bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3597: Malignancy of bladder cancer cells is enhanced by tumor associated fibroblasts through a cytokine-chemokine loop
Authors: Grimm Susanne , Jennek Susanne , Bratsch Jens , Wohlmann Andreas , Mohta Vrinda , Junker Kerstin , Friedrich Karlheinz 
Published: May 2015

Abstract
The tumor microenvironment plays an important role in cancer progression. In particular, stromal fibroblasts have profound influence on migration and invasiveness of tumor cells. We employ bladder carcinoma cell lines of different de-differentiation grade to pursue the hypothesis that tumor cell malignancy correlates with the cells' potential to recruit support by tumor associated fibroblasts (TAFs).
Bladder cancer cell lines RT112 and Cal29 were used as models for tumor cells of low or high de-differentiation grade, respectively. They were grown separately or in co-culture with tumor-associated fibroblast isolated from kidney carcinomas. Migratory and invasive cell properties were quantified by Boyden chamber (Matrigel) and wound healing assays. Cytokine production of cells was measured by intracellular staining followed by cytometry, by semi-quantitative PCR, or by subjecting conditioned medium to ELISA and Western blot analysis.
Migration and invasiveness of bladder cancer cell lines Cal29 and RT112 was enhanced by co-cultivation with TAFs. The low grade cell line RT112, however, was much more dependent on TAF-derived signals to increase the malignancy parameters. Conditioned medium from tumor cells induced the secretion of Matrix Metalloproteinases (MMPs) and the release of GM-CSF, HGF, and Monocyte Chemotactic Protein (MCP-1, CCL2) by TAFs. We could identify tumor cell-derived IL-1α and β as well as TGF-β as stimuli responsible for the upregulation of GM-CSF and MCP-1, respectively, in fibroblasts. MCP-1 is implicated in inflammatory disease states characterized by monocytic infiltrates and was shown to promote cell migration of both bladder cancer cell lines under investigation.
Employing pathway-specific reporter gene constructs we are currently addressing signal transduction reactions in both tumor cells and fibroblasts underlying this crosstalk.
Supported by: IZKF Jena, European Regional Development Fund ERDF.
Citation Format: Susanne Grimm, Susanne Jennek, Jens Bratsch, Andreas Wohlmann, Vrinda Mohta, Kerstin Junker, Karlheinz Friedrich. Malignancy of bladder cancer cells is enhanced by tumor associated fibroblasts through a cytokine-chemokine loop. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3597. doi:10.1158/1538-7445.AM2014-3597
Abstract 3609: Fibroblast influence on keratinocyte tumorigenic and metastatic potential in cutaneous squamous cell carcinoma
Authors: Gwilliam J Curtis , CVitan Alexander , Watson Nicholas , Falkenberg Lauren , Yousefi Nyousha , Alexander Hepzibha , Morin Adam , Wiest Jonathan S. , DeCicco-Skinner Katie L. 
Published: May 2015

Abstract
Skin cancer is the most common form of cancer in the United States, with an estimated two million cases diagnosed annually. Tpl2 (Map3k8) is a serine threonine protein kinase in the mitogen-activated protein kinase (MAPK) signal transduction cascade. Tpl2 was identified by our laboratory as having a tumor suppressive function in skin carcinogenesis, with Tpl2 knockout mice developing significantly more tumors than matched wildtype controls and Tpl2 knockout keratinocytes having greater angiogenic and metastatic potential. However, the microenvironmental contributions of underlying fibroblasts to the cancerous behavior seen in Tpl2 knockout keratinocytes has yet to be characterized. Therefore, we compared the tumorigenic, invasive, and metastatic potential of H-ras infected keratinocytes from wildtype and Tpl2 knockout mice with H-ras infected keratinocytes co-cultured with fibroblasts from wildtype and Tpl2 knockout mice. Using various techniques, including protein microarray, real-time polymerase chain reaction and Western blotting we found that fibroblasts in the microenvironment of H-ras-infected Tpl2 knockout keratinocytes contribute to an up-regulation of markers involved in growth and progression of squamous cell carcinoma. Additionally, Tpl2 knockout keratinocytes cultured in fibroblast conditioned media (FCM) have higher malignant conversion rates, colony formation and increased angiogenesis compared to co-cultured wildtype keratinocytes. In summary, our studies demonstrate that fibroblasts in the tumor microenvironment of Tpl2 knockout mice play a major role in the more aggressive,invasive, and metastatic character of knockout tumors than is seen in control animals.
Citation Format: J Curtis Gwilliam, Alexander CVitan, Nicholas Watson, Lauren Falkenberg, Nyousha Yousefi, Hepzibha Alexander, Adam Morin, Jonathan S. Wiest, Katie L. DeCicco-Skinner. Fibroblast influence on keratinocyte tumorigenic and metastatic potential in cutaneous squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3609. doi:10.1158/1538-7445.AM2014-3609
Abstract 3616: Hypoxia determines the fate of myeloid cell differentiation by controlling STAT3 activation in tumor site
Authors: Kumar Vinit 
Published: May 2015

Abstract
The abnormal differentiation of myeloid cells is the key element of altered immune response in cancer. Myeloid derived suppressor cells (MDSC) are a major component of immune suppressive network. MDSC accumulate in large numbers in peripheral lymphoid organs and inside tumors. Recent data indicated that the fate of MDSC depends on the localization of these cells. In contrast to peripheral lymphoid organs, inside tumors, MDSC rapidly differentiate to tumor associated macrophages with potent immune suppressive activity. Hypoxia is intricate part of tumor microenvironment. We have found that hypoxia plays a major role in regulation of MDSC differentiation to macrophages in tumor site. Hypoxia caused dramatic reduction in phosphorylated STAT3 in MDSC. These results were unexpected, since high level of STAT3 activity is a hallmark of MDSC present in blood and lymphoid organs. In our experiments spleen MDSC in tumor-bearing mice had much higher level of STAT3 phosphorylation than their counterparts in tumor site from the same mice. In cancer patients MDSC in tumor site also had substantially lower level of pSTAT3 than MDSC in peripheral blood. In vitro culture of spleen MDSC in hypoxia led to the suppression of STAT3 activation. Furthermore, mice with constitutively active STAT3 had shown the dramatic expansion of MDSC and down-regulation of macrophage population in tumor microenvironment, suggesting that STAT3 is a downstream target for hypoxia driven myeloid cells differentiation in cancer. We found that CD45 protein tyrosine phosphatase was a molecule that regulated STAT3 activation in hypoxia. The activity but not expression of CD45 phosphatase was up-regulated in tumor MDSC as compared to MDSC in spleens. The exposure of spleen MDSC to hypoxia led to the significant increase in CD45 activity. The siRNA mediated silencing of CD45 abrogated the effect of hypoxia on the down-regulation of STAT3 activity in MDSC. This suggests that hypoxia driven up-regulation of CD45 phosphatase activity suppresses STAT3 activation, which in turn plays a key role in differentiation of myeloid cells in tumor site. These unexpected findings may provide rationale for re-evaluation of therapeutic strategies targeting myeloid cells in cancer.
Citation Format: Vinit Kumar. Hypoxia determines the fate of myeloid cell differentiation by controlling STAT3 activation in tumor site. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3616. doi:10.1158/1538-7445.AM2014-3616
Abstract 3618: Ikkα modulates primary sclerosing cholangitis and intrahepatic cholangiocarcinoma
Authors: Jiang Qun , Xiao Zuoxiang , Back Timothy , Scarzello Anthony , Roan Scott , Willette-Brown Jami , Zhu Feng , Hu Yinling , Wiltrout Robert H. 
Published: May 2015

Abstract
Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer after hepatocellular carcinoma (HCC). The risk of ICC is higher in patients with primary sclerosing cholangitis (PSC). To date, the cellular and molecular mechanism underlying the pathological progression of PSC and ICC is poorly understood. IKKα is part of the IκB kinase (IKK) complex, which plays an important role in regulating inflammation- associated carcinogenesis through both NF-κB-dependent and independent pathways. Here, we show that IKKα mutant mice developed very serious PSC as early as four weeks of age. The ALT/AST and bilirubin levels were significantly increased in the serum of IKKα mutant mice along with lymphocytic and eosinophilic infiltration into the liver. Liver inflammation in the IKKα mutant mice, mediated by macrophages, neutrophils and CD4+ T cells, was associated with the death of cholangiocytes and hepatocytes, and obstruction of intrahepatic and extrahepatic bile ducts, which impeded bile flow and ultimately led to biliary fibrosis and cirrhosis. Additionally, upon activation of NOTCH signaling in the liver via hydrodynamic shear, we observed that NOTCH-induced ICC, with the PSC, developed significantly faster in IKKα mutant mice. To identify whether intrinsic IKKα dysfunction in hepatocytes promotes the NOTCH-induced ICC in IKKα mutant mice, we generated IKKα hep KO mice, in which IKKα is conditionally knocked out in hepatocytes. No biliary disease or liver injury was observed in these mice. We then established an accelerated ICC model utilizing hydrodynamic delivery of NICD and AKT expression vectors. Unexpectedly, ICC development was remarkably slower in the IKKα hep KO mice compared to the IKKα floxed control mice, and this delayed ICC development was associated with reduced activation or levels of AKT, NOTCH, MAPK/ERK and c-Myc. These data suggest that IKKα may play a protective role in PSC, while promote ICC derived from hepatocytes. In contrast to ICC, we also established the cMET/β-catenin-induced HCC model in the IKKα hep KO mice, which did not reveal any difference in tumor development between IKKα hep KO and control mice. Taken together, our findings suggest that IKKα plays complicated and important roles in PSC and ICC pathological progression.
Citation Format: Qun Jiang, Zuoxiang Xiao, Timothy Back, Anthony Scarzello, Scott Roan, Jami Willette-Brown, Feng Zhu, Yinling Hu, Robert H. Wiltrout. Ikkα modulates primary sclerosing cholangitis and intrahepatic cholangiocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3618. doi:10.1158/1538-7445.AM2014-3618
Abstract 3620: Neuropeptide signaling activates Type-1 immunity through the NK1 and NK2 receptors on human dendritic cells
Authors: Kitamura Hidemitsu , Ohtake Junya , Kaneumi Shun , Masuko Kazutaka , Sumida Kentaro , Terada Satoshi , Kishikawa Takuto , Ono Yosuke , Kita Toshiyuki , Kobayashi Hiroya 
Published: May 2015

Abstract
Dendritic cells (DCs), representative antigen-presenting cells, effectively induce antigen-specific immune responses through activation of CD4+T and CD8+T cells. In tumor microenvironments, DCs uptake tumor-derived antigens, and generate helper T cells and cytotoxic T lymphocytes (CTLs), which recognize and kill the target tumor cells in response to the antigens. Thus, proper regulation of DC function is important for tumor immunology.
Substance P (SP) and Neurokinin A (NKA), neurotransmitters, are widely distributed in both central and peripheral nervous system. Recently, we demonstrated that IFN-γ remarkably induced neurokinin-2 receptor (NK2R) expression human monocyte-derived dendritic cells. These findings strongly suggested that neuropeptide-signaling cascade might be closely related with regulation of DC-mediated immune responses. To elucidate the precise role of such neuro-immune crosstalk, we further investigated the effect of neuropeptide signaling on function of human DCs.
At first, we found that both neurokinin-1 receptor (NK2R) and NK2R mRNA expressions were significantly enhanced by IFN-β, IFN-γ, LPS, or poly I:C stimulation with human DCs generated from peripheral blood mononuclear cells (PBMCs). In addition, we confirmed that the upregulation of NK1R and NK2R mRNA expressions was induced in a STAT-1-dependent manner. Surface expression levels of HLA-DR and costimulatory molecules on DCs, augmented by poly I:C, were modulated in the presence of specific inhibitors against NK1R or NK2R. On the other hand, we confirmed that blockade of NK1R- and NK2R-mediated signaling cascade significantly suppressed cytokine productions by antigen-specific CD4+ T cells after the antigen stimulation with DCs. Finally, we found that human DCs also enhanced expression level of TAC-1 gene, which encodes SP and NKA, after the IFN-γ stimulation.
Thus, these findings indicate that NK1R- and NK2R-dependent neuropeptide signaling regulate Type-1 immunity through the activation of DC function, suggesting that such neuro-immune cross-talk through SP-NK1R and NKA-NK2R cascade might be involved in various diseases caused by excessive Type-1-dominant immunity including cancer and infection with chronic inflammation.
Citation Format: Hidemitsu Kitamura, Junya Ohtake, Shun Kaneumi, Kazutaka Masuko, Kentaro Sumida, Satoshi Terada, Takuto Kishikawa, Yosuke Ono, Toshiyuki Kita, Hiroya Kobayashi. Neuropeptide signaling activates Type-1 immunity through the NK1 and NK2 receptors on human dendritic cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3620. doi:10.1158/1538-7445.AM2014-3620
Abstract 3622: Total loss of HLA class I expression by two sarcomatoid hepatocellular carcinoma cell lines sHCC29 and sHCC63 was caused by a ∼49-kbp deletion at chromosome 15q15 across the β2m gene locus
Authors: Lei Wei-Yi , Hsieh Chin-Hsuan , Chang Chien-Chung , Wen Shao-Hsuan , Hu Chi-Tan , Liao Shuen-Kuei 
Published: May 2015

Abstract
Scanty information is available concerning the abnormalities in HLA class I antigen presentation in sarcomatoid hepatocellular carcinoma (sHCC), although this information can improve our understanding of sHCC-host interactions and assist in a rational design of T cell immunotherapy. In this study, we have established three sarcomatoid HCC cell lines, namely sHCC29, sHCC63, and sHCC74, from the tumor lesions in three patients; two of them were HBV carriers from which sHCC29 and sHCC63 were derived, while the other was a HCV carrier from which sHCC74 was generated. No viral DNA was detected in the culture supernatants or cell extracts from any of the three HCC cell lines by COBAS AmpliPre/TaqMan qPCR. In monolayer cell culture, both sHCC29 and sHCC74 showed similar elongated fibroblastic morphology, while sHCC63 exhibited a short-spindle shape and expanded in a density-sensitive manner. HLA class I molecules were abundantly expressed on sHCC74, but were not detected on sHCC29 and sHCC63 by cytofluorometric analysis. Cytoplasmic β2-microglobulin (β2m) was also undetected in the latter two cell lines, in spite of their baseline level of low-moderate cytoplasmic HLA class I heavy chain expression, which was 33-50% of that expressed by sHCC74 in terms of % positive cells. The observed HLA class I-loss phenotype was IFN-γ (300 μg/ml, 48 h)-uncorrectable. Inactivation of the β2m genes in both sHCC29 and sHCC63 was supported by the failure to detect their β2m mRNA transcripts and by the identification of a large (∼49,187 bp) deletion across the β2m gene locus at chromosome 15q15. This deletion, the largest reported to date, explains the loss of β2m protein and lack of assembly of HLA class I molecules in the sHCC29 and sHCC63 cell lines. Mapping of the deletion breaking point and validation by immunohistochemistry of β2m loss in the corresponding tumor lesions from which sHCC29 and sHCC63 were derived are currently underway.
Citation Format: Wei-Yi Lei, Chin-Hsuan Hsieh, Chien-Chung Chang, Shao-Hsuan Wen, Chi-Tan Hu, Shuen-Kuei Liao. Total loss of HLA class I expression by two sarcomatoid hepatocellular carcinoma cell lines sHCC29 and sHCC63 was caused by a ∼49-kbp deletion at chromosome 15q15 across the β2m gene locus. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3622. doi:10.1158/1538-7445.AM2014-3622
Abstract 3634: Postoperative neutrophil-lymphocyte ratio as a surrogate marker for recurrence of gastric cancer
Authors: Tanaka Hiroaki , Muguruma Kazuya , Sakurai Katsunobu , Toyokawa Takahiro , Kubo Naoshi , Ohira Masaichi , Hirakawa Kosei 
Published: May 2015

Abstract
Tumor markers such as carcinoembryonic antigen (CEA) and CA 19-9 are both widely used in the follow up of patients with gastric cancer. However, there are no definitive predictive markers for postopetrative recurrence. It has been reported that preoperative neutrophil lymphocyte ratio (NLR) was associated with prognosis of patients with gastrointestinal cancer. The aim of this study was to examine the impact of postoperative NLR on prediction for postoperative recurrence of gastric cancer. We retrospective reviewed data from 133 patients underwent curative surgical resection for gastric cancer between 2006 and 2010. Patients treated with preoperative chemotherapy or multiple cancers were excluded. We found 52 recurrent diseases which mainly included 29 cases with peritoneal dissemination, 13 with distant lymph node metastasis and 7 with liver metastasis. In univariate analysis, histological diffuse type, mascroscopical type 4 cancer, regional lymph node metastasis, lymphatic invasion in primary tumor, tumor diameter, elevated CEA or CA19-9, and elevated NLR. By ultivariate analysis, we found that elevated CA19-9 and NLR were independent predictive markers. Increased NLR was associated with diffuse type, peritoneal ddissemination, elevated CEA and preoperative NLR but not the presence or absence of adjuvant chemotherapy. In conclusion, postoperative NLR might be one of the surrogate markers for recurrence after curative surgery for gastric cancer.
Citation Format: Hiroaki Tanaka, Kazuya Muguruma, Katsunobu Sakurai, Takahiro Toyokawa, Naoshi Kubo, Masaichi Ohira, Kosei Hirakawa. Postoperative neutrophil-lymphocyte ratio as a surrogate marker for recurrence of gastric cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3634. doi:10.1158/1538-7445.AM2014-3634
Abstract 3638: Vaccine-draining lymph nodes of cancer patients for generating anticancer antibodies
Authors: Shukla Girja S. , Pero Stephanie C. , Sun Yu-jing , Carman Chelsea L. , Olson Walter C. , Slingluff Craig L. , Krag David N. 
Published: May 2015

Abstract
Human vaccine studies regularly demonstrate vaccine-induced antibodies in the blood. Characterization of these antibodies shows that they exhibit an extensive range of bioactive mechanisms. Unfortunately, there is not yet a reliable method by which to produce these important antibodies. Human antibodies may not require any molecular modification for therapeutic application and have a much faster pipeline to clinical studies than animal-derived antibodies. Lymph nodes are the primary destination of tumor or vaccine antigens. In the node, B cells undergo clonal expansion and somatic hypermutation, leading to the affinity-matured populations of effector B cells secreting antibodies that bind to the tumor or vaccine. The cancer vaccine-draining node is the ideal source of B cells that produce anticancer antibodies. Despite multiple human cancer vaccine studies, very little research has been done to recover the B cells responsible for vaccine-induced anticancer antibodies. In the present study, we used mononuclear cells from surgically removed vaccine-draining lymph nodes of melanoma patients vaccinated with 6 melanoma peptides derived from cancer-testis antigens and from melanocytic differentiation proteins for generating anti-vaccine peptide antibodies. The lymph node draining the vaccination site was localized by lymphatic mapping with a radiotracer. The development and maturity of B cells were assessed by determining phenotypic characters of lymph node cells using multicolor flow cytometry. The results showed that these vaccine-draining nodes contain high numbers of class-switched (CD19+CD27+IgD-IgM-) B cells and plasmablasts (CD19+CD38+IgM-). B-cell ELISpot assay was used to quantify the proportion of B cells in vaccine-draining lymph nodes that secrete anti-melanoma peptide antibodies. Positive ELISpot responses were observed in patients who also showed serum antibody-reactivity towards the vaccine peptides. The identification of lymph node cell samples exhibiting strong B-cell responses allows efficient generation and screening of hybridomas that secrete antibodies against cancer vaccine antigens. This study establishes a step-wise protocol for generating anti-cancer antibodies from vaccine-draining lymph nodes. We anticipate that the ability to reliably generate in vitro the same antibodies observed in the blood of vaccinated patients will further stimulate research to understand mechanisms of human antibody activity.
Citation Format: Girja S. Shukla, Stephanie C. Pero, Yu-jing Sun, Chelsea L. Carman, Walter C. Olson, Craig L. Slingluff, David N. Krag. Vaccine-draining lymph nodes of cancer patients for generating anticancer antibodies. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3638. doi:10.1158/1538-7445.AM2014-3638
Abstract 364: Mechanical properties of growing melanocytic nevi and the progression to melanoma
Authors: Taloni Alessandro , Alemi Alexander A. , Ciusani Emilio , Sethna James , Zaperi Stefano , Laporta Caterina Anna Maria 
Published: May 2015

Abstract
Melanocytic nevi are benign proliferations that sometimes turn into malignant melanoma in a way that is still unclear from the biochemical and genetic point of view. Diagnostic and prognostic tools are then mostly based on dermoscopic examination and morphological analysis of histological tissues. To investigate the role of mechanics and geometry in the morpholgical dynamics of melanocytic nevi, we study a computation model for cell proliferation in a layered non-linear elastic tissue. Numerical simulations show that the morphology of the nevus is correlated to the initial location of the proliferating cell starting the growth process and to the mechanical properties of the tissue. Our results also show that melanocytes are subject to compressive stresses that fluctuate widely in the nevus and depend on the growth stage. This result is important because mechanical stress has an effect that differs for primary and metastatic melanoma. We show experimentally that osmotic stress and collagen inhibit growth in primary melanoma cells while the effect is much weaker in metastatic cells. Knowing that morphological features of nevi might also reflect geometry and mechanics rather than malignancy could be relevant for diagnostic purposes.
Citation Format: Alessandro Taloni, Alexander A. Alemi, Emilio Ciusani, James Sethna, Stefano Zaperi, Caterina Anna Maria Laporta. Mechanical properties of growing melanocytic nevi and the progression to melanoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 364. doi:10.1158/1538-7445.AM2014-364