate cancer. These findings are very encouraging, however the specific mechanisms of response and reasons why only a subset of patients respond to therapy are not known. The extraordinary potential of immune therapies in cancer suggests that human-relevant mouse models of cancer are required to understand mechanisms of response and to optimize therapy so that a larger proportion of patients respond.

We have generated and evaluated a series of genetically engineered mouse models driven by alterations in Braf, Pten, Cdkn2a, beta catenin, p53, and Nras that reflect the genetic diversity of the vast majority of human melanoma. We have backcrossed 6 of these models to C57Bl/6, generated tumors, and isolated a series of over 20 congenic mouse melanoma cell lines. Several independent lines have been isolated for each melanoma genotype generated. Injection of these lines into C57Bl/6J mice results in reproducible tumor formation in the absence of co-injection of Matrigel. We are in the process of characterizing the tumor microenvironment in each of the models using flow cytometry and a novel quantitative imaging approach of histological sections, which produces flow cytometry-like data, but retains spatial and morphologic information that can be utilized to characterize much smaller tumors. This series of congenic lines represents an ideal set of models for the study of cancer immunology and responses to immune therapies.

Citation Format: Katrina M. Meeth, William E. Damsky, Marcus Bosenberg. A comprehensive system of congenic mouse melanoma models for evaluation of immune therapies. [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 5141. doi:10.1158/1538-7445.AM2015-5141
Abstract 5148: NR5A2 is essential for pancreas development and affects pancreas carcinogenesis
Authors: Nissim Sahar , Weeks Olivia , Hedgepeth John , Wucherpfennig Julia , Wang Xiao-Xu , Kimmelman Alec , Goessling Wolfram
Published: August 2015

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal cancers today. A genome-wide association study (GWAS) has implicated the nuclear receptor NR5A2 in modulating risk for pancreatic cancer. While a role for this gene in pancreatic cancer has not been previously recognized, it plays an important role in stem cell pluripotency and metabolism. Using the zebrafish model, we demonstrate that genetic or pharmacologic inhibition of nr5a2 activity results in absence of exocrine pancreas, while leaving endocrine pancreas unaffected, indicating an essential role in pancreas organ formation. Further, we localize nr5a2 expression to the endodermal bud that gives rise to the exocrine pancreas, consistent with a role for nr5a2 in regulating differentiation of pancreas progenitors. In resected human PDAC specimens and in vitro cell lines, we find that NR5A2 expression is altered compared to normal ductal epithelium. Moreover, shRNA knockdown of NR5A2 dramatically alters proliferation of PDAC cell lines, supporting the hypothesis that NR5A2 is involved in pancreatic carcinogenesis. These data support a model that NR5A2 regulates the proliferation and differentiation of exocrine pancreas progenitor cells during development. We postulate that loss-of-function of NR5A2 results in dysregulation of these processes in the adult pancreas by driving acinar cells to acquire properties of progenitor cells, and therefore is an important step in pancreas oncogenesis.

Note: This abstract was not presented at the meeting.

Citation Format: Sahar Nissim, Olivia Weeks, John Hedgepeth, Julia Wucherpfennig, Xiao-Xu Wang, Alec Kimmelman, Wolfram Goessling. NR5A2 is essential for pancreas development and affects pancreas carcinogenesis. [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 5148. doi:10.1158/1538-7445.AM2015-5148
Abstract 5175: Preclinical monitoring of prostate cancer lesions in bone by intravital multiphoton microscopy
Authors: Dondossola Eleonora , Alexander Stephanie , Alexander Steve , Holzapfel Boris , Logothetis Christopher , Hutmacher Dietmar , Friedl Peter
Published: August 2015

Bone metastases are a major complication in patients with advanced prostate cancer (PCa) and frequent cause of therapy resistance. Besides cell intrinsic mechanisms, signals received from the bone microenvironment critically contribute to support PCa cell survival and growth. Recent advances in intravital microscopy enable to investigate this complex PCa cell-microenvironment interaction and allow preclinical examination of tumor responses to both conventional and targeted therapy.

We developed a window model amenable to intravital multiphoton microscopy (MPM) to longitudinally monitor PCa lesions in humanized neobone established in the dermis of the mouse. Tissue engineered bone constructs, TEBCs, were generated by functionalizing polymeric polycaprolactone scaffolds with human mesenchymal stem cells differentiated to osteoblasts, and characterized in vitro for cell distribution and calcium deposition. After in vivo implantation, TEBC maturation was monitored by μCT, MPM and histological analysis over time to generate a miniaturized-neobone with defined cortical thickness (50-60 μm) suitable for intravital imaging through a body window. PCa (PC3) lesions, after implantation into the bone cavity, were longitudinally monitored for growth and niche development, using multi-parameter recording of: collagen/bone matrix (SHG), bone surface (THG), blood vessels/stromal phagocytes (fluorescent dextran), and PC3 cells (nuclear H2BeGFP, cytoplasmatic DsRed2).

By combining innovative tissue engineering with optical windows, state-of-the-art fluorescence reporter technology and intravital MPM, this model will provide mechanistic and applied insight into the therapy response of bone metastasis.

Citation Format: Eleonora Dondossola, Stephanie Alexander, Steve Alexander, Boris Holzapfel, Christopher Logothetis, Dietmar Hutmacher, Peter Friedl. Preclinical monitoring of prostate cancer lesions in bone by intravital multiphoton microscopy. [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 5175. doi:10.1158/1538-7445.AM2015-5175
Abstract 5176: Chemotherapy alters the natural history of metastatic progression
Authors: Aiello Nicole M. , Bajor David L. , Pham Minh N. , Vonderheide Robert H. , Stanger Ben Z.
Published: August 2015

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with a 5-year survival rate of less than 6%. Metastatic disease accounts for a majority of PDAC-related deaths, even for patients with no evidence of metastasis at the time of resection. In this study we sought to understand the natural history of metastatic colonization and the cellular mechanisms governing the metastatic cascade. Using an autochthonous PDAC mouse model combined with a lineage-labeling approach, we have tracked and characterized the various stages of metastatic progression, from single cells to macroscopic lesions. PDAC tumors primarily metastasize to the liver, where small lesions of 10 cells or less reside closest to portal veins and exhibit a high frequency of epithelial-mesenchymal transition (EMT). Larger metastatic lesions exhibit reduced EMT and are hypovascular, resembling primary PDAC tumors. Metastases gradually accumulate desmoplasia as they grow, which consists of myofibroblasts, leukocytes and extracellular matrix components including collagen, hyaluronic acid, fibronectin and SPARC. Treatment with gemcitabine and nab-paclitaxel reduces overall metastatic burden and shifts the size distribution of metastases toward small lesions of 10 cells or less. Single seeded cells in particular seem to be protected from chemotherapy-induced killing, while larger lesions that have accumulated a desmoplastic stroma are more susceptible. Our results demonstrate that chemotherapy has direct measurable effects on metastatic burden, which may explain the improved outcomes for PDAC patients who receive adjuvant chemotherapy.

Citation Format: Nicole M. Aiello, David L. Bajor, Minh N. Pham, Robert H. Vonderheide, Ben Z. Stanger. Chemotherapy alters the natural history of metastatic progression. [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 5176. doi:10.1158/1538-7445.AM2015-5176
Abstract 5184: Synaptonemal complex protein 3 is associated with lymphangiogenesis in non-small cell lung cancer
Authors: Kitano Haruhisa , Chung Joon-Yong , Hanaoka Jun , Inoue Shuhei , Yoshinori Doki , Fukuoka Junya , Hewitt Stephen M.
Published: August 2015

Lymphangiogenesis is a key component for tumor growth and metastasis in non-small cell lung cancer (NSCLC), and is a promising target in cancer therapy strategy. The axis of vascular endothelial growth factor-C (VEGF-C)/VEGF-D/VEGFR-3 is considered to be a major driver of lymphangiogenesis but the detailed mechanism of this process remains unclear. We recently showed that the expression of synaptonemal complex protein 3 (SCP3) is associated with poor prognosis and linked with lymph node metastasis in NSCLC. To investigate the possible lymphangiogenic significance of SCP3 in NSCLC, we assessed SCP3, VEGF-A, VEGF-B, VEGF-C, and VEGF-D expressions in archival tumor tissues from 89 NSCLC patients with lymph node metastasis by immunohistochemical staining. The positive staining of SCP3, VEGF-A, VEGF-B, VEGF-C and VEGF-D expressions were detected in 24 (27.0%), 22 (24.7%), 27 (30.3%), and 24 cases (27.0%), respectively. Notably, we have identified that the SCP3 expression is positively correlated with VEGF-C and VEGF-D (for both, p < 0.001) whereas its expression is negatively correlated with VEGF-A and VEGF-B (p = 0.029 and p = 0.026, respectively). Furthermore, we have also detected the linkage between SCP3 and VEGF-C or VEGF-D expressions in human lung cancer cell lines. In multivariate analysis of patients with lymph node metastasis, SCP3 expression displayed the worse overall survival (HR = 1.86, p = 0.008). These data suggest that SCP3 is linked with lymphangiogenesis and provide insight into the SCP3-VEGF-C/VEGF-D axis based anti-lymphangiogenic therapy strategy.

Citation Format: Haruhisa Kitano, Joon-Yong Chung, Jun Hanaoka, Shuhei Inoue, Doki Yoshinori, Junya Fukuoka, Stephen M. Hewitt. Synaptonemal complex protein 3 is associated with lymphangiogenesis in non-small cell lung 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 5184. doi:10.1158/1538-7445.AM2015-5184
Abstract 5195: A novel, multimodal theranostic nanoprobe is effectively incorporated into melanoma brain metastatic cells
Authors: Aasen Synnøve N. , Eichler Tilo W. , Hruby Martin , Pospisilova Aneta , Stepanek Petr , Spriet Endy , Jirak Daniel , Skaftnesmo Kai Ove , Thorsen Frits
Published: August 2015

Metastatic cancer is a significant cause of death worldwide, despite continuing advances in research on diagnostics and therapy. To overcome unsuccessful drug delivery to the tumor tissue, several different nanoprobes have been designed, of which some have combined diagnostic and therapeutic properties. It is an advantage to use non-toxic, biodegradable materials in the design of nanoprobes, as such compounds are easily eliminated from the body. In this work we report for the first time the use of a recently developed nanoprobe to label melanoma metastatic cell lines, for multimodal imaging of cellular uptake and drug delivery. Our nanoprobe consisted of a backbone of glycogen, where the red fluorescent marker Dyo-615 for fluorescence microscopy and Gadolinium for MRI were incorporated.

Fluorescence microscopy showed effective labeling of human melanoma metastatic cells over 24 hours and cell viability was not affected by the labeling. The nanoprobe was found in the cytosol of the cells, and a gradual degradation of the probe inside the lysosomes could be observed. In vitro MRI relaxivity measurements showed significant reduction in T1 mapping times, compared to unlabeled cells. In vivo MRI studies showed that subcutaneous melanoma tumors in mice could be effectively visualized with T1 weighted MRI after intravenous injection of the nanoprobe, and the contrast enhancement was comparable to what was seen after using standard Omniscan contrast agent.

In summary, our biodegradable glycogen nanoprobe shows a high potential to be used further as a theranostic entity. The nanoprobe may offer additional advantages over MRI contrast agents, as tumor uptake of pharmaceuticals attached to the nanoprobe can be traced in real-time in vivo.

Citation Format: Synnøve N. Aasen, Tilo W. Eichler, Martin Hruby, Aneta Pospisilova, Petr Stepanek, Endy Spriet, Daniel Jirak, Kai Ove Skaftnesmo, Frits Thorsen. A novel, multimodal theranostic nanoprobe is effectively incorporated into melanoma brain metastatic 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 5195. doi:10.1158/1538-7445.AM2015-5195
Abstract 5196: Histologic and whole slide quantitative image analysis of lung metastases in a mouse breast cancer model using carboplatin and Nutlin 3A alone and in combination
Authors: Strack Margaret , Tonsing-Carter Eva , Sinn Tony , Spragin Tiaishia , Peterson Kacie , Bailey Barbara , Pollok Karen , Sandusky George
Published: August 2015

Breast cancer will affect 1 in 8 women in their lifetime. Approximately 20-30% of all breast cancer cases become metastatic. This study was designed to evaluate the effect of tumor met knockdown with individual and combination drug therapy. In this mouse model, with 3 per group, we used the following chemotherapeutic agents: a vehicle control, carboplatin (20mg/kg i.p.), Nutlin-3a (200mg/kg p.o.) and a combination group with the same doses. Nutlin-3 is a small-molecule inhibitor that binds to the p53-binding pocket of MDM2, and activates the p53 pathway. This has been shown to be effective in mouse models of cancer that have a wild-type p53, including prostate, leukemia and multiple myeloma.

The lungs were removed and examined for metastatic lung lesions, fixed in 10% neutral buffered formalin (NBF), processed into a paraffin block, microtomed, and slides were stained with H&E. Ki67 immunostain was performed to clearly define the met lesions and the whole slide was imaged with an Aperio Digital Imaging System. The Image Scope positive pixel algorithm was used to quantitate the met lesions in the lungs. The data revealed upon drug treatment the number of mets in the lungs were unchanged in the carboplatin and nutlin-3a individual treatment groups compared to the vehicle group. The lungs in the combination group had the lowest number of tumors with both visual and quantitative image analysis compared to the vehicle control group.

The Aperio automated positive pixel count for Ki67 immunostain was consistent with the H&E pathology read. The values decreased with combination treatment (control vs. combination 20mg/kg carboplatin, 200mg/kg nutlin 3a) and were 6% and 1.7% respectively.

It was concluded that mice treated with carboplatin and Nutlin-3a combination therapy had fewer metastatic breast cancer mets in the lungs. The individual drug doses were low compared to previous studies and did not have an effect as was expected. This finding suggests that the combination of a traditional chemotherapy with a small protein inhibitor could inhibit the spread of metastatic breast cancer.

Citation Format: Margaret Strack, Eva Tonsing-Carter, Tony Sinn, Tiaishia Spragin, Kacie Peterson, Barbara Bailey, Karen Pollok, George Sandusky. Histologic and whole slide quantitative image analysis of lung metastases in a mouse breast cancer model using carboplatin and Nutlin 3A alone and in combination. [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 5196. doi:10.1158/1538-7445.AM2015-5196
Abstract 5202: Magnolin suppresses cell migration by abrogation of ERK-mediated RSK2/NF-κB signaling pathway
Authors: Lee Cheol-Jung , Lee Mee-Hyun , Song Ji-Hong , Yoo Sun-Mi , Cho Yong-Yeon
Published: August 2015

Our previous study demonstrated that RSK2 directly phosphorylated IκBα at ser 32, resulted in its degradation which leads to promote NF-κB activation and might be an important role in metastasis. Therefore, it is reasonable the discovery of chemotherapeutic and/or chemopreventive agent. Magnolin, an ingredient of magnolia species, suppressed EGF-induced neoplastic cell transformation by targeting of ERK 1 and 2. In this study, we found that magnolin inhibited cell migration and invasion of normal cell and human lung cancer cells. Further, we demonstrated that magnolin suppressed epidermal growth factor-induced ERK/RSK2 signaling pathway, resulting in suppression of NF-κB activity in JB6Cl41 cells. The EGF-induced NF-κB activity was abrogated up-regulation of COX-2 mRNA expression and COX-2 protein levels by co-treatment of EGF and magnolin. Interestingly, magnolin suppressed ERK-mediated MMP-2 and NF-κB-mediated MMP-9 expression. In addition, we found that the protein expression of N-cadherin involved in metastasis was markedly attenuated upon treatment with magnolin. Notably, magnolin suppressed the migration and invasion of lung cancer cells in a dose-dependent manner. Taken together, these results demonstrated that magnolin is beneficial for the anti-invasion and -migration in cancer metastasis.

Citation Format: Cheol-Jung Lee, Mee-Hyun Lee, Ji-Hong Song, Sun-Mi Yoo, Yong-Yeon Cho. Magnolin suppresses cell migration by abrogation of ERK-mediated RSK2/NF-κB signaling pathway. [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 5202. doi:10.1158/1538-7445.AM2015-5202
Abstract 524: Assessing the interaction between acetylenic tricyclic bis-(cyano enone) and cysteine residues of actin to inhibit non-small cell lung cancer cell migration
Authors: Chan Eddie , Saito Akira , Honda Tadashi , Di Guglielmo John
Published: August 2015

During epithelial-to-mesenchymal transition (EMT), epithelial cells lose their cell-cell junctions, apical-basal polarity, and reorganize their actin cytoskeleton. These changes, among others, increase the motility of individual cells and promote a highly invasive phenotype. The actin cytoskeleton is also key in cell migration and invasion, as the polymerization of actin at the leading edge provides the necessary driving force for translocation and protrusions. Therefore, the actin cytoskeleton is a potential target for inhibiting tumour progression. In this study we characterized a novel tri-cyclic synthetic triterpenoid derivative, TBE-31, in its ability to associate with actin and inhibit cell migration. Using a pull-down approach, we demonstrate that TBE-31 binds directly to actin. Furthermore, TBE-31 inhibited branched and linear actin polymerization in vitro, and stress fibre formation in cultured cells. Actin cytoskeleton reorganization and stress fiber formation was also inhibited in TGFβ-dependent EMT of non-small cell lung cancer cells. We next used SwissDock software to investigate the potential molecular interaction between TBE-31 and actin. Swissdock predicted that TBE-31 would have a high affinity for binding to the same cleft as Cytochalasin D, a bona-fide actin polymerization inhibitor. As triterpenoids associate with cysteine residues in target proteins, we generated cysteine to alanine mutants of actin and verified TBE-31 binding using pull-down assays. Finally, we observed that TBE-31 inhibited fibroblast and non-small cell lung tumor cell migration with an IC50 of 1.0 and 2.5 μM, respectively. Taken together, our results suggest that TBE-31 targets actin polymerization to alter cell morphology and inhibit cell migration.

Citation Format: Eddie Chan, Akira Saito, Tadashi Honda, John Di Guglielmo. Assessing the interaction between acetylenic tricyclic bis-(cyano enone) and cysteine residues of actin to inhibit non-small cell lung 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 524. doi:10.1158/1538-7445.AM2015-524
Abstract 528: PP4C plays a braking role in Akt-driven cancer migration/invasion
Authors: Ho Ming-Yi , Liang Shu-Mei , Liang Chi-Ming
Published: August 2015

Treatment of lung cancer cells with growth factors and PGE2 activates phospho-AktS473 to transiently up-regulate phosphorylation of IKK and NF-κB within 1 hour, resulting in the elevation of cellular COX-2/PGE2, E-cadherin suppressor Snail and epithelial-mesenchymal transition (EMT) 24-31 hours later. In this study, we further found that in the cells treated with PGE2 or growth factors, the activated phospho-AktS473 declined to basal level 2 hours after treatment and increased again to reach a plateau 24-31 hours later but this resurgence of phospho-AktS473 was no longer associated with phosphorylation of IKK and NF-κB. To identify which factors or proteins cause dephosphorylation of IKK and NF-κB during phospho-AktS473 resurgence, we used immunoprecipitation, immunoblotting and activity assay approaches. Our results showed that during 12-31 hours of PGE2 treatment, the activity of protein phosphatase (PP) 4C and its association with IKK was increased. In comparison, there was no association between PP1, PP2A and PP5 with IKK. PP4C knockdown in PGE2 treated lung cancer cells increased their IKK/IκB/NF-κB signaling leading to the sustainment of COX-2 and Snail for much longer time. Knockdown of PP4C also increased the cellular EMT, migration and invasion of the lung cancer cells significantly. These findings demonstrated that PP4C downregulates IKK/IκB/NF-κB signaling to play a braking role in the Akt-driven lung cancer migration/invasion.

Citation Format: Ming-Yi Ho, Shu-Mei Liang, Chi-Ming Liang. PP4C plays a braking role in Akt-driven cancer migration/invasion. [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 528. doi:10.1158/1538-7445.AM2015-528
Abstract 5282: A 3q amplification gene signature associated with triple negative breast cancer lung-specific metastasis
Authors: Qian Jun , Chen Heidi , Massion Pierre P.
Published: August 2015

Triple negative breast cancers (TNBC), devoid of expression of the estrogen, progesterone and HER2 receptors, are among the most aggressive and deadly breast cancers, with high rates of tumor recurrence and metastatic spread. New prognostic markers are needed to identify patients who are at the highest risk for developing metastases, which might assist in tailoring treatment strategies. We previously identified 20 candidate driver genes in Chromosome 3q26-29, a critical amplified region in many human cancers including breast and lung cancer. The aim of this study was to assess the prognostic power of this 20-gene signature in TNBC patients. 855 breast tumors combined from four breast cancer cohorts (GSE12276, GSE2603, GSE2034 and NKI295) were used to evaluate the signature for prediction of organ-specific distant metastasis free survival (DMFS). The predictive ability of the 20-gene signature was evaluated by Cox proportional hazards model and was compared against other published breast cancer gene signatures including MammaPrint (70-gene), Veridex (76-gene), PAM50, 54 lung metastasis signature genes, HIF1 hypoxia pathway (8-gene), TGF β pathway (153-gene), BACH1 pathway metastasis gene signature (25-gene) and Gene Expression Prognostic Index Using Subtypes (GENIUS). The 20-gene signature was strongly associated with worse lung-specific metastasis-free survival (HR, 2.0, 95%CI, 1.4 to 2.87, P=0.00014) in TNBC patients (n = 131), independently of intrinsic subtype or published gene signatures and less so in non-TNBC patients (n = 724, HR, 1.3, 95%CI, 1.01 to 1.69, P=0.04). In conclusion, we discovered an amplified 20-gene signature in 3q26-29 that are associated with lung metastasis in patients with TNBC, suggesting on a potential role of 3q genes in the mechanism of lung-specific metastasis and incriminating cancer driver genes in this location as potential attractive novel therapeutic targets in this disease. The work is supported by the NIH Grant R01 CA102353 to PPM.

Citation Format: Jun Qian, Heidi Chen, Pierre P. Massion. A 3q amplification gene signature associated with triple negative breast cancer lung-specific metastasis. [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 5282. doi:10.1158/1538-7445.AM2015-5282
Abstract 5286: Expression of cancer type amino acid transporter LAT1 is a prognosis prediction factor in breast carcinoma: comparison between triple-negative and non-triple-negative types
Authors: Ichinoe Masaaki , Mikami Tetsuo , Hana Kiyomi , Yanagisawa Nobuyuki , Endou Hitoshi , Okayasu Isao , Murakumo Yoshiki
Published: August 2015

Purpose: Amino acid transporters play important roles in cell survival and proliferation. L-Type amino acid transporter 1 (LAT1) is one of the major Na+-independent neutral amino acid transporters. LAT1 has an aspect of oncofetal protein. Recently, high levels of LAT1 expression have been reported in various tumors, such as prostate cancer, pancreas cancer, bile duct cancer, lung cancer and stomach cancer.We revealed LAT1 expression is an independent prognosis marker in stomach cancer, prostate cancer and pancreas cancer. The purpose of the present study was to investigate the LAT1 expression in breast carcinoma and relationship between LAT1 expression and prognosis.

Materials & Methods: We enrolled 290 patients with invasive breast carcinoma surgically resected between 2006 and 2010 at Kitasato University Hospital and Toho University Omori Medical Center, Japan. We investigated LAT1 expression in the enrolled breast carcinoma cases by immunohistochemical staining. All cases were divided into two groups, 78 cases of triple negative carcinoma (TNBC) and 212 cases of non-triple negative carcinoma (NTNBC), with immunohistochemistry of estrogen and progesterone receptors and Her2. After LAT1 expression was compared between TNBC and NTNBC, correlations among LAT1 expression, Ki-67 labeling index (LI), CD98 expression, and clinicopathological findings was assessed.

Results: TNBC showed significantly higher LAT1 expression than NTNBC. TNBC also showed significantly higher CD98 expression rather than NTNBC. LAT1 expression of extraductal invasive cancer cells was significantly higher than those of intraductal carcinoma cells. A significantly positive correlation was observed between LAT1 expression and CD98 expression. Further, a positive correlation between LAT1 expression and Ki-67 LI was found. Log-rank test in overall breast cancers revealed that high LAT1 expression group showed significantly poor prognosis, compared to low LAT1 group. In NTNBC, high LAT1expression cases possessed poorer prognosis than low LAT1 expression cases.

Conclusion: In invasive ductal carcinoma of breast, LAT1 expression has a potential for a prognosis prediction factor. LAT1 offers a potential target for anti-cancer therapy by its inhibitor JPH203 which has been developed by us.

Citation Format: Masaaki Ichinoe, Tetsuo Mikami, Kiyomi Hana, Nobuyuki Yanagisawa, Hitoshi Endou, Isao Okayasu, Yoshiki Murakumo. Expression of cancer type amino acid transporter LAT1 is a prognosis prediction factor in breast carcinoma: comparison between triple-negative and non-triple-negative types. [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 5286. doi:10.1158/1538-7445.AM2015-5286
Abstract 5290: Identification of novel prognostic genes associated with risk of recurrence in early breast cancer patients
Authors: Kwon Mi Jeong
Published: August 2015

Introduction : In order to determine the need for adjuvant chemotherapy in early estrogen receptor-positive breast cancer, especially luminal A, it is important to identify patients at high risk of recurrence. This study was performed to identify novel prognostic genes associated with the recurrence risk of patients with lymph node-negative luminal A (LN−/LumA) breast cancer.

Methods : RNA was extracted from formalin-fixed, paraffin-embedded (FFPE) breast cancer tissues. The expression levels of 16 candidate genes identified from our previous study were measured using quantitative real-time reverse transcription-PCR in 926 FFPE samples. We used Cox proportional hazards models to assess the prognostic value of clinical and gene variables for recurrence.

Results : Of the 796 patients with recurrence-free survival (RFS) and 826 patients with overall survival (OS), 24.4% (194/796) and 24.0% (198/826), respectively, had LN−/LumA breast cancer. Univariate analysis showed that clinical variables such as tumor size, involvement of lymph nodes, and tumor stage were not significantly associated with RFS and OS of patients with this subtype. In contrast, there was a significant association between gene expression and patient outcomes. Higher expression of 4 proliferation-related genes (F4, M4, R3, and U4) was significantly correlated with shorter OS and RFS. Importantly, in multivariate analyses, high expression of R3 and U4 remained significantly associated with RFS. While not significantly related to RFS in univariate analysis, immune response-related B5 expression had a significantly lower harzard ratio in the multivariate analysis (P = 0.004), indicating it was an independent positive prognostic factor. Higher expression of M4 (P = 0.014) and R3 (P = 0.01) was independently associated with shorter OS. When tested using gene variables, prognostic models for risk of recurrence included M4, R3, U4, and B5. Moreover, patients with low B5 and high U4 expressions had a significantly shorter RFS than those with high B5 and low U4 expressions.

Conclusions : Novel prognostic genes identified in our study give more significant information on the risk of recurrence than clinical variables in patients with LN−/LumA breast cancer. Multigene expression signatures, including immune response-related genes, can be used to identify patients at high risk of recurrence in this subtype of breast cancer.

Citation Format: Mi Jeong Kwon. Identification of novel prognostic genes associated with risk of recurrence in early breast cancer patients. [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 5290. doi:10.1158/1538-7445.AM2015-5290