bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1568: Detection of HBV-host junction DNA sequences in urine of patients with HBV-infected HCC
Authors: Lin Selena , Song Benjamin , Trauger Evan , Hoffman Malcolm , Thompson Emilie , Zhou Rebecca , Jain Surbhi , Song Wei , Su Ying-Hsiu
Published: August 2015

Hepatitis B virus (HBV) related liver diseases often progresses from hepatitis to cirrhosis and eventually to hepatocellular carcinoma (HCC), which has a poor prognosis, mostly due to late detection. Chronic HBV infections are associated with >50% of HCC cases worldwide, and up to 90% of HBV-related HCCs (HBV-HCC) were found to contain integrated HBV DNA. During infection, HBV DNA integrates in the host chromosome at variables sites, generating a unique HBV-host junction sequence (HBV-JS) that can serve as a DNA finger print of an infected hepatocyte. During hepatocarcinogenesis, tumorigenic hepatocyte(s) undergo uncontrolled clonal expansion resulting in one to few particular HBV-JS's becoming dominant. Thus, we hypothesize that the appearance of expanded HBV-JS species is a marker of clonal expansion and can be a potential marker for early detection of HBV-HCC.

A PCR-based enrichment assay and next generation sequencing (NGS) approach were developed for the HBV DR1-2 integration hotspot region to explore the feasibility of detecting HBV-JS's in the circulation (i.e. urine) of patients with HBV-HCC. HBV-HCC tissue DNA samples underwent library construction and were enriched for DNA containing the HBV DR1-2 sequences. Enriched DNA fragments were then cloned and sequenced, from which 5 HBV-JS's were discovered at Chr5 (hTERT promoter), Chr7 (AOAH intronic transcript 1), Chr12 (KCN2), Chr19 (miRNA 512-1,-2), and Chr22 (LARGE) in four patient samples. These sites were further confirmed in matched urine DNA samples by designing primers to amplify the HBV-JS. The NGS approach, which includes HBV-HCC (n = 18), HBV-hepatitis (n = 5), and HBV-cirrhosis (n = 8) urine samples to identify HBV-JS’s, is in progress. Overall, this would be the first study both to demonstrate unambiguous detection of HBV-JS's in the urine of HBV-HCC patients and to suggest the potential of detecting expanded HBV-JS species in urine as a marker for HBV-HCC screening. Furthermore, if successful, the high-throughput detection of HBV-JS's in urine by NGS offers a promising noninvasive HBV-HCC screening platform, as well as provides insight into the carcinogenic mechanisms of HBV DNA integration.

Citation Format: Selena Lin, Benjamin Song, Evan Trauger, Malcolm Hoffman, Emilie Thompson, Rebecca Zhou, Surbhi Jain, Wei Song, Ying-Hsiu Su. Detection of HBV-host junction DNA sequences in urine of patients with HBV-infected HCC. [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 1568. doi:10.1158/1538-7445.AM2015-1568
Abstract 1600: A multiplexed marker-based algorithm for diagnosis of carcinoma of unknown primary using circulating tumor cells
Authors: Matthew Elizabeth M. , Zhou Lanlan , Vijayvergia Namrata , Dicker David T. , Gustafson Karen S. , Cooper Harry S. , Ross Eric A. , Lim Bora , Harouaka Ramdane , Zheng Si-Yang , Lamparella Nicholas E. , Drabick Joseph J. , Truica Cristina I. , Yang Zhaohai , El-Deiry Wafik S.
Published: August 2015

Abstract: Real-time, single-cell multiplex immunophenotyping of circulating tumor cells (CTCs) is hypothesized to inform diagnosis of tissue of origin in patients with carcinoma of unknown primary (CUP). In 20 to 50% of CUP patients, the primary site remains unidentified, presenting a challenge for clinicians in diagnosis and treatment. Invasive tissue biopsy is the gold standard for diagnosis but improved methodology is needed. We developed a post-CellSearch CTC assay using multiplexed Q-dot or DyLight conjugated antibodies with the goal of detecting multiple markers in single cells within a CTC population. We adapted our approach to size-based CTC enrichment protocols for capturing CTCs and subsequent immunofluorescence (IF) using a minimal set of markers to predict the primary sites for common metastatic tumors. The carcinomas are diagnosed with cytokeratin 7 (CK7), cytokeratin 20 (CK20), thyroid transcription factor 1 (TTF-1), estrogen receptor (ER) or prostate-specific antigen (PSA) as the minimal set of informative markers. IF has been optimized in cultured tumor cells with individual antibodies, then with conjugated antibodies to form a multiplex antibody set. With IF, we evaluated antibodies specific to these 5 markers in lung, breast, colorectal, and prostate cancer cell lines and blood samples from metastatic prostate and breast cancer patients. Our technological advance provides a noninvasive, inexpensive, diagnostic blood test as an adjunct to routine tissue biopsy. Its further implementation requires prospective clinical testing.

Citation Format: Elizabeth M. Matthew, Lanlan Zhou, Namrata Vijayvergia, David T. Dicker, Karen S. Gustafson, Harry S. Cooper, Eric A. Ross, Bora Lim, Ramdane Harouaka, Si-Yang Zheng, Nicholas E. Lamparella, Joseph J. Drabick, Cristina I. Truica, Zhaohai Yang, Wafik S. El-Deiry. A multiplexed marker-based algorithm for diagnosis of carcinoma of unknown primary using circulating tumor 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 1600. doi:10.1158/1538-7445.AM2015-1600
Abstract 1665: Design, synthesis, biological evaluation of cyclopenta[d]pyrimidines as antitubulin agents and discovery of N-(4-methylthiophenyl) and N-(4-dimethylaminophenyl) substituted N,2-dimethyl-cyclopenta[d]pyrimidines as long acting and poten
Authors: Gangjee Aleem , Xiang Weiguo , Mooberry Susan L. , Hamel Ernest
Published: August 2015

Compounds that bind to the colchicine site of microtubules have promising antitumor potential. Some of these agents also possess the ability to overcome multiple drug resistance mechanisms, including those mediated by the expression of P-glycoprotein (Pgp) or expression of the tubulin β-III isotype. We have reported that compounds 1 and 2 were potent antiproliferative agents that bind to the colchicine site of tubulin. The previous structure-activity relationship studies of cyclopenta[d]pyrimidine based antitubulin agents demonstrated the importance of the para-methoxyphenyl moiety for antiproliferative activity. However, the literature suggests that the major metabolism of the methoxyphenyl moiety was likely O-demethylation. In order to increase the potency and in vivo stability of 1, we focused our attention on the variation of the para-methoxy aniline moiety. Compound 3 with a 4′-methylthiophenyl substitution was designed to decrease potential metabolic demethylation. This S-methyl analog was a potent inhibitor of proliferation with an IC50 of 4.6 ± 0.5 nM in MDA-MB-435 cells. The possible sulfoxide metabolite, 4 was synthesized and had an IC50 = 7.9 ± 0.5 nM. Each of these depolymerized microtubules. In addition, the 4′-dimethylamino moiety compound 5 also demonstrated potent antitubulin and antiproliferative activity (MDA-MB-435: IC50 = 8.2 ± 0.6 nM). The possible N-demethylated metabolite of 5, para-methylamino 6 was also a highly potent antiproliferative agent (MDA-MB-435: IC50 = 20 ± 1.9 nM). The parent compounds 3 and 5, as well as their probable metabolites 4 and 6, inhibited the binding of [3H]colchicine to tubulin, which showed that these compounds bind to the colchicine site. The potent antiproliferative activities of the parent compounds and the potential metabolites of compounds 3 and 5 suggest the utility of 3 and 5 as long acting, potent antimitotic agents. Finally, the in vivo efficacy of the S-methyl analog 2 was demonstrated against MDA-MB-435 triple negative breast cancer murine xenograft model, indicating the potential utility of these analogs as antitumor agents.

Citation Format: Aleem Gangjee, Weiguo Xiang, Susan L. Mooberry, Ernest Hamel. Design, synthesis, biological evaluation of cyclopenta[d]pyrimidines as antitubulin agents and discovery of N-(4-methylthiophenyl) and N-(4-dimethylaminophenyl) substituted N,2-dimethyl-cyclopenta[d]pyrimidines as long acting and poten [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 1665. doi:10.1158/1538-7445.AM2015-1665
Abstract 1672: Structural basis for inhibition of ligand-dependent and -independent ErbB3 activation by KTN3379
Authors: Alvarado Diego , Lee Sangwon , Greenlee Etienne , Ligon Gwenda F. , Lillquist Jay S. , Natoli Edward J. , LaVallee Theresa M. , Schlessinger Joseph
Published: August 2015

ErbB3 signaling plays an important role in the progression of a number of solid tumor types, and is the target of several therapeutic monoclonal antibodies. In cancer, ErbB3 activation can be driven by its ligand Neuregulin, or in its absence, through high overexpression of its co-receptor ErbB2. Here, we report that the ErbB3 monoclonal antibody KTN3379, currently in phase 1 clinical development, robustly inhibits both mechanisms of ErbB3 activation in different tumor model settings. The crystal structure of a KTN3379 Fab in complex with the full ErbB3 extracellular domain reveals that the antibody binds with very high affinity to a novel and unique epitope in the boundary of domains 2 and 3, and locks the receptor in an inactive conformation. Thus, KTN3379 interferes with the first step of ErbB3 activation, which is required for both ligand-dependent and ligand-independent signaling. The mechanism of inhibition described here should be also applicable to EGFR and ErbB4.

Citation Format: Diego Alvarado, Sangwon Lee, Etienne Greenlee, Gwenda F. Ligon, Jay S. Lillquist, Edward J. Natoli, Theresa M. LaVallee, Joseph Schlessinger. Structural basis for inhibition of ligand-dependent and -independent ErbB3 activation by KTN3379. [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 1672. doi:10.1158/1538-7445.AM2015-1672
Abstract 168: MALAT1 is crucial for epithelial-mesenchymal transition of breast cancer cells in acidic microenvironment
Authors: Gupta Subash C. , Mo Yin-Yuan
Published: August 2015

Metastasis Associated Lung Adenocarcinoma Transcript 1 (MALAT1), a long non-coding RNA (lncRNA), is found up-regulated in several tumors. However, little is known about the role of MALAT1 in breast cancer, which is one of the highly invasive tumors among women worldwide. In this study, we found that MALAT1 was highly expressed in both breast cancer cell lines and breast cancer specimens in comparison to normal specimens. We found that low extracellular pH (pHe-acidosis), a major hallmark of solid tumors, further up-regulates MALAT-1 in several breast cancer cell lines in a time-dependent manner. Furthermore, acidosis induces epithelial to mesenchymal transition (EMT)-associated vimentin and Twist 1 expression but suppresses E-cadherin levels in breast cancer cells. Moreover, breast cancer cells under low pHe adopt stem-ness property as revealed by an increase in CD44+ and ESA+ population, mammosphere formation, and expression of OCT-4, SOX-2 and Nanog. Acidosis promotes translocation of β-catenin from cytoplasm to nucleus. The antisense oligonucleotides mediated gene silencing of MALAT-1 prevents acidosis induced nuclear translocation of β-catenin. This is in consistent with our previous finding that breast cancer cells cultured under low pHe exhibit an increased invasion activity. These results suggest that acidosis-induced MALAT-1 trigger an EMT program, leading to cell invasion and metastasis. Therefore, MALAT-1 represents a potential target for breast cancer therapy. Studies are underway to delineate the mechanistic association between MALAT-1, β-catenin, and EMT in acidic microenvironment.

Citation Format: Subash C. Gupta, Yin-Yuan Mo. MALAT1 is crucial for epithelial-mesenchymal transition of breast cancer cells in acidic microenvironment. [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 168. doi:10.1158/1538-7445.AM2015-168
Abstract 1732: Effect of 19-substituted benzoquinone ansamycin Hsp90 inhibitors on Hsp90/Cdc37/co-chaperone complexes and casein kinase 2 (CK2) activity
Authors: Chang Chuan-Hsin , Ross David , Siegel David , Moody Christopher J , Kitson Russell
Published: August 2015

Benzoquinone ansamycin (BQA) Hsp90 inhibitors such as 17-DMAG and 17-AAG have off target toxicities including hepatotoxicity. Mechanisms underlying the toxicity of BQAs are a function of their ability to redox cycle and/or arylate cellular nucleophiles at the unsubstituted 19-position of the molecule. Therefore, we designed 19-substituted BQAs (19BQAs) to prevent conjugation with glutathione and protein thiols as an approach to reduce the hepatotoxicity and minimize off target effects of the BQA class of Hsp90 inhibitors. Our data demonstrated that 19BQAs had reduced toxicity to liver cells relative to their parent quinones while maintaining Hsp90 inhibitory activity, validating the overall approach. 19BQAs depend on tumor cell levels of NQO1 (NAD(P)H: quinone oxidoreductase 1) for optimal activity. We examined 19-phenyl-17-DMAG as a model compound in human isogenic breast cancer cell lines MDA468 (NQO1 null) and MDA468/NQ16 (NQO1 overexpressing) and in NQO1expressing BT474 breast cancer cells. 19-phenyl-17-DMAG induced growth inhibition, apoptosis and the molecular signature of Hsp90 inhibition including decreases in Hsp90 client protein levels and compensatory induction of Hsp70. In addition, treatment of MD468/NQ16 and BT474 cell lines with 19-phenyl-17-DMAG or 17-DMAG also caused the dissociation of Hsp90 from the co-chaperone Cdc37. In these studies we also observed that treatment with19-phenyl-17-DMAG or 17-DMAG resulted in inhibition of Cdc37 phosphorylation. A major kinase responsible for phosphorylation of Cdc37 is casein kinase 2 (CK2) and inhibitors of CK2 are currently under development as anticancer drugs. CK2 activity assays, immunoprecipitation and western blot analysis demonstrated that both 19-phenyl-17-DMAG and 17-DMAG inhibited CK2 kinase activity and reduced CK2α protein subunit expression in NQO1-expressing MD468/NQ16 and BT474 breast cancer cells. Decreased phosphorylation of Cdc37 led to dissociation of the Hsp90/Cdc37/client complex and resulted in the degradation of multiple kinase clients including Raf-1, Cdk4, Akt and HER2. These data suggest that inhibition of both Hsp90 and CK2 by Hsp90 inhibitors may play a role in their antitumor effects. (Supported by CA51210).

Citation Format: Chuan-Hsin Chang, David Ross, David Siegel, Christopher J Moody, Russell Kitson. Effect of 19-substituted benzoquinone ansamycin Hsp90 inhibitors on Hsp90/Cdc37/co-chaperone complexes and casein kinase 2 (CK2) activity. [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 1732. doi:10.1158/1538-7445.AM2015-1732
Abstract 1752: 6-Methoxyethylamino-numonafide (MEAN) inhibits hepatocellular carcinoma as a single agent or in combination with sorafenib
Authors: Liu Yanning , Lou Guohua , Norton John , Wang Chen , Kandela Irawati , Tang Shuai , Huang Min , Avram Michael , Green Richard , Mazar Andrew , Appella Daniel , Chen Zhi , Huang Sui
Published: August 2015

Hepatocellular carcinoma (HCC) is the third leading form of cancer worldwide and the incidence is increasing rapidly in the United States, tripling over the past 3 decades. Unfortunately, chemotherapeutic treatment strategies against localized and metastatic HCC are very ineffective, leading to a high mortality from the disease. Sorafenib is the sole FDA approved chemotherapeutic currently used clinically for the disease and it shows limited efficacy and substantial toxicity. We have developed a small molecule, 6-methoxyethylamino-numonafide (MEAN), which is highly effective in two murine xenograph models of human HCC. MEAN is more effective in tumor growth inhibition and less toxic than sorafenib at the same concentration. MEAN, at efficacious doses, does not significantly affect animal body weight and does not significantly induce liver damage as determined by the serum levels of liver enzymes at the experimental endpoint. NCI60 cell assay analyses using the COMPARE algorithm indicate that, at IC50 concentration, MEAN does not significantly correlate with any of the small molecules in the 60 cell assay database (correlation<0.5), suggesting novel modes of action in tumor inhibition. MEAN suppresses some of the well-known oncogene expression and enhances the expression of a subset of tumor suppressors. MEAN remains effective against cancer cells that are drug resistant due to expression of P-glycoprotein drug efflux pumps and has a favorable single dose pharmacokinetic profile. In summary, MEAN is effective against HCC tumor growth and is an excellent candidate for future development as therapeutic agent for management of HCC.

Citation Format: Yanning Liu, Guohua Lou, John Norton, Chen Wang, Irawati Kandela, Shuai Tang, Min Huang, Michael Avram, Richard Green, Andrew Mazar, Daniel Appella, Zhi Chen, Sui Huang. 6-Methoxyethylamino-numonafide (MEAN) inhibits hepatocellular carcinoma as a single agent or in combination with sorafenib. [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 1752. doi:10.1158/1538-7445.AM2015-1752
Abstract 1776: Synergistic anticancer effects of triptolide with celastrol, two main compounds from Thunder God Vine
Authors: Jiang Qi-Wei , Cheng Ke-Jun , Wei Xing , Shi Zhi
Published: August 2015

Objective: Triptolide and celastrol are two most widely studied and promising compounds isolated from Thunder God Vine (Trypterigium wilfordii Hook F) with promising anticancer activity. However, their clinical application was limited by toxicity. We aim to investigate the synergistic anti-proliferative effects of triptolide with celastrol in cancer and the molecular mechanism.

Methods: The effects of triptolide and celastrol in cancer cells and the molecular mechanism were assessed by using MTT assay in vitro, xenografts mouse model in vivo, flow cytometry, fluorescence microscope and Western blot analysis.

Results: The combination of triptolide with celastrol induced strong cell growth inhibition, cell cycle arrest at G2/M phase and apoptosis with the increasing intracellular ROS accumulation in cancer cells. Pretreatment with ROS scavenger N-acetyl-L-cysteine totally blocked the apoptosis induced by triptolide with celastrol. Treatment with celastrol alone decreased many Hsp90 client proteins including Akt, Cdks, EGFR and Raf1, but lead to stress-induced increase of Hsp70 and Hsp27. Adding triptolide to celastrol treatment abrogated this phenomenon and induced more apoptosis. In xenografts mouse model, the lower-dose combination of triptolide with celastrol significant inhibited the growth of tumors without obvious toxicity.

Conclusions: Triptolide in combination with celastrol showed outstanding synergistic anticancer effect, suggesting that this beneficial combination may offer a promising treatment option for cancer patients.

Citation Format: Qi-Wei Jiang, Ke-Jun Cheng, Xing Wei, Zhi Shi. Synergistic anticancer effects of triptolide with celastrol, two main compounds from Thunder God Vine. [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 1776. doi:10.1158/1538-7445.AM2015-1776
Abstract 1791: Targeting radiation-inducible cell surface GRP78 using GIRLRG peptide as a novel imaging and therapeutic strategy for tumors
Authors: Kapoor Vaishali , Dadey David , Nguyen Kim , Li Hua , Rogers Buck , Thotala Dinesh , Hallahan Dennis
Published: August 2015

Traditional radiation therapy is often associated with significant toxicity to normal cells that could limit the success of cancer therapy. The greater understanding in the molecular differences between cancer cells and normal cells has lead to targeted therapies in cancer treatment. We have discovered novel radiation-inducible antigens in cancer using phage-display peptide libraries. One such inducible-antigen is Glucose regulated protein 78kDa (GRP78) that was shown to bind the hexapeptide GIRLRG. GRP78 is known to regulate cellular stresses, including hypoglycemia, hypoxia and the ER stress response. It is over-expressed in different cancer subtypes and has been correlated to their poor prognosis.

In this study, we evaluated the cell surface induction of GRP78 post irradiation (IR) on lung cancer (A549 and LLC), glioma (D54 and GL261) and endothelial (HUVEC and 3B11) cell lines using flow cytometry. Significantly higher surface expression of GRP78 was observed in all cell lines. We determined the specificity of GIRLRG to bind tumors in vivo using nano SPECT technology. GIRLRG was conjugated to a 40KDa PEG (for longer circulation time) and radiolabelled with 111Indium (111In) using diethylene triamine pentaacetic acid (DTPA) as the chelator. The A549 tumor bearing nude mice were irradiated (3Gy x 3 times) or sham irradiated prior to tail vein injections of 111In labeled GIRLRG. The mice were imaged 48h and 96h post injection using nano CT-SPECT imager. The SPECT images revealed that GIRLRG specifically bound to the irradiated A549 tumors while little or no binding was seen in the sham irradiated tumors. The post-SPECT imaging bio-distrubution also revealed maximum uptake in irradiated tumors. We further evaluated tumor binding of GIRLRG in Glioma (D54), esophageal cancer (OE33), cervical cancer (HT3), and pancreatic cancer (BxPC3). The nano-SPECT imaging showed that GIRLRG specifically bound to all the irradiated tumors tested. Phosphor images of the tumor sections showed that GIRLRG specifically bound to the tumors and not to normal tissues.

In conclusion, GIRLRG peptide has high affinity to GRP78 in vitro and in vivo. Radiolabeled GIRLRG is a novel tool for imaging tumors and may be developed further as a therapeutic agent to deliver cancer specific drugs or therapeutic radioisotopes.

Citation Format: Vaishali Kapoor, David Dadey, Kim Nguyen, Hua Li, Buck Rogers, Dinesh Thotala, Dennis Hallahan. Targeting radiation-inducible cell surface GRP78 using GIRLRG peptide as a novel imaging and therapeutic strategy for tumors. [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 1791. doi:10.1158/1538-7445.AM2015-1791
Abstract 1793: Novel radiation mitigators and anticancer drugs
Authors: Schiestl Robert H. , Davoren Michael , Rivina Yelena
Published: August 2015

The possibility of a radiation disaster from a nuclear detonation or accident has existed for over 50 years and spawned much of the basic research in radiobiology in the 1950-60s. The recent Fukushima accident was yet another reminder that there remains a dire need to develop novel therapies against radiation-induced toxicities. Here we report on the development of two novel radiation countermeasure therapies: Yel001 and Yel002. These small, biologically active, drug-like molecules were uncovered in the DEL high throughput assay reducing radiation-induced cyto- and geno-toxicity in yeast. Radiation-modulating activity was further confirmed in yeast plate-based DEL Assay: addition of either Yel001 or Yel002 to irradiated cultures reduced cell death and genomic instability. Further, Yel compounds increases survival to 75% in vivo following an LD100/30 dose of ionizing radiation (IR) with the first therapeutic injection administered 24 hours post exposure followed by injections at 48,72,96, and 120 hours. Additionally, treatment with Yel001 and Yel002 compounds reduces radiation-induced leukemia from 90% to to 50% and 40% respectively. Of note, treatment with either Yel001 or Yel002 reduced spontaneous leukemia rate from 10% to 0%. Furthermore, Yel002 significantly prolonged the life of Atm deficient mice. Treatment with Yel002 following IR accelerates the recovery of the hematopoietic cells and protects stem cells after sub-lethal exposures. In addition, treatment with Yel002 reduces I131, EMS, MMS, UV, cigarette smoke extract as well as nitrogen mustard induced toxicity as well as genotoxicity showing a broad application spectrum. Proteomics shows that Yel002 induces DNA repair mediated by Atm signaling, homologous recombination, nonhomologous end joining, base excision repair, DNA damage binding proteins and chromosome segregation functions. Toxicity has not been observed in neither in vitro or in vivo administrations, even in a two generation teratogenicity assay. Overall, Yel compounds have much potential as stockpile therapies for radiation-induced lethality and cancer as well as spontaneous cancer: they are highly effective when administered up to 24hours post exposure, they reduce radiation-induced sequelae such as leukemia, and appear to have an acceptable toxicity profile.

Citation Format: Robert H. Schiestl, Michael Davoren, Yelena Rivina. Novel radiation mitigators and anticancer drugs. [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 1793. doi:10.1158/1538-7445.AM2015-1793
Abstract 180: Expanding the definition of core binding factor leukemia
Authors: Fischer John A. , Rossetti Stefano , Datta Arani , Beghini Alessandro , Sacchi Nicoletta
Published: August 2015

The core binding factor (CBF) transcription factor complex regulates coding and non-coding genes that play a critical role in hematopoiesis. Chromosomal rearrangements involving the two CFB subunits, RUNX1 and CFB beta, are common in acute myeloid leukemia (AML). The fusion proteins resulting from these rearrangements deregulate the transcription of RUNX1-target genes, including microRNAs critical for KIT-mediated proliferation (e.g. miR-221) and myeloid differentiation (e.g. miR-223). We found that overexpression of miR-17, which downregulates RUNX1 level by targeting RUNX1-UTR, recapitulates the biological effects of CBF-AML fusion proteins by affecting the transcription of common coding and non-coding RUNX1-targets. Consistently, increased levels of miR-17 can be detected in AML patient samples displaying increased KIT level, but without evidence of CBF mutations. Based on these finding, we propose that the definition of CBF-AML leukemia can be expanded to accommodate leukemia induced not only by genetic factors but also epigenetic factors, such as microRNAs targeting RUNX1.

Acknowledgements: Funding for this study was provided by a Roswell Park Alliance Foundation award, the University of Rochester-RPCI pilot grant, and the Mark Diamond Research Fund.

Citation Format: John A. Fischer, Stefano Rossetti, Arani Datta, Alessandro Beghini, Nicoletta Sacchi. Expanding the definition of core binding factor leukemia. [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 180. doi:10.1158/1538-7445.AM2015-180
Abstract 1804: The role of adipose derived stromal cells for reversal of radiation fibrosis
Authors: Zhao Xiao , Lee Ju Hee , Yip Kenneth , Ailles Laurie , Liu Fei-Fei
Published: August 2015

Hypothesis: Up to 70% of patients after radiotherapy develop radiation fibrosis (RF), an irreversible scarring of normal tissue resulting in functional morbidity and increased risk of surgical complications. Adipose-derived stromal cells (ADSCs) have been used effectively for the treatment of complex wounds in animal models and clinical trials. We hypothesize that ADSCs may reverse RF by repopulating mesenchymal precursor cells and by providing angiogenic, anti-inflammatory, and matrix remodeling factors.

Methods: A RF animal model was developed and ADSC isolation was confirmed by immunophenotype and differentiation assay. GFP and luciferase labelled ADSCs were used to assess biodistribution after transplantation. To determine the therapeutic effect of ADSC transplantation for RF, we assessed functional changes to leg contracture, oxygen saturation and perfusion and molecular changes to inflammation, vascularization, and matrix remodeling. To determine the mechanism of ADSC-mediated fibrosis reversal, we assessed transcriptomic and epigenetic changes to RF tissue.

Results: A RF model was created by radiating the hind limb of C3H mice. This model showed a dose dependent leg contracture and histological findings of fibrosis. We confirmed the immunophenotype of isolated ADSC and their ability to differentiate into adipogenic, chondrogenic, and osteogenic lineages. ADSC transplantation showed a statistically significant trend towards improved leg contracture (2-way ANOVA, p<0.05). Biodistribution studies confirmed the presence of ADSCs in the subdermis of RF tissue with persistence for at least 18 days post-transplantation. Preliminary RNA-seq analysis using unsupervised hierarchical clustering showed ADSC treated radiation tissue was more similar to normal tissue than control. In depth RNA-seq data analysis is underway.

Conclusions: ADSC transplantation may be an effective treatment for the reversal of radiation fibrosis. As cancer survivorship increases, the prevalence of radiation fibrosis will rise and necessitate increased focus on effective treatment strategies for this condition.

Citation Format: Xiao Zhao, Ju Hee Lee, Kenneth Yip, Laurie Ailles, Fei-Fei Liu. The role of adipose derived stromal cells for reversal of radiation fibrosis. [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 1804. doi:10.1158/1538-7445.AM2015-1804
Abstract 1819: Profiling of extracellular matrix proteins in pancreatic ductal adenocarcinoma progression and metastasis
Authors: Tian Chenxi , Ohlund Daniel , Clauser Karl , Carr Steven A. , Tuveson David A. , Hynes Richard O.
Published: August 2015

Pancreatic ductal adenocarcinoma (PDAC) is one of the most common causes of cancer-related death, and is characterized by accumulation of excessive extracellular matrix (ECM). Although the ECM of tumors, such as PDAC, has long been known to have major effects on behavior of both tumor and stromal cells, including invasion, angiogenesis, metastasis, and resistance to therapy, the detailed composition of the ECM in PDAC tumor progression and metastasis are not understood. To study the involvement of ECM proteins in PDAC primary tumor growth and the tissue-specific metastasis program, we modified the comprehensive ECM proteomic approach by Mass Spectrometry (MS) recently developed in our lab to profile the ECM changes during PDAC progression in both the LSL-KrasG12D/+; LSL-Trp53R127H/+; Pdx-1-Cre (KPC) mouse models of PDAC and human patient samples. We will compare ECM profiles from normal pancreas, pancreatitis, early and late pancreatic intraepithelial neoplasia (PanIN) stages, and PDAC. Initial MS identified between 100 and 140 ECM proteins in samples representing each stage. The changes identified include changes previously associated with PDAC (e.g., upregulation of Tnc, Lox) plus novel, previously unreported changes. The combination of mouse models and human samples will allow us to uncover critical ECM changes during PDAC progression and metastasis and, furthermore, to investigate and understand the functional consequences of such changes. The ongoing studies will characterize the ECM of PDAC in detail and provide novel insights into diagnosis, prognosis and treatments of this very difficult disease.

Citation Format: Chenxi Tian, Daniel Ohlund, Karl Clauser, Steven A. Carr, David A. Tuveson, Richard O. Hynes. Profiling of extracellular matrix proteins in pancreatic ductal adenocarcinoma progression and metastasis. [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 1819. doi:10.1158/1538-7445.AM2015-1819
Abstract 1833: KRAS-regulated P4HA1 in pancreatic tumor and its hydroxylated peptide as a serum biomarker for early diagnosis
Authors: Deng Zaian , Mann Karen M. , Koay Eugene J. , Ferrari Mauro , Wu Xifeng , Chiao Paul J. , Hu Tony
Published: August 2015

Protein posttranslational modification (PTM) plays a pivotal role in cancer progression through regulating gene expression, protein activities and stabilization. These chemistry events are regarded as potential biomarkers for cancer diagnosis, prognosis, and therapeutic efficacy evaluation. Hydroxylated proline is the most common PTM amino acid residue in the proteome. However, this modification is often overlooked. Here, we demonstrate the expression level of prolyl-4-hydroxylase alpha-1(P4HA1) in tumor tissues and the ratio of its catalyst (hydroxyprolyl-3-bradykinin, Hyp-BK) to its substrate (bradykinin, BK) in blood can serve as biomarkers for early pancreatic cancer diagnosis. First, we found that the P4HA1 expression level responds to the activation of KRAS which is commonly observed in pancreatic cancer cells. Immunohistochemistry testing for tumor tissue array reveals that pancreatic cancer tumor tissues, even in stage I tumor, present higher P4HA1 expression levels than tissues with chronic pancreatitis and healthy control. Peptidomic studies on clinical blood samples revealed that the Hyp-BK/BK value was 2.3 fold higher in pancreatic cancer patients than chronic pancreatitis patients and control group. The results were further validated in an independent sample cohort with blind information. We combined CA19-9 and the Hyp-BK/BK value as a biomarker panel obtaining an area under the curve (AUC) of 0.742 and 0.813 in the comparison of pancreatic cancer to pancreatitis patients and controls, respectively. Our results demonstrate that the measurement of Hyp-BK/BK value in blood, which reflects the hydroxylation level regulated by tumor-localized P4HA1, can be used as a biomarker for early detection of pancreatic cancer.

Citation Format: Zaian Deng, Karen M. Mann, Eugene J. Koay, Mauro Ferrari, Xifeng Wu, Paul J. Chiao, Tony Hu. KRAS-regulated P4HA1 in pancreatic tumor and its hydroxylated peptide as a serum biomarker for early diagnosis. [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 1833. doi:10.1158/1538-7445.AM2015-1833
Abstract 1855: Structural requirements for the association of the ETS domain transcription factor Elk1 and the androgen receptor in enabling the growth of prostate cancer cells
Authors: Rosati Rayna , Chari Venkatesh , Patki Mugdha , Ratnam Manohar
Published: August 2015

The ETS domain transcription factor Elk1 tethers the androgen receptor (AR) to chromatin, enabling sustained activation of a set of genes critical for cell growth in established prostate cancer cell lines. This role of Elk1 is independent of the ability of Elk1 to transiently activate immediate early genes in response to phosphorylation by ERK. We compared the structural requirements for the association of AR and Elk1 with that for activation of Elk1 by phosphorylation using mammalian one- and two-hybrid assays. The critical polypeptide segments were mapped by systematic deletion mutagenesis. The amino-terminal A/B domain of AR, which lacks the ligand-binding domain (LBD), was adequate for association with Elk1 and to activate Elk1-AR target genes in LNCaP prostate cancer cells. The AR A/B domain also supported hormone-independent growth of LNCaP cells reflecting the ability of overexpressed natural splice variants of AR, which also lack LBD, to support prostate tumor growth. The association of AR with Elk1 did not require ERK1/2 or serum response factor (SRF), which are the known binding partners of Elk1. We identified two sites on Elk1 that are critical for its association with the AR A/B domain as well as the whole AR molecule. One of those sites spans amino acid residues 297 to 317, overlapping one of two ERK docking sites. The other site on Elk1 mapped to amino acid residues 387 to 397, adjacent to the downstream ERK docking site. The results suggest that the association of AR with Elk1 in situ is direct rather than through the Elk1 ternary complex and that the association may be disrupted by small molecules to selectively inhibit growth signaling in prostate cancer cells.

Citation Format: Rayna Rosati, Venkatesh Chari, Mugdha Patki, Manohar Ratnam. Structural requirements for the association of the ETS domain transcription factor Elk1 and the androgen receptor in enabling the growth of prostate 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 1855. doi:10.1158/1538-7445.AM2015-1855
Abstract 1868: Expression of MAS1 in triple-negative breast cancer
Authors: Luo Yi , Kitadai Yasuhiko , Nishiguchi Yukiko , Fujiwara Rina , Sasaki Takamitsu , Ohmori Hitoshi , Kuniyasu Hiroki
Published: August 2015

MAS1 is a receptor for angiotensin 1-7 (A1-7), which is derived from angiotensin II (A-II) by the action of angiotensin converting enzyme (ACE) 2. We examined the role of MAS1 in 132 cases of invasive ductal carcinoma (IDC) of the breast by using immunohistochemistry. While normal mammary tissues expressed MAS1 at high levels, MAS1 expression was attenuated in all IDCs, especially in scirrhous IDCs. The decrease in MAS1 expression was associated with tumor growth, lymph node metastasis, and grade. MAS1 expression was inversely associated with the proliferation index and epidermal growth factor receptor and human epidermal growth factor receptor-2 expression. Of the 132 cases, 12 (9.1%) were triple-negative breast cancer (TNBC) cases. Using the mouse TNBC cell line 4T1, which expresses MAS1, we found that cell growth, anti-apoptotic survival, and invasion were suppressed by A1-7 treatment and enhanced by MAS1 knockdown. Combination treatment with cisplatin, an ACE2 activator, and an A-II type 1 receptor blocker showed synergic effects on tumor growth inhibition. These findings suggest that MAS1 might be a pivotal molecular target for TNBC.

Note: This abstract was not presented at the meeting.

Citation Format: Yi Luo, Yasuhiko Kitadai, Yukiko Nishiguchi, Rina Fujiwara, Takamitsu Sasaki, Hitoshi Ohmori, Hiroki Kuniyasu. Expression of MAS1 in triple-negative 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 1868. doi:10.1158/1538-7445.AM2015-1868
Abstract 1890: Dietary intake of 502 polyphenols and risk of colorectal cancer in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort study
Authors: Zamora-Ros Raul , Romieu Isabelle , Scalbert Augustin ,   EPIC Study group
Published: August 2015

Introduction. Epidemiological evidence on the association between polyphenol intake and colorectal cancer (CRC) risk is still inconsistent. Case-control studies suggest inverse associations, but these were not confirmed in cohort studies so far. Moreover, to date, only some polyphenol classes (flavonoids and lignans) have been studied. We aimed to explore the association between polyphenol intake and CRC risk in the European Prospective Investigation into Cancer and Nutrition (EPIC) study.

Design. The study included 477,386 subjects (29.8% men) aged 35-70 years from 10 European countries. Both lifestyle and dietary questionnaires were collected at enrollment. Dietary polyphenol intakes were estimated using dietary questionnaires and the Phenol-Explorer database, including data on 502 individual polyphenols in 452 plant-based foods. Retention factors for processed and cooked food from the Phenol-Explorer database were also applied.

Results. After a mean follow-up of 11 years, 4,701 validated incident CRC cases (2,979 colon cancer and 1,722 rectal cancer) were identified and included in the analysis. A wide range of total polyphenol intake and a South-to-North geographical gradient (ranging from 584mg/d in Greek women to 1,786mg/d and Danish men) were observed. Phenolic acids (52.6%) were the main contributors to total polyphenol intake, followed by flavonoids (42.4%), alkylphenols (2.7%), tyrosols (1.0%), lignans, stilbenes and other minor polyphenols (<0.3%). Coffee, tea and fruits were the most important food sources of total polyphenols. Cox proportional hazards model will be used to calculate hazard ratios and 95% confidence intervals of CRC for categories of dietary polyphenols intake (total and by classes and subclasses). All analyses will be done separately for CRC and by anatomical subsite: colon, proximal and distal colon, and rectum. Several risk factors known or suspected to influence CRC risk will be considered as potential confounders.

Conclusions. This will be the first study evaluating the association between all polyphenol classes and CRC risk. EPIC is a convenient study to assess these relationships due to the large number of CRC cases, its longitudinal study design, and the high variability in the polyphenol intake amongst the participants across Europe.

Citation Format: Raul Zamora-Ros, Isabelle Romieu, Augustin Scalbert, EPIC Study group. Dietary intake of 502 polyphenols and risk of colorectal cancer in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort study. [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 1890. doi:10.1158/1538-7445.AM2015-1890
Abstract 1891: Butyric acid reverts DRP1-mediated cell proliferation and migration in colorectal cancer cells
Authors: Tailor Dhanir M. , Hahm Eun Ryeong , Singh Shivendra V. , Singh Rana P.
Published: August 2015

Colorectal cancer (CRC) is fourth most commonly diagnosed cancer in the world and more than 1 million new cases diagnosed annually. Human gastro-intestinal (GI) tract carries more than 100 billion microbial cells of over 1000 different species. These gut microbiota have direct correlation with dietary habits and health. Butyric acid (BA) is the byproduct of anaerobic microbial fermentation inside the GI tract that could reach up to 20 mM, and has been shown to inhibit the growth of various cancers. Recently, we observed that BA induced G2/M phase arrest and apoptosis, and down regulated the expression of DRP1. Further, we evaluated the effect of DRP1 expression on cell proliferation and migration and its response to BA treatment. DRP1 overexpression increased cell proliferation rate by 20% when compared to control vector transfected HCT116 cells. BA treatment (1 mM) for 12 h to DRP1 overexpressed cells reverted cell proliferation by 11% and induced cell cycle arrest in S and G2/M phase, BA caused significant G2/M arrest in control cells. DRP1 overexpression led to pro-survival signaling including activation of ERK1/2 and Akt, and concomitantly decreased the levels of cell cycle inhibitory and pro-apoptotic molecules including p53, p21 and Bax. BA treatment reverted the pro-survival signaling and induced the expression of p53 and p21 as well as enhanced the interaction between DRP1 and p53. DRP1 also led to an increase in cell migration and invasion by 28% and 40%, respectively which was associated with decreased expression of E- cadherin. In presence of BA, DRP1 overexpressing cells loss their ability to migrate up to 70% and increased the expression of E-cadherin. These results suggest that regulation of DRP1-dependent mitochondrial fission mediated the cell proliferation and migration and thus could be a novel target to regulate colon cancer growth, progression and metastasis. BA is product of anaerobic fermentation of dietary fibers which is directly related to the nutritional practices of individuals; thus healthy dietary practices may help to reduce the incidence of colorectal cancer.

Citation Format: Dhanir M. Tailor, Eun Ryeong Hahm, Shivendra V. Singh, Rana P. Singh. Butyric acid reverts DRP1-mediated cell proliferation and migration in colorectal 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 1891. doi:10.1158/1538-7445.AM2015-1891
Abstract 1892: Curcumin induces apoptosis in H-Ras transformed human mammary epithelial cells: Cysteine 259 of STAT3 as a putative target
Authors: Hahn Young-Il , Kim Su-Jung , Choi Bu-Young , Na Hye-Kyung , Surh Young-Joon
Published: August 2015

Signal transducer and activator of transcription 3 (STAT3) is a transcription factor that is latent but constitutively activated in many types of cancers. It is well known that STAT3 plays a key role in inflammation-associated tumorigenesis. Curcumin, a natural compound isolated from the traditional Indian spice turmeric (Curcuma longa.,Zingiberaceae), has anti-tumor, anti-proliferative and anti-inflammatory properties. This compound has been extensively used in traditional herbal medicine over the centuries mainly due to its pronounced anti-inflammatory effects. Many studies suggest that curcumin regulates various cancer-related intracellular signaling pathways without damaging normal cells and tissues. Here, we report that curcumin effectively inhibits activation of STAT3 in dose- and time-dependent manners and causes apoptosis of H-Ras transformed breast epithelial cells (H-Ras MCF10A). Specific cysteine residues present in STAT3 appear to be critical for the activity as well as conformation of this transcription factor. We identified the cysteine residue 259 as a putative target of curcumin. We found that the α,β-unsaturated carbonyl moiety of curcumin appears to be essential in its direct binding to STAT3. Tetrahydrocurcumin that lacks the electrophilic α,β-unsaturated carbonyl moiety failed to inhibit STAT3 activation and to induce apoptosis in the same cell line. Taken together, our findings suggest that curcumin can abrogate persistently activated STAT3 signaling through direct interaction, thereby inhibiting STAT3-mediated mammary carcinogenesis.

Citation Format: Young-Il Hahn, Su-Jung Kim, Bu-Young Choi, Hye-Kyung Na, Young-Joon Surh. Curcumin induces apoptosis in H-Ras transformed human mammary epithelial cells: Cysteine 259 of STAT3 as a putative target. [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 1892. doi:10.1158/1538-7445.AM2015-1892