bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4546: Depleting cysteine: A novel approach to tumor therapy
Authors: Hanigan Marie H. , Burgett Anthony W. G. , Wakeham Nancy , Terzyan Simon S.
Published: August 2015

A cure remains elusive for many types of tumors that initially respond to chemotherapy but recur with drug-resistant disease. Cysteine, (or its oxidized form cystine) is an essential amino acid for tumors. Enhanced uptake of cysteine and/or cystine is required for tumors to maintain their intracellular glutathione (GSH) levels and redox balance. In addition, some stem cells within epithelial tumors express CD44v8-10, which stabilizes the cystine transporter, increasing cystine uptake into the cell and enhancing resistance to pro-oxidant therapy. Depleting cysteine during treatment has a profound effect on sensitizing the tumor to therapy. We have a novel strategy that reduces the cysteine (and cystine) concentration in serum and extracellular fluid. The strategy exploits the finding that a severe cysteine deficiency develops in mice lacking the enzyme gamma-glutamyl transpeptidase (GGT). This enzyme reclaims the cysteine from GSH as the blood is filtered through the kidney. Inhibiting GGT activity results in excretion of GSH in the urine and a precipitous drop in the concentration of cysteine and cystine in serum. The inhibitors of GGT that have been evaluated in the clinic are glutamate analogs and are too toxic for clinical use. We have discovered and are developing a new class of GGT inhibitors that are not glutamate analogs. These inhibitors are orders of magnitude less toxic than the glutamate analogs. We have determined the crystal structure of human GGT. In the current study we have co-crystallized human GGT with a series of inhibitors. We report the identification of residues within the active site that are critical for inhibitor binding. We have identified changes in the conformation of the active site upon substrate binding. We have also delineated movement within the active site during substrate cleavage. Site-directed mutagenesis and functional assays have confirmed the role of the key residues identified in the structural studies. These data provide new insights for continued development of a potent, specific, non-toxic inhibitor of GGT that can be used clinically in cancer therapy. Supported by an Institutional Development Award (IDeA) from the National Institutes of Health under grant P20GM103640.

Citation Format: Marie H. Hanigan, Anthony W. G. Burgett, Nancy Wakeham, Simon S. Terzyan. Depleting cysteine: A novel approach to tumor therapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4546. doi:10.1158/1538-7445.AM2015-4546
Abstract 4547: Preparation of peptide-based Shp2 substrates with phosphatase activity-dependent fluorescence
Authors: Gunawan Steven , Luo Yunting , Ren Yuan , Lawrence Harshani R. , Wu Jerry , Lawrence Nicholas J.
Published: August 2015

The Shp2 protein tyrosine phosphatase mediates signal transduction of growth factor receptors and regulates cellular activities critical to tumor growth and metastasis. Shp2-activating mutations are associated with several types of leukemias and solid tumors, making Shp2 an attractive target for anticancer development. While there are reports of Shp2 inhibitors, there is a need for compounds with higher potency and improved selectivity. One approach to the discovery of new Shp2 inhibitors is via high throughput screening (HTS) of small molecule libraries. The small molecule probe 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) is currently a widely used substrate for phosphatases and has been used to assay Shp2 activity. However, DIFMUP binds only the catalytic site of the phosphatase and is unlikely to reveal compounds that are selective for Shp2 in an HTS. There is clearly a need to develop probes that are more sensitive and more specific for a particular phosphatase to identify Shp2 selective inhibitors and to measure Shp2 activity. We have developed a new phosphopeptide-like Shp2 substrate that becomes fluorescent upon enzyme-mediated hydrolysis, based on a phosphopeptide sequence that has high affinity for Shp2. We will report details of the novel Shp2 activity probe and its use as a reagent for HTS and determination of Shp2 inhibitor potency. We will also report on its suitability for HTS and preliminary results from screening and its potential for the discovery of novel Shp2 inhibitors. We will discuss the binding mode of the probe, based on novel co-crystal structures of related peptides with Shp2.

Citation Format: Steven Gunawan, Yunting Luo, Yuan Ren, Harshani R. Lawrence, Jerry Wu, Nicholas J. Lawrence. Preparation of peptide-based Shp2 substrates with phosphatase activity-dependent fluorescence. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4547. doi:10.1158/1538-7445.AM2015-4547
Abstract 4549: A comparative efficacy study of Saquinavir in combination with cisplatin or paclitaxel in platinum-resistant ovarian cancer cells
Authors: Waite Terry-Ann E. , Reboe Monique , Levy Arkene , Dhandayuthapani Sivanesan , Rathinavelu Appu
Published: August 2015

Background and Objective: Ovarian cancer is the 7th most common cancer affecting women Worldwide which has a 5-year survival rate of 45%. First line therapy, platinum and taxol compounds, produces 60% to 80% response rates in platinum sensitive ovarian cancer. However, incidence of resistance to this therapy is high resulting in poor prognosis. The present study evaluated the comparative efficacy of Saquinavir (SAQ) in combination with Cisplatin (CIS) or Paclitaxel (TAX) versus first line therapy, CIS+TAX, in platinum-resistant OVCAR-3 cells.

Method: OVCAR-3cells were treated with varying concentrations of SAQ (20μM, 40μM, 60μM and 100μM). OVCAR-3 cells were treated with SAQ (100μM), SAQ+CIS(26μM), SAQ+TAX(1.5nM) and CIS+TAX. Cells were treated over a 24hr period in triplicate. Media only and DMSO only treated cells were used as controls. The cytotoxic effect of each treatment was determined using the automated Trypan Blue exclusion protocol and DNA fragmentation assay was performed to evaluate the mechanisms of induced cell death.

Results: Dose dependent and statistically significant (p <0.05) cytotoxic effects were seen with SAQ in OVCAR-3 cells. An IC50 value of 84μM for SAQ was determined in OVCAR-3 cells. Significant cytotoxic effects (p<0.05) in OVCAR-3 treated cells were observed with SAQ, SAQ+TAX, SAQ+CIS AND CIS+TAX combinations when compared to controls. However, SAQ+CIS was the only combination that showed more significant cytotoxicity when compared to CIS+TAX (p = 0.041). SAQ+CIS combination produced an average percentage cell death of 73.25% in OVCAR-3 cells. Cell death by apoptosis was confirmed from observed fragmented DNA in OVCAR-3 cells treated with SAQ+TAX, SAQ+CIS and CIS+TAX.

Conclusion: SAQ+CIS produced more significant cytotoxic effect than first line therapy, CIS+TAX, in platinum-resistant ovarian cancer cells. This data supports further evaluation of SAQ for its potential utilization in the treatment of platinum-resistant ovarian cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Terry-Ann E. Waite, Monique Reboe, Arkene Levy, Sivanesan Dhandayuthapani, Appu Rathinavelu. A comparative efficacy study of Saquinavir in combination with cisplatin or paclitaxel in platinum-resistant ovarian cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4549. doi:10.1158/1538-7445.AM2015-4549
Abstract 4554: An evaluation of the role of Mucin in drug resistant ovarian cancer
Authors: Richards Steven M. , Campbell Robert B.
Published: August 2015

MUC1 is a transmembrane glycoprotein that is overexpressed in epithelial ovarian cancer. Due to extensive O-glycosylation, mucins form a physical barrier that limits the intracellular accumulation and cytotoxic effect of chemotherapeutic drugs. Our investigation represents a key step in determining if drug resistance in ovarian cancer develops due to altered MUC1 expression and glycosylation. Immunoblotting and ELISA techniques were utilized to characterize the extent of membrane-bound and shed MUC1 expression in drug-resistant and drug-sensitive ovarian cancer cells. An (anti-MUC1) antibody-based in vitro model was employed to determine if the inhibition of O-glycosylation enhanced functional access of small molecule drugs using FACS and fluorescence microscopic analyses. Expression of MUC1 was detected in both the drug resistant ovarian cancer cell line SK-OV-3 (MDR) and the parent cell line SK-OV-3 (WT), with the highest levels observed in the drug resistant sub-clone. The greatest improvement in functional anti-MUC1 antibody access following inhibition of O-glycosylation was observed in the drug-resistant ovarian cancer cells. Our results provide evidence to support drug resistance in ovarian cancer due in part to a mucin barrier mesh. The inhibition of mucin O-glycosylation may be a viable strategy to improve drug delivery and efficacy of small molecule delivery to ovarian carcinomas.

Citation Format: Steven M. Richards, Robert B. Campbell. An evaluation of the role of Mucin in drug resistant ovarian cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4554. doi:10.1158/1538-7445.AM2015-4554
Abstract 4557A: Development of the proteasome deubiquitinase inhibitor VLX1570 for treatment of multiple myeloma
Authors: Wang Xin , Mohanty Chitralehka , D'Arcy Padraig , Olofsson Maria Hägg , Bossler Felicitas , Chanan-Khan Asher , Linder Stig
Published: August 2015

Inhibitors of the proteasome-ubiquitin system have been found to be effective in the clinical management of multiple myeloma. We here describe the compound VLX1570, an optimized lead of the 19S deubiquitinase inhibitor b- AP15. VLX1570 was shown to inhibit the enzymatic activities of both USP14 and UCHL5 in vitro, and was shown to induce accumulation of high molecular weight polyubiquitinated proteins in multiple myeloma cells. The compound showed limited inhibitory activity of a panel of kinases. VLX1570 induces apoptosis and cell death of multiple myeloma cell lines at submicromolar concentrations. Importantly, a multiple myeloma cell line selected for bortezomib resistance was found to be sensitive to VLX1570. VLX1570 was found to be able to overcome apoptosis resistance by these cells. Induction of the phosphorylation of stress-activated kinases (JNK and p38) and inhibition of ERK phosphorylation was observed in multiple myeloma cells. We finally showed that VLX1570 confers antineoplastic activity in two different myeloma xenograft models and conclude that VLX1570 is a candidate agent for treatment of multiple myeloma.

Citation Format: Xin Wang, Chitralehka Mohanty, Padraig D'Arcy, Maria Hägg Olofsson, Felicitas Bossler, Asher Chanan-Khan, Stig Linder. Development of the proteasome deubiquitinase inhibitor VLX1570 for treatment of multiple myeloma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4557A. doi:10.1158/1538-7445.AM2015-4557A
Abstract 4561: Cancer preventive effect of antioxidants for spontaneous liver cancer in XPA(-/-) mice
Authors: Fu Ying , Silverstein Shana , Dyba Marcin , Coia Heidi , Sinclair Elizabeth , Pan Jishen , Kallakury Bhaskar , Johnson Michael D. , Chung Fung-Lung
Published: August 2015

γ-Hydroxy-1, N2-propanodeoxyguanosine (γ-OHPdG), a widely studied acrolein-derived cyclic adduct arisen from oxidized ω-3 and ω-6 polyunsaturated fatty acids (PUFAs), is a mutagenic lesion that is repaired by nucleotide excision repair (NER) pathway. C3H/HeN xeroderma pigmentosum group A (XPA) knockout mice which are deficient on NER pathway showed a significant elevation in liver tumors. We hypothesized that accumulation of γ-OHPdG is playing a role in liver tumorigenesis of XPA(-/-) mice; suppressing the formation of γ-OHPdG by dietary antioxidants should result in the prevention of liver cancer in this model. In this work, we investigated the chemoprotective efficacy of three antioxidants: α-lipoic acid, polyphenon E and vitamin E against liver tumorigenesis in XPA(-/-) mice. Autopsy at 19 months of age revealed that all the antioxidants exhibited significant inhibition effects on the development of liver tumors. Polyphenon E showed the most potent effects with 80% of the mice showing complete protection. The formation γ-OHPdG was measured in the livers of XPA(-/-) mice. An age-dependent increase of γ-OHPdG was found in the liver DNA of XPA(-/-) mice, but not in lungs (a non-target tissue). All the antioxidants decreased the formation of γ-OHPdG. Immunohistochemistry (IHC) for Ki-67 (proliferation index marker) was performed on all the liver samples, and the results showed that dietary antioxidants significantly decrease the proliferation in the liver tumors. α-Lipoic acid and vitamin E showed significant inhibitory effect, although to a lesser extent than Polyphenon E, against the spontaneous liver carcinogenesis in XPA gene-deficient mice model. A correlation between the cancer preventive efficacy and suppression of γ-OHPdG adduct level is observed, indicating a possible role of γ-OHPdG in the liver cancer development in this model. We also explored the feasibility of next-generation sequencing (NGS) to validate the mutation frequency changes as a result of feeding the antioxidant diets. This work was supported by the NCI grant: RO1-CA-134892. Y.F. thanks the Prevent Cancer Foundation for his fellowship (Marcia and Frank Carlucci Charitable Foundation Award in Cancer Prevention and Early Detection)

Note: This abstract was not presented at the meeting.

Citation Format: Ying Fu, Shana Silverstein, Marcin Dyba, Heidi Coia, Elizabeth Sinclair, Jishen Pan, Bhaskar Kallakury, Michael D. Johnson, Fung-Lung Chung. Cancer preventive effect of antioxidants for spontaneous liver cancer in XPA(-/-) mice. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4561. doi:10.1158/1538-7445.AM2015-4561
Abstract 457: Depletion of glioma infiltrating myeloid derived suppressor cells promotes anti-tumor T cell responses
Authors: Kamran Neha , Li Youping , Moreno-Ayala Mariela , Assi Hikmat , Candolfi Marianela , Dzaman Marta , Lowenstein Pedro , Castro Maria
Published: August 2015

MDSCs represent a population of immature myeloid cells at various stages of differentiation that have the potential to inhibit anti-tumor T cell immunity. We demonstrate the accumulation of MDSCs in GL26 and M7-induced glioma (GBM) bearing mice. Absolute numbers of Ly-6G+ (Gr-1high) MDSCs showed a 200 fold increase within the tumor microenvironment (TME) 28 days post-tumor implantation. The numbers of Ly-6C+ (Gr-1low) MDSCs also showed a similar trend within the TME. While this massive influx of MDSCs was noted within intracranial tumors, MDSC levels did not increase in the dLNs, spleen or bone marrow (BM) of intracranial tumor bearing mice. MDSC numbers were significantly elevated in the blood of GL26 and M7 intracranial tumor bearing mice at 28 days. While both Gr-1high and Gr-1low MDSCs isolated from the TME of GL26 intracranial tumor bearing mice inhibited antigen-specific T cell proliferation, Gr-1low MDSC were found to be more efficient. Gr-1high or Gr-1low MDSCs from the bone marrow of intracranial tumor bearing mice failed to suppress antigen-specific T cell proliferation suggesting that TME derived factors may activate MDSCs to exert their immune-suppressive properties. In vivo, depletion of Gr-1+ cells enhanced the median survival of GBM bearing mice. Furthermore, when combined with Ad-TK + Ad-FLT3L immune-gene therapy, Gr-1+ depletion significantly enhanced the frequency of tumor-specific T cells within the TME and spleen and increased IFN-γ production by splenic T cells. Our data therefore indicates that inhibiting the accumulation of MDSCs with in the GBM TME promotes the generation of robust anti-tumor immunity. Preliminary experiments to determine the mechanism of MDSC trafficking to the TME point towards the receptor CXCR2 and its ligand CXCL1. Microarray analysis of glioma cell lines showed elevated levels of CXCL1 mRNA. Additionally a culture of primary mixed glial cells also produced CXCL1 when stimulated with GBM cell lysates. SB225002, a CXCR2 inhibitor suppressed the migration of MDSCs towards GBM cells in an in vitro migration assay. Overall, our data suggests that strategies that inhibit MDSC recruitment to the GBM TME and/or block their activity could enhance the T cell mediated tumor clearance and provide survival benefit.

Work supported by grants from NIH-NINDS

Citation Format: Neha Kamran, Youping Li, Mariela Moreno-Ayala, Hikmat Assi, Marianela Candolfi, Marta Dzaman, Pedro Lowenstein, Maria Castro. Depletion of glioma infiltrating myeloid derived suppressor cells promotes anti-tumor T cell responses. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 457. doi:10.1158/1538-7445.AM2015-457
Abstract 4573: 6-C-(E-phenylethenyl)-naringenin suppresses colorectal cancer growth by inhibiting cyclooxygenase-1
Authors: Li Haitao , Zhu Feng , Chen Hanyong , Cheng Ka Wing , Zykova Tatyana , Oi Naomi , Lubet Ronald A. , Bode Ann M. , Wang Mingfu , Dong Zigang
Published: August 2015

Recent clinical trials raised concerns regarding the cardiovascular toxicity of selective cyclooxygenase-2 (COX-2) inhibitors and cyclooxygenase-1 (COX-1) is now being reconsidered as a target for chemoprevention. Our aims were to determine whether selective COX-1 inhibition could delay or prevent cancer development and also clarify the underlying mechanisms. Data clearly showed that COX-1 was required for maintenance of malignant characteristics of colon cancer cells or tumor promoter-induced transformation of preneoplastic cells. We also successfully applied a ligand-docking computational method to identify a novel selective COX-1 inhibitor, 6-C-(E-phenylethenyl)-naringenin (designated herein as 6CEPN). 6CEPN could bind to COX-1 and specifically inhibited its activity both in vitro and ex vivo. In colorectal cancer cells, it potently suppressed anchorage-independent growth by inhibiting COX-1 activity. 6CEPN also effectively suppressed tumor growth in a 28-day colon cancer xenograft model without any obvious systemic toxicity. Taken together, COX-1 plays a critical role in human colorectal carcinogenesis, and this specific COX-1 inhibitor merits further investigation as a potential preventive agent against colorectal cancer.

Citation Format: Haitao Li, Feng Zhu, Hanyong Chen, Ka Wing Cheng, Tatyana Zykova, Naomi Oi, Ronald A. Lubet, Ann M. Bode, Mingfu Wang, Zigang Dong. 6-C-(E-phenylethenyl)-naringenin suppresses colorectal cancer growth by inhibiting cyclooxygenase-1. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4573. doi:10.1158/1538-7445.AM2015-4573
Abstract 4577: Mitagation of potassium dichromate micronuclei induction and kidney damage by ethanol extract of Moringa oleifera in Swiss albino rats
Authors: Akinwum Kazeem A. , Adedoja Ayobami W. , Osifeso Osifeso O. , Adegboyega Adenike M. , Aralamo Deborah O. , David Olaitan O. , Akinwumi Kazeem A.
Published: August 2015

Chromate (VI) compounds are human carcinogen. Exposure by inhalation or ingestion causes lung and intestinal cancers. Current therapy has side effects, necessitating the search for antidote from medicinal plant such as Moringa oleifera (M.oleifera). This study was undertaken to evaluate the effect of ethanol extract of M. oleifera on potassium dichromate (K2Cr2O7) induced clastogenicity and kidney damage. Leaves of M.oleifera were air dried, powdered, extracted with ethanol and phytochemical analysis carried out. Thirty male rats were grouped into 6 of 5 animals each. Negative and positive control rats had water and 12mg/kg K2Cr2O7 once per week respectively. Test rats were exposed to 3.5 and 7.0 body weight M.oleifera alone for 35 days and / or 12 mg/kg body wt K2Cr2O7 once a week before they were sacrificed. The frequency of micronucleated polychromatic erythrocytes (mPCEs) was monitored in the bone marrow while creatinine and urea levels were assessed in the serum. Kidney superoxide dismutase (SOD), glutathione S-transferase (GST) and malondialdehyde (MDA) levels were also determined. K2Cr2O7 significantly (p <0.05) increased mPCEs and urea while creatinine was decreased when compared with control. MDA was also markedly increased, while SOD and GST were significantly reduced. Severe kidney damage was observed in the same sets of animals. However, M.oleifera mitigated the toxicity by reversing the markers towards that of control. M.oleifera is rich in flavonoids and phenolics which may account for the observed .protection. The results suggest that extract of M.oleifera has valuable potentials in the treatment/management of chromate (VI) toxicity

Note: This abstract was not presented at the meeting.

Citation Format: Kazeem A. Akinwum, Ayobami W. Adedoja, Osifeso O. Osifeso, Adenike M. Adegboyega, Deborah O. Aralamo, Olaitan O. David, Kazeem A. Akinwumi. Mitagation of potassium dichromate micronuclei induction and kidney damage by ethanol extract of Moringa oleifera in Swiss albino rats. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4577. doi:10.1158/1538-7445.AM2015-4577
Abstract 4580A: Metabolic activation of 3-nitrobenzanthrone by human aldo-keto reductases (AKR1C1-AKR1C4)
Authors: Murray Jessica R. , Huang Meng , Zang Tianzhu , Arlt Volker M. , Schmeiser Heinz H. , Penning Trevor M.
Published: August 2015

In 2012, the International Agency for Research on Cancer (IARC) classified diesel exhaust as a Group 1 carcinogen due to sufficient evidence that exposure is associated with increased risk for lung cancer in humans. However, only a subset of individuals exposed to diesel exhaust develops cancer, indicating the need to identify the genes involved in metabolic activation of these compounds and their genetic variants. Nitro-polycyclic aromatic hydrocarbons (NO2-PAH) are a major component of diesel exhaust and require metabolic activation to exert their carcinogenic activity. A representative NO2-PAH, 3-nitrobenzanthrone (3-NBA), is metabolically activated to 3-aminobenzanthrone (3-ABA) via a 6-electron nitroreduction catalyzed by NQO1 and POR. The reaction leads to the formation of 3-aminobenzanthrone (3-ABA) derived DNA adducts which promote G to T transversions. Building upon previous data that shows human aldo-keto reductase 1C3 (AKR1C3) contains nitroreductase activity towards chemotherapeutic agents (Guise, C.P., Abbattista, M.R., et al., Cancer Res, 70(4), 2010), we chose to examine the nitroreductase activity of AKR1C1-AKR1C4 towards NO2-PAH. We have demonstrated here for the first time that AKR1C enzymes catalyze the nitroreduction of 3-NBA to 3-ABA. We monitored reactions with reverse phase HPLC coupled to in-line photo-diode-array detection (PDA) and fluorescence detection (FLD) to quantify 3-NBA and 3-ABA levels. Fluorescence and UV spectroscopy were used to validate the identity of the compounds. This method was adapted for discontinuous enzymatic assays to measure steady state kinetic parameters for the nitoreductase activity of AKR1C1-AKR1C4 and NQO1. Results indicate that the NQO1 catalyzed reduction of 3-NBA has a higher specific activity, but the combined specific activities of AKR1C catalyzed reduction may play a more significant role in the overall production of 3-ABA. These results suggest that the relative expression of NQO1 and AKR1C enzymes will determine their respective contribution to 3-NBA reduction, especially since all the aforementioned enzymes are inducible by the Nrf2-Keap1 system. This work is supported by P30E513508 and RO1 CA39504 to TMP.

Citation Format: Jessica R. Murray, Meng Huang, Tianzhu Zang, Volker M. Arlt, Heinz H. Schmeiser, Trevor M. Penning. Metabolic activation of 3-nitrobenzanthrone by human aldo-keto reductases (AKR1C1-AKR1C4). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4580A. doi:10.1158/1538-7445.AM2015-4580A
Abstract 4583: Computational analysis predicts unbalanced IDH1/IDH2 expression associate with 2-HG-inactivating beta-oxygenation pathway in colorectal cancer
Authors: Koseki Jun , Colvin Hugh , Fukusumi Takahito , Nishida Naohiro , Konno Masamitsu , Kawamoto Koichi , Tsunekuni Kenta , Doki Yuichiro , Mori Masaki , Ishii Hideshi
Published: August 2015

Bioinformatics and computational modeling offer innovative approaches to investigate cancer metabolism and predict the secondary and tertiary cellular responses. Dysregulation of metabolism has also been implicated in the pathophysiology of cancer. A significant proportion of patients with glioblastoma and hematological malignancies harbor the mutated forms of the oxidative phosphorylation (OxPhos) enzymes, isocitrate dehydrogenase (IDH) 1 or 2. The mutated forms of IDH1 and IDH2 produce an oncogenic metabolite, D-2-hydroxyglutarate (D2HG). A recent study of breast cancer patients showed that D2HG can also be produced in the absence of mutated IDH, through an alternative route involving over-activated MYC signaling. We developed a novel methodology to computationally analyze gene expression in colorectal cancer, and identified novel sets of genes that are associated with patient survival. The study of OxPhos-related genes revealed that an imbalance between the expression of IDH1 and IDH2, defined as overexpression of one isoform in relation to the other, was associated with worse prognosis in colorectal cancer patients. This effect was further accentuated by reduced expression of the beta-oxygenation enzyme, 3-D-hydroxyacyl-CoA dehydratase (HCDH) 4, which has been reported to contribute to metabolism of intracellular D2HG. The present computational analysis revealed a novel and potential mechanism of colorectal cancer development, through over-production of D2HG when there is an imbalance between IDH1 and IDH2 expression, resulting in decreased clearance of D2HG when the beta-oxidization pathway is diminished. Additional validation analysis with other gene expression dataset has resulted in that IDH1/2 imbalanced expression had a shorter DFS compared with balanced expression. Altogether, these findings provide a strong rationale for studying this mechanism further in order to discover novel therapeutic targets for the treatment of colorectal cancer.

Citation Format: Jun Koseki, Hugh Colvin, Takahito Fukusumi, Naohiro Nishida, Masamitsu Konno, Koichi Kawamoto, Kenta Tsunekuni, Yuichiro Doki, Masaki Mori, Hideshi Ishii. Computational analysis predicts unbalanced IDH1/IDH2 expression associate with 2-HG-inactivating beta-oxygenation pathway in colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4583. doi:10.1158/1538-7445.AM2015-4583
Abstract 459: Targeting TIGIT and PD-1 to increase the expansion and function of tumor antigen-specific CD8+ T cells in melanoma patients
Authors: Chauvin Joe-Marc , Pagliano Ornella , Fourcade Julien , Sun Zhaojun , Sanders Cindy , Kirkwood John M. , Chen Tseng-hui T. , Maurer Mark , Korman Alan , Zarour Hassane
Published: August 2015

T-cell Immunoglobulin and ITIM Domain (TIGIT) is an inhibitory receptor expressed by activated T cells, regulatory T cells and NK cells. Here, we show that tumor antigen (TA)-specific CD8+ T cells and CD8+ tumor-infiltrating lymphocytes (TILs) upregulate TIGIT and most often co-express TIGIT and PD-1 in melanoma patients. CD8+ TILs downregulate CD226, supporting an imbalance of TIGIT/CD226 expression in metastatic melanoma. TIGIT is a marker of early T cell activation, which is further upregulated by T cells upon PD-1 blockade and by dysfunctional PD-1+Tim-3+ TA-specific CD8+ T cells. PD-1+TIGIT+, PD-1−TIGIT+ and PD-1+ TIGIT− CD8+ TILs exhibit similar functional capacities ex vivo, suggesting that TIGIT upregulation alone or together with PD-1 is not a marker of T cell dysfunction. However, in the presence of TIGIT ligand-expressing cells, TIGIT blockade adds to PD-1 blockade to increase the proliferation, cytokine production and degranulation of TA-specific CD8+ T cells and CD8+ TILs. Collectively, our findings show that TIGIT together with PD-1 regulate the expansion and function of TA-specific CD8+ T cells and CD8+TILs in melanoma patients. They also support the use of dual TIGIT and PD-1 blockade to stimulate potent antitumor CD8+ T cells responses in patients with advanced melanoma.

Citation Format: Joe-Marc Chauvin, Ornella Pagliano, Julien Fourcade, Zhaojun Sun, Cindy Sanders, John M. Kirkwood, Tseng-hui T. Chen, Mark Maurer, Alan Korman, Hassane Zarour. Targeting TIGIT and PD-1 to increase the expansion and function of tumor antigen-specific CD8+ T cells in melanoma patients. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 459. doi:10.1158/1538-7445.AM2015-459
Abstract 4594: Serum microRNA signatures predict recurrence and survival in Caucasian patients with early stage non-small cell lung cancer
Authors: Zhang Liren , Roth Jack A. , Lian Jie , Ye Yuanqing , Gu Jian , Wu Xifeng
Published: August 2015

Early stage non-small cell lung cancer (NSCLC) patients can be curatively treated by surgery alone or with adjuvant chemotherapy. However, about half of patients develop recurrence and eventually die of this disease. The purpose of this study is to identify circulating microRNAs (miRNA) as minimally invasive biomarkers for the prognosis of early stage NSCLC that may allow personalized clinical management of early stage NSCLC patients. Using a large series of 171 stage I and II NSCLC Caucasian patients, we screened and validated serum miRNA signatures predictive of recurrence and survival in these patients. We identified six miRNA expression ratios that were associated with recurrence and three miRNA expression ratios that were associated with overall survival. In Receiver Operating Characteristic (ROC) curve analysis, the area under the curve (AUC) for clinical variable-only model was 0.714 for recurrence and 0.678 for survival; however, the AUC increased to 0.878 for recurrence and 0.796 for survival when the miRNA ratio signatures were added to the recurrence and survival models respectively. The patients in the high risk group as defined by the 6 miRNA ratio signature exhibited an approximately 5-fold increased risk of develop recurrence compared with low risk patients. In survival analysis, the death rate increased over 4-fold in the high risk group compared with the low risk group. Our results suggest that the miRNA ratio signatures could serve as prognostic biomarkers for early stage NSCLC.

Citation Format: Liren Zhang, Jack A. Roth, Jie Lian, Yuanqing Ye, Jian Gu, Xifeng Wu. Serum microRNA signatures predict recurrence and survival in Caucasian patients with early stage non-small cell lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4594. doi:10.1158/1538-7445.AM2015-4594
Abstract 4596: Genetic variants associated with longer telomere length are associated with increased lung cancer risk among never-smoking women in Asia: A report from the Female Lung Cancer Consortium in Asia
Authors: Machiela Mitchell J. , Hsiung Chao A. , Shu Xiao-Ou , Seow Wei J. , Wang Zhaoming , Matsuo Keitaro , Hong Yun-Chul , Seow Adeline , Wu Chen , Hosgood H Dean , Chen Kexin , Wang Jiu-Cun , Wen Wanqing , Wu Tangchun , Wong Maria P. , Wu Yi-Long , Yang Pan-Chyr , Zhou Baosen , Shin Min-Ho , Fraumeni Joseph F. , Zheng Wei , Lin Dongxin , Chanock Stephen J. , Rothman Nathaniel , Lan Qing
Published: August 2015

Recent evidence from several relatively small nested case-control studies in prospective cohorts shows an association between longer telomere length measured phenotypically in peripheral white blood cell (WBC) DNA and increased lung cancer risk. We sought to further explore this relationship by examining a panel of 7 telomere-length associated genetic variants in a large study of 5,457 never-smoking female Asian lung cancer cases and 4,493 never-smoking female Asian controls using data from a previously reported genome-wide association study. Using a group of 1,536 individuals with phenotypically measured telomere length in WBCs in the prospective Shanghai Women's Health study, we demonstrated the utility of a genetic risk score (GRS) of 7 telomere-length associated variants to predict telomere length in an Asian population. We then found that GRSs used as instrumental variables to predict longer telomere length were associated with increased lung cancer risk (OR = 1.51 (95% CI = 1.34-1.69) for upper vs. lower quartile of the weighted GRS, P-value = 4.54×10−14) even after removing rs2736100 (P-value = 4.81×10−3), a SNP in the TERT locus robustly associated with lung cancer risk in prior association studies. Stratified analyses suggested the effect of the telomere-associated GRS is strongest among younger individuals. We found no difference in GRS effect between adenocarcinoma and squamous cell subtypes. Our results indicate that a genetic background that favors longer telomere length may increase lung cancer risk, which is consistent with earlier prospective studies relating longer telomere length with increased lung cancer risk.

Citation Format: Mitchell J. Machiela, Chao A. Hsiung, Xiao-Ou Shu, Wei J. Seow, Zhaoming Wang, Keitaro Matsuo, Yun-Chul Hong, Adeline Seow, Chen Wu, H Dean Hosgood, Kexin Chen, Jiu-Cun Wang, Wanqing Wen, Tangchun Wu, Maria P. Wong, Yi-Long Wu, Pan-Chyr Yang, Baosen Zhou, Min-Ho Shin, Joseph F. Fraumeni, Wei Zheng, Dongxin Lin, Stephen J. Chanock, Nathaniel Rothman, Qing Lan. Genetic variants associated with longer telomere length are associated with increased lung cancer risk among never-smoking women in Asia: A report from the Female Lung Cancer Consortium in Asia. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4596. doi:10.1158/1538-7445.AM2015-4596
Abstract 4597: A novel variant in DNA repair gene GTF2H4 is associated with lung cancer risk: A reanalysis of GWAS datasets from the TRICL consortium
Authors: Wei Qingyi , Liu Hongliang , Liu Zhensheng , Amos Christopher I. , Doherty Jennifer A. , Bickeboller Heike , Hung Rayjean J. , Brennan Paul , Houlston Richard , Landi Maria Teresa , Caporaso Neil E. , Christiani David
Published: August 2015

DNA repair pathways maintain genomic integrity and stability, and dysfunction of DNA repair is involved in cancer. We hypothesize that genetic variants in DNA repair genes are associated with risk of lung cancer. To test this hypothesis, we performed a large-scale meta-analysis of 123,371 SNPs in 169 DNA repair genes obtained from six datasets of previously published genome-wide association studies (GWASs), consisting of 12,160 lung cancer cases and 16,838 controls. We evaluated functional relevance of the identified, previously unpublished important and significant SNPs. Across the six GWASs, 14 SNPs were found to be significantly associated with lung cancer risk with a false discovery rate (FDR) of P < 0.05, of which rs114596632 in GTF2H4 at 6q31.33 was the most statistically significant SNP (OR = 1.19, 95% CI = 1.12-1.25, Pcombined = 5.40 × 10−10). The SNP rs114596632 was not strongly correlated with the previously reported GWAS hit MSH5 rs3131379 in that region (r2 = 0.383). Further mapping of the human leukocyte antigen (HLA) region identified that rs114596632 was partially tagged by HLA-B*0801 (r2 = 0.592) and HLA-C*0701 (r2 = 0.675). Importantly, the rs114596632 T allele was significantly associated with decreased DNA repair capacity (DRC) measured in peripheral T lymphocytes and decreased mRNA expression levels of GTF2H4 in lymphoblastoid cell lines. We also validated two previously known SNPs in DNA repair genes MSH5 and XRCC4 associated with lung cancer risk. Subsequent pathway analysis with gene relationships across implicated loci (GRAIL) revealed multiple significant connections for GTF2H4, ERCC2, and MMS19. Our results provide evidence that genetic variants of GTF2H4 confer susceptibility to lung cancer and suggest the involvement of GTF2H4 in the etiology of lung cancer. (Supported by U19CA148127, R01CA CA074386, R01CA092824 and the start-up funds from Duke Cancer Institute, Duke University Medical Center)

Citation Format: Qingyi Wei, Hongliang Liu, Zhensheng Liu, Christopher I. Amos, Jennifer A. Doherty, Heike Bickeboller, Rayjean J. Hung, Paul Brennan, Richard Houlston, Maria Teresa Landi, Neil E. Caporaso, David Christiani. A novel variant in DNA repair gene GTF2H4 is associated with lung cancer risk: A reanalysis of GWAS datasets from the TRICL consortium. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4597. doi:10.1158/1538-7445.AM2015-4597
Abstract 460: Novel compound elicits anti-tumor macrophages associated with tumor regression in breast cancer
Authors: Ponichtera Holly E. , Guerriero Jennifer L. , Sotayo Alaba O. , Letai Anthony
Published: August 2015

Cancer immuno-therapies targeting components of the immune system have shown great promise for the treatment of various forms of cancer. Most of the immuno-therapies currently under trial intervene in adaptive immune cell pathways; however, the innate immune system has also recently been shown to play a potent immunomodulatory role in the tumor microenvironment. Tumor associated macrophages (TAMs) compose up to 50% of tumor volume. Presently, we demonstrate that administration of a novel compound stimulates rapid tumor regression that is associated with the infiltration of F4/80+ macrophages into tumors in the MMTV-PyMT murine model of breast cancer. A significant increase in tumor-infiltrating macrophages was visible by immunohistochemistry after five days of treatment and macrophages were necessary for tumor regression as demonstrated by cell depletion assays in vivo. Of note, macrophage infiltration correlated with an increase in tumor cell death as shown by elevated expression of the apoptotic protein cleaved-caspase-3 (CC3); conversely, cell proliferation factor Ki67 was decreased. Additionally, compound-treated animals exhibited striking changes in tumor vascularization, with a marked decrease in overall endothelial cell marker expression as displayed by immunohistochemistry. Taken together, these findings reveal a novel role for macrophages in breast tumor regression during treatment with a novel compound that alters the tumor microenvironment.

Citation Format: Holly E. Ponichtera, Jennifer L. Guerriero, Alaba O. Sotayo, Anthony Letai. Novel compound elicits anti-tumor macrophages associated with tumor regression in breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 460. doi:10.1158/1538-7445.AM2015-460
Abstract 4618: The risk prediction for esophageal cancer by drinking, smoking, and the polymorphisms of ALDH2 and ADH1B
Authors: Ito Hidemi , Oze Isao , Hosono Satoyo , Watanabe Miki , Tanaka Hideo , Matsuo Keitaro
Published: August 2015

Background: The impact of drinking and smoking on the risk of esophageal cancer has been established worldwide (IACR, 2012 updated). In our previous study, we found that the polymorphisms ofalcohol dehydrogenases 1B (ADH1B - rs1229984) and aldehyde dehydrogenases 2 (ALDH2 - rs671) were associated with esophageal cancer risk in a Japanese population (Matsuo K, 2001 & Oze I, 2009). However, the extent to which these environmental and genetic risk factors contribute as predictors of esophageal cancer among Japanese is unknown. The objective of our study was to evaluate the performance of a risk assessment model that incorporates both environmental and genetic risk factors in a Japanese population.

Methods: We conducted a case-control study with 265 cases and age- and sex- matched 530 controls within the framework of the Hospital-based Epidemiological Research Program at Aichi Cancer Center (HERPACC). We fit regression models with the risk factors. For discrimination assessment, we compared the area under the curves (AUC) in the receiver - operating curve (ROC) analysis of Environmental model (E-model), Genetic model (G-model), and Inclusive model. In addition, we evaluated contribution to risk prediction for each risk factor by calculating the integrated discrimination improvement (IDI) index.

Results: We created a risk prediction model for esophageal cancer by polymorphisms of ADH1B and ALDH2, drinking and smoking. The AUCs were 0.873 (0.847-0.898) for E-model and 0.747 (0.711-0.782) for G-model. The Inclusive model including all risk factors, with an AUC of 0.931 (0.912-0.949), showed promise for stratifying people into different risk groups. We found that each risk factor statistically significantly contributed to risk prediction for esophageal cancer (p< 0.001, the IDI indexes of 0.064 for rs1229984, 0.066 for rs671, 0.220 for drinking and 0.101 for smoking).

Conclusion: The risk prediction model that includes these environmental and genetic factors could be useful to classify Japanese into relevant risk groups of esophageal cancer for personalized prevention programs.

Citation Format: Hidemi Ito, Isao Oze, Satoyo Hosono, Miki Watanabe, Hideo Tanaka, Keitaro Matsuo. The risk prediction for esophageal cancer by drinking, smoking, and the polymorphisms of ALDH2 and ADH1B. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4618. doi:10.1158/1538-7445.AM2015-4618
Abstract 462: Cytotoxic CD8+ T cells and immunosuppressive T regulatory cells are associated with aggressive breast cancer subtypes
Authors: Khramtsova Galina F. , Nanda Rita , Khramtsova Ekaterina A. , Sveen Lise , Olugbile Sope , Olopade Olufunmilayo I.
Published: August 2015

Background: An increase in tumor infiltrating lymphocytes (TILs) indicates an immune response to tumor antigens. CD8+ TILs play a vital role in cancer progression, and may have tumor suppressive effects. FoxP3+ T regulatory cells (Tregs), however, suppress a variety of immune cells - including CD4+ TILs, CD8+ TILs, dendritic cells, B cells, and macrophages - and thus may inhibit the beneficial effect these cells have on tumor suppression. Specifically, CD8+ TILs - which are a critical component of tumor-specific adaptive immunity - are a major target of Tregs. This study was aimed at understanding how these TILs correlate with breast cancer molecular subtypes in diverse population of breast cancer patients.

Methods: Tissue Microarrays (TMAs) of breast cancer cases from 1992-2010 were obtained from the University of Chicago breast cancer biospecimen bank under IRB approved protocols. Molecular subtype was assigned based on immunohistochemical (IHC) staining into the following groups: luminal A (ER+ and/or PR+, HER2-), luminal B (ER+ and/or PR+, HER2+), HER2-like (ER/PR-, HER2+) and basal-like (ER/PR/HER2-, EGFR+ and/or CK5/6+). IHC for CD8 and FoxP3 was performed on TMAs using DAKO, cat # M7103 antibody, and Abcam antibody cat #ab20034. The amount of CD8+ and FoxP3+ cells presents within the neoplastic tissue were counted manually in two high-powered fields (x400) by two independent pathologists, and recalculated for 1 mm2. The ratio of FoxP3+/CD8+ was calculated and correlated with molecular subtype.

Results: Of the 292 cores evaluated, 268 were evaluable representing 134 individual cases. There were 28 basal-like (20.9%), 14 HER2 (10.4%), 78 luminal A (58.3%), and 14 luminal B (10.4%). Basal-like tumors showed the highest infiltration of both Tregs and CD8+ TILs when compared to other breast cancer subtypes, and the ratio of FoxP3+: CD8+ TILs in basal-like tumors were 1.7, 2.5, and 4.5-fold higher than in HER2, Luminal A, and Luminal B tumors respectively (p<0.05 for all correlations).

Summary: FoxP3+ and CD8+ TIL infiltration density were significantly different among molecular subtypes of breast cancer. Future studies will examine the genomic determinants and prognostic significance of TILs in aggressive breast cancer subtypes, especially basal-like breast cancers.

Citation Format: Galina F. Khramtsova, Rita Nanda, Ekaterina A. Khramtsova, Lise Sveen, Sope Olugbile, Olufunmilayo I. Olopade. Cytotoxic CD8+ T cells and immunosuppressive T regulatory cells are associated with aggressive breast cancer subtypes. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 462. doi:10.1158/1538-7445.AM2015-462