bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3228: High salt induces anti-inflammatory MΦ2-like phenotype in peripheral macrophages
Authors: Amara Suneetha , Mehta Toral R. , Whalen Margaret , Tiriveedhi Venkataswarup
Published: July 2016

Macrophages play a critical role in inflammation and antigen-presentation. Abnormal macrophage function has been attributed in autoimmune diseases and cancer progression. Recent evidence suggests that high salt tissue micro-environment causes changes in macrophage activation. In our current report, we studied the role of extracellular sodium chloride (NaCl) on phenotype changes in peripheral circulating monocyte/macrophages collected from healthy donors. High salt (0.2 M NaCl) treatment resulted in a decrease in MΦ1 macrophage phenotype (CD11b+CD14highCD16low) from 77.4±6.2% (0.1 M) to 29.3±5.7% (0.2 M, p<0.05), while there was an increase in MΦ2 macrophage phenotype (CD11b+ CD14lowCD16high) from 17.2±5.9% (0.1 M) to 67.4±9.4% (0.2 M, p<0.05). ELISA-based cytokine analysis demonstrated that high salt treatment induced decreased expression of in the MΦ1 phenotype specific pro-inflammatory cytokine, TNFα (3.3 fold), IL-12 (2.3 fold), CCL-10 (2 fold) and CCL-5 (3.8 fold), but conversely induced an enhanced expression MΦ2 phenotype specific anti-inflammatory cytokine, IL-10, TGFβ, CCL-17 (3.7 fold) and CCR-2 (4.3 fold). Further high salt treatment significantly decreased phagocytic efficiency of macrophages and inducible nitric oxide synthetase expression. Taken together, these data suggest that high salt extracellular environment induces an anti-inflammatory MΦ2 macrophage phenotype with poor phagocytic and potentially reduced antigen presentation capacity commonly found in tumor microenvironment.

Citation Format: Suneetha Amara, Toral R. Mehta, Margaret Whalen, Venkataswarup Tiriveedhi. High salt induces anti-inflammatory MΦ2-like phenotype in peripheral macrophages. [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 3228.
Abstract 3232: NF-κB regulates suppression of macrophage activity during tumor initiation
Authors: Ratnam Nivedita , Wang David , Guttridge Denis
Published: July 2016

Constitutive activation of NF-κB is characteristic of most cancer cells. However, how NF-κB functions during tumor initiation is not well established. Recent findings from our laboratory showed that one mechanism by which NF-κB functions to promote tumor initiation is by suppressing immune surveillance. Here we investigated how this occurs. Initiating tumors are characterized by a massive infiltration of immune cells, especially macrophages that exhibit anti-tumor activity through the secretion of pro-apoptotic factors like tumor necrosis factor (TNF) and nitric oxide (NO). Using co-culture assays with conditioned media from Ras transformed tumor cells in the presence of NF-κB, we find that the tumor cells are able to protect themselves from the pro-apoptotic activity of the macrophages by secreting a neutralizing factor. RNA sequencing analysis identified this factor as macrophage inhibitory cytokine-1 (MIC-1)/ growth and differentiation factor-15 (GDF-15). Our results show that MIC-1/GDF-15 is a direct transcriptional target of NF-κB. Furthermore, xenografts in SCID mice show delayed tumor initiation in p65+/+ Ras tumor cells that are depleted for MIC-1/GDF-15 compared to wild type p65+/+ Ras tumor cells. Moreover, we find that MIC-1/GDF-15 inactivates macrophages directly by suppressing their ability to produce TNF and NO. This suppression occurs through the inhibition of NF-κB signaling in the macrophages. Interestingly, expression of MIC-1/GDF-15 is increased in patients with pancreatic ductal adenocarcinoma. In the pancreatic cancer cell line, Panc02, we show that knocking down MIC-1/GDF-15 prevents macrophage inhibition thereby increasing their sensitivity to macrophage-mediated killing both in vitro and in vivo. Thus, NF-κB functions during pancreatic cancer initiation to suppress the anti-tumor activity of infiltrating macrophages by regulating the expression of MIC-1/GDF-15.

Citation Format: Nivedita Ratnam, David Wang, Denis Guttridge. NF-κB regulates suppression of macrophage activity during tumor initiation. [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 3232.
Abstract 3233: Macrophages orchestrate early dissemination of HER2+ cancer cells
Authors: Linde Nina , Mortha Arthur , Saenger Nicole , Sosa Maria Soledad , Harper Kathryn , Tardio Ethan , Fehm Tanja , Karn Thomas , Merad Miriam , Aguirre-Ghiso Julio A.
Published: July 2016

Metastatic dissemination was proposed to occur only in invasive cancer. However, detection of cancer cells in the bone marrow of patients with pre-invasive breast ductal carcinoma in situ (DCIS) showed that dissemination could, through unknown mechanisms, take place earlier. Late evolved invasive breast cancer cells disseminated by recruiting circulating monocyte-derived tumor associated macrophages (TAMs). We therefore investigated whether macrophages might actively participate in early dissemination. We show that pre-malignant cancer cells in MMTV-HER2 mice produced CCL2 in an NFkB dependent manner and thereby recruited resident mammary tissue macrophages into the duct, months before monocyte-derived TAMs arrived. Intra-epithelial mammary tissue macrophages induced an epithelial-to-mesenchymal transition in early HER2+ cells in a Wnt-dependent manner and thereby drove early dissemination. Consequently, depletion of macrophages from pre-malignant MMTV-HER2 mice significantly reduced early dissemination of pre-malignant cancer cells as measured by reduction in levels of circulating cancer cells and disseminated cancer cells in lungs. Interestingly, macrophage depletion during pre-malignant stages alone was sufficient to reduce the onset of late metastasis in MMTV-HER2 mice, indicating that early disseminating cancer cells contributed to late metastasis formation. In humans DCIS samples we frequently found intra-epithelial macrophages inside of ducts and their presence correlated with reduced E-Cadherin levels. Finally, a first pilot study indicated that those patients whose DCIS lesions contained macrophage +/E-Cadherin low microenvironments frequently had DCCs in the bone marrow. Taken together, we reveal that resident mammary tissue macrophages can promote early dissemination, and unravel a mechanism how early cancer spread might proceed in breast cancer patients and in other cancers where metastasis are diagnosed and the primary tumor is never detected (cancer of unknown primary). We also propose that early DCCs play a long-term causal role in metastasis development when patients are diagnosed in late stages, implying that the heterogeneity of metastases is higher than we might anticipate.

Citation Format: Nina Linde, Arthur Mortha, Nicole Saenger, Maria Soledad Sosa, Kathryn Harper, Ethan Tardio, Tanja Fehm, Thomas Karn, Miriam Merad, Julio A. Aguirre-Ghiso. Macrophages orchestrate early dissemination of HER2+ cancer cells. [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 3233.
Abstract 3234: Genetic deletion of macrophage migration inhibitory factor reduces oral carcinogenesis
Authors: Oghumu Steve , Knobloch Thomas , Terrazas Cesar , Varikuti Sanjay , Bollinger Claire , Iwenofu Hans , Weghorst Christopher , Satoskar Abhay
Published: July 2016

Oral cancer kills about 1 person every hour, each day in the United States and is the 6th most prevalent cancer worldwide. The pro-inflammatory cytokine ‘macrophage migration inhibitory factor’ (MIF) has been shown to be expressed in oral cancer patients, yet its precise role in oral carcinogenesis is not clear. In this study, we examined the impact of global Mif deletion on the cellular and molecular process occurring during oral carcinogenesis using a well-established mouse model of oral cancer with the carcinogen 4-nitroquinoline-1-oxide (4NQO). C57BL/6 Wild-type (WT) and Mif knock-out mice were administered with 4NQO in drinking water for 16 weeks, then regular drinking water for 8 weeks. Mif knock-out mice displayed fewer oral tumor incidence and multiplicity, accompanied by a significant reduction in the expression of pro-inflammatory cytokines Il-1β, Tnf-α, chemokines Cxcl1, Cxcl6, Cxcl8 and Ccl3 and other molecular biomarkers of oral carcinogenesis Mmp1 and Ptgs2. Further, recruitment of myeloid-derived tumor promoting immune cells was inhibited in Mif knock-out mice. Our results demonstrate that genetic Mif deletion reduces the incidence and severity of oral carcinogenesis, by inhibiting the expression inflammatory cytokines and infiltration of tumor promoting immune cells. Thus, targeting MIF is a promising strategy for the prevention or therapy of oral cancer.

Citation Format: Steve Oghumu, Thomas Knobloch, Cesar Terrazas, Sanjay Varikuti, Claire Bollinger, Hans Iwenofu, Christopher Weghorst, Abhay Satoskar. Genetic deletion of macrophage migration inhibitory factor reduces oral carcinogenesis. [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 3234.
Abstract 3235: IDH mutant gliomas escape natural killer cell immune surveillance by downregulation of NKG2D ligands
Authors: Rao Aparna , Zhang Xiaoran , Deibert Christopher , Sette Paola , Chan Tim , Grandi Paola , Amankulor Nduka
Published: July 2016

Background: Diffuse gliomas are fatal primary brain tumors that are poorly immunogenic. The basis for insufficient anti-tumor immunity in diffuse gliomas is not understood. Gain of function mutations in isocitrate dehydrogenases (IDH1 and IDH2) promote diffuse glioma formation through epigenetic reprogramming of a number of genes, including immune-related genes. Here, we identify epigenetic dysregulation of natural killer (NK) cell ligand genes as significant contributors to immune escape in glioma. Methods: We analyzed the TCGA database for immune gene expression patterns in IDH mutant or wild-type gliomas and identified differentially expressed immune genes. NKG2D ligands expression levels and NK cell-mediated lysis were measured in IDH mutant and wild-type patient-derived glioma stem cells and in genetically engineered astrocytes. Finally, we assessed the impact of hypomethylating agent 5-aza- 2’deoxycytodine (Decitabine) as a potential NK cell sensitizing agent in IDH mutant cells. Results: All IDH mutant glioma stem-like cell lines exhibited significantly lower expression of NKG2D ligands compared with IDH wild-type cells. Consistent with these findings, IDH mutant glioma cells and astrocytes were resistant to NK cell- mediated lysis. The hypomethylating agent Decitabine increased NKG2D ligand expression, and restored NK-mediated lysis of IDH mutant cells in an NKG2D- dependent manner. Conclusions: IDH mutant glioma cells acquire resistance to NK cells through epigenetic silencing of NKG2D ligands ULBP1 and ULPB3. Decitabine-mediated hypomethylation restores ULBP1 and ULBP3 expression in IDH mutant glioma cells and may provide a clinically useful method to sensitize IDH mutant gliomas to NK cell- mediated immune surveillance in patients with IDH mutated diffuse gliomas.

Citation Format: Aparna Rao, Xiaoran Zhang, Christopher Deibert, Paola Sette, Tim Chan, Paola Grandi, Nduka Amankulor. IDH mutant gliomas escape natural killer cell immune surveillance by downregulation of NKG2D ligands. [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 3235.
Abstract 3247: Granulocytic MDSCs (CD11b+CD15+CD14-HLADR-) in peripheral blood of mesothelioma patients are elevated and suppress T cell proliferation and function via reactive oxygen species
Authors: Khanna Swati , Mussai Francis , Thomas Anish , Middleton Gary , Yuan Constance , Morrow Betsy , Zhang Jingli , Pastan Ira , Stetler-Stevenson Maryalice , Hassan Raffit
Published: July 2016

The role of myeloid derived suppressor cells (MDSCs) in mediating tumor immunosuppression in patients with malignant mesothelioma has not been well characterized. The goal of our study was to analyze for the presence of both monocytic (Mo) and granulocytic (Gr) MDSCs in blood of mesothelioma patients, and characterize their mechanism of suppression of T cells responses. We evaluated the peripheral blood of patients (n = 23) for the presence of Gr-MDSC (CD11b+CD15+CD14-HLADR-) and Mo-MDSC (CD11b+CD14+HLADR-). Gr-MDSC (62.3±13.9% vs. 47.8±17.8%; p = 0.005) as well as monocytic MDSC (0.2±0.4% vs. 0.1±0.3%; p = 0.01) were significantly elevated in peripheral blood of mesothelioma patients as compared to healthy donors (n = 21). The frequency of Gr-MDSC was higher than Mo-MDSC. Thus, Gr-MDSCs appeared to be the main immunosuppressive population and were investigated further.Gr-MDSC from mesothelioma patients were found to inhibit the proliferation of both autologous CD4 and CD8 T cells (mean inhibition of proliferation of 61.9% for CD4 and 75.5% for CD8 at 1:2 T: Gr-MDSC ratio) and was accompanied by a decrease in IFN-γ released by T cells in the co-culture supernatants (92.4 pg/mL at 1:2 ratio). To determine the mechanism by which Gr-MDSC inhibit T cells, we tested the levels of reactive oxygen species (ROS), and nitrite oxide species and arginase activity in Gr-MDSC sorted from peripheral blood. Both arginase (0.3±0.3 vs. 0.2±0.3 μM) and nitrite levels (1.8±2.5 vs. 0 μM) in Gr-MDSC were below detection limit in both patients and donors and were marginally affected by the addition of their respective inhibitors (nor-NOHA and L-NMMA). However, the levels of ROS in Gr-MDSCs derived from mesothelioma patients were on average 3 folds higher than the Gr-myeloid cells from healthy donors (average MFI ∼15000 vs. ∼5000). In summary our results show that the major immunosuppressive cell population in mesothelioma patients are Gr-MDSCs and they suppress T cell proliferation and activation through release of reactive oxygen species.

Citation Format: Swati Khanna, Francis Mussai, Anish Thomas, Gary Middleton, Constance Yuan, Betsy Morrow, Jingli Zhang, Ira Pastan, Maryalice Stetler-Stevenson, Raffit Hassan. Granulocytic MDSCs (CD11b+CD15+CD14-HLADR-) in peripheral blood of mesothelioma patients are elevated and suppress T cell proliferation and function via reactive oxygen species. [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 3247.
Abstract 3249: Regulation of interferon regulatory factor-8 (IRF8) during myelopoiesis is a critical checkpoint for the formation of defective myeloid cells in cancer
Authors: Netherby Colleen S. , Messmer Michelle N. , Abrams Scott I.
Published: July 2016

Aberrant myelopoiesis is commonly observed in patients with solid cancers. This results in the production of myeloid-derived suppressive cells (MDSCs) which promote tumor growth and metastasis. However, the molecular mechanisms underlying MDSC development have remained poorly understood. The transcription factor IRF8 is integral for overseeing myelopoietic fate; high IRF8 expression during normal myelopoiesis favors monocytic differentiation and directly inhibits granulocytic differentiation. We recently showed that tumor-derived STAT3/5-activating cytokines downregulate the expression of IRF8 in myeloid cells leading to the accumulation of MDSCs in the periphery, which are primarily granulocytic. However it is unclear if MDSC production is a consequence of IRF8 downregulation in the periphery or upstream in the bone marrow during myelopoiesis. To that end, we utilized a novel mouse model expressing an IRF8-EGFP fusion protein which allowed us to investigate changes in IRF8 expression during both orthotopic and spontaneous mammary tumor growth and progression. Our results showed that: 1) the total granulocyte-monocyte progenitor (GMP) fraction, as with the peripheral MDSC population, was responsive to tumor burden and greatly expanded with increasing tumor size, and 2) IRF8 expression within the total GMPs significantly decreased during tumor growth, reflecting the expansion of a newly defined primarily IRF8lo granulocyte progenitor (GP) population, and 3) tumor-induced GPs were hyper-responsive not only to G-CSF, but also M-CSF to generate a ‘biased’ granulocytic MDSC-like phenotype. Expression of IRF8 in other progenitor populations was not affected, suggesting that IRF8lo GMPs/GPs are a key source of the peripheral MDSC response. Lastly, genetically enforced IRF8 expression in myeloid progenitors of the bone marrow constrained aberrant myelopoiesis during tumor growth, resulting in delayed autochthonous tumor growth and reduced spontaneous lung metastasis. Altogether, these data reinforce the notion that modulation of IRF8 in myeloid progenitors is an early consequence of the neoplastic process and a potential target for therapeutic intervention.

NIH R01CA140622

NIH T32CA085183

Citation Format: Colleen S. Netherby, Michelle N. Messmer, Scott I. Abrams. Regulation of interferon regulatory factor-8 (IRF8) during myelopoiesis is a critical checkpoint for the formation of defective myeloid cells in cancer. [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 3249.
Abstract 3260: Differential effects of antiangiogenesis treatments on betel-nuts-related head and neck squamous cell carcinoma in Taiwan
Authors: Chen Jo-Pai , Chang Jui-Ying , Kuo Sung-Hsin , Hong Ruey-Long
Published: July 2016

Background:

The treatment of head and neck squamous cell carcinoma(HNSCC) in Taiwan is very challenging and might be related to betel-nuts use. Betel-nuts chewing might contribute to (1)strong inflammation, invasion, and angiogenesis; (2)easy recurrence or metastasis; (3) poor response to chemotherapy, radiation, and epidermal growth factor receptor(EGFR) inhibitors.

Purpose:

We try to prove different kinds of anti-angiogenesis treatments will lead to different response on betel-nuts related HNSCC in Taiwan.

Methods:

Different anti-angiogenesis treatments, such as axitinib(VEGFR2 inhibitor), nintedanib(VEGFR2/FGFR inhibitor), and regorafenib(VEGFR2/FGFR/ more other signals inhibitor) were first used to treat (1)HUVEC; (2)HNSCC cell lines(SCC4, SCC9, SCC15, SCC25, FaDu, SAS, KB, Cal27, and TW2.6, betel-nuts related) to evaluate (a) invasion capacity by wound healing; (b) drug sensitivity by MTT assay; (c)synergistic effect with chemotherapy, EGFR inhibitor, and polo-like kinase inhibitor. Western blotting was also used to test signal change by treatments. TW2.6 had already been proved to possess defective p53, p16 loss, and increased Bcl2.

Results:

In our previous study, TW2.6 was resistant to chemotherapy, radiation, EGFR inhibitors, and ani-angiogenesis treamtents, such as axitinib and sunitinib. Axitinib, pure VEGFR2 inhibitor, was found to suppress HUVEC more prominently than nintedanib or regorafenib did. Invasion capacity of all HNSCC cell lines were all blocked by the three drugs but regorafenib did mostly well. However, axitinib had no effect on TW2.6; but nintedanb and regorafenib had moderate response on TW2.6. Besides, nintedanib and regorafenib both would resensitize TW2.6 to respond to chemotherapy, EGFR inhibitor, and polo-like kinase inhibitor again. Western blotting showed mesenchaml differentiation markers(slug, Twist, snail, Axl, c-MET, Vimentin) decreased after nintedanib and regorafenib use, too.

Conclusion:

TW2.6 might reflect treatment refractoriness of betel-nuts related HNSCC in Taiwan. In addition to PI3K/mTOR dual inhibition and polo-like kinase inhibitor with radiation(our studies shown in AACR and ESMO2015), VEGFR2/FGFR dual blockage might be effective on TW2,6 and resensitize TW2.6 to EGFR inhibitor, polo-like kinase inhibitor, & chemotherapy, maybe through the inhibition of mesenchymal transformation. VEGFR2/FGFR dual blockage will be one future combination backbone of betel-nuts related HNSCC.

Citation Format: Jo-Pai Chen, Jui-Ying Chang, Sung-Hsin Kuo, Ruey-Long Hong. Differential effects of antiangiogenesis treatments on betel-nuts-related head and neck squamous cell carcinoma in Taiwan. [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 3260.
Abstract 3263: Dietary supplement 3, 3’-diindolylmethane (DIM) as an antiangiogenic agent in breast cancer
Authors: Ben Rahoma Ghada M. , Tuli Neha Y. , Bednarczyk Robert B. , Maniyar Rachana R. , Mittelman Abraham , Geliebter Jan , Tiwari Raj
Published: July 2016

Tumor angiogenesis refers to the sprouting and cooption of proliferating endothelial cells (EC’s) from adjacent pre-existing host vasculature, and is a key target of cancer therapy. Tumor cells exploit their microenvironment by releasing cytokines and growth factors to promote and support angiogenesis. Within this complex tumor microenvironment, we and others have shown that tumors can recruit bone marrow derived endothelial progenitor cells that differentiate into mature bone marrow-derived endothelial cells and incorporate into sprouting tumor neovessels. Under pathological circumstances, such as breast cancer, a clear association between estrogen receptor expression by EC’s, angiogenic activity, and/or tumor invasiveness has been made. Approximately, 80% of breast cancers are hormone-receptor-positive cancers, thus enabling tamoxifen as the mainstay of breast cancer therapy. The roles of the anti-estrogens fulvestrant (ICI) and the dietary supplement 3, 3’-diindolylmethane (DIM) on cell-cell interaction and angiogenesis have not been fully elucidated. This study is designed to evaluate and compare the effect of these antiestrogens on angiogenesis at the cellular and molecular levels using tube formation of human umbilical vein endothelial cells (HUVEC) as an in vitro angiogenesis model. HUVEC cells were treated with serial dilutions of either DIM or ICI in presence and absence of (3nM) estrogen, and subjected to in vitro tube formation, proliferation, migration, and angiogenesis antibody array assays. We report that HUVEC cells are more sensitive to DIM than ICI. At 25 μM concentration, DIM significantly inhibited the crucial steps of angiogenesis including HUVEC cells proliferation, migration, cytokine release, and tube formation in an estrogen independent manner. On the other hand, at 1 μM concentration, ICI significantly exerted an antiangiogenic effects inhibiting HUVEC cells proliferation, migration, and tube formation, but this effect was totally dependent on the presence of estrogen. These results are validated by our observation that HUVEC cells express estrogen receptor beta (ER-β) and not estrogen receptor alpha (ER-α). A correlative effect between the antiangiogenic activity of DIM and ER-β upregulation was noted. We believe that the anti-estrogenic activity of DIM is mediated through the genomic and non-genomic activity of ER-β in endothelial cells predicting a new target for DIM to manifest its antiangiogenic effect.

Citation Format: Ghada M. Ben Rahoma, Neha Y. Tuli, Robert B. Bednarczyk, Rachana R. Maniyar, Abraham Mittelman, Jan Geliebter, Raj Tiwari. Dietary supplement 3, 3’-diindolylmethane (DIM) as an antiangiogenic agent in breast cancer. [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 3263.
Abstract 3267: Eltd1 as an anti-angiogenic therapy against glioma tumors
Authors: Ziegler Jadith , Smith Nataliya , Saunders Debra , Evans Blake , Battiste James D. , Sughrue Michael , Tompkins Paul , Sutton Jake , Lerner Megan , Coutinho de Souza Patricia , Fung Kar-Ming , Wren Jonathan , Towner Rheal
Published: July 2016

Gliomas consist of up to 80% of malignant brain tumors that are invasive and typically resistant to radiotherapy and chemotherapy. Finding biomarkers to high-grade gliomas can enable better diagnosis and therapeutic intervention for this disease. We have identified ELTD1 as a biomarker for high-grade human gliomas. Here, we report our findings in vivo using anti-ELTD1 (at two different concentrations), Bevacizumab, and IgG antibodies on human G55 xenograft glioma models. Using MRI, we investigate tumor growth, perfusion, tumor blood flow, and microvessel density changes in mice. In addition, survival rates were measured.

Mice were implanted with human G55 xenograft glioma cells and were left untreated, administered anti-ELTD1, (1 mg/kg or 2 mg/kg every 2-3 days) Bevacizumab (2 mg/kg every 2-3 days) or IgG (1 mg/kg every 2-3 days). MRI experiments were performed to assess tumor growth and calculate tumor volumes. In order to assess angiogenesis, representative histology slides were obtained and stained for blood vessels using CD34 antibody and microvessel density (MVD) was then calculated. Finally, cerebral blood flow rates from MR perfusion imaging was obtained as well as MR angiography to determine tumor blood volume from mice in each group.

Our results show a significant decrease in tumor volumes and increase in percent survival for mice treated with 1 mg/kg and 2 mg/kg of ELTD1 antibody compared to untreated mice and IgG treated mice. Mice also had an increase in perfusion, decrease in tumor blood volume and decrease in MVD. Anti-ELTD1 antibody therapy reduced tumor volumes, prolonged life, and overall decreased angiogenesis in our mouse model. Anti-ELTD1 therapy may be an ideal anti-angiogenic treatment for high-grade gliomas.

Citation Format: Jadith Ziegler, Nataliya Smith, Debra Saunders, Blake Evans, James D. Battiste, Michael Sughrue, Paul Tompkins, Jake Sutton, Megan Lerner, Patricia Coutinho de Souza, Kar-Ming Fung, Jonathan Wren, Rheal Towner. Eltd1 as an anti-angiogenic therapy against glioma tumors. [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 3267.
Abstract 3269: Recapitulating the BETH adjuvant breast cancer trial (NCT00625898) using clinically accurate orthotopic surgical resection models
Authors: Shiels Liam S. , Miller Ian S. , Morgan Clare , Cremona Mattia , Kerbel Robert , Hennessey Bryan , O’Donovan Norma , Crown John , Byrne Annette T.
Published: July 2016

Background: An overwhelming majority of phase III clinical trials in the oncology setting fail as promising results observed in the ‘pre-clinic’ are not successfully translated to patients. As such, improved models which more accurately predict outcomes are now required. Herein, we have established orthotopic surgical resection models of Her2+ breast cancer, which replicate the phenotype of clinical post-resection metastasis [1,2]. Using these models we have recapitulated the BETH adjuvant breast cancer trial (NCT00625898) where the addition of bevacizumab (BVZ) to chemotherapy plus trastuzumab (TRAST) failed to provide additional benefit in the adjuvant setting. Methods: SCID mice were orthotopically implanted with bioluminescent Her2+ MDA-MB-231or HCC1954 cells and palpable tumors resected approximately 5 weeks later. 3 weeks after resection, mice were treated with 10 mg/kg TRAST + 5mg/kg paclitaxel IP once weekly for 6 cycles with or without weekly BVZ (5mg/kg IP). Metastasis was monitored by weekly bioluminescence imaging. Results: Tumor growth and imaging data confirmed that the addition of BVZ to adjuvant TRAST + chemotherapy provided no additional benefit compared with TRAST + chemotherapy alone. Previous pre-clinical studies using inappropriate non-resection models failed to predict this response. Furthermore, Reverse Phase Protein Array analysis of treated tumors implicated HER2, VEGF, mTOR and the intrinsic mitochondrial cell death apoptotic pathways in treatment resistance. Conclusion: In summary, our data provides evidence for pre-clinical models which better predict clinical outcome in the breast cancer adjuvant setting, and which represent an important resource for interrogating resistance pathways and identifying novel biomarker signatures. [1]. Breast. 2013 Aug;22 Suppl 2:S57-65; [2]. Cancer Res. 2013 May 1;73(9):2743-8.This work was performed under Irish HPRA Authorization AE18982 and was supported by the Clinical Cancer Research Trust. ATB receives funding from the Irish Cancer Society Collaborative Cancer Research Centre BREAST-PREDICT Grant (CCRC13GAL).

Citation Format: Liam S. Shiels, Ian S. Miller, Clare Morgan, Mattia Cremona, Robert Kerbel, Bryan Hennessey, Norma O’Donovan, John Crown, Annette T. Byrne. Recapitulating the BETH adjuvant breast cancer trial (NCT00625898) using clinically accurate orthotopic surgical resection models. [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 3269.
Abstract 3270: Myriocin, an inhibitor of serine palmitoyltransferase activity, has the anti-angiogenic effect by attenuating VEGF-induced angiogenesis signaling pathway
Authors: Lee Sun Hee , Jung Yuk Dong , Lee Hye-Eun , Lee You Mie
Published: July 2016

Myriocin is a powerful serine palmitoyl transferase inhibitor, by inhibiting of sphingosine biosynthesis. Myriocin is also known to have immunosuppressive activity. Here, we demonstrate the anti-angiogenic effect of myriocin. Myriocin decreased vascular endothelial cell growth factor (VEGF)-induced angiogenic process, that include proliferation, migration, invasion, tube formation and cell adhesion in human umbilical vein endothelial cells (HUVECs). Myriocin down-regulated VEGF-induced VEGFR2, focal adhesion kinase, AKT kinase, rho-associated coiled-coil-containing protein kinase 1 (ROCK1) and matrix metalloproteinase-2 (MMP2) expression. Myriocin greatly reduced new vessel formation in chick chorioallantoic membrane and vessel sprouting in mouse aortic ring assay. Furthermore, VEGF-induced phalloidin expression was inhibited by miyriocin in HUVECs. Our result indicated that myriocin inhibited VEGF-induced proliferation, migration, invasion, tube formation and cell adhesion in vitro by inhibiting VEGFR2-mediated activation of FAK, AKT, ROCK1 and MMP2. Taken together, we suggest that myriocin may act as significant angiogenic inhibitor by inhibiting VEGF-induced signaling pathway.

Citation Format: Sun Hee Lee, Yuk Dong Jung, Hye-Eun Lee, You Mie Lee. Myriocin, an inhibitor of serine palmitoyltransferase activity, has the anti-angiogenic effect by attenuating VEGF-induced angiogenesis 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 3270.
Abstract 3277: Prion-like protein “Doppel” is a selective therapeutic target for tumoral angiogenesis
Authors: Al-Hilal Taslim , Yang Yoosoo , Kim In-San , Byun Youngro , Ahsan Fakhrul
Published: July 2016

Regulating growth factor signaling in a controlled and site-specific manner is the main hurdle that current antiangiogenic treatment modalities face. Here we show that the prion-like protein, doppel, is uniquely found in tumoral endothelial cells (TEC) but not in the normal ECs. Tissue sections containing primary human tumor (lung and colon) and normal tissues were used to study doppel protein expression. Strong expression and clear co-localization of doppel protein with CD34, a classical endothelial marker, was observed in both lung and colon cancer tissues compared to normal tissues. To evaluate doppel expression at the molecular level, we isolated TECs from different tumors derived from mouse xenografts. Extensive analysis showed that doppel was highly and ubiquitously expressed in the vasculature of squamous, breast, lung, and colon cancers. To elucidate the role of doppel in TEC during tumor angiogenesis, we generated a gain-of-function EC model to express doppel in ECs. Inducible overexpression of doppel in ECs correlates with high vessel formation in a spheroid-based EC transplantation technique. To integrate doppel expression with the canonical angiogenic circuits, we studied the mechanism by which doppel regulates angiogenesis. We evidently observed that doppel co-localized and formed complexes with vascular endothelial growth factor receptor 2 (VEGFR2). Also, doppel prolonged the surface residency of VEGFR2 and amplified the responsiveness of VEGF to VEGFR2. Genetic deletion of doppel in TEC not only depleted membrane residency but also induced internalization and degradation of VEGFR2. This indicates that doppel blocking can lead to effective control of VEGF-signaling in TECs and selective inhibition of tumor angiogenesis. Based on the functional role of doppel in angiogenesis and tumor growth inhibition, we generated doppel knockout mice on immunogenic C57BL6 mice. Tumor growth was inhibited in both hetero (Dpl+/-) and homo (Dpl-/-)-type mice compared to their wild-type littermate controls when syngeneic cancer cell lines including melanoma (B16f10) and lymphoma (EL4) or allogeneic colon cancer CT26 cell line were inoculated. Moreover, we developed doppel targeting monoclonal antibodies. Anti-doppel antibodies, 5C7 and 4D6, were effective against both murine CT26 and human HCT116 colon cancer cell lines. Our studies thus demonstrate the success of doppel targeting in inhibiting pathological tumor growth and establish doppel-VEGFR2 interactions as a specific molecular and therapeutic target in tumoral angiogenesis.

Citation Format: Taslim Al-Hilal, Yoosoo Yang, In-San Kim, Youngro Byun, Fakhrul Ahsan. Prion-like protein “Doppel” is a selective therapeutic target for tumoral angiogenesis. [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 3277.
Abstract 3278: Dopamine inhibits growth of multiple myeloma
Authors: Sarkar Chandrani , Chakroborty Debanjan , Basu Sujit
Published: July 2016

Introduction: Multiple myeloma (MM), progressive plasma cell tumor leading to overproduction of monoclonal immunoglobulins, osteolytic bone lesions, renal disease and immunodeficiency, is the second most prevalent hematologic malignancy in the United States. Vascular endothelial growth factor-A (VEGF-A) is a critical cytokine responsible for the induction of angiogenesis in several solid and hematological malignancies like MM. It is likely that VEGF-A induced bone marrow (BM) angiogenesis promotes the survival, growth and proliferation of MM .Therefore targeting BM angiogenesis in MM may be effective for the treatment of MM patients

Objective/Hypothesis: Bone marrow stromal cells (BMSC) are believed to play a central role in progression of MM pathogenesis. Along with other factors, these cells overproduce IL-6 that in turn stimulates growth, survival and progression of MM cells. Since it has been shown that VEGF-A can stimulate IL-6 secretion by these cells and because we have demonstrated that neurotransmitter dopamine (DA) can inhibit VEGF-A functions by acting through its D2 receptors (D2R) and BMSC express DAD2R, we therefore hypothesize that DA or D2R agonists have a role in inhibiting VEGF-A mediated IL-6 secretion by BMSC and thereby, MM progression.

Results: Our results indicate that VEGF-A can significantly increase the secretion of IL6 by isolated BMSC cells in vitro. Addition of DA or D2R agonist, quinpirole, to VEGF-A treated BM cells results in suppression of IL-6 production. However treatment with D2R antagonist, eticlopride, prior to treatment with DA results in abrogation of the effect of DA. Similar effects of DA on VEGF-A induced IL-6 secretion was also observed in HS-5 human BM stromal cells. Our results further indicate that VEGF-A induced VEGFR1 phosphorylation is suppressed in HS-5 cells on treatment with D2R agonist. Furthermore, DA also inhibits the proliferation of MM1.S human MM cells in vitro.

Conclusion: DA may be a new and an effective approach to retard the progression of MM. Since anti-angiogenic monotherapy cannot eradicate the disease completely, strategies using DA with more traditional modalities to eradicate the disease at the time of initial treatment might be beneficial. Finally, DA or its D2R agonists are being used in the clinics for several years for the treatment of other disorders, therefore these inexpensive drugs (in comparison to presently available drugs for the treatment of MM) with known and manageable side effects can be used in the clinics in near future for the treatment of both early and late stage MM.

Citation Format: Chandrani Sarkar, Debanjan Chakroborty, Sujit Basu. Dopamine inhibits growth of multiple myeloma. [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 3278.
Abstract 3286: Targeting mir101/ EZH2/NFkb axis by FGFR1 inhibitor in mantle cell lymphoma
Authors: Sehgal Lalit , Jain Neeraj , Khashab Tamer , wang xin , Neelapu Sattva , Samaniego Felipe
Published: July 2016

Mantle cell lymphoma is classified as distinct non-Hodgkin's lymphoma subtype characterized by the t(11;14)(q13;q32) translocation, which results in overexpression of Cyclin D1. The clinical presentation often includes extra-nodal involvement, of the bone marrow and gut. The prognosis of patients with mantle cell lymphoma (median overall survival, 3-5 years) is poorest among B-cell lymphoma patients. However, MCL pathogenesis has been delayed until the recent development of a tissue culture system, using stromal cells and adipocytes, suitable for propagating primary MCL cells. We hypothesized that tumor microenvironment might activate survival-signaling pathways essential for survival and maintenance of MCL, and targeting the identified signaling pathways might be an ideal for designing of curative treatment strategies. We developed an in-vitro culture system, mimicking bone marrow microenvironment that can be exploited to understand the biology of MCL resistance to death pathways. Primary MCL cells grown in co-culture with bone marrow stromal cells grew faster than cells cultured alone. We identified the soluble factors responsible for survival of MCL cancer stem cells (MCL-IC) that can be targeted to prevent relapse. We found the IL6 secretion triggered FGF-2 autocrine loop in MCL cells contributing to cell survival and growth by regulating mir101/EZH2/NFkb axis. The death signaling pathways inactive in MCL are restored upon therapeutic intervention by specific pathways inhibitors. In conclusion, we delineated three different signaling arm regulated in MCL cells to promote cell survival and resistance. We defined the factors supporting MCL and MCL-ICs survival essential for forming new targeting treatment strategies. To overcome MCL resistance to death pathways we have outlined the hMSCs-mediated support of MCL and identified drug targets to sensitize MCL to apoptosis.

Citation Format: Lalit Sehgal, Neeraj Jain, Tamer Khashab, xin wang, Sattva Neelapu, Felipe Samaniego. Targeting mir101/ EZH2/NFkb axis by FGFR1 inhibitor in mantle cell lymphoma. [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 3286.
Abstract 3300: Gal-GalNAc overexpressed in colorectal carcinoma mediates attachment and colonization of Fusobacterium nucleatum utilizing the Fap2 lectin
Authors: Abed Jawad H. , Emgård Johanna , Chaushu Stella , Garrett Wendy , Bachrach Gilad
Published: July 2016

Recent reports revealed overabundance of oral Fusobacterium nucleatum (Fn) in colorectal carcinoma (CRC) compared to adjacent healthy tissue. This study aims to investigate the molecular mechanisms that mediate CRC colonization by Fn.

Using the orthotopic Colon-26 colorectal cancer model, we found that intravascular injected Fn colonizes CRC more than adjacent healthy tissue. Fusobacterial localization in the colon is tumor-dependent, as injected fusobacteria were undetected in the colons of tumor-free mice. CRC colonization is species specific as the gram-negative, anaerobe and periodontitis-related pathogen Prophyromonas gingivalis, which was shown to be abundant in oral tumors failed to colonize CRC.

Higher levels of D-galactose-β(1-3)-N-acetyl-D-galactosamine (Gal-GalNAc) are known to be detected in CRC. We recently identified the fusobacterial Fap2 outer-surface protein as a galactose binding lectin, suggesting that it might promote binding of fusobacteria to CRC. A fluorescent-labeled Peanut Agglutinin lectin (PNA) detected more Gal-GalNAc in human and mouse CRC compared to adjacent healthy tissues. Binding of FITC-labeled Fn corresponded with the levels of Gal-GalNAc and colocalized with Gal-GalNAc in the tumor. Flow cytometry revealed that Fap2-deficient Fn mutants are impaired in binding to CRC cell lines compared with their WT parent. Soluble GalNAc inhibited the binding of PNA and WT Fn to CRC cells in a dose-dependent manner, but did not affect the residual CRC binding of fap2 mutated Fn. Furthermore, in the colorectal cancer murine model, tumor colonization by Fap2-deficient Fn mutants was significantly lower than that of the Fap2 sufficient parental strain.

Our results demonstrate that the fusobacterial Fap2 lectin binds to Gal-GalNAc on CRC and mediates the tumor-specific attachment of Fn. Understanding the interaction of Fn with CRC, may enable future development of novel diagnostic and therapeutic agents for this disease.

Citation Format: Jawad H. Abed, Johanna Emgård, Stella Chaushu, Wendy Garrett, Gilad Bachrach. Gal-GalNAc overexpressed in colorectal carcinoma mediates attachment and colonization of Fusobacterium nucleatum utilizing the Fap2 lectin. [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 3300.
Abstract 3301: Breast cancer subtypes have distinct microbial and immune cytolytic transcriptomic signatures
Authors: Pedamallu Chandra Sekhar , Meyerson Matthew , Ojesina Akinyemi
Published: July 2016

Background: The role of genetic mutations in breast cancer is well documented and has formed the basis for novel therapies. However, the impact of viral and bacterial infection in the pathogenesis of breast cancer is not well understood.

Methods: We analyzed RNA sequencing (RNASeq) data generated by the Cancer Genome Atlas (TCGA) from over 900 pairs of breast tumor and adjacent normal tissues, using our pathogen discovery tool, PathSeq. We developed a novel metric for quantifying and normalizing microbial sequence abundance from RNASeq data by correcting for both the sequencing coverage and relative genome sizes of bacteria in the dataset. We also performed linear discriminant analyses (LDA) to test the hypothesis that the differences between the canonical breast cancer subtypes can be modeled on microbial abundance data. In addition, we interrogated the human gene expression data derived from the RNASeq dataset to determine the relationships between the breast cancer subtypes and immune cytolytic activity (CYT index; defined as the geometric mean of the expression levels of perforin and granzyme).

Results: Viral sequences were rare but we identified bacterial sequences to varying degrees of abundance. Intriguingly, we demonstrated that microbial abundance data can be used to recapitulate the well-established PAM50 gene expression-based subsets. Comparison of tumors and adjacent normal tissues also revealed subtype-associated microbial profiles. In addition, we observed that the basal and Her2 subtypes of breast cancer had a higher frequency of tumors with high CYT index compared with luminal A and B subtypes.

Conclusion: This study has revealed that the various breast cancer subtypes have unique microbial transcriptomic signatures as well as differential immune cytolytic activity profiles. These data suggest the potential for differential responses of breast cancer subsets to immunotherapy.

Citation Format: Chandra Sekhar Pedamallu, Matthew Meyerson, Akinyemi Ojesina. Breast cancer subtypes have distinct microbial and immune cytolytic transcriptomic signatures. [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 3301.
Abstract 331: Beta-catenin nuclear translocation in colorectal cancer cells is suppressed by PDE10A inhibition, cGMP elevation, and activation of PKG
Authors: Lee Kevin J. , Lindsey Ashley S. , da Silva Luciana Madeira , Trinh Alisa , Gary Bernard , Andrews Joel , Ramirez-Alcantara Veronica , Keeton Adam B. , Chang Wen-Chi , Clapper Margie , Piazza Gary A.
Published: July 2016

Phosphodiesterase 10A (PDE10A) is a cGMP and cAMP degrading PDE isozyme that is highly expressed in the brain striatum where it appears to play an important role in cognition and psychomotor activity. PDE10 inhibitors are being developed for the treatment of schizophrenia and Huntington's disease and are generally well tolerated, possibly because of low expression levels in most peripheral tissues. We recently reported high levels of PDE10 in colon tumors and that genetic silencing of PDE10 by siRNA or inhibition with small molecule inhibitors can suppress colon tumor cell growth with a high degree of selectivity over normal colonocytes (Li et al., Oncogene 2015). These observations suggest PDE10 may have an unrecognized role in tumorigenesis. Here we report that the concentration range by which the highly specific PDE10 inhibitor, Pf-2545920 (MP-10), inhibited colon tumor cell growth parallels the concentration range required to increase cGMP and cAMP levels, and activate PKG and PKA, respectively. Moreover, PDE10 knockdown by shRNA reduced the sensitivity of colon tumor cells to the growth inhibitory activity of Pf-2545920. Pf-2545920 also inhibited the translocation of β-catenin to the nucleus, thereby reducing β-catenin mediated transcription of survivin, which resulted in caspase activation and apoptosis. This was determined to be through a PKG mediated pathway through the use of small molecule inhibitors of PKG and PKA. PDE10 mRNA was also found to be elevated in colon tumors compared with normal tissues in a cDNA array. We also report the increase in PDE10 mRNA in 50% of a small collection of human clinical specimens collected at the Mitchell Cancer Institute (n = 13). In addition, novel PDE10 inhibitor, MCI-030, reduced tumor size and activated PDE10 signaling mechanisms in vivo. These findings suggest that PDE10 can be targeted for cancer therapy or prevention whereby inhibition results in cGMP elevation and PKG activation to reduce β-catenin-mediated transcription of survival proteins leading to the selective apoptosis of cancer cells.

Citation Format: Kevin J. Lee, Ashley S. Lindsey, Luciana Madeira da Silva, Alisa Trinh, Bernard Gary, Joel Andrews, Veronica Ramirez-Alcantara, Adam B. Keeton, Wen-Chi Chang, Margie Clapper, Gary A. Piazza. Beta-catenin nuclear translocation in colorectal cancer cells is suppressed by PDE10A inhibition, cGMP elevation, and activation of PKG. [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 331.