bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3210: An in vitro approach to study T cell exhaustion
Authors: Hastings William , Dranoff Glenn , Cao Alexander 
Published: July 2016

Abstract
Immune checkpoint inhibition is now a validated modality in the treatment of multiple cancers. In vitro immune cell assays are a key tool in the study of pharmacological agents for immune modulation. Assays often involve isolating lymphocytes from human peripheral blood followed by a T cell stimulus such as anti-CD3, staph enterotoxin B, or allogeneic APCs. While this approach is useful to probe the events of immune stimulation, it likely reflects a primary or secondary immune activation, rather than a state of exhaustion. As intra-tumoral T cells are thought to be exhausted by chronic/repeated exposure to self-antigen, we sought to define experimental conditions that would reflect this state in vitro. We reasoned that repeated stimulation of T cells using anti-CD3/anti-CD28 coated microbeads would induce an exhausted phenotype. We found that, using PBMC from normal healthy donors, frequent and repeated stimuli resulted in a loss of T cells’ ability to produce IL-2, TNFa, and IFNg as measured by intracellular cytokine staining. Interestingly, only one restimulation was necessary to confer this phenotype, as repeated stimuli over a longer time course did not result in further suppression. We also found that repeat stimulation conferred a surface marker phenotype suggestive of exhausted cells. Finally, we used cell death inhibitors z-7AAD-cmk and EGTA and found no effect on cytokine loss, supporting the hypothesis that cells are exhausted and not deleted by activation induced cell death. Thus we have defined experimental conditions that possibly reflect an exhausted T cell phenotype in vitro. This may be useful for the pharmacological evaluation of different therapeutic agents for their immune modulatory activities in cancer.
Citation Format: William Hastings, Glenn Dranoff, Alexander Cao. An in vitro approach to study T cell exhaustion. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3210.
Abstract 3222: Preclinical development of a humanized blocking antibody targeting the CD39 immune checkpoint for cancer immunotherapy
Authors: Augier Severine , Perrot Ivan , Dejou Cecile , Joly Rachel , Delahaye Stephane , Rispaud Blanc Helene , Denis Caroline , Gauthier Laurent , Bensussan Armand , Eliaou Jean-francois , Morel Yannis , Bonnefoy Nathalie , Bastid Jeremy , Paturel Carine 
Published: July 2016

Abstract
CD39 (ENTPD1) is a cell membrane ectonucleotidase that hydrolyzes extracellular immunoactivating ATP and ADP into AMP, which can be further hydrolyzed by ectonucleotidase CD73 into immunosuppressive adenosine. Within the tumor microenvironment, adenosine accumulation causes immune suppression and dysregulation of immune cell infiltrates resulting in tumor spreading. The role of CD39 expression on both Tregs and on tumor cells in promoting immunosuppression has been demonstrated in several reports. Blockade of CD39 may promote anti-tumor immunity by directly accumulating immunostimulating ATP and indirectly by reducing adenosine accumulation. Here, we describe the discovery and preclinical development of an anti-huCD39 blocking antibody for cancer immunotherapy. Parental anti-huCD39 mouse monoclonal antibody was humanized. The humanized mAb specifically binds huCD39 protein, but not CD39-like proteins. Nanomolar affinities for human CD39 were measured in SPR studies on recombinant CD39 protein and in flow cytometry titration studies on CD39 expressing transfectants and tumor cell lines. The humanized mAb blocked human CD39 ATPase activity in vitro in the nanomolar range, as demonstrated using transfected cells, CD39-expressing tumor cell lines, as well as human PBMC and ex-vivo isolated fresh tumor samples. The humanized mAb cross-reacted on cynomolgus CD39 and blocked ATPase activity on cynomolgus PBMC with similar efficacy as on human PBMC. Finally, treatment with blocking anti-CD39 mAb inhibited tumor growth in vivo in mouse tumor models. Taken together, these data support the clinical development of anti-CD39 neutralizing mAb for cancer immunotherapy.
Citation Format: Severine Augier, Ivan Perrot, Cecile Dejou, Rachel Joly, Stephane Delahaye, Helene Rispaud Blanc, Caroline Denis, Laurent Gauthier, Armand Bensussan, Jean-francois Eliaou, Yannis Morel, Nathalie Bonnefoy, Jeremy Bastid, Carine Paturel. Preclinical development of a humanized blocking antibody targeting the CD39 immune checkpoint for cancer immunotherapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3222.
Abstract 3238: Imbalance of circulating monocyte subsets in patients with squamous cell carcinoma of the head and neck
Authors: Sakakura Koichi , Takahashi Hideyuki , Motegi Sei-ichiro , Chikamatsu Kazuaki 
Published: July 2016

Abstract
Introduction: Human circulating monocyte is classified as three populations according to CD14 and CD16 expression: CD14+CD16- classical monocytes, CD14+CD16+ intermediate monocytes and CD14dimCD16+ nonclassical monocytes. Recent studies have unveiled the specific surface molecules and functions of each subset; however their dynamics in patients with cancer still remains unclear. In this study, we evaluated the proportions and the expressions of immunological/angiogenic molecules of the monocyte subsets in peripheral circulation in patients with squamous cell carcinoma of the head and neck (SCCHN).
Materials and Methods: Peripheral blood mononuclear cells were isolated from 9 patients with SCCHN and age-matched 4 healthy donors. The 3 monocyte subsets were identified according to CD14/CD16 expressions using multi-color flow cytometry. Mean fluorescence intensities of various HLA molecules and angiogenic markers were analyzed.
Results: Percentage of CD14+CD16+ intermediate monocyte among total monocyte tended to decrease in SCCHN patients than that in normal donors (p = 0.079). Higher surface expression of immunosuppressive HLA-G molecule on circulating classical (p = 0.045) and intermediate (p = 0.052) monocyte was observed in patients with SCCHN. An immunosuppressive receptor ILT4 showed tendency to increase on nonclassical monocyte (p = 0.061). Moreover, intracytoplasmic level of milk fat globule-EGF factor 8 (MFG-E8) in association with phagocytosis and angiogenesis tended to increase in intermediate (p = 0.064) and nonclassical (p = 0.052) monocytes of SCCHN patients.
Discussion and Conclusion: CD16+ intermediate and nonclassical monocytes have been reported as mature, proinflammatory and angiogenic population in periphery. In the present study, we showed possible alternations of these subsets in patients with SCCHN. Furthermore, increasing surface expressions of immunosuppressive molecules HLA-G and ILT4 on circulating monocytes implied immunological deterioration in cancer patients. However the sample size is still small, our preliminary data strongly suggests proportional and functional changes of peripheral monocyte subsets in SCCHN patients. Further analyses in more patients are currently underway.
Citation Format: Koichi Sakakura, Hideyuki Takahashi, Sei-ichiro Motegi, Kazuaki Chikamatsu. Imbalance of circulating monocyte subsets in patients with squamous cell carcinoma of the head and neck. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3238.
Abstract 3239: Cancer-associated fibroblasts induce immunosuppressive macrophages in head and neck squamous cell carcinoma
Authors: Takahashi Hideyuki , Sakakura Koichi , Chikamatsu Kazuaki 
Published: July 2016

Abstract
Background
Cancer-associated fibroblasts (CAFs) are one of main elements in the tumor microenvironment (TME). Among the innate and adaptive immune cells in the TME, tumor-associated macrophages (TAMs) are particularly abundant and play a protumoral role. However, an interaction between CAFs and TAMs in head and neck squamous cell carcinoma (HNSCC) still remains unclear. In the present study, we investigated whether CAFs skewed macrophage function toward the immune suppressive phenotype.
Materials and Methods
In vitro assay: CAFs were prepared from surgical tissue of HNSCC patients, and culture supernatants of CAFs were collected. Using this supernatants and peripheral blood mononuclear cells collected from healthy donors, immunological interactions between CAFs and TAMs was investigated.
In vivo assay: The immunohistochemistry (IHC) of 73 patients with HNSCC was performed. Anti-α-smooth muscle actin, CD68, and CD163 antibodies were used to identify CAFs and TAMs, respectively. The relationship between immunohistochemical parameters and clinical parameters was evaluated.
Results
The expression levels of CD68, CD163, CD14, CD80, CD86, and HLA-G were up-regulated in CD14+ cells co-cultured with the supernatants of CAFs (CAF-educated cells) compared with control cells. Moreover, the gene expressions of ARG1, IL6, IL10 and TGFB1 were also increased in CAF-educated cells. The CAF-educated cells suppressed T-cell proliferation more intensively than control cells, and neutralization of TGF-β and IL-10 led to significant restoration of T-cell proliferation. In IHC, the intensity of CAFs correlated with the number of CD68-positive macrophages, especially with CD163-positive macrophages. It also correlated with several clinical parameters such as lymphatic invasion, vascular invasion, nodal status, tumor stages and recurrence rate. Moreover, the high intensity of CAFs correlated with poor PFS and OS. The high intensity of CAFs and M2 macrophage infiltration were independent prognostic factors of PFS and OS in HNSCC.
Conclusion
The present study suggests that CAFs can induce an immunosuppressive phenotype of macrophages and contribute to establishing the immunosuppressive networks in TME, resulting poor prognosis in HNSCC patients.
Citation Format: Hideyuki Takahashi, Koichi Sakakura, Kazuaki Chikamatsu. Cancer-associated fibroblasts induce immunosuppressive macrophages in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3239.
Abstract 3241: M1 macrophages promote ovarian cancer metastasis through NF-κB/TWIST axis signaling pathway
Authors: Jo Un-Tek , Song. YongSang 
Published: July 2016

Abstract
The importance of tumor microenvironment in cancer progression has been recognized recently. However, factors associated with the tumor microenvironment are diverse and the molecular mechanisms are needed to be further defined for therapeutic intervention. Here we focused on the interaction between obesity related inflammatory cell, macrophage type 1 (MФ-1), and ovarian cancer cells in relation to metastasis. Two types of ovarian cancer cell lines; PA-1 and SKOV3, and monocyte-derived macrophages were used in this study. In the treatment with MФ-1 conditioned media, only SKOV3 cells were affected in proliferation activity. However migration and invasion were promoted by treatment with MФ-1 conditioned media in both ovarian cancer cells. In parallel, MФ-1 conditioned media treatment increased TWIST, EMT related gene. Interestingly, TWIST expression was correlated with protein expression levels of nuclear NF-κB (p65 and p50). Upon MФ-1 conditioned media treatment, nuclear translocation level and transcriptional activity of the NF-κB increased. Also, TPCK, NF-κB inhibitor suppressed invasion activity by MФ-1 conditioned media, in parallel with down-regulation of TWIST expression. Taken together, MФ-1 promoted EMT related metastatic potential in ovarian cancer cells through NF-κB signaling pathway. These results provide a new insight into the obesity related tumor microenvironment in ovarian cancer cell progression.
Citation Format: Un-Tek Jo, YongSang Song. M1 macrophages promote ovarian cancer metastasis through NF-κB/TWIST axis signaling pathway. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3241.
Abstract 3245: IL-15 superagonist/IL-15RαSushi-Fc fusion complex (IL-15SA/IL-15RαSu-Fc; ALT-803) markedly enhances specific subpopulations of NK and memory CD8+ T cells, and mediates potent anti-tumor activity of murine breast and colon carcinomas
Authors: Kim Peter S. , Kwilas Anna R. , Xu Wenxin , Alter Sarah , Jeng Emily K. , Wong Hing C. , Schlom Jeffrey , Hodge James W. 
Published: July 2016

Abstract
Interleukin (IL)-15-N72D superagonist-complexed with IL-15RαSushi-Fc fusion protein (IL-15SA/IL-15RαSu-Fc; ALT-803) has been reported to exhibit significant anti-tumor activity in murine myeloma, rat bladder cancer, and murine glioblastoma models. In this study, we examined the immunomodulatory and anti-tumor effects of IL-15SA/IL-15RαSu-Fc in tumor-free and highly metastatic tumor-bearing mice. Here, IL-15SA/IL-15RαSu-Fc significantly expanded NK and CD8+ T cells. In examining natural killer (NK) cell subsets, the greatest significant increase was in highly cytotoxic and migrating (CD11b+, CD27hi; high effector) NK cells, leading to enhanced function on a per-cell basis. CD8+ T cell subset analysis determined that IL-15SA/IL-15RαSu-Fc significantly increased IL-15 responding memory (CD122+, CD44+) CD8+ T cells, in particular those having the innate (NKG2D+, PD1-) phenotype. In 4T1 breast tumor-bearing mice, IL-15SA/IL-15RαSu-Fc induced significant anti-tumor activity against spontaneous pulmonary metastases, depending on CD8+ T and NK cells, and resulting in prolonged survival. Similar anti-tumor activity was seen in the experimental pulmonary metastasis model of CT26 colon carcinoma cells, particularly when IL-15SA/IL-15RαSu-Fc was combined with a cocktail of checkpoint inhibitors, anti-CTLA-4 and anti-PD-L1. Altogether, these studies showed for the first time that IL-15SA/IL-15RαSu-Fc (1) promoted the development of high effector NK cells, (2) enhanced function of NK cells, and (3) played a vital role in reducing tumor metastasis and ultimately survival, especially in combination with checkpoint inhibitors.
Citation Format: Peter S. Kim, Anna R. Kwilas, Wenxin Xu, Sarah Alter, Emily K. Jeng, Hing C. Wong, Jeffrey Schlom, James W. Hodge. IL-15 superagonist/IL-15RαSushi-Fc fusion complex (IL-15SA/IL-15RαSu-Fc; ALT-803) markedly enhances specific subpopulations of NK and memory CD8+ T cells, and mediates potent anti-tumor activity of murine breast and colon carcinomas. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3245.
Abstract 3250: Caspase-1 from MDSC promote MyD88 dependent carcinogenesis that is T-cell independent
Authors: Zeng Qi , Qualliotine Jesse R. , Blosser Lee , Pardoll Drew , Kim Young J. 
Published: July 2016

Abstract
Myeloid-derived-suppression cells (MDSC) are functionally defined to suppress T-cells in their primary means of immune evasion mechanism in cancer. However, we have discovered that MDSC can have direct, T-cell independent pro-carcinogenic effect on tumor cells. Sorted monocytic CD14+/CD11b+/HLA-DRlow MDSCs from head and neck squamous cell carcinoma (HNSCC) patients were found to increase the proliferation index of HNSCC cells. This induction of tumor proliferation was not dependent on MDSC-tumor cell contact. Exome analysis showed that these human MDSC from cancer patients expressed high levels of inflammasome complex, and supernatant from the MDSCs were found to secrete IL-1b and IL-18. When we probed the functional significance of these inflammasome complex, we found that MDSC's promotion of tumor proliferative index of the tumor was found to be Caspase-1 dependent. To test this in vivo, T-cell depleted caspase-1 null mice showed significant decrease in tumor growth rate. To confirm the importance of myeloid inflammasome signaling in carcinogenesis, we suppressed MyD88 gene in tumor cell line Cal27 and found that the ability of MDSC to promoting tumor proliferation is diminished. Taken together, our findings demonstrate that tumor infiltrating MDSCs may play a prominent role in chronic inflammation associated carcinogenesis.
Citation Format: Qi Zeng, Jesse R. Qualliotine, Lee Blosser, Drew Pardoll, Young J. Kim. Caspase-1 from MDSC promote MyD88 dependent carcinogenesis that is T-cell independent. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3250.
Abstract 3259: LY3127804, a novel anti-Angiopoietin-2 antibody in combination with an anti-VEGFR2 antibody potently inhibits angiogenesis, tumor growth and metastasis
Authors: Chintharlapalli Sudhakar R. , Croy Johnny E. , Leung Donmienne , Gerald Damien , Lu Jirong , Iversen Philip W. , Lee Linda N. , Huber Lysiane , Tetreault Jonathan , Almonte-Baldonado Rowena , Xu Jianghuai , Ramamurthy Bharathi , Pereira Jennifer A. , Chow Chi-Kin , Hanauske Axel-Rainer , Wacheck Volker , Benjamin Laura , Liu Ling 
Published: July 2016

Abstract
Angiopoeitin-2 (Ang2) is released from endothelial cells only in response to stimulus (e.g. wound healing, tumor growth) and facilitates blood vessel sprouting and inhibits pericyte-endothelial cell interaction via Tie2 signaling. Combination of an anti-Ang2 antibody and aflibercept, a VEGF trap, has been shown to inhibit tumor growth and decrease tumor vascularity in mouse xenograft tumor models (Daly et al., Cancer Res (2013) 73(1):108). Multiple investigational anti-Ang2 antibody therapies are currently in clinical trials. LY3127804 is a humanized and engineered IgG4 isotype antibody that selectively binds to Ang2 with high affinity and neutralizes Ang2 induced phospho-Tie2. LY3127804 inhibits sprouting angiogenesis and increases pericyte coverage in a mouse developmental retinal angiogenesis model and in mice bearing PC3 xenograft tumors. Combination of LY3127804 and DC101, a potent anti-VEGFR2 antibody, exhibits enhanced efficacy when compared to monotherapy in multiple patient derived xenograft models including NSCLC and ovarian cancers. Anti-Ang2 antibody monotherapy alone resulted in marginal reduction of tumor growth and improved overall survival, while DC101monotherapy had greater reduction in tumor volume with no survival benefit in MDA-MB-231 breast orthotopic model. Combination of anti-Ang2 antibody with anti-VEGFR2 antibody shows reduction in tumor volume and improved overall survival. This robust pre-clinical evidence supports testing the combination of anti-Ang2 and anti-VEGFR2 antibodies in the clinic. LY3127804 is currently in Phase 1 clinical trials (NCT02597036)
Citation Format: Sudhakar R. Chintharlapalli, Johnny E. Croy, Donmienne Leung, Damien Gerald, Jirong Lu, Philip W. Iversen, Linda N. Lee, Lysiane Huber, Jonathan Tetreault, Rowena Almonte-Baldonado, Jianghuai Xu, Bharathi Ramamurthy, Jennifer A. Pereira, Chi-Kin Chow, Axel-Rainer Hanauske, Volker Wacheck, Laura Benjamin, Ling Liu. LY3127804, a novel anti-Angiopoietin-2 antibody in combination with an anti-VEGFR2 antibody potently inhibits angiogenesis, tumor growth and metastasis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3259.
Abstract 3292: Manipulating osteoblast differentiation in order to inhibit protection of AML cells within the bone marrow
Authors: Sterner Rosalie , Kremer Kimberly , Dudakovic Amel , Kaufmann Scott , Westendorf Jennifer , van Wijnen Andre , Hedin Karen 
Published: July 2016

Abstract
While standard chemotherapeutics can initially induce remission in patients with acute myelogenous leukemia (AML), patients often relapse in part due to the protective bone marrow microenvironment, which promotes the survival of AML cells. Using an in vitro cell culture model of the bone marrow, our lab recently showed that differentiating osteoblasts potently protect AML cells from apoptosis. In addition, we found that differentiating osteoblasts treated with histone deacetylase inhibitors (HDACi) fail to protect AML cells in this model. Since HDACi decreased the expression of genes required for osteoblast differentiation, we hypothesize that blocking specific stage(s) of osteoblast differentiation may make AML cells within the bone marrow more susceptible to chemotherapy. We are currently disrupting specific osteoblast differentiation stages via molecular or therapeutic drug manipulation and exploring the impact on AML cell survival both in cell culture models of the bone marrow microenvironment and in a mouse model of AML. The results of our studies will characterize novel bone marrow molecular and cellular interactions that promote the survival of AML cells. Targeting these interactions in combination therapies may more completely eradicate the AML cell reservoir and better prevent relapse.
Citation Format: Rosalie Sterner, Kimberly Kremer, Amel Dudakovic, Scott Kaufmann, Jennifer Westendorf, Andre van Wijnen, Karen Hedin. Manipulating osteoblast differentiation in order to inhibit protection of AML cells within the bone marrow. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3292.