bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4620: Signaling cascade of metastasis-associated macrophages determines breast cancer distal metastasis
Authors: Qian Bin-Zhi 
Published: July 2017

Abstract
Macrophages are abundantly found in the tumor microenvironment and enhance malignancy. At distal metastatic sites, our previous studies identified a distinct population of metastasis-associated macrophages (MAMs) that promote tumor cell extravasation, seeding and persistent growth. These macrophages were derived from inflammatory monocytes recruited by CCL2/CCR2 chemokine signaling and directly promote tumour cell extravasation through VEGF production. Our recent studies identified two subsequent signaling pathways that mediate the retention and function of MAMs after recruitment that are critical for tumour cell survival and persistent growth. Specifically, activation of CCR2 signaling turns on CCL3 expression in MAMs. Genetic deletion of CCL3 or its receptor CCR1 in macrophages reduces lung metastasis, and MAM accumulation in tumor-challenged lung. Adoptive transfer of wild type monocytes restores lung metastasis in Ccl3 deficient mice. Mechanistically, Ccr1 deficiency prevents MAM retention in the lung by reducing direct MAM-tumour cell interactions. On the other hand, MAMs turn on cell surface expression of FMS-like tyrosine kinase 1 (FLT1) upon differentiation from monocytes. Using several genetic models of Flt1 deficiency, we show that macrophage specific FLT1 signaling is critical for breast tumor distal metastatic potential. FLT1 specific inhibitory antibody significantly inhibits the metastatic seeding and persistent growth but does not affect the recruitment or retention of MAMs. Furthermore, we identified that FLT1 signaling regulates a set of downstream inflammatory response genes including Colony Stimulating Factor 1 (CSF1) a central regulator of macrophage biology. Using a genetic gain-of-function approach we show that CSF1 mediated autocrine signaling in MAMs is downstream of FLT1 and can restore the tumor-promoting activity in MAMs even when FLT1 is inhibited. Together, our data indicates CCL3/CCR1 and FLT1 signaling pathways are critical in regulating MAMs and promoting breast cancer distal metastasis, and suggests the therapeutic potential of targeting these pathways in treating metastatic disease.
Note: This abstract was not presented at the meeting.
Citation Format: Bin-Zhi Qian. Signaling cascade of metastasis-associated macrophages determines breast cancer distal metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4620. doi:10.1158/1538-7445.AM2017-4620
Abstract 4623: Transcriptional analysis of the functional differences between M1 and M2 macrophages to identify new targets for myeloid cell modulation in pancreatic tumors
Authors: Yang Kimberline R. , Armstrong Todd D. , Jaffee Elizabeth M. 
Published: July 2017

Abstract
Macrophages exhibit dynamic plasticity in their function in response to the accumulating genetic alterations that occur during tumorigenesis, resulting in the establishment of an immune tolerized tumor microenvironment. However, depending on their environmental cues, they can switch between two functional phenotypes: pro-inflammatory/anti-tumorigenic (M1) or anti-inflammatory/pro-tumorigenic (M2). The importance of these two macrophage subsets is well-recognized; however, identifying and distinguishing M1 from M2 macrophages in patient tumors still remains a challenge. Prior studies have utilized transcriptome profiling of artificially skewed M1 and M2 macrophages from healthy donors to identify individual genes or gene clusters unique to each macrophage subset. However, these studies are limited to surface marker expression and gene expression profiles alone to distinguish subsets and lack functional validation of M1 and M2 macrophages. Surface marker and gene expression profiling of human M1 and M2 macrophages provide an incomplete characterization, as functionality is an important feature distinguishing these macrophage subsets. To improve the characterization of M1 and M2 macrophages, we performed a thorough three-step validation of artificially skewed M1 and M2 macrophages from healthy donor monocytes using surface marker expression, gene expression profiling, and multi-analyte ELISA analyses. Using our validation assays, we tested previously published protocols for their efficacy in generating M1 and M2 macrophages. While M1 macrophages using these protocols generated as expected M1 characteristics, M2 macrophages lacked hallmark functional features, particularly IL-10 secretion. This necessitated the generation of an optimized protocol to skew M1 and M2 macrophages that exhibited distinctive surface marker expression, gene expression profiles, and cytokine and chemokine production for their respective subset. We then performed RNA-sequencing to interrogate the transcriptional landscape of functional M1 and M2 macrophages and have found 1,472 transcripts that have at least a two-fold change in differential expression between M1 and M2 macrophages across 4 different donors. These results represent the most in-depth characterization of human M1 and M2 macrophages and provide insights into better biomarkers and targets for novel immunotherapy approaches.
Citation Format: Kimberline R. Yang, Todd D. Armstrong, Elizabeth M. Jaffee. Transcriptional analysis of the functional differences between M1 and M2 macrophages to identify new targets for myeloid cell modulation in pancreatic tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4623. doi:10.1158/1538-7445.AM2017-4623
Abstract 4626: Macrophages alter heparan sulfate-modifying enzyme levels, growth factor activity and cancer cell growth in 3D hydrogels
Authors: Costa Fabio Henrique Brasil da , Navone Nora M. , Farach-Carson Mary C. , Carson Daniel D. 
Published: July 2017

Abstract
Tumor progression in both primary and metastatic niches often involves the activation of associated or “reactive” stroma similar to that seen in a wound healing response. The stromal participants include fibroblasts, endothelial and immune cells, including macrophages. Cancer-associated fibroblasts (CAFs) and tumor-associated macrophages communicate with local cells including cancer cells and the surrounding stroma to induce epithelial-to-mesenchymal transition, immunosuppression and increased production of extracellular matrix (ECM) components, growth factors, and ECM-remodeling enzymes. Heparan sulfate (HS)-modifying enzymes produced by tumor-associated stroma play key roles in the delivery of heparin-binding growth factors (HBGFs) and cytokines, serving to enhance tumorigenesis, foster growth and recruit other bystander cells to sites of metastasis. Through the use of a novel, biomimetic, 3D collagen/hyaluronic acid hydrogel system developed in our labs, this study focused on interactions between primary human macrophages and cancer cells, specifically on how this dialogue impacts the production of the secreted HS-modifying enzymes, heparanase (HPSE) and endosulfatase 1 (SULF1). We studied the impact of co-cultures containing macrophages and endometrial or prostate cancer cell lines, as well as patient-derived xenografts, on the levels of HPSE and SULF1. We also assessed cancer growth characteristics in the presence of macrophages. Taken together, our observations indicate cancer cell responses to macrophages are cell type specific, and we demonstrated significant functional consequences of interactions between macrophages and HS-mediated signaling. We are currently determining if macrophage polarization to classically activated (M1) or alternatively activated (M2) phenotypes alters cancer cell responses in 3D. This system is being used to determine novel approaches to intervene in HBGF delivery to limit cancer growth in a more physiologically relevant in vitro system.
Citation Format: Fabio Henrique Brasil da Costa, Nora M. Navone, Mary C. Farach-Carson, Daniel D. Carson. Macrophages alter heparan sulfate-modifying enzyme levels, growth factor activity and cancer cell growth in 3D hydrogels [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4626. doi:10.1158/1538-7445.AM2017-4626
Abstract 4627: Prognostic significance and population shift of peripheral monocytes in patients with oropharyngeal squamous cell carcinoma
Authors: Takahashi Hideyuki , Sakakura Koichi , Chikamatsu Kazuaki 
Published: July 2017

Abstract
Evidence has accumulated that chronic inflammation plays a critical role in carcinogenesis, tumor growth, and metastasis. To date, several inflammatory biomarkers are considered as potential prognostic factors in various cancers. The aim of the present study was to investigate the prognostic significance of the pretreatment levels of inflammatory biomarkers in patients with oropharyngeal squamous cell carcinoma (OPSCC). A total of 67 patients newly diagnosed as OPSCC was finally included. The neutrophil, lymphocyte, monocyte and platelet counts recorded before treatment was initiated, then lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were calculated. These inflammatory biomarkers and clinical parameters including age, differentiation, T-stage, N-stage, TNM-stage, smoking, and drinking were compared with progression-free survival (PFS) and overall survival (OS) using cox regression. Elevated monocyte count and advanced T-stage were significantly associated with poor PFS; in addition, these were independent prognostic factors for PFS. Elevated monocyte count and low LMR were significantly associated with poor OS; moreover, low LMR was an independent prognostic factor for OS. Of note, monocyte count and LMR were associated with prognosis more intensively in patients with p16-negative OPSCC. Following these results, we evaluated the peripheral blood mononuclear cells of 14 patients with OPSCC collected before treatments. CD14+ CD86+ HLR-DR+ cells were gated as monocytes, then divided into three subsets according to the expression of CD14 and CD16. The proportion of intermediate monocytes was lower and that of classical monocytes was higher in OPSCC patients than in healthy donors, especially in patients with elevated monocyte count. Furthermore, the expression level of PD-L1 was higher in patients than in healthy donors on all subsets. In conclusion, our data suggests that elevated monocyte count and low LMR seem to be useful biomarkers for predicting prognosis of OPSCC; furthermore, the population shift of monocytes may relate to poor prognosis of OPSCC.
Citation Format: Hideyuki Takahashi, Koichi Sakakura, Kazuaki Chikamatsu. Prognostic significance and population shift of peripheral monocytes in patients with oropharyngeal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4627. doi:10.1158/1538-7445.AM2017-4627
Abstract 4630: Development of selective and potent CDK8 inhibitors that increase NK cell activity, which translates in tumor surveillance
Authors: Hofmann Marco H. , Engelhardt Harald , Carotta Sebastian , Arnhof Heribert , Scharn Dirk , Kerenyi Marc , Mayer Moritz , Gmaschitz Gerhard , Egger Georg , Engelhardt Christian , Sanderson Michael , Impagnatiello Maria A. , Schnitzer Renate , Pearson Mark , McConnell Darryl , Kraut Norbert , Moll Jürgen 
Published: July 2017

Abstract
Background: Cyclin-dependent kinase 8 (CDK8) is part of the mediator complex that can either positively or negatively influence transcription. CDK8 is known to phosphorylate signal transducer and activator of transcription 1 (STAT1) at the position Ser727. STAT1 activity is regulated by JAK-mediated phosphorylation of tyrosine701 which leads to dimerization, nuclear translocation and IFN-γ induced phosphorylation mediated by CDK8. Introduction of an alanine mutation at the phosphorylation site STAT1-S727 results in enhanced NK cell cytotoxicity accompanied by increased levels of perforin and granzyme B (Putz et al. 2013).
Method: Here we present the discovery and development of potent and selective CDK8 inhibitors guided by crystallography. The inhibitory effect of optimized compounds BI 9811 and BI 1347 on STAT1 phosphorylation and perforin release was investigated in the human NK cell line NK-92MI. Direct effects on cancer cells were furthermore analyzed in a broad panel of cell lines. The compound BI 1347 was profiled in vivo in the orthotopic B16-F10 melanoma mouse model.
Results: Highly potent and selective CDK8 inhibitors were identified with an IC50 of below 10 nM in a biochemical kinase assay, which translated in a potent down regulation of the STAT1- Ser727 signal and in increased perforin and granzyme B secretion. BI 9811 and BI 1347 were highly selective for CDK8, as tested in a broad kinase panel and showed no cytotoxic activity on NK cells and most cancer cell lines, which distinguishes this compound class from published CDK8 inhibitors. A representative molecule out of this compound class demonstrated in vivo biomarker modulation and survival increase in the murine B16-F10 melanoma mouse model.
Conclusion: We developed potent CDK8 inhibitors that show activation of NK cells that translates into biomarker modulation (pSTAT1Ser727) and in vivo efficacy.
Citation Format: Marco H. Hofmann, Harald Engelhardt, Sebastian Carotta, Heribert Arnhof, Dirk Scharn, Marc Kerenyi, Moritz Mayer, Gerhard Gmaschitz, Georg Egger, Christian Engelhardt, Michael Sanderson, Maria A. Impagnatiello, Renate Schnitzer, Mark Pearson, Darryl McConnell, Norbert Kraut, Jürgen Moll. Development of selective and potent CDK8 inhibitors that increase NK cell activity, which translates in tumor surveillance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4630. doi:10.1158/1538-7445.AM2017-4630
Abstract 4632: Exosome-mediated transfer of miR-183-5p from tumor cells to macrophages contributes to regulate TAMs phenotypes and promote tumor progression and metastasis
Authors: Guo Jian , Wang Wei , Liu Yan , He Huiwen , Zhang Chen , Chen Chong , Luo Yunping 
Published: July 2017

Abstract
Tumor-associated macrophages (TAMs) perform supportive roles in promoting tumor progression and metastasis. In tumor microenvironment, the phenotypes of TAMs are regulated by a variety of factors, such as cytokines, ligends and products of metabolism. Recently, Exosome-mediated transfer of miRNAs is proved to be a significant way for intercellular communications. Whether exosomal miRNAs derived from tumor cells contribute to TAMs’ phenotypes remains unclear. In this study, we aim to identify the miRNAs in exosomes transferred from tumor cells to macrophages and explore how these miRNAs regulate the TAMs phenotypes. In the beginning we transferred CY3-labled miRNA NC to 4T1 mouse breast cancer cells and then co-cultured with bone marrow-derived macrophages (BMDMs). The CY3 fluorescence could be observed in BMDMs after 48 hour, suggesting that miRNAs could be transferred from tumor cells to macrophages. Next, we performed miRNAs sequencing in either BMDM or exosomes derived from 4T1 cancer cells. We analyzed and compared the miRNAseq data by bio- informatics and found that there were 30 of the most abundant miRNAs in 4T1 exosomes, and nearly all of which were rich in BMDMs. However, it was found interestingly that miR-183-5p was extremely low in BMDMs. Furthermore, we treated BMDMs with exosomes isolated form 4T1 cell conditioned medium, and then we found importantly the miR-183-5p in BMDMs was increased significantly, while cultured with exosome-depleted 4T1 conditioned medium, the expression of miR-183-5p in BMDMs was not changed. Meanwhile, we isolated TAMs from 4T1 mouse breast tumor while spleen macrophages from normal mouse as a control. The expression of miR-183-5p in TAMs was much higher than that in spleen macrophages. Moreover, it was determined that expression of pro-inflammatory cytokine TNF and IL-6 in BMDM were increased significantly in BMDMs treated with 4T1-derived exosomes. Correspondingly, transferred miR-183-5p mimic into BMDMs, the expression of TNF and IL-6 were also increased. Finally we co-cultured 4T1 cells with BMDMs transferred with miR-183-5p, and the migration and invasion of 4T1 cells were significantly enhanced. Taken together, our study demonstrate that miR-183-5p that high expression in 4T1 exosomes can transferred into macrophages to regulated their phenotypes and promote tumor progression and metastasis in breast tumor microenvironment.
Citation Format: Jian Guo, Wei Wang, Yan Liu, Huiwen He, Chen Zhang, Chong Chen, Yunping Luo. Exosome-mediated transfer of miR-183-5p from tumor cells to macrophages contributes to regulate TAMs phenotypes and promote tumor progression and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4632. doi:10.1158/1538-7445.AM2017-4632
Abstract 4636: p53 governs the tumoricidal activity of M1 macrophages in lung cancer
Authors: Hsiao Yi-Jing , Chen Hsuan-Yu , Su Kang-Yi , Ho Bing-Ching , Chen Jeremy J.W. , Yang Pan-Chyr , Yu Sung-Liang 
Published: July 2017

Abstract
Macrophages in a tumor microenvironment have been characterized as M1- and M2-polarized subtypes. M1 macrophage is associated with favorable clinical outcome of lung cancer patients, but the detail molecular mechanism is not thoroughly understood. Herein, we elucidate the anti-tumor mechanism of M1 macrophages by in vitro and in vivo assays. M1 conditioned media (CM) significantly reduced the tumor size in the xenograft mouse model via intratumoral administration. The M1-altered differential expression was profiled by microarray and the pathway analysis indicated that p53 was a critical regulator. Lung cancer cells harboring wild-type p53 (wt-p53) went apoptosis after M1 CM culture, but cells with mutant p53 did not. Additionally, M1 CM enhanced wt-p53 accumulation through reducing p53 ubiquitination and increasing protein stability. Moreover, we identified a p53 interacting protein STAT1 involved in M1-induced apoptosis and p53 accumulation. The M1 CM not only enhanced the interaction between STAT1 and p53 but also up-regulated and activated STAT1. We found that STAT1-Y701 phosphorylation is required for the p53 stability. The interaction of STAT1-Y701F and p53 was reduced compared to wild-type STAT1. To reveal the upstream signaling, we found that knockdown of IFNAR2, JAK1 and TYK2 attenuates M1-induced apoptosis. Taken together, STAT1 signaling is essential for M1 CM-induced apoptosis and p53 accumulation. Our findings imply that p53 and M1 macrophages play important roles in the immune surveillance in lung cancers.
Citation Format: Yi-Jing Hsiao, Hsuan-Yu Chen, Kang-Yi Su, Bing-Ching Ho, Jeremy J.W. Chen, Pan-Chyr Yang, Sung-Liang Yu. p53 governs the tumoricidal activity of M1 macrophages in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4636. doi:10.1158/1538-7445.AM2017-4636
Abstract 4650: Mutation detection by target sequence analyses using tissue-specific panels in esophageal squamous cell carcinoma
Authors: Iwaya Takeshi , Endo Fumitaka , Kume Kohei , Sasaki Yasushi , Tokino Takashi , Nishizuka Satoshi 
Published: July 2017

Abstract
Whole-exome sequencing (WES) studies on esophageal squamous cell carcinoma (ESCC) have revealed that only a small number of frequently mutated genes, such as TP53, NOTCH1, NRF2, and ZNF750, are present in individual tumors, whereas hundreds of others less frequently mutated genes are accumulated. The mutation detection rate by target sequence analyses is an important factor for clinical sequence in the course of ESCC treatment. This study aimed to evaluate the mutation detection rates of several target sequence analyses for ESCC. Ten pairs of surgically or endoscopically acquired tumor tissues and corresponding peripheral blood mononuclear cells were obtained from ESCC patients. We used three types of target sequence panels for mutation screening in 10 ESCC patients as follows: 1) Cancer Hotspot Panel (CHPv2), targeting mutation hotspot regions in the most common 50 human cancer genes that are frequently mutated (panel size: 22,027 bp), for five patients; 2) TP53 Panel, covering all exons of TP53 (panel size: 2,359 bp), for five patients; and 3) ESCC Panel, originally designed to target 31 frequently mutated genes in ESCC (panel size: 210,570 bp), for seven patients. Four missense and two nonsense mutations in TP53 were detected in CHPv2 and the TP53 Panel. Furthermore, one missense mutation in FBXW7 and one intronic mutation in the vicinity of exon 8 in TP53 were also observed in CHPv2 and the TP53 Panel, respectively. In the ESCC Panel, 47 mutations in 17 genes were detected in seven patients. The mutation detection rates by the TP53 Panel, CHPv2, and ESCC Panel were 593.0/Mb, 63.5/Mb, and 30.4/Mb, respectively. These mutation detection rates were more than 20-fold higher than that of WES, which has been reported to be 3.1/Mb for ESCC. These results suggest that tissue-specific cancer-related gene panels are a highly effective approach for mutation detection in ESCC.
Citation Format: Takeshi Iwaya, Fumitaka Endo, Kohei Kume, Yasushi Sasaki, Takashi Tokino, Satoshi Nishizuka. Mutation detection by target sequence analyses using tissue-specific panels in esophageal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4650. doi:10.1158/1538-7445.AM2017-4650