bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2318: Genetic and functional diversity of propagating cells in glioblastoma
Authors: Piccirillo Sara Grazia Maria , Colman Sue , Potter Nicola E. , van Delft Frederik W. , Lillis Suzanne , Carnicer Maria-Jose , Kearney Lyndal , Watts Colin , Greaves Mel 
Published: August 2015

Abstract
Glioblastoma (GBM) is a lethal malignancy whose clinical intransigence has been linked to extensive intraclonal genetic and phenotypic diversity and the common emergence of therapeutic resistance. This interpretation embodies the implicit assumption that cancer stem cells or tumor-propagating cells are themselves genetically and functionally diverse. To test this, we screened primary GBM tumors by SNP array to identify copy number alterations (a minimum of three) that could be visualized in single cells by multicolor fluorescence in situ hybridization. Interrogation of neurosphere-derived cells (from four patients) and cells derived from secondary transplants of these same cells in NOD-SCID mice allowed us to infer the clonal and phylogenetic architectures. Whole-exome sequencing and single-cell genetic analysis in one case revealed a more complex clonal structure.This proof-of-principle experiment revealed that subclones in each GBM had variable regenerative or stem cell activity, and highlighted genetic alterations associated with more competitive propagating activity in vivo.
Citation Format: Sara Grazia Maria Piccirillo, Sue Colman, Nicola E. Potter, Frederik W. van Delft, Suzanne Lillis, Maria-Jose Carnicer, Lyndal Kearney, Colin Watts, Mel Greaves. Genetic and functional diversity of propagating cells in glioblastoma. [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 2318. doi:10.1158/1538-7445.AM2015-2318
Abstract 2325: Activated glucocorticoid signaling in pancreatitis contributes to acinar-to-ductal metaplasia and KrasG12D-driven tumorigenesis
Authors: Lin Pei-Yu , Su Chien-Chang , Hsieh Chi-Che , Shen Chia-Ning 
Published: August 2015

Abstract
Acinar-to-ductal metaplasia (ADM), an inflammatory lesion associated with replacement of acinar cells by tubular complexes, is thought to represent a condition with increased risk of neoplasia. However, the direct evidence linking acinar-to-ductal metaplasia and development of pancreatic neoplasm is still lacking. Glucocorticoid-mediated signaling pathway is known to be a part of the feedback control machinery of immune system that acts to suppress excessive immune activity. In the present study, we explored the role of GR in development of ADM and pancreatic intraepithelial neoplasia (PanIN) in mice with pancreatitis. We initially identified that evaluated levels of activated glucocorticoid receptors (GR) were observed in patients of chronic pancreatitis and pancreatic ductal adenocarcinomas (PDAC). Utilizing mice conditionally expressed KrasG12D in acinar cell with or without loss of GR, we demonstrated that GR activation was required for generation of metaplastic ductal lesion and development of pancreatic intraepithelial neoplasia (PanIN) in mice with experimental pancreatitis. Administration of dexamethasone, a potent synthetic glucocorticoid, was sufficient to induce development of PanINs and PDACs in KrasG12D mice that harbored single-allele deletion of p53. We further revealed that non-genomic activation of GR was a key action that led to evaluate c-Src signaling and trigger a pathological threshold of Ras activity necessary for neoplastic transformation. In contrast, treatment of dasatinib, a Src kinase inhibitor, suppressed development of PanINs and PDACs induced by dexamethasone. Importantly, Cre-lox based lineage-tracing mice was applied and identified that GR activation directly transformed GFP-tagged KrasG12D-expressing acinar cells to tumorigenic cells. The findings explained why inflammatory ductal reprogramming predisposes to development of pancreatic neoplasm.
Citation Format: Pei-Yu Lin, Chien-Chang Su, Chi-Che Hsieh, Chia-Ning Shen. Activated glucocorticoid signaling in pancreatitis contributes to acinar-to-ductal metaplasia and KrasG12D-driven 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 2325. doi:10.1158/1538-7445.AM2015-2325
Abstract 2326: Sox10 promotes both stem-like and EMT properties in mammary cells
Authors: Dravis Christopher , Wahl Geoffrey M. 
Published: August 2015

Abstract
Molecular pathways that guide cell proliferation and programming events during embryonic development may be aberrantly re-activated in the cancer state to drive tumor progression. To identify mechanisms that drive progression of aggressive breast cancers, we identified molecules that are present in the fetal mammary stem cells (fMaSCs) that drive early development of the mammary gland. These analyses revealed that Sox10 is one of the most highly expressed transcription factors within fMaSCs, and prompted us to investigate the function of Sox10 in the mammary gland and in subtypes of cancer originating from it. Our studies described here indicate that Sox10 is specifically expressed in mammary cells that exhibit higher levels of stem or progenitor cell functions using in vitro or in vivo assays. This includes fetal and adult mammary cells in vivo, as well as mammary cells cultured in vitro as organoids, and for the first time, male fetal mammary stem cells. Sox10 appears to contribute to these stem/progenitor behaviors, as the genetic deletion of Sox10 limits stem/progenitor activities, while the overexpression of Sox10 significantly expands clonogenic behavior. Intriguingly, ectopic Sox10 expression is also able to elicit an EMT-like response in mammary organoids. Sox10 thus represents a single molecule capable of directly mediating both stem-like and EMT-like properties in mammary cells. Furthermore, by modulating Sox10 levels in 3D mammary organoids, we are able to induce cells into sequential motile and stem/progenitor states. This provides a striking in vitro surrogate for metastatic behavior. Consistent with these findings, we find that Sox10 is highly expressed in basal-like and claudin-low subtypes of breast cancer, which are the most stem-like and EMT-like manifestations of the disease, respectively. Finally, we show that Sox10 expression is regulated through a feedback loop involving FGF signaling, and that inhibition of FGF signaling can block EMT-like cell behaviors mediated by Sox10. These findings have important implications in how stem-like and metastatic properties may be specified in mammary cells.
Citation Format: Christopher Dravis, Geoffrey M. Wahl. Sox10 promotes both stem-like and EMT properties in mammary 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 2326. doi:10.1158/1538-7445.AM2015-2326
Abstract 2345: The collagen-dense tumor microenvironment increases neutrophil recruitment in mouse mammary carcinoma
Authors: Garcia Mendoza Maria Gracia , Inman David , Ponik Suzanne M. , Keely Patricia J. 
Published: August 2015

Abstract
The breast tumor microenvironment (TME) is a diverse area where complex interactions between the extra-cellular matrix (ECM) and multiple cell types (tumor epithelial, endothelial, and inflammatory cells, fibroblasts, and adipocytes) occur resulting in tumor promotion, invasion and metastasis. Changes in breast tissue density have been correlated with a 4-6-fold increased risk for developing breast cancer, and are associated with increased stromal deposition of the ECM protein, collagen I. In a transgenic mouse model of increased collagen (Col1A1Tm1Jae), there is a 3-fold increase in MMTV-PyMT tumor formation and lung metastases compared to control mice. Of the non-malignant cells in the TME, tumor-associated macrophages (TAMs) and tumor-associated neutrophils (TANs) have been shown by several investigators to alter the TME and ECM, enhance tumor cell migration and invasion, stimulate angiogenesis, and suppress antitumor immunity. The direct interaction of both increased collagen and immune (myeloid) cell recruitment leading to increased cancer progression and metastasis has not been characterized. We hypothesized that a collagen-dense tumor microenvironment recruits an abnormal population of myeloid cells, which promotes tumor progression and metastasis. Flow cytometry analysis of cells from collagen-dense mouse tumors showed an increase in Ly6G+ TAN recruitment compared to wild type tumors, while TAM recruitment did not seem to be affected. The increased Ly6G+ neutrophil recruitment is a global immune response also observed in the spleens and circulating blood. Using a cytokine/chemokine ELISA plate array, we found that the collagen-dense TME cytokine cross talk supports neutrophil and monocyte recruitment and maturation. In addition, many classic inflammatory signals that support lymphocyte recruitment and activation, B and T-cell proliferation, and natural killer cell and monocyte activation are downregulated in the collagen-dense TME. Further experiments will determine the time point where neutrophil recruitment is critical for tumor progression. Experiments are underway to study the possible suppression of natural killer cell activity in the collagen-dense TME.
This work is being supported by NIH grants (NIH-CA-114462 and NIH-U54-CA163131) to P.J.K. and NSF Graduate Research Fellowship (2011129283) to M.G.G.M.
Citation Format: Maria Gracia Garcia Mendoza, David Inman, Suzanne M. Ponik, Patricia J. Keely. The collagen-dense tumor microenvironment increases neutrophil recruitment in mouse mammary 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 2345. doi:10.1158/1538-7445.AM2015-2345
Abstract 2347: Multiplexed quantitative assessment and prognostic value of TIL subtypes in non-small cell lung cancer
Authors: Schalper Kurt A. , Hoyt Cliff , Wang Chichung , Rimm David 
Published: August 2015

Abstract
Background: The presence of high levels of tumor-infiltrating lymphocytes (TILs) is associated with favorable outcome in diverse human neoplasms including non-small cell lung cancer (NSCLC). Previous studies suggest that different TIL subtypes have distinct biological roles in cancer progression and their contribution to prognosis remains unclear. Here, we simultaneously quantified TIL subtypes using 2 different automated, objective methods in human NSCLC and determined their association with tumor characteristics and outcome.
Methods: Using a sequential multiplexed immunofluorescence staining protocol with tyramide signal amplification and heat-inactivation steps, we simultaneously visualized all cell nuclei (DAPI), epithelial cells (Cytokeratin AE1/AE3, Dako), T helper cells (CD4-368, Leica), Cytotoxic T cells (CD8, C8/144B, Dako), B-lymphocytes (CD20, L26, Leica) and Regulatory T cells [Tregs], (Foxp3, D608R, CST) in 202 stages I-IV NSCLC represented in tissue microarray format. The preparations were digitalized and the number of each TIL subpopulation was counted using multispectral fluorescence analysis and a trainable tissue/cell image segmentation algorithm with the inForm software (PerkinElmer). Markers were also measured using the AQUA method of quantitative immunofluorescence (QIF).
Results: NSCLC samples show a highly variable TIL content ranging from 0 to 2717 lymphocytes per tumor histospot. The median TIL content was 23.9 T-helper cells, 55.8 Cytotoxic T cells, 59.7 B-lymphocytes and 6.1 Tregs. Although the amount of CD4+, CD8+ and CD20+ cells was comparable between the histological NSCLC subtypes, the average number of Tregs (CD4+/Foxp3+ cells) was significantly higher in squamous cell carcinomas than in adenocarcinomas (10.4 and 4.9 cells per histospot, respectively, P = 0.03). Increased numbers of CD8+ or CD20+ TILs were significantly associated with longer overall survival (log-rank P&lt;0.05), but the amount of CD4+ and Foxp3+ cells were not. Correlations were seen between multiplexed counting methods and QIF.
Conclusions: This strategy allows simultaneous and objective quantification of diverse TIL subpopulations in lung tumor samples. Our results indicate that increased Cytotoxic T cells and B-lymphocytes, but not T helper cells or Tregs have prognostic value in NSCLC. Accurate quantification of TIL subpopulations in the tumor microenvironment could be useful to predict response and study the local effects of novel anti-cancer immunostimulatory therapies.
Citation Format: Kurt A. Schalper, Cliff Hoyt, Chichung Wang, David Rimm. Multiplexed quantitative assessment and prognostic value of TIL subtypes 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 2347. doi:10.1158/1538-7445.AM2015-2347
Abstract 2349: EGFR is transferred from triple negative breast cancer cells to immune cells via trogocytosis and expression of EGFR on immune cells is associated with high tumor grade of triple negative breast cancer patients
Authors: Suzuki Eiji , Yamaguchi Ayane , Kataoka Tatsuki R. , Hirata Masahiro , Kawaguchi Kosuke , Nishie Mariko , Toi Masakazu 
Published: August 2015

Abstract
We and others previously reported that HER2 was transferred from HER2 overexpressing breast cancer cells to CD14+ monocytes via trastuzumab dependent trogocytosis. In the current study, we report that EGFR of MDA-MB-231 (triple negative human breast cancer cell line) is also transferred to immune cells via trogocytosis and those immune cells express EGFR on the cell surface without antibody administration. Interestingly, the EGFR-trogocytosed MDA-MB-231 cells reduced the expression of EGFR and changed their morphology to the spindle shape (well known as mesenchymal transition) and acquired greater cell motility. These cancer cell properties are representative as higher malignant potentials in breast cancer patients. Thus, we hypothesized that EGFR expression on tumor infiltrating lymphocytes (TILs), which probably is caused via EGFR trogocytosis by TILs, is correlated with higher tumor grade. We evaluated the EGFR expression of TILs in 36 triple negative breast cancer specimens and found the correlation between EGFR expression status on TILs and higher tumor grade (P = 0.02). These findings may explain the reason why triple negative breast cancer with greater proportion of TILs exhibit higher tumor grade and poor prognosis.
Citation Format: Eiji Suzuki, Ayane Yamaguchi, Tatsuki R. Kataoka, Masahiro Hirata, Kosuke Kawaguchi, Mariko Nishie, Masakazu Toi. EGFR is transferred from triple negative breast cancer cells to immune cells via trogocytosis and expression of EGFR on immune cells is associated with high tumor grade of triple negative 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 2349. doi:10.1158/1538-7445.AM2015-2349
Abstract 2374: Mir-29/Hsp47 regulate breast cancer progression by modulating the ECM transcription network
Authors: Zhu Jieqing , Xiong Gaofeng , Xu Ren 
Published: August 2015

Abstract
The extracellular matrix (ECM), an essential component of tumor microenvironment, play crucial roles in cancer development and progression. Using gene co-expression network analysis, we have identified an ECM transcription network from hundreds of human breast cancer. In this report, we show how the molecular chaperone protein Hsp47 functions as a nodal hub in regulating an ECM gene transcription network. We found that Hsp47 expression was activated during breast cancer development and progression. Hsp47 silencing reprogrammed human breast cancer cells to form growth-arrested and/or non-invasive structures in 3D cultures, and to limit tumor growth in xenograft assays by reducing deposition of collagen and fibronectin. Co-expression network analysis also showed that levels of microRNA-29b and 29c were inversely correlated with expression of Hsp47 and ECM network genes in human breast cancer tissues. We found that miR-29 repressed expression of Hsp47 along with multiple ECM network genes. Ectopic expression of miR-29b suppressed malignant phenotypes of breast cancer cells in 3D culture. Clinically, increased expression of Hsp47 and reduced levels of miR-29b and 29c were associated with poor survival outcomes in breast cancer patients. Our results show that Hsp47 is regulated by miR-29 during breast cancer development and progression, and that increased Hsp47 expression promotes cancer progression in part by enhancing deposition of ECM proteins.
Citation Format: Jieqing Zhu, Gaofeng Xiong, Ren Xu. Mir-29/Hsp47 regulate breast cancer progression by modulating the ECM transcription network. [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 2374. doi:10.1158/1538-7445.AM2015-2374
Abstract 2380: Integrin αv is necessary for skin tissue generation and SCC tumor invasion, but dispensable for tissue maintenance
Authors: Duperret Elizabeth K. , Dahal Ankit , Ridky Todd W. 
Published: August 2015

Abstract
Integrin heterodimers play crucial roles in epithelial adhesion, proliferation, differentiation, wound healing and tumorigenesis. However, the effects of acute loss of many of the 26 integrin subunits in epidermis have not been determined. This information is critical since many inhibitors for specific integrin heterodimers are entering clinical trials for various cancers. We performed an shRNA-based multiplexed screen to define roles for each integrin subunit in a 3-dimensional in vitro organotypic skin model. In this screen, we transduced individual populations of human keratinocytes with shRNAs against each integrin subunit. We then quantitatively determined the fitness of each cell population in a competition assay in reconstituted human skin. This screen identified the integrin αv class of heterodimers as essential for generation of human skin tissue: the population of keratinocytes lacking αv was selected against 125-fold compared to non-silencing control cells (p&lt;0.001). An identical screen in human melanocytes showed no change in fitness upon loss of integrin αv, indicating cell-intrinsic functions for this subunit. Loss of integrin αv alone in keratinocytes leads to cell cycle arrest with a 4-5 fold increase in the G1/S ratio. Surprisingly, we found that integrin αv is not localized within focal adhesions in human keratinocytes and does not control focal adhesion dynamics in this cell type. Instead, it forms an independent signaling complex at the cellular membrane that controls keratinocyte growth through a FAK c-myc signaling axis. Both FAK and c-myc total protein levels are lost upon αv knock down, and re-expression of c-myc in αv-null keratinocytes is sufficient to completely rescue the cell cycle defects and skin tissue phenotypes. The effects of integrin αv loss are dependent on αv's binding partners β5 and β6, but not β1 or β8. Using a doxycycline-inducible system, we show that αv loss plays a crucial role in tissue generation and tumor invasion, but is not necessary for epidermal tissue maintenance. Furthermore, both β5 and β6 binding partners are required for tumor invasion, thus identifying αvβ5 and αvβ6 as therapeutic targets for metastatic squamous cell carcinoma.
Citation Format: Elizabeth K. Duperret, Ankit Dahal, Todd W. Ridky. Integrin αv is necessary for skin tissue generation and SCC tumor invasion, but dispensable for tissue maintenance. [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 2380. doi:10.1158/1538-7445.AM2015-2380
Abstract 2391: Microglia-stimulated glioblastoma cell invasion is dependent on the EGFR ligand amphiregulin
Authors: Desai Bhavika , Rath Uttama , Segall Jeffrey E. , Coniglio Salvatore J. 
Published: August 2015

Abstract
Glioblastoma Multiforme (GBM) is one of the deadliest human cancers with a median survival rate of only one year following diagnosis. The highly infiltrative nature of GBM is one of the main reasons this cancer is so difficult to completely remove surgically. Therefore increasing our knowledge of the mechanism GBM cells use to invade normal brain is of critical importance in designing novel therapies. We recently showed that tumor associated microglia stimulate GBM cell invasion and this process is mediated by CSF-1R signaling. In this study, we seek to identify factors that are upregulated in microglia in a CSF-1R-dependent manner and ascertain whether they are involved in mediating invasion of GBM cells. We assayed cDNA and protein from microglia treated with conditioned media from the murine GBM cell line GL261, and discovered that the EGFR ligand amphiregulin (AREG) is strongly upregulated. This upregulation is blocked by addition of a pharmacological CSF-1R inhibitor. Using RNA interference, we show that microglia depleted of AREG can not promote invasion of GL261 cells into matrigel-coated invasion chambers as effectively. In addition, an AREG blocking antibody strongly attenuates the ability of THP1 macrophages to activate human GBM cell line U87 invasion. Furthermore, we have identified a signaling pathway which involves CSF-1R signaling through MAPK ERK to upregulate AREG expression in microglia and interfering with ERK using pharmacological inhibitors also prevents microglia-stimulated GL261 invasion. Interestingly, microglia that have been activated by GBM cells appear to require cell-cell contact in order to promote invasion. This data highlights AREG as a key factor in produced by tumor associated microglia in promoting glioblastoma invasion.
Citation Format: Bhavika Desai, Uttama Rath, Jeffrey E. Segall, Salvatore J. Coniglio. Microglia-stimulated glioblastoma cell invasion is dependent on the EGFR ligand amphiregulin. [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 2391. doi:10.1158/1538-7445.AM2015-2391