bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 316: Development of a novel 3D cancer model by cultivation of malignant effusions in a 3D cell culture system
Authors: Amann Arno , Zwierzina Marit , Huber Julia M. , Gamerith Gabriele , Koeck Stefan , Bitsche Mario , Lorenz Edith , Zwierzina Heinz 
Published: August 2015

Abstract
Introduction: Metastatic spread and the occurrence of tumour cell resistance to therapeutic interventions can be induced by different molecular processes. Among those, tumour stroma interactions play a crucial role. So far only few cell culture models exist to study these cell processes.
Therefore, we describe the development of a hanging drop technology for 3D culture of primary cells isolated from malignant effusion obtained from cancer patients.
Methods: A 96-well hanging drop microtiter plate (InSphero AG, Zürich, Switzerland) was applied for the 3D cultivation of cells. Primary cells were isolated either from malignant ascites or pleura effusion of cancer patients. Cells were seeded either into 2D cell culture plates or into hanging drops for an incubation period of ten days. Apart from conventional histology (H&E), protein expression was displayed immunohistochemically (IHC) and by flow cytometry. The applied antibodies displayed the expression of e-cadherin, vimentin, Ki67, CD31, CD45, Epcam, Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5) and α-smooth muscle actin (α-SMA). In addition, viability of all samples was determined by an AnnexinV/Propidium Iodide staining using flow cytometry.
Results: Microtissue aggregation differed significantly depending on whether the malignant cell effusions displayed expression of Epcam. Effusions consisting of a high number of Epcam positive cells formed solid and viable microtissues. The viability of microtissues remained high (&gt;80%) similar to cells cultivated in 2D cell culture plates. Tumour cells, defined as e-cadherin and Epcam positive by flow cytometry were still detected in microtissues after ten days of incubation in 3D.
Furthermore, endothelial cells, defined as CD31 and Lgr5 positive could also be discriminated in microtissues.
Conclusion: We demonstrate that our method is a promising tool for the generation of tumour microtissues consisting of primary isolated cells from malignant effusions. Therefore, our technology represents a promising method to investigate tumour stroma interactions in an in vitro system that reflects more closely the in vivo environment.
Citation Format: Arno Amann, Marit Zwierzina, Julia M. Huber, Gabriele Gamerith, Stefan Koeck, Mario Bitsche, Edith Lorenz, Heinz Zwierzina. Development of a novel 3D cancer model by cultivation of malignant effusions in a 3D cell culture system. [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 316. doi:10.1158/1538-7445.AM2015-316
Abstract 3166: The role of CD4 T cells in murine model of NASH-promoted HCC
Authors: Ma Chi , Felsher Dean , Greten Tim 
Published: August 2015

Abstract
Aim: Non-alcoholic steatohepatitis (NASH), a common condition in obese patients, is an important risk factor for hepatocellular carcinoma (HCC). However, only little information exists about the immunological mechanisms of how NASH promotes HCC. This study was designed to better understand the role of adaptive immunity during the process of NASH-promoted hepatocarcinogenesis.
Experimental procedure: Methionine choline deficient (MCD) diet is widely used to induce NASH. In this study we fed Lap-tTA-myc mice, which develop HCC, with MCD diet. Choline-deficient and amino acid-defined (CDAA) diet and high fat (HF) diet was also used to induce NASH in B6 mice. GK1.5 antibody was used to deplete CD4 T cells. Gas chromatography-mass spectrometry was performed to identify free fatty acids. Difference immune cell subsets were identified by flow cytometry. Cell death was measured by 7AAD/Annexin V staining.
Results: MCD diet accelerated tumor development in MYC-on mice. A robust selective decrease of hepatic CD4 but not CD8 T cells was observed in NASH mice, which was exacerbated by MYC transgene expression. The selective decrease of hepatic CD4 T cells was also observed in B6 mice treated with CDAA or HF diet suggesting it's a general phenomena in NASH. As expected higher level of cell death was detected in hepatic CD4 T cells after NASH. Phenotypic analysis demonstrated that these CD4 T cells had a CD44highCD62Llow effector memory phenotype and produced more IFNγ. Selective CD4 T cell depletion further accelerated HCC development suggesting an important role of these cells for tumor development. Next, we studied the cause of CD4 T cell death. In co-culture experiments we demonstrated that lipid-laden hepatocytes from mice on MCD diet induced selective CD4 but not CD8 T cells death through a contact-independent mechanism. FFAs were found from hepatocyte culture medium to be responsible for T cell cytotoxicity, and linoleic acid recapitulates the selective toxicity to CD4 T cells. Blocking ROS abolished linoleic acid-induced CD4 T cell death. In vivo study showed that N-acetyl cysteine reversed the NASH-induced hepatic CD4 T cell decrease and delayed NASH-promoted tumor development.
Conclusion: Our results suggest the critical role of CD4 T cells in the disease progression of NASH to HCC, and provide a new link between lipid metabolism dysfunction with impaired anti-tumor surveillance.
Citation Format: Chi Ma, Dean Felsher, Tim Greten. The role of CD4 T cells in murine model of NASH-promoted HCC. [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 3166. doi:10.1158/1538-7445.AM2015-3166
Abstract 3172: CCL9 induction in myeloid cells of the premetastatic niche enhances tumor cell survival and metastatic colonization
Authors: Yan Hannah HY , Pang Yanli , Hunter Kent , Khanna Chand , Yang Li 
Published: August 2015

Abstract
Tumor cell survival in the hostile distant organ is a rate-limiting step in cancer metastasis. Bone marrow-derived myeloid cells can form a premetastatic niche and provide a tumor-promoting microenvironment. However, it is unclear whether these myeloid cells in the premetastatic site have any direct effect on tumor cell survival. Here we report that chemokine CCL9 was highly induced in Gr-1+CD11b+ immature myeloid cells and in premetastatic lung of mice bearing 4T1 mammary tumor and B16 melanoma. Knockdown of CCL9 in myeloid cells decreased tumor cell survival and metastasis. The expression level of CCL23, the human orthologue for CCL9, in peripheral blood mononuclear cells correlated with progression and survival of cancer patients. Our studies identify CCL9 as a key pro-survival factor for cancer cells in the premetastatic organ.
Interestingly, the expression of CCL9 in myeloid cells is regulated by TGF-β signaling as overexpression of CCL9 in myeloid cells lacking TGF-β signaling rescued the tumor metastasis defect observed in mice with myeloid-specific deletion of TGF-β signaling. These data together with our previous report of a fundamental role of myeloid TGF-β signaling in host compromised immune surveillance (Cancer Discovery 2013); demonstrate that myeloid-specific TGF-β signaling constitutes an essential component of the metastasis-promoting puzzle of TGF-β. This finding is different from TGF-β signaling in fibroblasts and a variety of epithelial cells, as well as T cells in which the TGF-β signaling functions as a tumor suppressor. This new understanding of TGF-β at the interface of tumor and host may provide insight into how TGF-β switches from a tumor suppressor to a tumor promoter, a long-standing challenge in cancer biology field. Our study demonstrates that CCL9 could serve as a good candidate for anti-metastasis treatment by targeting the rate-limiting step of cancer cell survival. Additionally, targeting CCL9 may avoid the adverse effects of TGF-β-targeted therapy.
Citation Format: Hannah HY Yan, Yanli Pang, Kent Hunter, Chand Khanna, Li Yang. CCL9 induction in myeloid cells of the premetastatic niche enhances tumor cell survival and metastatic colonization. [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 3172. doi:10.1158/1538-7445.AM2015-3172
Abstract 3174: Retinoblastoma protein 1 implication in the monocytic - granulocytic transition
Authors: Mastio Jerome , Condamine Thomas , Gabrilovich Dmitry 
Published: August 2015

Abstract
Alterations in the myeloipoiesis during cancer lead to production of myeloid-derived suppressor cells (MDSC). MDSC are comprised of two subsets: monocytic MDSC (M-MDSC) and polymorphonuclear MDSC (PMN-MDSC). Both subsets contribute to the immunosuppressive tumor microenvironment. Therefore elimination of MDSC could potentially improve the clinical success of cancer immune therapy. Direct targeting of MDSC progenitors is an attractive possibility. Our laboratory has recently discovered that M-MDSC from the bone-marrow can give rise to PMN-MDSC and that this phenomenon is mediated by the loss of retinoblastoma protein 1 (Rb1) expression. However, a full phenotypic and molecular characterization of these MDSC progenitors has not been established. To identify the subpopulation able to give rise to PMN-MDSC within the M-MDSC population, we used Rb1-GFP reporter mice to track Rb1 expression in vivo. We found that Rb1 low M-MDSC express the stem cell marker c-Kit and low levels of CD115. These Rb1 low c-Kit + M-MDSC are substantially expanded in tumor-bearing mice. Interestingly, this population is also present in naive mice, albeit at very low frequency. Rb1 low c-Kit + monocytic cells from naïve and tumor bearing mice can give rise to granulocytic cells in vitro. These preliminary results indicate that these cells are indeed granulocytic progenitor and are expanded during tumor development. Although this monocyte-granulocyte conversion is associated with a decrease in Rb1 expression, it seems that a minimal Rb1 level is required to ensure this process, at least in naïve mice. Further characterization of the phenotype and molecular profiles of this new MDSC progenitor could lead to the development of new ways to target MDSC.
Citation Format: Jerome Mastio, Thomas Condamine, Dmitry Gabrilovich. Retinoblastoma protein 1 implication in the monocytic - granulocytic transition. [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 3174. doi:10.1158/1538-7445.AM2015-3174
Abstract 3175: CD38-expressing myeloid-derived suppressor cells promote tumor growth in a murine model of esophageal cancer
Authors: Karakasheva Tatiana A. , Waldron Todd J. , Eruslanov Evgeniy , Lee Ju-Seog , O'Brien Shaun , Basu Devraj , Singhal Sunil , Malavasi Fabio , Rustgi Anil K. 
Published: August 2015

Abstract
Myeloid derived suppressor cells (MDSCs) are an immunosuppressive population of immature myeloid cells found in advanced stage cancer patients and mouse tumor models. MDSCs suppress T cell-mediated tumor clearance by production of inducible nitric oxide synthase (iNOS) and arginase, as well as through other mechanisms. The purpose of this study was to identify the factor(s) contributing to the pro-tumorigenic role of MDSCs in esophageal squamous cell carcinoma (ESCC).
We have used an unbiased approach to search for changes in global gene expression profile of MDSCs derived from conditional p120-catenin knockout mice (L2-cre;p120ctnf/f, a model of oral-esophageal cancer; Stairs D.B., et al Cancer Cell 2011), compared to immature myeloid cells from age-matched wild-type mice. We have detected a dramatic increase in expression of the Cd38 gene in MDSCs, compared to control cells. CD38 belongs to the ADP-ribosyl cyclase family and possesses both ectoenzyme and receptor functions. It has been described to function in lymphoid and early myeloid cell differentiation, cell activation and neutrophil chemotaxis. We find that CD38 expression in MDSCs is evident in several mouse tumor models, and CD38high MDSCs are more immature than MDSCs lacking CD38 expression, suggesting a potential role for CD38 in the maturation halt found in MDSC populations. CD38high MDSCs also possess a greater capacity to suppress activated T cells, and promote tumor growth to a greater degree than CD38low MDSCs, likely as a result of increased iNOS production. We have identified novel tumor-derived factors, including IL-6, IGFBP-3 and CXCL16, which induce CD38 expression by MDSCs ex vivo. Finally, we have demonstrated that monoclonal antibody-mediated targeting of CD38 results in suppressed tumor growth in vivo in a subcutaneous transplant model of ESCC.
In conclusion, we have shown that expression of CD38 plays an important role in MDSC biology by promoting an enhanced immunosuppressive capacity and a more immature phenotype in vivo. We have also detected an expansion of CD38-positive MDSCs in peripheral blood of advanced stage cancer patients and propose targeting CD38 in vivo as an approach to cancer therapy.
Supported by NCI P01CA098101.
Citation Format: Tatiana A. Karakasheva, Todd J. Waldron, Evgeniy Eruslanov, Ju-Seog Lee, Shaun O'Brien, Devraj Basu, Sunil Singhal, Fabio Malavasi, Anil K. Rustgi. CD38-expressing myeloid-derived suppressor cells promote tumor growth in a murine model of esophageal 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 3175. doi:10.1158/1538-7445.AM2015-3175
Abstract 3176: Linking suppressive activity and ER-Stress in Myeloid Derived Suppressor Cells
Authors: Condamine Thomas C. , Kumar Vinit , Gabrilovich Dmitry I. 
Published: August 2015

Abstract
In recent years, immunotherapies have shown great promise for the treatment of cancer patients. However, the presence of a very potent immunosuppressive microenvironment in the tumor site remains one of the major obstacles in fully harnessing the potential of immunotherapeutic strategies. Myeloid Derived Suppressor cells (MDSC) represent one of the main suppressive populations inside the tumor. These cells can be divided in two sub-populations: PMN-MDSC and Monocytic-MDSC. They suppress immune response using various well-known mechanisms and accumulate to a large extent in tumor-bearing hosts. Despite the increasing body of literature on MDSC, which mechanisms drives their suppressive capacity remains unclear. Our group recently reported that the ER-Stress response is activated in MDSC isolated from tumor bearing mice and cancer patients. In this study, we investigated the possibility that the ER-stress response could be responsible for the immune suppressive capacities of MDSC. To test this hypothesis, we induce ER-Stress using well-known chemical inducer into non suppressive myeloid cells (PMN and monocyte) and assess their ability to suppress T cells function. Our results show that, following ER-Stress induction, normal myeloid cells were able to suppress T cells response (in murine and human system). To better understand how the ER-Stress response can induce suppressive capacities, we performed microarray analysis following treatment with ER-Stress inducer. Interestingly, we found that arginase-1, one the major enzyme implicated in MDSC suppressive capacities, was induced by more than 5 fold. Altogether, our results suggest that activation of the ER-Stress response in MDSC could be responsible for the up-regulation of arginase-1 and the suppressive phenotype of MDSC. These results suggest that targeting the ER-Stress response in cancer patients could inhibit the suppressive activity of MDSC and improve the efficacy of current immunotherapies.
Citation Format: Thomas C. Condamine, Vinit Kumar, Dmitry I. Gabrilovich. Linking suppressive activity and ER-Stress in Myeloid Derived Suppressor 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 3176. doi:10.1158/1538-7445.AM2015-3176
Abstract 3185: Complement C5 promotes male bias of pancreatic neuroendocrine tumor metastasis
Authors: Contractor Tanupriya , Chan Chang , Kobayashi Shinta , Clausen Richard , Sun Yvonne , da Silva Edaise , Vosburgh Evan , Levine Arnold J. , Tang Laura , Harris Chris R. 
Published: August 2015

Abstract
Among patients with neuroendocrine tumors, males are more likely than females to develop liver metastatic disease. We show that this male bias toward liver metastasis is also observed in a mouse model for pancreatic neuroendocrine tumorigenesis (PanNETs). Interestingly, primary tumors from mice with metastatic disease display hepatomimicry, expressing genes normally expressed in liver. Both male and female primary tumors from mice with metastatic disease show hepatomimicry, although one of the liver genes, complement C5, is only induced in males. In mice knocked out for the complement C5 gene, the frequency of liver metastasis dropped significantly; moreover, males and females lacking a gene for complement C5 developed liver metastasis at the same frequency. Liver metastasis was also prevented by PMX53, a small molecule antagonist of C5aR1/CD88, which is the receptor for complement C5a. Thus complement C5 is epistatic to metastasis and promotes gender bias of liver metastatic disease.
Citation Format: Tanupriya Contractor, Chang Chan, Shinta Kobayashi, Richard Clausen, Yvonne Sun, Edaise da Silva, Evan Vosburgh, Arnold J. Levine, Laura Tang, Chris R. Harris. Complement C5 promotes male bias of pancreatic neuroendocrine tumor 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 3185. doi:10.1158/1538-7445.AM2015-3185
Abstract 32: The role of Lrig1 signaling in mammary gland development and tumorigenesis
Authors: Simion Catalina , Chen Qian J. , Wilkerson Charles L. , Rafidi Hanine , Borowsky Alexander D. , Sweeney Colleen 
Published: August 2015

Abstract
Lrig1 is a transmembrane leucine-rich repeat protein and a negative regulator of oncogenic receptor tyrosine kinases (RTKs) such as the ErbB family members. The Sweeney lab recently demonstrated that under-expression of Lrig1 predicts poor prognosis in breast cancer. Breast cancer patients with high Lrig1 expression show significantly longer relapse-free survival, identifying Lrig1 as a new prognostic marker. Recently, Lrig1 was found to be a tumor suppressor in intestinal tissue. It was shown that Lrig1 null mice develop ErbB receptor-dependent duodenal adenomas. Based on Lrig1's function as a negative regulator of RTKs and its reported tumor suppressor activity in intestinal epithelium, we hypothesized that Lrig1 null mice would be susceptible to mammary tumorigenesis. To characterize the function of Lrig1 in the mammary gland, we compared mammary development in Lrig1 wild type (+/+) and null (-/-) mice at regular intervals from 4 to 12 weeks. Mice were screened for evidence of hyperplasia, pre-malignant lesions or tumors by whole mount analysis across age and the intensity of specific proteins was surveyed by western blot. Organotypic growth of the mammary epithelial cells from wild type and null mice was examined in three-dimensional cell culture to examine whether any phenotypes are inherent to the epithelium. Since Lrig1 has been defined as a stem cell marker in the intestine, its role in stem cell maintenance was determined by examining the impact of Lrig1 loss on mammary stem cells. Although these studies are ongoing, early results indicate an increase in the amount of Lrig1-target proteins by western blot in the Lrig1-/- mice as opposed to the Lrig1+/+ mice mammary gland tissue. Epithelium from Lrig1-/- mice present an exaggerated branching response when stimulated with TGFα in comparison to Lrig1+/+ epithelium. Our studies also indicate that Lrig1-/- mammary glands are enriched in stem cells when compared to Lrig1+/+ mammary glands. In addition, a number of proliferative lesions were encountered in the mammary gland whole mounts of the Lrig1-/- mice, but not that of the Lrig1+/+ mice, which strongly suggests that Lrig1 plays a growth suppressor role in the mammary gland.
Citation Format: Catalina Simion, Qian J. Chen, Charles L. Wilkerson, Hanine Rafidi, Alexander D. Borowsky, Colleen Sweeney. The role of Lrig1 signaling in mammary gland development and tumorigenesis. [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 32. doi:10.1158/1538-7445.AM2015-32
Abstract 3204: Tissue engineered models of metastatic bone disease for the study of prostate cancer cell dormancy
Authors: Liao Yanwan , Tan Jun Jie Aaron , Tamilselvan Sahaana , Chan Jerry , Ge Ruowen , Lee Lui Shiong , Chong Mark 
Published: August 2015

Abstract
Cellular dormancy refers to a state of quiescence during metastatic progression, in which disseminated tumor cells enter cell cycle arrest. Such dormant cells are thought to influence cancer drug sensitivity, and strategies have hence emerged to target dormant cells for novel therapeutic approaches. Mechanisms of cellular dormancy remain poorly understood, with current evidence suggesting a major role of the metastatic microenvironment in deciding cell fate. Research in this area, however, is hampered by the lack of appropriate experimental models of cancer cell dormancy. Here, we described the use of tissue engineering methods to generate an in vitro model of bone-metastatic disease. Briefly, human fetal mesenchymal stem cell (hfMSC) and human umbilical vein endothelial cell (HUVEC) were co-cultured to respectively represent the osteo-progenitor and vascular compartments, and subsequently induced to form a vascularized bone marrow model. Metastatic prostate cancer PC3 cells were then added to simulate metastatic dissemination and the co-culture was maintained for 28 days. Reporter genes were employed to facilitate live-monitoring of HUVEC and PC3 cells in the co-culture. Using this system, we observed single cell dormancy, particularly when osteogenic induction was performed. Additionally, PC3 cells were observed to exert osteo-modulatory effects on the engineered bone microenvironment. Immunostaining for cellular markers indicated low expression of proliferation marker Ki-67 and various mesenchymal markers, indicating absence of Epithelial-Mesenchymal Transition (EMT). Dormancy was functionally assessed by retrieving the PC3 cells and subsequently replating under normal culture conditions to rescue cellular proliferation. Finally, drug sensitivity assays demonstrated reduced susceptibility of dormant cells to conventional chemotherapeutic drugs. This in vitro model of prostate cancer cell dormancy in the engineered bone marrow may thus be useful for furthering our understanding of cellular dormancy, and may further provide a useful tool for drug testing.
Citation Format: Yanwan Liao, Jun Jie Aaron Tan, Sahaana Tamilselvan, Jerry Chan, Ruowen Ge, Lui Shiong Lee, Mark Chong. Tissue engineered models of metastatic bone disease for the study of prostate cancer cell dormancy. [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 3204. doi:10.1158/1538-7445.AM2015-3204