bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3129: miR-340 is a modulator of oncogenic signaling in melanoma
Authors: Poenitzsch Strong Ashley M. , Adochio Emily G. , Setaluri Vijayasaradhi , Spiegelman Vladimir S. 
Published: August 2015

Abstract
Melanoma is responsible for only 4% of skin cancer diagnoses; nonetheless 80% of skin cancer deaths are attributed to this aggressive skin cancer each year. Growing evidence points to an important role miRNAs may play in the aberrant cellular signaling of cancers, including melanoma. microRNAs (miRNAs) are highly conserved 22-25nt non-coding RNAs involved in regulating RNA stability and translation and are predicted to regulate ∼60% of the human transcriptome. We are investigating the tumor suppressor role miR-340 plays in the development and progression of melanoma. A limited number of miR-340 targets were previously characterized, however, it is unlikely the effects of miR-340 on the melanoma phenotype are the result of directly targeting a few mRNAs, as miRNAs are likely to target upwards of 100 mRNAs each. Using an in silico target identification and pathway mapping system, we identified a multitude of potential miR-340 mRNA targets that encode proteins distributed amongst a variety of cellular signaling pathways. In silico analysis revealed a high proportion of miR-340 predicted mRNA targets that encode for members of the MAPK signaling pathway. Our results show that miR-340 modulated most of the assayed mRNAs that encode MAPK components. We also demonstrate that miR-340 inhibits the tumorigenic phenotype of melanoma cells. We recently developed a novel method to identify direct targets of miRNAs. Using this method, we demonstrate miR-340 directly targets mRNAs that encode protein kinases of the MAPK pathway. Further direct target identification is crucial toward understanding the role miR-340-directed regulation plays in triggering pleiotropic effects during melanoma development and progression. Elucidating the roles aberrant gene regulatory networks play in melanoma is critical for understanding the disease and developing treatment options to improve the melanoma prognosis.
Supported by NIH grants ES007015 (to A.M.P.S.), AR063361 (to V.S.S.) and CA191550 (to V.S.S.).
Citation Format: Ashley M. Poenitzsch Strong, Emily G. Adochio, Vijayasaradhi Setaluri, Vladimir S. Spiegelman. miR-340 is a modulator of oncogenic signaling in melanoma. [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 3129. doi:10.1158/1538-7445.AM2015-3129
Abstract 3141: Restricting aerobic glycolysis provides functional improvement of chimeric antigen receptor (CAR)-modified T cells
Authors: Smith Jillian P. , Long Adrienne H. , Mackall Crystal L. 
Published: August 2015

Abstract
Chimeric antigen receptor (CAR) T cell therapy targeting CD19 has proven effective at treating hematologic malignancies, however similar success has not been observed using CARs targeting other antigens. We have observed that several CARs, including a GD2-targeted CAR, are limited in efficacy due to T cell exhaustion induced by tonic, antigen-independent signaling. This phenomenon is triggered by oligomerization of scFvs on the surface of transduced T cells, leading to chronic CAR zeta signaling and early exhaustion characterized by a diminished proliferative potential, poor cytokine production, and overexpression of inhibitory receptors including PD1, TIM3 and LAG3. Early CAR T cell exhaustion is also associated with an altered cellular metabolism that is more highly dependent on glycolysis compared to CD19 CAR T cells or untransduced, activated control cells. Using a Seahorse Extracellular Flux Analyzer, we found that the rate of glycolysis (ECAR) in GD2 CAR T cells was twice that of CD19 CAR T cells or untransduced controls, and no deficiencies in oxidative phosphorylation in GD2 CAR T cells were observed (normal OCR and spare respiratory capacity). To evaluate whether increased glycolytic metabolism contributes to the development of CAR T cell exhaustion, we sought to restrict glycolysis by culturing GD2 CAR T cells with the hexokinase inhibitor 2-deoxyglucose (2DG) or in media supplemented with galactose instead of glucose. Restricting CAR T cell glycolysis was associated with decreased TIM3 and LAG3 expression and enhanced cytokine production of GD2 CAR T cells but a modest decrease in cytokine production of CD19 CAR T cells. Given the previously reported role of mTOR on promoting glycolytic metabolism, we used rapamycin (sirolimus) in the cell culture during transduction and expansion. Culturing GD2 CAR T cells with rapamycin similarly resulted in slower acidification of cell culture medium and diminished expression of TIM3 and LAG3. Rapamycin treated GD2 CAR T cells retain specific lytic ability of GD2-positive tumor cells compared to non-treated GD2 CARs as measured by chromium release, and these cells persist longer than non-treated cells in a murine xenograft model of GD2-positive 143b osteosarcoma. These results suggest that modulation of T cell metabolism may provide functional benefit and enhance antitumor activity of adoptively transferred tumor antigen specific T cells by diminishing the susceptibility of highly activated T cells to exhaustion.
Citation Format: Jillian P. Smith, Adrienne H. Long, Crystal L. Mackall. Restricting aerobic glycolysis provides functional improvement of chimeric antigen receptor (CAR)-modified T 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 3141. doi:10.1158/1538-7445.AM2015-3141
Abstract 3161: Predictive Immunotherapy Models: Overcoming the challenge of T cells gene transfer
Authors: Bouillé Pascale , Duthoit Christine , Gayon Régis , Feuillet Vincent , Auffray Cédric 
Published: August 2015

Abstract
The initiation of an appropriate T cell response is a key event in the establishment of efficient anti-tumoral response. In a context of tumor development, different T cell populations are mobilized and understanding the pathways involved in their functions and in their interactions is a major field of research in cancer immunotherapy. Thus, while the activation of antigen-specific naive CD4+ T cells that can differentiate into appropriate effector T cells and home to the tumor might be highly beneficial, regulatory CD4+ T cells are considered as deleterious. Many efforts are thus made to understand the behavior of both CD4+ T cells subsets and to decipher their functions within the tumor micro-environment. A key step to reach this goal is the development of relevant predictive cancer models including genetic modifications of T cell populations in vitro and in vivo, either for gene target studies, or to create new tools for sensitive readouts.
While human and activated mouse T cells are easily transduced with lentiviral vectors, transduction of naive and regulatory murine T cells is a technical challenge since these target cells are poorly permissive to transfection or viral transduction. Here, extensive in vitro transduction data show that the use of highly concentrated (1E9 TU/mL) and pure (1E8 TU/mg of proteins) lentiviral vectors increases significantly the transduction level of naive and regulatory CD4+ T cells. Furthermore, transduced T cells exhibit an original cell phenotype without any changes of T cells specific markers expression (CD44, CD69, CD25, Ly-6C) attesting that the composition of this pure batch of lentiviral vector does not have any impact on T cells phenotype. Transduced T cells were then assessed for their properties once adoptively transferred into recipient mice. Analysis of these mice show that not only the genetically modified T cells/regulatory cells can be easily followed thanks to the fluorescent reporter but also their phenotype is not modified by the transduction, as shown in vitro.
Our data bring out that the composition of lentiviral vectors in terms of titer, specific activities and purity are the success keys to ensure transduction of murine T cells. This major technological leap will allow to tightly control expression of various modifiers of these cells and thus paves the way to design original tools and develop cell-based cancer.
Citation Format: Pascale Bouillé, Christine Duthoit, Régis Gayon, Vincent Feuillet, Cédric Auffray. Predictive Immunotherapy Models: Overcoming the challenge of T cells gene transfer. [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 3161. doi:10.1158/1538-7445.AM2015-3161
Abstract 3183: Role of danger signals in tumor elicited inflammation
Authors: Francescone Ralph , Vendramini-Costa Debora , Dmitrieva Oxana , Hou Vivi , Posocco David , Grivennikov Sergei 
Published: August 2015

Abstract
Spontaneous colon cancer (CRC) is traditionally thought to arise from a series of genetic mutations to key tumor suppressors and oncogenes that drive its progression. Recently, however, the importance of the immune system and inflammation in exacerbating CRC development has been demonstrated, as seemingly non-inflammatory tumors have massive infiltration of immune cells. This phenomenon has been deemed “Tumor elicited inflammation” (TEI), whereby the tumor cells recruit immune cells to the tumor, promoting tumor development. Danger Associated Molecular Patterns (DAMPs), proteins that are released during cellular stress or oncogenic transformation, promote a robust immune response and are often up-regulated in cancer. Thus, DAMPs may serve as an initial cue to induce TEI. The precise mechanisms of these so-called “danger signals” in CRC development and TEI have not been fully elucidated.
Using a number of in vitro and in vivo models, we sought to uncover the major cellular producers of danger signals in the tumor microenvironment, how these danger signals recruit various immune cells to the tumor, and finally their effects on tumor progression. We found an up-regulation of three danger signals, S100A8, S100A9, and HMGB-1, in tumor tissue from mouse models of spontaneous CRC, at the RNA and protein levels. In addition, using FACS, the myeloid compartment of sorted immune cells demonstrated the highest expression levels of S100A8 and S100A9, as measured by qPCR. In order to further explore role of danger signals, lentiviral overexpression of S100A8, S100A9, and HMGB-1 was performed in MC38 colon cancer cells. Subsequently, these new cell lines were injected subcutaneously into C57BL/6 mice and tumor development was followed. S100A8 and S100A9 overexpressing tumors showed accelerated development and a greater influx of myeloid cells into the tumor when compared to GFP overexpressing control cells. Future work will focus on the crosstalk between cancer cells and immune cells, the key cellular pathways affected in cancer cells and immune cells, and finally if blockade of these signals has any therapeutic benefit in the treatment of CRC.
Citation Format: Ralph Francescone, Debora Vendramini-Costa, Oxana Dmitrieva, Vivi Hou, David Posocco, Sergei Grivennikov. Role of danger signals in tumor elicited inflammation. [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 3183. doi:10.1158/1538-7445.AM2015-3183
Abstract 3189: Tunneling nanotube formation is significantly upregulated in invasive cancer cells
Authors: Desir Snider , Subramanian Subbaya , Steer Clifford , Lou Emil 
Published: August 2015

Abstract
Introduction: Intercellular communication is critical for cancer growth and progression. Research to date has focused on gap junctions, micro-vesicles/exosomes, and diffusible factors. There is limited understanding of how efficient cell-to-cell communication occurs between distant cells in the complex tumor microenvironment. Tunneling nanotubes (TNTs) are thin, non-adherent actin filament-based, open conduits that facilitate direct sharing of various cellular cargos between cells. TNTs can be 50-800 nanometers in diameter and up to several hundred micrometers in length in vitro. Our lab was the first to provide in vivo evidence of TNTs in human solid tumors. We hypothesized that TNT formation is significantly upregulated in human cancers compared to their non-malignant counterparts. In this study, we examined the rate of TNT formation in cell lines from multiple human cancers.
Methods: Cell lines from four cancers (colon, mesothelioma, pancreatic, and ovarian cancer) were plated at similar density in 6-well tissue culture plates. The number of TNTs and cells in 10 randomly chosen fields of each well were quantified every 24 hrs for up to 96 hrs and representative images were taken. Experiments were performed in duplicate for each cell line.
Results: The number of TNTs per cell (TNTs/cell), which we have titled “TNT index,” was calculated in order to control for cell proliferation. The ratio of TNTs formed between malignant colorectal cells compared benign adenoma ranged from 20:1 to 100:1 by 96 hrs.
For mesothelioma, malignant:mesothelial TNTs/cell ratio ranged from 8:1 to 78:1 over 72 hrs. The mesothelioma cells were too confluent at 96 hrs to quantify TNTs. The pancreatic malignant:stellate human pancreatic ductal epithelial TNTs/cell ratio ranged between 2:1 to 63:1. Interestingly, we found that non-malignant ovarian epithelial (IOSE) cells had a significantly higher rate of TNT formation than malignant ovarian cells.
Conclusions: We have demonstrated that malignant cells have significantly higher rates of TNT formation compared to stromal or premalignant cells of their corresponding tumor. TNTs potentially provide malignant cells a direct route for transmitting influential messages across long distances in the complex tumor microenvironment. We believe that upregulation of TNTs in cancer may amplify cellular cross-talk and thus increase rates of tumor recurrence and invasion.
Citation Format: Snider Desir, Subbaya Subramanian, Clifford Steer, Emil Lou. Tunneling nanotube formation is significantly upregulated in invasive 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 3189. doi:10.1158/1538-7445.AM2015-3189
Abstract 3193: Expression analysis reveals candidate genes involved in highly invasive high-hyaluronan binding subpopulations of prostate cancer cell lines
Authors: Leith Sean J. , Chambers Ann F. , McCarthy James B. , Chin Joseph L. , Turley Eva A. 
Published: August 2015

Abstract
Tumour heterogeneity is one of the hallmarks of aggressive cancers, which can be detected by differential gene expression, DNA mutation patterns, and ligand binding profiles. Hyaluronan (HA) is an extracellular matrix polysaccharide whose elevated accumulation is associated with tumour recurrence in colon and breast carcinomas. Previously, we reported that triple-negative breast cancer cell lines exhibit heterogeneous binding of HA, with high and low binding subpopulations showing differences in proliferation and invasion. Here we assessed whether or not a similar binding heterogeneity detects prostate cancer cell subsets with differential invasive capability. Flow cytometry HA-binding profiles reveal heterogeneous binding of a fluorescent HA probe to the PC3M-LN4 prostate cancer cell line. Fluorescence-activated cell sorting (FACS) was used to isolate low and high hyaluronan binding subpopulations. Sorted cells were cultured and, after one week of growth, were analyzed for cell proliferation and invasion using alamar blue, transwell invasion and gelatin degradation assays. Results confirmed that HAhigh tumor cells displayed reduced growth but increased invasion and degradation relative to HAlow subpopulations. We then employed human gene 2.0 expression arrays to assess transcriptome changes between these two subpopulations. The expression of 7 genes was significantly elevated or lowered in HAhigh vs. HAlow cells, and these were linked to inflammation, angiogenesis, MMP9 regulation and metastasis. These results identify a novel form of heterogeneity common to both breast and prostate cancer cell lines and suggest that subpopulations binding high levels of HA can be used as indicators of aggressive tumors.
Citation Format: Sean J. Leith, Ann F. Chambers, James B. McCarthy, Joseph L. Chin, Eva A. Turley. Expression analysis reveals candidate genes involved in highly invasive high-hyaluronan binding subpopulations of prostate cancer cell lines. [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 3193. doi:10.1158/1538-7445.AM2015-3193
Abstract 3208: Stromal up-regulation of gene expression in prostate cancer cells: Role of microenvironment
Authors: Ashida Shingo , Kawada Chiaki , Inoue Keiji , Shuin Taro 
Published: August 2015

Abstract
Recently, tumor microenvironment has been reported to be important in carcinogenesis and tumor progression. Especially in prostate cancer (PC), it has been suggested that tumor microenvironment plays an important role in progression, acquisition of androgen independence, and distant metastasis. However, little is known about the genetic basis of human PC microenvironment. To clarify the mechanism of PC progression and metastasis, we previously reported the analysis of genome-wide gene expression profiles using human PC cells (LNCaP), normal human prostate epithelial cells (PrEC), and normal human prostate stromal cells (PrSC). In that study, we identified genes that were up-regulated in LNCaP when co-cultured with PrSC, which included alpha-methylacyl-CoA racemase (AMACR), homeobox B13 (HOXB13), and UDP glucuronosyltransferase 2 family, polypeptide B17 (UGT2B17). We often confront the discrepancy in the difference of gene expression between cancer cells and tissues. Our data reveals that stromal cells could affect on PC cells to increase their gene expression, which might contribute to aggressiveness of PC cells. We then focused on secreted proteins that can mutually affect in this experimental design and identified genes that were overexpressed only in stromal cells. They included chemokine (C-C motif) ligand 2 (CCL2), hepatocyte growth factor (HGF), insulin-like growth factor binding protein 5 (IGFBP5), wingless-type MMTV integration site family, member 5A (WNT5A), and collagens (COL1A1, COL1A2, COL3A1, COL6A3, COL15A1). We further investigated the influence of these genes on the expression of AMACR, HOXB13, and UGT2B17 in PC cells to identify crucial signaling pathways in this process. This study provides potential candidate genes the products of which might serve as microenvironmental targets for the treatment of PC.
Citation Format: Shingo Ashida, Chiaki Kawada, Keiji Inoue, Taro Shuin. Stromal up-regulation of gene expression in prostate cancer cells: Role of microenvironment. [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 3208. doi:10.1158/1538-7445.AM2015-3208
Abstract 3209: The expression of SLC7A11 transporter in lung and pancreatic cancer tissues at different stages of development
Authors: Moreno Jonathan A. , Lambros Maria P. 
Published: August 2015

Abstract
Purpose: The purpose is to investigate the expression of the SLC7A11 transporter in human biopsy samples from patients with lung or pancreatic cancer at different stages of development. The SLC7A11 is a membrane antiporter highly specific for cystine. It is a major transporter for the uptake of cystine in exchange of intracellular glutamate. It plays a role in the growth and progression of cancers because they need extracellular cystine for growth. The expression of SLC7A11 was compared between normal and malignant tissues.
Methods: Lung and pancreatic tissue array slides were purchased from US Biomax, Inc. Pancreatic samples were categorized into adenocarcinoma, normal and adjacent to cancer tissues. Lung cancer samples were categorized into: adenocarcinoma or squamous cell carcinoma, and normal tissue. The slides were hydrated with a series of washes using xylene and graded alcohols. The tissue arrays underwent blocking of endogenous peroxidase and albumin. The tissue arrays were processed with a primary antibody (1:500) dilution of a rabbit polyclonal antibody against SLC7A11, and a secondary antibody (1:3000) dilution of goat anti-rabbit conjugated to horseradish peroxidase (HRP). The slides were counterstained with hematoxylin and images were taken with the EVOS® FL Cell Imaging System. The software ImageJ was used to determine proportion of stained cells. The staining intensity was determined by comparing the samples to their negative control supplied on the tissue arrays. The immune-reactive score (IRS) was calculated as SI x PP and it ranged from 0 to 12. An IRS within 0-3 indicated no, or low expression, 4-7 medium expression and 8-12 high expression. The distribution of the IRS score frequency in the different stages was compared to that of the normal tissues and compared using the chi square test for trend.
Results: The SLC7A11 is significantly overexpressed on all cancer tissues compared to normal tissues or adjacent to cancer tissues. Pancreatic and lung adenocarcinomas as well as lung squamous cells carcinomas have significantly different distributions of IRS score staining for SLC7A11 compared to normal tissues. However, there is no significant difference between the different cancer stages and the distributions of the IRS staining for SLC7A11.
Conclusion: The results indicate that expression of the SLC7A11 transporter is significantly overexpressed in cancer compared to the normal tissues, but there is no significant difference between the expression levels of SLC7A11 at different stages of lung or pancreatic cancer.
Citation Format: Jonathan A. Moreno, Maria P. Lambros. The expression of SLC7A11 transporter in lung and pancreatic cancer tissues at different stages of 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 3209. doi:10.1158/1538-7445.AM2015-3209