Abstract Chronic inflammation increases the susceptibility of the colon to neoplasia and cancer through establishment of an inflammatory microenvironment and persistent release of reactive oxygen species (ROS), inducing genome damage. Interleukin -1beta (IL-1beta), a pro-inflammatory cytokine, plays a pivotal role in the pathogenesis of acute and chronic intestinal inflammation, regulating both innate and adaptive immune responses. IL-1beta has been shown to promote accumulation of IL-17A through lymphoid and Th17 infiltration in the colon tumor microenvironment, supporting increased inflammation and correlating directly with poor patient prognosis. New insights into the signaling pathways supported by IL-1beta in the pathogenesis of colon cancer, including angiogenesis and metastasis, may provide downstream targets for treatments that minimally perturb intestinal immunity. Prior studies from our group have demonstrated significant up-regulation of IL-1, dual oxidase 2 (DUOX2) and its partner maturation factor, DUOXA2, in surgically resected colon cancer specimens compared with adjacent normal colonic epithelium. To elucidate a direct link between IL-1 family cytokines, DUOX2/DUOXA2 derived ROS, and colorectal cancer, we examined human colon cancer cells (HT29, Ls513, T84 and Colo205) stimulated with IL-1beta, in cooperation with IL-6. Co-stimulation resulted in a dramatic, synergistic up-regulation of a hydrogen peroxide producing (Amplex Red oxidizing) DUOX2 enzyme complex and was directly associated with enhanced histone H2AX phosphorylation (γH2AX), a marker of DNA double strand breaks. Interestingly, this concentration- and time-dependent induction of expression and oxidative response was not mediated by other IL-1 family members (IL-18, IL-33 or IL-37). Investigations with the interleukin-1 receptor antagonist anakinra established that signaling for IL-1beta/IL-6 co-treatments proceeded through the IL-1 receptor for all cell lines, though minimal receptor expression is present in T84 and Ls513 cells. Similarly, exposure to tocilizumab, an IL-6R antagonist, demonstrated that the IL-1beta/IL-6 synergistic increase in DUOX2/DUOXA2 complex expression is IL-6 receptor dependent. Perturbation of the IL-1 and IL-6 signaling pathway elements MYD88, IRAK1, STAT1, and STAT3 by siRNA knockdown demonstrated their significant contribution to DUOX2/DUOXA2 up-regulation, while dependence on RELA was absent. Knockdown facilitated by MAPK14 siRNA and SB203580 p38 alpha/beta inhibition also demonstrated a role for p38alpha in the regulation of DUOX2/DUOXA2 expression by the IL-1beta/IL-6 signaling axis. Current studies are focused on elucidating the DUOX2 promoter transcription factor binding site(s) responsible for DUOX2 up-regulation through chromatin immunoprecipitation. Citation Format: Jennifer L. Meitzler, Becky A. Diebold, David J. Mallick, Yongzhong Wu, Smitha Antony, Mariam M. Konaté, Krishnendu Roy, James H. Doroshow. IL-1 beta/ IL-6 co-stimulation synergistically mediates DUOX2/DUOXA2 complex up-regulation via JAK/STAT and p38 signaling events [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 3319.

Abstract Dual oxidase 2 (DUOX2), one of the seven NADPH oxidases family members, plays a critical role in both host defense and chronic inflammation-associated cancer in the gastrointestinal system. In vitro, pro-inflammatory cytokines, such as IFN- y, IFNα/β, IL-4 and IL-17A, enhance DUOX2/DUOXA2 expression through activation of STATs and NF-kB signaling pathway proteins; in vivo, DUOX protein and mRNA levels are substantially upregulated in chronic pancreatitis, pre-malignant pancreatic intraepithelial neoplasms and the early stages of pancreatic cancer patients compared to histologically normal pancreatic tissues. In pancreatic adenocarcinoma, increased DUOX2 expression is adversely correlated with overall patient survival. Heat-Shock Protein 90 (Hsp90), an important molecular chaperone involved in tumorigenesis, invasion and metastasis of cancer cells, is critical in folding, maturation and stability of many oncogenic client proteins, including kinases such as AKT and JAK1/2, and transcription factors, such as STAT3 and HIF-1α. Several STAT family members, along with Hsp90, are overexpressed in human pancreatic carcinomas. Using a panel of human pancreatic cancer cell lines (BxPC-3, AsPC-1 and CFPAC-1), we found that two different Hsp90 inhibitors, Tanespimycin (17-AAG) and Ganetespib (STA-9090), inhibit JAK1 and JAK2 kinases, blocking cytokine-induced, JAK-regulated STAT phosphorylation. Additionally, these Hsp90 inhibitors suppress cytokine-induced DUOX2, VEGF-A, MMP-7 and PD-L1 expression in human pancreatic cancer cell lines with varying sensitivity. Furthermore, the JAK1/2 inhibitor Ruxolitinib inhibits IL-4 induced and JAK-mediated STAT6 phosphorylation, and DUOX2 mRNA and protein expression in BxPC-3 cells. Similar results were observed with JAK1, STAT1, 2 and STAT6 specific siRNA knockdown. However, simultaneous knockdown of both isoforms of Hsp90, Hsp90α and Hsp90β, with specific siRNA did not inhibit JAK1 activity, cytokine-induced DUOX2 mRNA or protein expression. Either remaining Hsp90 protein or other isoforms of Hsp90 in cells may compensate decreased Hsp90 function after siRNA knockdown. Our data suggests that Hsp90 inhibitors, through blocking the cytokine-activated JAK-STATs oncogenic signaling pathway and their downstream genes such as DUOX2, VEGF-A, MMP-7 and PD-L1expression, may be a valuable therapeutic approach for inflammation-associated pancreatic cancer. Citation Format: Yongzhong Wu, David J. Mallick, Allan Di, Smitha Antony, Jennifer Meitzler, Mariam M. Konaté, Becky Diebold, Krishnendu Roy, James H. Doroshow. Heat shock protein inhibitors suppress cytokine-induced DUOX2 mRNA and protein expression in human pancreatic cancer cells in a JAK, STAT dependent manner [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6578.

Abstract Chronic pancreatitis increases the risk of developing pancreatic ductal adenocarcinoma (PDAC) and is associated with enhanced expression of the NADPH oxidase (NOX) isoform dual oxidase 2 (DUOX2). Interleukin-1 (IL-1) is a major upstream pro-inflammatory cytokine secreted by malignant or microenvironmental cells that supports pancreatic inflammation, tumorigenesis, invasiveness, and tumor heterogeneity. Previously, our laboratory demonstrated that the expression of DUOX2, one of seven NADPH oxidase (NOX) family members, is significantly enhanced by a wide variety of pro-inflammatory cytokines, and that DUOX2-related ROS production may contribute to the development of an oxidative, pro-angiogenic microenvironment in PDAC. In the current studies, we have shown that DUOX2 is upregulated and expressed along with its maturation factor, DUOXA2, in several human pancreatic cancer cell lines following exposure to both IL-1α and IL-1ß, two major agonists of IL-1 signaling. Administration of the anti-inflammatory antibody anakinra (an IL-1R1 antagonist) either in conjunction with or after prolonged treatment with IL-1 in vitro ablated induction of DUOX2/DUOXA2 and of IL-8 (CXCL8), a chemokine essential for neutrophil recruitment and a potent promoter of angiogenesis. Increased expression of other NOX family members following IL-1 exposure was not observed. Using CRISPR/Cas9-mediated DUOX2 knockout models, we report that DUOX2 is the major contributor to IL-1-mediated ROS production in PDAC cells. In a panel of PDAC lines, siRNA knockdown of proximal IL-1 signaling nodes, such as MYD88, significantly decreased expression of DUOX2/DUOXA2 and of CXCL8. However, knockdown of IRAK1 or IRAK4, two critical signaling kinases downstream of MYD88, as well as known downstream mediators of IL-1-related transcription (such as NF-κB, STAT1, MEK/ERK), did not affect IL-1-induced DUOX2/DUOXA2 upregulation, suggesting that alternative pathways of IL-1 signaling might be involved in the enhancement of DUOX2 expression by the cytokine. Because IL-1 signaling is known to affect various acetylation/methylation events in epithelial cells, we evaluated the effect of administering the histone acetyltransferase (HAT) inhibitor A-485 with IL-1; we found that A-485 significantly diminished cytokine-induced DUOX2/DUOXA2 upregulation; however, CXCL8 induction was unchanged. These studies suggest that epigenetic modulation of PDAC cells by IL-1 could contribute to DUOX2-induced ROS production and ROS-related inflammatory stress in the PDAC microenvironment. Citation Format: David J. Mallick, Yongzhong Wu, Mariam M. Konaté, Becky Diebold, Smitha Antony, Jennifer L. Meitzler, Guojian Jiang, Jiamo Lu, Krishnendu Roy, James H. Doroshow. Interleukin-1 upregulates dual oxidase 2 expression and ROS production in human pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5327.

Pancreatic inflammation contributes to increased risk of pancreatic adenocarcinoma (PDAC) development through the release of pro-inflammatory cytokines and generation of reactive oxygen species (ROS). Interleukin-1 (IL-1) is a major upstream pro-inflammatory cytokine secreted by malignant or microenvironmental cells that supports pancreatic tumorigenesis, invasiveness, and tumor heterogeneity. Previously, our laboratory has established that dual oxidase 2 (DUOX2), one of seven NADPH oxidase (NOX) family members, plays a critical role in H2O2-mediated host defense and chronic inflammation in the gastrointestinal tract. DUOX2 is upregulated and expressed along with its maturation factor, DUOXA2, in several human pancreatic cancer cell lines following exposure to various pro-inflammatory cytokines, including IFN-γ, IL-4, and IL-17A. Using a panel of PDAC cells lines, we report the induction of DUOX2/DUOXA2 mRNA and protein expression by both IL-1α and IL-1ß, two major agonists of IL-1. In BxPC-3, CFPAC-1 and Hs 766T (HTB-134) cells, prolonged treatment with IL-1α or IL-1ß results in transient activation of STAT1 and NF-kB signaling followed by prolonged induction of DUOX2/DUOXA2 mRNA and protein expression. Stimulation by IL-1α or IL-1ß results in a specific increase in the expression of DUOX2/DUOXA2; increased expression of other NOX family members was not observed. Activation of canonical IL-1 signaling in our PDAC cell lines was supported by the observation of IL-1-related upregulation of IL-8 (CXCL8) mRNA, a chemokine essential for neutrophil recruitment and a potent promoter of angiogenesis, in IL-1-treated PDAC cells. Moreover, when the anti-inflammatory compound anakinra (an IL-1R antagonist) was co-administered in vitro with IL-1, cytokine-induced DUOX2/DUOXA2 and CXCL8 induction was significantly suppressed. Finally, xenografts established with BxPC-3 cells demonstrated slower growth and significant downregulation of IL-1ß and DUOX2 when the immunosuppressive corticosteroid dexamethasone was administered. Given our previous findings that DUOX2 is highly expressed in the pancreas during episodes of chronic pancreatitis or pancreatic cancer, and contributes significantly to extracellular ROS formation, these studies suggest that IL-1 may play a role in promoting inflammation and tumorigenesis in pancreatic malignancies through upregulation of DUOX2. Citation Format: David J. Mallick, Yongzhong Wu, Mariam M. Konaté, Becky A. Diebold, Smitha Antony, Jennifer L. Meitzler, Guojian Jiang, Jiamo Lu, Krishnendu Roy, James H. Doroshow. Interleukin-1 promotes enhanced functional expression of dual oxidase 2 in human pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1318.

Abstract Inflammatory bowel diseases are states of long-term inflammation of the colon resulting from an interplay of environmental factors and dysregulated immune responses. As inflammatory exposure increases over the time frame of active disease, patients with these chronic conditions are at risk of developing colorectal cancers. Chronic inflammation is thought to contribute to cancer initiation through persistent release of reactive oxygen species (ROS), leading to genome damage. Accumulating evidence suggests inflammatory cells contribute to this process, as the secretion of cytokines and growth factors in response to gut inflammation also supports cancer cell progression. Specifically, IL-1 family members play a pivotal role in the pathogenesis of acute and chronic intestinal inflammation, regulating both innate and adaptive immune responses. To elucidate a direct link between IL-1 family cytokines, ROS generation and colorectal cancer, we observed that treatment of HT29 or T84 cells with IL-1α or IL-1β, in cooperation with IL-6, resulted in oxidant production through up-regulation of both NOX1 and DUOX2 enzymes. Up-regulation of the hydrogen peroxide generating DUOX2/DUOXA2 enzyme complex was directly associated with enhanced histone H2AX phosphorylation (γH2AX), a marker of DNA double strand breaks. Interestingly, this concentration and time-dependent induction of expression and oxidative response was absent in Caco2 cells and was not mediated by other IL-1 family members (IL-18, IL-33 or IL-37). Prior studies from our group have demonstrated significant up-regulation of DUOX2 and DUOXA2 in surgically resected colon cancer specimens compared with adjacent normal colonic epithelium. Future studies will focus on cytokine stimulation of colon organoids derived from patient tumor and adjacent non-malignant tissues to evaluate enzymatic expression and oxidant level changes in a novel patient-derived colon model system. Citation Format: Jennifer L. Meitzler, Yongzhong Wu, Agnes Juhasz, Annamaria Rapisarda, Petreena Campbell, Becky A. Diebold, Smitha Antony, Guojian Jiang, Jiamo Lu, David J. Mallick, Mariam M. Konaté, Krishnendu Roy, James H. Doroshow. Synergistic, cytokine mediated stimulation (IL-1α/β + IL-6) of colon cancer cells reveals a novel mechanism of DNA damage facilitated by up-regulation of NOX1 and DUOX2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2915.

Dual oxidase 2 (DUOX2) is a transmembrane protein that produces hydrogen peroxide in the extracellular environment, mediates innate immunity at mucosal surfaces, contributes to chronic inflammation-related tissue injury and angiogenesis, and can promote the initiation of epithelial-derived cancers. In pancreatic adenocarcinomas (PDAC), the composition of the tumor microbiome significantly affects patient outcomes. However, it remains unclear how microbes in the tumor microbiome affect PDAC on a cellular level. In recent years, the cGAS-STING signaling pathway has been identified as a crucial mediator of anti-viral defense by recognizing the presence of DNA in the cytosol and subsequently producing Type I interferons (IFNs) as well as a host of other anti-viral genes. Using Western blot and qPCR, we show that the introduction of exogenous DNA into the cytosol significantly upregulates DUOX2 expression in human PDAC cells after activating cGAS-STING signaling. Expression of DUOX2 occurs most strongly 2 days post-transfection of DNA, while cGAS-STING signaling is activated within hours. Treatment of BxPC-3 cells with exogenous cyclic GMP-AMP (cGAMP) for 24 hrs increases DUOX2 expression in a concentration-dependent manner, suggesting the necessity of cGAS activity for DUOX2 induction. Furthermore, Western analysis of phospho-NF-κB p65 in BxPC-3 cells demonstrates activation of NF-κB that mirrors the time-dependent induction of DUOX2. In CFPAC-1 cells, another PDAC line, Jak-Stat signaling is activated along with the components of cGAS-STING. These data suggest that DNA-induced DUOX2 upregulation in PDAC cells may be mediated in a cell context-specific fashion, beyond the canonical components of the cGAS-STING pathway. Upregulation of DUOX2 by exogenous DNA is not observed in colon cancer cell lines. In summary, these data suggest that extracellular DNA could contribute to a DUOX2-induced, H2O2-mediated pro-inflammatory milieu that specifically contributes to the pathogenesis of PDAC, in part through activation of the cGAS-STING pathway. Citation Format: Stephen L. Wang, Yongzhong Wu, Smitha Antony, Agnes Juhasz, Jennifer Meitzler, Goujian Jiang, Iris Dahan, Jiamo Lu, Krishnendu Roy, James Doroshow. Exogenous DNA upregulates DUOX2 expression in pancreatic cancer via activation of the cGAS-STING signaling pathway [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3348.

Colorectal cancer (CRC) develops through a multistep process that is enhanced by chronic inflammation and includes complex interactions between the colonic epithelium and pro-inflammatory cytokines capable of stimulating cancer initiation and progression due, in part, to enhanced production of reactive oxygen species (ROS). Expression of colonic DUOX2, a H2O2 generator, and NOX1, a superoxide producer, is increased in patients with inflammatory bowel disease and adenomatous polyps. For these experiments, we investigated the effect of prolonged treatment with pro-inflammatory cytokines known to be associated with CRC (IL-4, IL-6, and IL-17A alone and in combination) on NADPH oxidase (NOX) expression in human colon cancer lines, and early passage, two-dimensional cultures of conditionally-reprogrammed patient-derived colon cancer cells (PDCs). HT-29 colon cancer cells were treated with IL-4, IL-6, IL-17A, IL-4+IL-17A, and IL-6+IL-17A at a concentration of 25ng/ml each for 4, 6, 8, 15, and 30 days. We measured the effect of these cytokines on expression of NOX family members NOX1, DUOX2, and DUOXA2 by quantitative RT-PCR, and by Western analysis. The production of ROS by NOX proteins was measured with chemiluminescence (superoxide) and Amplex Red assays (H2O2), respectively. DUOX2 protein, which is absent at baseline in HT-29 cells, was easily detectable after 4 days, and increased as a function of the duration of cytokine exposure and for cytokine combinations. In addition to HT-29 cells, similar results were obtained using Ls513 and DLD-1 colon cancer lines. Six PDC lines were also studied: T 280-R, F725, F1126, CK7962, CK7354 and 027-R1. Modest increases in NOX1 expression were observed for 2 lines (T 280-R, CK7962) following 6 days of treatment with IL-6+IL-17A or IL-4; however, NOX1 was highly expressed in 4 cell lines following exposure to the combination of IL-4+IL-17A. Moreover, DUOX2 mRNA and protein were highly expressed after IL-6+IL-17A treatment for 6 days in the T 280-R and F725 cell lines, and very highly expressed after IL-4+IL-17A treatment in all 6 PDCs. Enhanced NOX1 and/or DUOX2 expression in CRC cells was accompanied by concomitant enhancement of NOX-specific ROS. RNA-Seq analysis of cytokine-modulated NOX expression in HT-29 cells and a NOX1 shRNA inhibited subline of HT-29 cells, suggests that both NF-κB and Stat signaling pathways are involved in the control of DUOX2 expression in both established and early passage human CRC cells. These experiments suggest that multiple pro-inflammatory cytokines known to be present in the CRC microenvironment may contribute to the development of a hyperoxidant milieu through the up-regulation of the two NOX species, NOX1 and DUOX2, expressed in the colon.Citation Format: Agnes N. Juhasz, Mariam Konate Monnard, Guojian Jiang, YongZhong Wu, Abdalla Amr Abdelmaksoud, Jinqiu Chen, Xialong Luo, Noemi Kedei, Smitha Antony, Jennifer L. Meitzler, Jiamo Lu, Iris M. Dahan, Krishnendu Roy, James H. Doroshow. Long-term treatment with IL-4, IL-6, and/or IL-17 increases the expression of the NADPH oxidase DUOX2 in human colon cancer cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5240.

CD40 stimulation together with IL4 has been shown to promote growth of normal B cells in vitro. In contrast to this, CD40 stimulation in several B cell lymphoma cell lines has been demonstrated to be growth inhibitory. Though the mechanism is not completely known, CD40-stimulated cells exhibit increased levels of reactive oxygen species (ROS). In B lymphocytes, a major source of ROS production is the NADPH oxidase (NOX) that is comprised of a membrane-bound Nox2 catalytic subunit and four cytosolic regulatory components, with the p67phox cytosolic protein being the activator factor. Upon stimulation, by either cross linking of the B-cell receptor or following treatment with PMA, the active NADPH oxidase generates superoxide. We have investigated the role of NADPH oxidase in CD40-mediated growth inhibition in Burkitt lymphoma (BL) cells. For this study, we first evaluated the effect of CD40 stimulation on the expression of Nox2 and p67phox in BL cells that demonstrate different levels of endogenous Nox2 and p67phox. Accordingly, Daudi and Raji cell lines, with endogenously high Nox2 and low p67phox, NAMALWA cells which have high Nox2 and no p67phox expression, and Ramos cells that contain no Nox2 or p67phox were analyzed. Treatment with anti-CD40 or CD40 ligand (CD40L) for 24 h induced up-regulation of p67phox in Daudi, Ramos, and NAMALWA cells, but not in the Raji line. No significant change in the expression of Nox2 was observed. Additionally, Daudi, Ramos and NAMALWA cells showed higher production of ROS after CD40 stimulation and activation with anti-IgM or PMA, in a chemiluminescence assay. To determine whether the change in ROS level was related to the upregulation of p67phox, Daudi cells that express either stable p67phox shRNA or control shRNA were generated. Treatment with CD40L for 24 h demonstrated significant ROS production only in control shRNA and not in the p67shRNA cells, confirming that the ROS production is NOX2-p67phox mediated. This enhanced ROS production was observed following treatment with CD40L alone and was enhanced further in conjunction with anti-IgM or PMA. However, cells with the p67phox shRNA showed detectable level of ROS only after activation with anti IgM, albeit to much lower levels than the control. Co-treatment of Daudi cells with anti-CD40 or CD40L and IL4 further increased the expression of p67phox. Although the basal level of ROS in the cells was unchanged upon co-treatment, the ROS generated following stimulation with anti-IgM or PMA was significantly enhanced. Preliminary MTT studies confirmed the growth inhibition of CD40-stimulated Daudi cells but not of Raji cells, suggesting a role for p67phox-dependent, ROS-related inhibition of lymphoma cell proliferation. Experiments are currently ongoing to delineate further the role of CD40 stimulation and Nox2-p67phox mediated ROS in affecting the growth of Burkitt lymphomas and signaling pathways downstream of CD40.Citation Format: Mazal Iris Dahan, Smitha Antony, Agnes Juhasz, Guojian Jiang, Mariam Konate, Jiamo Lu, Jennifer Meitzler, YongZhong Wu, Krishnendu Roy, James H. Doroshow. CD40-induced growth inhibition of Burkitt lymphoma: A possible role for NADPH oxidase by upregulation of p67phox [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 876.

Abstract NADPH oxidase (NOX)-derived reactive oxygen species (ROS) contribute significantly to inflammation-associated carcinogenesis. Expression of dual oxidase 2 (Duox2), one of seven members of the NOX gene family, is up-regulated in inflammatory bowel disease, chronic pancreatitis, and in many human malignancies including carcinomas of the colon, pancreas, prostate, lung, and breast. Previously, we demonstrated that Stat1 and/or NF-kB play a critical role in modulating the enhanced expression of Duox2, and its cognate maturation factor DuoxA2, by the pro-inflammatory mediators IFN-γ and lipopolysaccharide in human pancreatic cancer cells. This cytokine-mediated increase in Duox2 expression was responsible for a significant increase in tumor cell H2O2 production, DNA damage, and HIF-1α and VEGFA upregulation in pancreatic cancer cell lines. Using the T84 human colon cancer line, and BxPC-3, AsPC-1, and HTB134 pancreatic cancer cell lines, we now report that the pro-inflammatory cytokine IL-17A, pivotal in the adaptive immune response against various pathogens, and in the promotion of colon and pancreatic cancer progression, strongly induces Duox2/DuoxA2 mRNA and protein expression (alone or in combination with IL-4 or IFN-γ). In BxPC-3 cells, IL-17A enhances the expression of Duox2/DuoxA2 in a time- and concentration-dependent manner, which leads to a significant increase in the generation of extracellular H2O2. When IL-17A is combined with IL-4 in either T-84 or AsPc-1 lines, Duox2-mediated ROS formation is sufficient to produce a DNA damage response, indicated by the strong induction of γH2AX expression; moreover, in AsPc-1 cells, Duox2-specific siRNA significantly attenuated cytokine-induced ROS production and γH2AX phosphorylation. IL-17A in combination with IL-4 also significantly increases Stat3 phosphorylation and promotes proliferation in AsPC-1 cells. Ongoing cellular experiments are focused on whether the cytokine-mediated upregulation of Duox2/DuoxA2 is responsible for Stat3 activation and cell growth promotion in the K-ras mutated AsPC-1 cells. Bio-informatic analysis with the TCGA data set, as well as RT-PCR using surgically-resected human colon cancer samples compared to adjacent, uninvolved colonic epithelium revealed that Duox2 was significantly upregulated in human colon cancers. These studies, in concert with our previous data demonstrating high-level Duox protein expression in pancreatitis, pancreatic intra-epithelial neoplasia (PanIN), and frank pancreatic cancer, strongly suggest that ROS, produced as a consequence of cytokine-related Duox2 up-regulation, may be associated with the development of inflammation-related gastrointestinal malignancies. Citation Format: James H. Doroshow, Yongzhong Wu, Jiamo Lu, Smitha Antony, Jennifer Meitzler, Agnes Juhasz, Guojian Jiang, Iris Dahan, Mariam Konate, Krishnendu Roy. IL-17A alone or in combination with IL-4/IFN-γ induces Duox2/DuoxA2 expression and reactive oxygen production in human colon and pancreatic cancer cells [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 2698. doi:10.1158/1538-7445.AM2017-2698

This workshop will provide details on how to plan and run non-residential, week-long computing summer camps for 4th-12th grade students. Georgia Tech has been offering computing summer camps since 2004. These camps are financially self-sustaining and effective. Items used in the camps include: CS Unplugged, LightBot, Scratch, Alice, LEGO robots (WeDo, NXT, EV3, and Tetrix), EarSketch, and App Inventor. Georgia Tech helped start other computing camps at eleven other colleges and universities in Georgia from 2007 to 2010 as part of Georgia Computes! NSF BPC grant. Since 2014, as part of the Expanding Computing Education Pathways (ECEP) NSF grant we have also helped institutions start or expand summer computing camps in South Carolina, Massachusetts, and California. The workshop will include presentation about application forms, a timeline, sample agendas, sample flyers, budget plans, a planning checklist, suggested projects, surveys, pre and post-tests, evaluation results, lessons learned, and more. Laptops are recommended for this workshop.

A step-by-step introductory guide to mobile app development with App Inventor 2About This BookGet an introduction to the functionalities of App Inventor 2 and use it to unleash your creativityLearn to navigate the App Inventor platform, develop basic coding skills and become familiar with a blocks based programming languageBuild your very first mobile app and feel proud of your accomplishmentFollow tutorials to expand your app development skillsWho This Book Is ForApp Inventor 2 Essentials is for anyone who wants to learn to make mobile apps for Android devices no prior coding experience is necessary.What You Will LearnPerform technical setup and navigate the App Inventor platformUtilize the interactive development environment by pairing a mobile device with a computer using Wi-Fi or USBBuild three apps: a game, an event app and a raffle appCreate the user interface of the app in the Designer and program the code in the Blocks EditorIntegrate basic computer science principles along with more complex elements such fusion tables and listsTest and troubleshoot your applicationsPublish your apps on Google Play Store to reach a wide audienceUnleash your creativity for further app developmentIn DetailApp Inventor 2 will take you on a journey of mobile app development. We begin by introducing you to the functionalities of App Inventor and giving you an idea about the types of apps you can develop using it. We walk you through the technical set up so you can take advantage of the interactive development environment (live testing). You will get hands-on, practical experience building three different apps using tutorials. Along the way, you will learn computer science principles as well as tips to help you prepare for the creative process of building an app from scratch. By the end of the journey, you will learn how to package an app and deploy it to app markets. App Inventor 2 Essentials prepares you to amass a resource of skills, knowledge and experience to become a mobile app developerStyle and approachEvery topic in this book is explained in step-by-step and easy-to-follow fashion, accompanied with screenshots of the interface that will make it easier for you to understand the processes.

App Inventor is one of the most popular block-based programming environments. Currently, there are limited instructional resources that guide students to create tangible apps using App Inventor. In this positional statement we make a case for the need for more App Inventor instructional resources related to tangible apps. We also present our proposal to address this need.

An adult male patient got hospitalized with fever, anorexia, weight loss, generalized weakness, and visual problem. Examination revealed solitary axillary lymphadenopathy and soft tissue swellings over scalp and left shoulder. Tests detected severe anemia and raised erythrocyte sedimentation rate (ESR), lactate dehydrogenase, and uric acid. Magnetic resonance imaging (MRI) brain showed irregular soft tissue swellings both outside and inside the skull with pressure effect on brain. Fundoscopy revealed bilateral choroidal metastatic deposits with papilledema. Histopathology findings of biopsy from scalp mass and bone marrow were consistent with metastatic high grade neuroblastoma. No primary lesion was found on computed tomography (CT) scan of thorax, abdomen, and pelvis. There was also suggestion of paraneoplastic neuropathy. The patient responded fairly to chemotherapy. Neuroblastoma is usually known to be a childhood malignancy that often presents with vague symptoms or symptoms due to tumor mass or with metastatic or paraneoplastic features. Most common metastatic sites are bone, lymph node, liver, cranium, and chest. But in this case apart from bony and dural metastases there was also choroidal metastasis which is previously not reported in adults. Likewise, non-detection of primary tumor is also uncommon.

This workshop will provide details on the projects that we recommend for non-residential computing summer camps for 4th -- 12th grade students. Georgia Tech has been offering computing summer camps since 2004. These camps are financially self-sustaining and effective at improving attitudes towards computing, especially for females and African-American students. Items used in the camps include: CS Unplugged, LightBot, Scratch, Alice, LEGO robots (WeDo, NXT, and Tetrix), EarSketch, and App Inventor. Georgia Tech has also helped eleven other colleges and universities in Georgia start or expand computing summer camps. Some of these other institutions also offer camps on GameSalad, GameMaker, and web development. These other camps have also improved attitudes towards computing. The workshop will include an overview of the projects that we have found to be successful and include hands-on work with App Inventor, GameSalad, LEGO NXT robots, and LEGO EV3 robots. laptop optional

Abstract The NADPH oxidase (NOX) gene family (NOX1-5 and Duox1/2) plays an important role in host defense and the development of inflammation-induced carcinogenesis. We recently found that two iodonium-class NOX inhibitors, diphenyleneiodonium (DPI) and di-2-thienyliodonium (DTI), block NOX-dependent reactive oxygen species (ROS) formation, NOX1 mRNA expression, and growth of human colon cancer cells in vitro and in vivo (Free Rad. Biol. Med. 57: 162, 2013). To improve the efficacy and bioavailability of DPI and DTI, we synthesized 48 new analogs of these compounds (21 DPI analogs, 12 DTI analogs, and 15 DPI and/or DTI hybrids; structures to be presented at the meeting) and prioritized the agents for further study based on inhibition of: HT-29 growth (MTT and clonogenic assay), ROS generation (by chemiluminescence and flow cytometry), and NOX1 mRNA expression (by qRT-PCR). The compounds were also characterized for lack of altered mitochondrial ROS production, and inhibitory effects on ROS generated by cell-based assays specific for NOX1, NOX2, NOX4, NOX5, and Duox2. DPI and five DPI analogs, NSCs 740104, 734428, 742837, 751140, 737392, strongly inhibited HT-29 cell growth in a dose dependant manner (IC50's after 48 hr drug exposure of 300, 50, 74, 87, 148, and 202 nM, respectively). Treatment for 24 hr with 10 nM DPI, 740104, 734428, or 751140, or with 100 nM 737392 significantly decreased ROS production in HT-29 cells. In addition, 60% or greater inhibition of NOX1 mRNA expression was observed following treatment with 250 nM 740104 and 734428 for 4 hr; 751140 and DPI for 24 hr; and 737392 for 48 hr. Analog 742837 demonstrated no significant inhibition of either ROS production or NOX1 expression in HT-29 cells; it was not analyzed further. Growth inhibitory properties of 734958, a DTI analog, were similar to the parent molecule. To evaluate compound specificity, a luminescence assay for O2•¯ was used with: HEK293 cells expressing all components of the NOX1 complex; fully differentiated HL-60 cells expressing active NOX2; and UACC257 melanoma cells that exclusively express active NOX5. H2O2 was measured by Amplex Red assay to determine NOX4 or Duox2 activity in fully reconstituted HEK293 cells. DPI, 734428, 740104, 737392, or 751140 exposure for 30 min inhibited NOX1, 2, 4, and 5 with low nM IC50's; analogs 740104, 734428, and 751140 were more potent inhibitors of NOX1, 2, and 4, respectively, than DPI; each was broadly active against these NOX isoforms. The DTI analog 734958 was more potent than its parent molecule. NSC737392 and 734428 were >10-fold more active than the other compounds examined against Duox2 (200-400 nM IC50's). In summary, we developed novel iodonium analogs with anticancer activity that possess enhanced potency against human NOX species, including the first agents with specificity against Duox2. These tool compounds will be evaluated in vivo for their therapeutic and anti-inflammatory activities in NOX-containing tumor xenograft models. Citation Format: Jiamo Lu, Prabhakar Risbood, Charles T. Kane, Md Tafazzal Hossain, Yongzhong Wu, Smitha Antony, Jennifer L. Meitzler, Agnes Juhasz, Han Liu, Guojian Jiang, Krishnendu Roy, James H. Doroshow. Development of potent NADPH oxidase inhibitors with significant activity against colon cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4750. doi:10.1158/1538-7445.AM2014-4750

Abstract Reactive oxygen species (ROS) produced by the family of NOX genes play a critical role in cytokine-mediated signal transduction in both normal and tumor tissue. Human colon cancers, compared to adjacent normal tissues, overexpress NOX1; and premalignant, chronic inflammatory conditions of the colon (ulcerative colitis, Crohn's disease) are associated with NOX up-regulation. However, a mechanistic understanding of how cytokines might increase ROS production and NOX1 expression in colon cancer remains unclear. We screened a panel of human colon cancer cells with a series of pro-inflammatory cytokines to evaluate their effects on NOX1 expression and function, as well as cell proliferation. Cytokine IL-4 significantly up-regulated NOX1 expression (but not other members of the NOX family) in HT-29 (as well as other) colon cancer cells while increasing O2• production and growth; decreasing NOX1 expression by 90% in stable, NOX1-shRNA-containing HT-29 clones blocked IL-4-related enhancement of cell growth, NOX1 expression, or ROS production. IL-4 up-regulated and activated the GATA3 transcription factor in addition to the Jak1-Stat6 pathway in HT-29 cells. GATA3 played an important role in the transcriptional up-regulation of NOX1 by IL-4; RNAi-mediated silencing of Jak1, Stat6, or GATA3 expression attenuated IL-4-induced NOX1 up-regulation in HT-29 cells. Transient overexpression of GATA3 cDNA in the presence of IL-4 resulted in a further increase in NOX1 expression with enhanced ROS activity beyond IL-4 treatment alone. Using an 1139 bp segment of upstream sequence from the transcription start site of the human NOX1 promoter as a promoter-reporter assay, we found that IL-4 treatment increased NOX1 reporter gene transcription 4-fold in HT-29 cells. DNA sequence analysis of this upstream promoter suggests the presence of four putative GATA3 binding, cis-active elements that may be sufficient for maximal IL-4-enahanced activity of the NOX1 promoter. Ongoing mutational analysis of the NOX1 promoter will further define the role of GATA3 in the control of NOX1 transcriptional up-regulation by IL-4. In summary, our results support a role for IL-4 in producing a NOX1-dependent pro-oxidant milieu that could increase the growth potential of colon cancer cells. Citation Format: Han Liu, Agnes Juhasz, Guojian Jiang, Smitha Antony, Jennifer L. Meitzler, Yongzhong Wu, Jiamo Lu, Krishnendu Roy, James H. Doroshow. Transcriptional upregulation of NADPH Oxidase 1 (NOX1) by Interleukin-4 (IL-4) enhances tumor cell proliferation and reactive oxygen production in human colon cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5406. doi:10.1158/1538-7445.AM2013-5406

This workshop will provide details on how to plan and run non-residential computing summer camps for 4th -- 12th grade students. Georgia Tech has been offering computing summer camps since 2004. These camps are financially self-sustaining and effective. Items used in the camps include: CS Unplugged, LightBot, Scratch, Alice, LEGO robots (WeDo, NXT, and Tetrix), EarSketch, and App Inventor. Georgia Tech has also helped start other computing camps at eleven other colleges and universities in Georgia. The workshop will include forms, a timeline, sample agendas, sample flyers, budget plans, a planning checklist, suggested projects, surveys, pre and post-tests, evaluation results, lessons learned, and more. Laptop Recommend.

Novice programming environments (NPEs) like Scratch and Alice witnessed tremendous growth in adoption and popularity in recent years. These NPEs have successfully lowered the barrier of initial entry to programming. They have also allowed novice computing students to exercise their creative freedom more meaningfully by enabling them to work on projects which have more real life context. Recent trends in adoption of smartphone and other mobile devices among our youth points to a time in near future when majority of them will be mobile device users. Hence, if the NPEs want to remain relevant among youth, they need to adapt to and cater to a mobile-device centric audience. Google and MIT's App Inventor for Android is one of the early NPEs that is trying to achieve this by offering a Scratch-like environment for mobile apps development. GameSalad is another free software primarily for Mac platform that can also offers a drag-n-drop rule-based environment for creating apps. In this paper we compare these two environments' suitableness for computing introduction. Our comparison is based on several logistical, instructional, and operational factors and points to the fact that both these mobile NPEs have their strong points as well as weaknesses. We believe the effectiveness and success of these mobile NPEs will vary depending on particular situations, and hence our work will aid someone looking for a mobile NPE in making a more judicious choice.

Google's App Inventor for Android (AIA) is the newest visual "blocks" programming language designed to introduce students to programming through creation of mobile applications (apps). AIA opens up the world of mobile apps to novice programmers. Success stories of using AIA to introduce college students to programming exist. We used AIA in computing summer camps for high school students that we offer at our university. This paper is an experience report about using AIA in our camps. We provide a detailed description of designing our camps with AIA including the process of selecting and setting-up an Android device and instructional materials that we developed and made available to everyone. We evaluated our camps through surveys to determine the effects on the students. We found that there was mostly a slight increase in the favorable disposition towards computing. We also share our successes with using AIA and what still needs improvement for wider use.