bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract CT-04: A phase II study of neoadjuvant metformin in prostatic carcinoma
Authors: Joshua Anthony M. , Zannella Vanessa , Bowes Barbara , Koritzinsky Marianne , Sweet Joan , Evans Andrew , Trachtenberg John , Jewett Michael , Finelli Antonio , Fleshner Neil , Pollak Michael
Published: August 2012

Background: Metformin is an inhibitor of the complex 1 in the respiratory chain, and is widely used in diabetes due to its effect on reducing insulin resistance. It has also been recently described to have effects via AMPK on inhibiting the mTOR kinase. Significant preclinical and epidemiological studies suggest its role in chemoprevention. These actions provide significant rationale to evaluate its utility in prostate cancer. We conducted a phase II single centre study of neoadjuvant metformin in localised prostate cancer. Methods: Men were required to have histologically confirmed prostate cancer involving at least 20% of at least 1 unfragmented biopsy core. Exclusion criteria included patients who on initial assessment are found to be on treatment with any drug used for the treatment of any form of diabetes, or patients that begin treatment for any form of diabetes during the course of the study. Pts were treated with up to 500mg tid of metformin. The primary objectives were to demonstrate safety and tolerability of neoadjuvant metformin administration in men with prostate cancer and to document changes in phospho-AKT signalling indices. Results: 24 patients were enrolled with 22 evaluable; median age was 64 yrs (range, 45-70 yrs). Baseline characteristics included median PSA 6 ng/mL (range, 3.22-36.11ng/mL). Median duration of drug treatment was 41 days (range 18-81). No grade 3 adverse events were reported, all patients underwent subsequent radical prostatectomy with adverse effects related to metformin. Significant pre-and post changes were noted in serum IGF1 (p=0.02), fasting glucose (p=0.03), BMI (p<0.01) and waist/hip ratio (p<0.01). There was a trend for a PSA reduction (p=0.08). There were no correlations between any metabolic, morphometric or cancer-related serum indices. Conclusions: Neoadjuvant metformin is well tolerated prior to radical prostatectomy. Data to date indicates promising effects on metabolic parameters, tissue results will be presented including proliferation indices and signaling pathway assessments.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr CT-04. doi:1538-7445.AM2012-CT-04
Abstract CT-05: A phase II trial of selumetinib in women with recurrent low-grade serous carcinoma of the ovary or peritoneum
Authors: Farley John H. , Brady William E. , Birrer Michael J. , Lankes Heather , Coleman Robert , Morgan Mark , Mannel Robert , Yamada Diane , Mutch David , Rodgers William H. , Gershenson David M.
Published: August 2012

Background: This study evaluated selumetinib (AZD6244, ARRY-142866), an inhibitor of MEK-1/2, and explored associations between RAS/RAF mutations with clinical outcome. Methods: Women with recurrent low-grade serous ovarian or peritoneal carcinoma were eligible and received selumetinib at 100 mg orally BID until progression or toxicity. Adverse events were assessed with CTCAE v 3.0. Primary measure of efficacy was tumor response by RECIST v 1.0. BRAF, KRAS, and NRAS mutational analysis was performed. Results: Between December 2007 and November 2009, 52 patients were enrolled. Fifty-eight percent of patients had received at least 3 prior chemotherapy regimens. Eight patients (15.4%) had complete (1) or partial (7) responses, and 34 (65%) had stable disease. The median PFS was 11.0 months. The median number of cycles received was 4.5, and 33% of patients received at least 12 cycles of selumetinib. Sixty-three percent of patients (33/52) had PFS> 6 months. Grade 4 toxicities included cardiac (1 patient), pulmonary (1), and pain (1). The most common grade 3 toxicities were gastrointestinal (13), and dermatologic (9). Thirty-four patients had sufficient DNA for mutational analysis: 6% BRAF, 41% KRAS, 15% NRAS mutations were found, and 38% had no detectable mutation. There were no statistically significant differences in the proportion of responses by any mutation. Conclusions: Selumetinib is well tolerated and is active in the treatment of recurrent low-grade serous carcinoma. In exploratory analyses, response to selumetinib did not appear to be related to RAS/RAF mutational status, however the sample size was small. The 81% disease control rate is encouraging and worthy of further evaluation of MEK inhibitors in this population.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr CT-05. doi:1538-7445.AM2012-CT-05
Abstract LB-102: Role of HOX genes in regulation of stem cell populations in normal and malignant colon tissue
Authors: Bhatlekar Seema , Czymmek Kirk , Viswanathan Vignesh , Gonye Greg , Boman Bruce
Published: August 2012

HOX genes are thought to play a role in regulating stem cells (SCs) during embryological development. Aberrant expression of HOX genes is seen in many cancers, including colorectal cancer (CRC). However, the mechanisms leading to aberrant HOX gene expression in CRC are not known. Because SCs are key to colon tumorigenesis, we hypothesized that (i) expression of specific HOX genes is different in malignant colonic SCs than in normal SCs and (ii) aberrant expression of these HOX genes is necessary for the tumor initiating ability of colon cancer SCs. Using real time PCR-based array analysis, we found that expression of many HOX genes are upregulated in colon carcinomas relative to normal colonic epithelium. We previously reported a ‘unique HOX gene signature’ for the SC enriched population; two-color microarray analysis showed that two HOX genes, HOXA4 and HOXD10 are expressed in the bottom part of the colonic crypt where SCs reside. Here, we confirmed this finding, and then, using immunohistochemistry, we validated it. HOXA4 and HOX D10 were expressed at the bottom part of the normal colonic crypt. Increased expression of these genes was also seen in colon carcinoma tissues. The results for colon carcinoma tissues were validated using real time PCR. As an independent method to validate the differential expression of HOX genes, we used flow cytometry and ALDEFLUOR to isolate, from a colon cancer cell line, SCs and non-SCs. HOXA4 and HOXD10 expression was then studied using real time PCR. HOXA4 was upregulated in SCs vs non-SCs. To determine whether these genes are differentially expressed in carcinomas relative to normal tissue, we co-stained tissues with antibodies against HOXA4 or HOXD10, and with SC markers, ALDH1 or CD166. We found that SCs express these HOX genes in the bottom part of the colonic crypt compared to non-SCs. In colon carcinoma tissues, not only was there co-staining, but also, the co-staining was increased relative to normal colonic epithelium. Our results implicate expression of HOXA4 and HOXD10 genes not only in the functioning of normal SCs, but also in the contribution of colon SCs to colon tumorigenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-102. doi:1538-7445.AM2012-LB-102
Abstract LB-104: Prostate-specific inactivation of Atbf1 causes mouse prostatic intraepithelial neoplasia
Authors: Sun Xiaodong , Fu Xiaoying , Li Jie , Xing Changsheng , Frierson Henry F. , Cardiff Robert D. , Wu Hao , Dong Jin-Tang
Published: August 2012

Cancer initiation and progression is triggered by loss of function of tumor suppressor genes. Previously, we have identified the AT-motif Binding Factor 1 (ATBF1) as a candidate tumor suppressor gene at 16q22 in the prostate. It undergoes frequent somatic mutations, reduced expression, or mislocalization in human cancers. However, its tumor suppressor role has not been examined in vivo. In the current study, we generated a conditional Atbf1 knockout mouse line using the Cre-loxP system. Tissue-specific inactivation of Atbf1 in prostatic epithelial cells caused hyperplasia in the dorsal lobes at as early as 4 months of age, which further progressed to mouse prostatic intraepithelial neoplasia (mPIN) in all four lobes. These precancerous lesions harbored molecular aspects similar to human PIN and prostate cancer, including increased cell proliferation, attenuated the basal cell layer, loss of the cell junction protein E-cadherin, and loss of an intact smooth muscle layer. Meanwhile, we observed elevated levels of Muc1 and p-Erk 1/2 in mPIN lesions. In addition, we identified and validated a number of genes that were disregulated in the Atbf1 knockout mice, and many of those genes encode secretory proteins. Supportively, an altered secretory protein profile was observed in the dorsal lobes. Taken together, our data demonstrated that loss of Atbf1 caused precancerous lesions in the prostate, thus providing in vivo evidence to a tumor suppressor role of the ATBF1 gene.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-104. doi:1538-7445.AM2012-LB-104
Abstract LB-105: Gli-IKBKE interactions in pancreatic tumorigenesis
Authors: Rajurkar Mihir , De Jesus-Monge Wilfredo E. , Driscoll David , Cotton Jennifer L. , Lewis Brian C. , Mao Junhao
Published: August 2012

Oncogenic mutations in KRAS are detected in a majority of human pancreatic malignancies, and are considered an initiating event in pancreatic ductal adenocarcinoma (PDAC). The downstream mechanisms through which KRAS drives tumorigenesis remain elusive. While recent in vitro studies have suggested that Gli proteins, the transcriptional effectors of Hedgehog signaling, may mediate KRAS oncogenic activity in PDAC cell lines, a requirement for Gli activation in pancreatic tumorigenesis has not been demonstrated in vivo. Here, we use genetically modified mouse models, and human PDAC cell lines to establish a requirement for Gli activation in KRAS initiation of pancreatic tumorigenesis, and identify IKBKE as a novel oncogenic target gene of Gli in PDAC. We show for the first time that inhibition of Gli transcription in the pancreatic epithelium suppressed the ability of activated KRAS to initiate tumorigenesis in vivo. Also, we find that expression of Gli1 along with oncogenic KRAS led to a dramatic acceleration in the formation of pancreatic intraepithelial neoplasia (PanIN) lesions, and increased mortality. Importantly, we identify IKBKE as a novel transcriptional target of Gli in PDAC cells, and also in the pancreatic epithelium in vivo. We also demonstrate that IKBKE is required for the survival and tumorigenicity of PDAC cells, thus implicating IKBKE as a novel pancreatic cancer oncogene. We have found that IKBKE activates the NF-kB pathway, and also activates Akt by phosphorylation in PDAC cells as part of its tumorigenic activity. Our studies demonstrate for the first time a requirement for Gli transcriptional activity in KRAS induced pancreatic tumorigenesis in vivo, and establish IKBKE as a novel Gli target and pancreatic cancer oncogene.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-105. doi:1538-7445.AM2012-LB-105
Abstract LB-11: Reduced cardiotoxicity and improved antitumor effect of the epirubicin-incorporating micelle NC-6300 in a hepatocellular carcinoma model
Authors: Yamamoto Yoshiyuki , Hyodo Ichinosuke , Takahashi Amane , Yasunaga Masahiro , Koga Yoshikatsu , Harada Mitsunori , Saito Hiroyuki , Tanaka Ryosuke , Hayashi Tatsuyuki , Kato Yasuki , Matsumura Yasuhiro
Published: August 2012

Background: Epirubicin, an anthracycline antitumor drug, has been approved for the treatment of various human tumors. However, epirubicin has significant dose-limiting cardiotoxicity, and there are no effective therapies to prevent or reduce this cardiotoxicity. Therefore, we investigated whether the epirubicin-incorporating micelle NC-6300 was more effective than epirubicin alone in reducing cardiotoxicity and improving the antitumor effect. Method: NC-6300 comprises epirubicin covalently bound to the polyaspartate chain of the polyethylene glycol-polyaspartate block copolymer via an acid-labile hydrazone bond. The conjugate spontaneously forms a micellar structure in an aqueous milieu. The diameter of NC-6300 is 60-70 nm, and the micellar formulation is stable under physiological condition. For evaluating the antitumor effects, NC-6300 (10 or 15 mg/kg) and epirubicin (10 mg/kg) were intravenously administered once a week for 3 weeks to C57BL/6 mice bearing human hepatocellular carcinoma (HCC) Hep3B xenografts implanted subcutaneously or orthotopically (n = 6-10). Cardiotoxicity was evaluated by using echocardiography to the mice that were given NC-6300 (10 or 15 mg/kg) or epirubicin (10 mg/kg) on days 1, 8, and 15 every 4 weeks, to a total of 9 administrations for 12 weeks (n = 6 for each evaluation point). Results: Compared to epirubicin, NC-6300 showed significant antitumor activity against Hep3B. Moreover, the survival rate in the orthotopic tumor model of Hep3B was significantly higher with NC-6300 treatment than with epirubicin treatment. With respect to cardiotoxicity, epirubicin-treated mice showed significantly deteriorated fractional shortening (FS) and ejection fraction (EF) 21 days after the beginning of administration. In contrast, no mice treated with NC-6300 experienced cardiotoxicity in terms of EF or FS during the observation period. Conclusion: The findings of this study imply that NC-6300 has a strong antitumor effect against HCC. Furthermore, NC-6300 significantly reduces the cardiotoxicity of epirubicin and warrants a clinical evaluation in patients with advanced solid tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-11. doi:1538-7445.AM2012-LB-11
Abstract LB-110: Establishment of a chordoma xenograft and in vivo testing of compounds identified by high-throughput screening
Authors: Trucco Matteo , Wilky Breelyn A. , Awad Ola , Shah Preeti , Gul Naheed , Xia Menghang , Huang Ruili , Loeb David M.
Published: August 2012

Chordoma is a rare primary bone malignancy that arises in the skull base, spine and sacrum and originates from remnants of the notochord. Therapy primarily consists of surgical resection and radiation. These tumors are typically resistant to conventional chemotherapy, and to date there are no FDA-approved agents for chordoma. The lack of in vivo models of chordoma has impeded the development of new therapies for this tumor. We have established a chordoma xenograft growing in NOD/SCID/IL-2R gamma-null mice. The histology of the xenograft is consistent with a dedifferentiated chordoma. The xenograft expresses the chordoma marker T brachyury, which is aberrantly localized in the cytoplasm. SNP array shows loss of heterozygosity at the p16 locus, and gene expression profiling shows high expression of numerous cancer-related genes, such as ROS1 and CPA4 The NIH Chemical Genomics Center performed high-throughput screening of 2,816 compounds. Two established chordoma cell lines, U-CH1 and UCH2B, were treated and cell viability measured by CellTiter-Glo assay. Cells were incubated for 48 hours with drug concentrations ranging from 0.5nM to 46uM. The screen yielded several compounds that showed activity and two were tested in the xenograft. Compared with mice treated with vehicle alone, both drugs showed a dramatic inhibition of tumor growth. In conclusion, we have established a xenograft model of dedifferentiated chordoma. High-throughput screening of compounds identified compounds that show activity against chordoma. In vivo testing of two identified compounds showed a dramatic reduction of tumor growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-110. doi:1538-7445.AM2012-LB-110
Abstract LB-113: Non-invasive analysis of drug sensitivity in homo and-heterotypic multicellular tumor spheroids
Authors: Drewitz Maren , Salomon Michael , Thirion Christian , Oberdanner Christian , Kelm Jens M.
Published: August 2012

To gain full benefit of more organotypic tumor culture systems, appropriate assays which enable high-throughput analysis over time without destroying the tissues have to be adopted. Especially read outs for co-culture systems which allow the discrimination of a drug effect on the cancer cell and stroma environment will be of advantage to foster drug testing using more complex 3D models. Here we report the development of microtissue models harboring either fluorescent and/or Luciferase for non-invasive drug sensitivity testing of homotypic and heterotypic tumor spheroids. Microtissues were assayed in microtissue assay plate which allows for biochemical assays and quantitative fluorescence of single multicellular tumor spheroids. Both the fluorescent and secreted luciferase reporter can be measured over time without destroying the tissue allowing for additional end point analysis of interest. HCT116, a colon cancer cell line, was transduced with the reporter genes under the control of a constitutive promoter. To assess the dependency of the fluorescence intensity in accordance to microtissue growth, HCT116-GFP microtissues were assayed over time in a multiplate fluorescence reader. The size profile correlated closely to the increase in fluorescence intensity. For further assay validation the IC50 value were determined by intra-tissue lactate dehydrogenase (LDH) and fluorescence decrease after 72h compound incubation. Reference compounds such as Taxol, Staurosprin and Chlorambucil were tested. The IC50 values from both assays resulted in comparable IC50 values. For non-invasive drug sensitivity testing in heterogenic tumor microtissues HCT116 harboring a secreted Luciferase were combined with a red fluorescent mouse fibroblasts (NIH3T3). Assaying Luciferase on the one hand and red fluorescence intensity on the other allowed discriminating effects on cancer cells and the non-proliferative. Discrimination between specific effects of drug treatment on tumor cells and fibrobast matrix cells is also possible using a two color heterotypic cell model composed of GFP-expressing HTC116 and RFP expressing NIH3T3 cells. Cytotoxic agents targeting either the total cell population or only proliferative cells were tested on the model discriminating the biological response on the two cell populations. To enable the implementation of organotypic cell culture systems at an early time point in the drug development process non-invasive assays. Here, we demonstrate for the first time such a dual cell tumor microtissues system with integrated reporter functions which allows to distinguish effects on different cell populations.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-113. doi:1538-7445.AM2012-LB-113
Abstract LB-153: The helicase antigen (HAGE) promotes tumor cell proliferation via up-regulation of Ras protein signalling.
Authors: McArdle Stephanie E.B. , Linley Adam A.J , Mathieu Morgan M.G , Miles Amanda A.K , Regad Tarik , Rees Robert C.
Published: August 2012

The discovery of cancer/testis (CT) antigens heralded a resurgence in interest in cancer immunotherapy. However, to date, therapy designed to target CT antigens in a variety of different tumours has had limited success and the function of these antigens remains for many still not completely understood. We believe that a better understanding of what possible processes CT antigens are involved in would help in the creation of additional treatment methods. The helicase antigen (HAGE) in a non-X-linked CT antigen shown to be expressed by a variety of tumours including melanoma, head and neck carcinoma and haematological cancers. It is classified as belonging to the DEAD-box family of RNA helicases, which are predicted to have roles in RNA metabolism including transcription, splicing, ribosome biogenesis and translation. These, along with tumour-restricted expression imply a definitive role in malignant cells. Here, we present data that strongly suggests a role for HAGE in promoting tumour cell line proliferation via increased Akt and Erk activation by increasing Ras protein expression. This work was funded by the John and Lucille van Geest Foundation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-153. doi:1538-7445.AM2012-LB-153
Abstract LB-156: The role of AKR1B10 in early tumorigenesis depends on AMP-activated protein Kinase-α checkpoint
Authors: Ma Jun , Cao Deliang
Published: August 2012

The role of AKR1B10 in early tumorigenesis depends on AMP-activated protein Kinase-α checkpoint Jun Ma and Deliang Cao 1Department of Medical Microbiology, Immunology, & Cell Biology, Simmons Cancer Institute, Southern Illinois University School of Medicine. 913 N. Rutledge Street, Springfield, IL 62794. Aldo-keto reductase 1B10 (AKR1B10) is upregulated in 71.4% ductal carcinoma in situ (DCIS) and 83.6% invasive breast cancer. Furthermore, AKR1B10 expression is correlated significantly with tumor size, node status and disease-related survival. In malignant cells, AKR1B10 promotes cell growth and survival via provoking cellular lipid synthesis and eliminating cytotoxic α, β-unsaturated carbonyl compounds. Therefore, AKR1B10 may play an oncogenic role in tumor growth and progression. This study is aimed to explore the role of AKR1B10 in early tumorigenesis. Interestingly, our data show that the expression of AKR1B10 in MCF10A cell, a normal breast epithelium cell line, led to elevation of p53 levels and premature senescence. However, in RAO-1 cells, an hTERT-immortalized breast epithelium cell line, AKR1B10 expression promoted glucose uptake, glycolysis and fatty acid synthesis, and subsequent cell growth instead. Further studies revealed that AMP-activated protein kinase-alpha (AMPKα), which is an important metabolic barrier during cell transformation, was activated upon AKR1B10 expression in both cell lines. On the other hand, down-regulation of AMPKα by shRNA rescued AKR1B10-induced senescence in MCF10A while partially abrogated pro-metabolism effect of AKR1B10 in RAO-1 cells and caused cell growth inhibition. In summary, the role of AKR1B10 in early tumorigenesis depended on AMPKα checkpoint. When this checkpoint is intact, AKR1B10 activates AMPKα and triggers p53 activation and consequential cellular senescence. On the contrary, when this checkpoint is dysfunctional, such as in cells with constitutive expression of telomerase, AKR1B10 activates AMPKα, enhances glucose and lipid metabolism and promotes cell proliferation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-156. doi:1538-7445.AM2012-LB-156
Abstract LB-164: IDH mutations and tumorgenicity
Authors: Yen Kate , Wang Fang , Choe Sung , Schalm Stefanie , Hansen Erica , Straley Kimberly , Popovici-Muller Janeta , Travins Jeremy , Yang Hua , Silverman Lee , Kaelin William , Gross Stefan , Dang Lenny , Salituro Frank , Saunders Jeff , Schenkein David , Su Michael , Biller Scott
Published: August 2012

IDH mutations and tumorgenicity Kate Yen1, Fang Wang1, Sung Choe1, Stefanie Schalm1, Erica Hansen1, Kimberly Straley1, Janeta Popovici-Muller1, Jeremy Travins1, Hua Yang1, Lee Silverman1, William Kaelin2, Stefan Gross1, Lenny Dang1, Frank Salituro1, Jeff Saunders1, David Schenkein1, Michael Su1, Scott Biller1 Mutations in the isocitrate dehydrogenase 1 (IDH1) and 2 (IDH2) genes are present in ∼16% of acute myeloid leukemia, and these mutations cause neomorphic enzyme activity that results in the production of 2-hydroxyglutarate (2HG). Mutational and epigenetic profiling of a large patient cohort of acute myeloid leukemia (AML) revealed that IDH1/2-mutant AMLs display global DNA hypermethylation and a specific hypermethylation signature. To further investigate the intrinsic effect of 2HG on hematopoietic differentiation, we studied erythroleukemia cell lines transfected with IDH1 and IDH2 mutant alleles which overexpress the mutant enzyme, have high levels of 2HG, and exhibit GM-CSF independent growth. Further investigation showed that GATA1, a transcriptional factor known to direct myeloid differentiation, is also down-regulated by IDH2m in these cells along with its downstream direct target SLC4A1. These results demonstrate that the IDH2m can perturb the expression of transcription factors that could lead to alterations in myeloid differentiation. These data suggest that an inhibitor of IDH1/2 mutations could correct for the altered gene expression patterns seen in IDH1/2 mutant AML tumors leading to a profound effect on hematopoietic differentiation, proliferation and tumor growth. We are currently studying the global effects of IDH1/2 mutant overexpression on the methylation of histones and DNA to have a broader understanding of the biological consequence of the IDH1/2 gain of function mutations. Furthermore, treatment of these cell lines with compounds specific for either IDH1 or IDH2 mutant enzymes has a profound effect on their biology.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-164. doi:1538-7445.AM2012-LB-164
Abstract LB-17: NKX2-1/TITF1/TTF-1-induced ROR1 is required to sustain EGFR survival signaling in lung adenocarcinoma
Authors: Yamaguchi Tomoya , Yanagisawa Kiyoshi , Sugiyama Ryoji , Hosono Yasuyuki , Shimada Yukako , Arima Chinatsu , Kato Seiichi , Tomida Shuta , Suzuki Motoshi , Osada Hirotaka , Takahashi Takashi
Published: August 2012

Emerging evidence, though currently sparse, suggests that “lineage-specific transcription factors” with developmental roles in normal progenitor cells of particular lineages may also confer dependency for survival to certain types of cancer cells. Accumulated evidence indicates that NKX2-1, a lineage-specific transcription factor also known as TITF1 and TTF-1, which is expressed in a major fraction of lung adenocarcinomas, plays essential roles in peripheral lung development. We previously identified NKX2-1 as a lineage-survival oncogene in lung adenocarcinoma, while other investigators reached similar conclusions through genome-wide searches for focal genomic aberrations. Previous findings including ours thus clearly indicate the requirement of sustained NKX2-1 expression for lung adenocarcinoma survival, while it remains elusive how NKX2-1 mediates survival signals. In the present study, we show that NKX2-1 induces the expression of the receptor tyrosine kinase-like orphan receptor 1 (ROR1), which in turn sustains a favorable balance between prosurvival PI3K-AKT and proapoptotic p38 signaling, in part through ROR1 kinase-dependent c-Src activation. Interestingly, ROR1 was also identified as a receptor tyrosine kinase with a “sustainer role” for the interaction of EGFR-ERBB3 and ERBB3 phosphorylation in a ROR1 kinase-independent manner, consequentially leading to PI3K activation. It was also of note that ROR1 knockdown effectively inhibited lung adenocarcinoma cell lines irrespective of their EGFR status, including those with resistance to the EGFR tyrosine kinase inhibitor gefitinib. Secondary EGFR mutation, MET amplification, and HGF overexpression may arise in lung adenocarcinomas in patients undergoing EGFR-TKI treatment, leading to treatment resistance. The existence of such diverse mechanisms makes it difficult to predict which should be targeted to prevent expansion of resistant clones. From a clinical point of view, it is thus of particular interest that ROR1 inhibition appears to be effective for treatment of gefitinib-resistant lung adenocarcinomas with various resistance mechanisms. Taken together, the present findings identify ROR1 as an “Achilles’ heel” in lung adenocarcinomas. Future development of novel therapeutic means including ROR1-specific antibodies and small molecules that inhibit both or either of the two distinct prosurvival signal-sustaining functions is greatly anticipated for attempts to reduce the intolerable death toll from currently “hard-to-cure” lung adenocarcinomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-17. doi:1538-7445.AM2012-LB-17
Abstract LB-173: Maraviroc, a CCR5 antagonist, prevents development of hepatocellular carcinoma in a mouse model
Authors: Ochoa-Callejero Laura , Pérez-Martínez Laura , Rubio-Mediavilla Susana , Oteo José A. , Martínez Alfredo , Blanco José R.
Published: August 2012

Chronic liver disease caused by infections or dietary toxins may result in liver fibrosis, cirrhosis and, eventually, hepatocellular carcinoma (HCC). The only curative treatments for patients with advanced liver disease are partial surgical resection and liver transplantation. However, organ shortage as well as an increasing organ demand call for early treatment of liver disease and prevention of fibrosis on high risk individuals. Stellate cells of the liver seem to be responsible for the fibrogenic response through the activation of an autocrine loop involving the chemokine receptor, CCR5. Since this receptor is also one of the main ports of entry for the human immunodeficiency virus (HIV), several CCR5 inhibitors are being used in the clinic to reduce viral load. We used one of these inhibitors, maraviroc (MVC), in a mouse model of diet-induced HCC to investigate whether this intervention would reduce disease progression. This diet was deficient in choline and was supplemented with ethionine in the drinking water (CDE diet). Animals receiving the CDE diet presented higher mortality, higher levels of markers for liver injury such as transaminases, alkaline phosphatase, and bilirubin, developed severe hepatic fibrosis, and were affected by numerous tumors by week 16, when compared with control mice receiving a regular diet. In addition, many cytokines, chemokines, and other matrix components including CCL2, CCL3, CCL4, CCL5, CCR5, CXCL10, IL-12, TGF-b1, or MMP9, were elevated by the CDE diet. Animals which received the CDE diet but were simultaneously treated with MVC presented significantly higher survival, less liver fibrosis, lower levels of liver injury markers and chemokines, and lower tumor burden than their counterparts receiving only the hepatotoxic diet. Animals treated with MVC on top of a normal control diet did not show any evidence of toxicity or any morphological change when compared with non-treated mice. This study suggests that MVC, a well tolerated and clinically characterized drug, may be used as a preventative treatment for HCC. Clinical studies are needed to demonstrate the efficacy of this drug, or other CCR5 inhibitors, in patients with high risk of developing HCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-173. doi:1538-7445.AM2012-LB-173
Abstract LB-182: Effects of methyl donor status on intestinal tumorigenesis in ApcMin/+ mice
Authors: Kadaveru Krishna , Protiva Petr , Greenspan Emily J. , Kim Young-In , Rosenberg Daniel W.
Published: August 2012

The goal of the present study is to determine the effects of folate deficiency on intestinal tumorigenesis in ApcMin/+ mice, a widely used animal model for human familial adenomatous polyposis (FAP). Since folate plays an important role in maintaining methyl donor status, dietary folate levels were reduced in combination with a panel of methyl donors, including choline, methionine and vitamin B12. Although the impact of folate on colorectal cancer (CRC) has been studied extensively, its role in cancer chemoprevention remains controversial. For example, folate supplementation has been widely discussed as a strategy for lowering cancer incidence. However, excess folate has also been associated with increased cancer risk. The following studies were designed to evaluate the effects of folate deficiency on intestinal tumorigenesis in ApcMin/+ mice. In Study 1, methyl donor sufficient diet (MDS) and methyl donor deficient diet (MDD) were fed to mice, beginning at either 5 or 10 weeks of age. Intestinal tumors were evaluated at 16 weeks of age. MDD diet suppressed intestinal tumor formation in ApcMin/+ mice by ∼80% when started at 5 weeks of age. Importantly, the protective effect was lost when the MDD diet was started at 10 weeks of age, indicating an important time-dependency on cancer suppression. Gene expression profiling of normal colonic mucosa after 11 weeks on MDD diet identified the most significantly down-regulated genes related to immune response and inflammation, data that are consistent with a recently published human colon expression profiling study (Protiva et al., 2011). The cancer suppression, however, was associated with reduced food intake and lowered body weight gain (up to 40%) throughout the experimental period. To control for this effect, Study 2 was performed in which ApcMin/+ mice were pair-fed with either methyl donor sufficient (MDS) or methyl donor deficient (MDD) diet, beginning at 5 weeks of age for a total of 11 weeks. A third group received MDS ad libitum. At 16 weeks of age, intestinal tumor formation was significantly reduced (54%, p<0.001) in the caloric-restricted MDS group. In the MDD group, however, tumor suppression was almost complete (∼96%, p<0.001). Associated with tumor protection were increased numbers of apoptotic cells within the villous epithelium of the small intestine, with a concomitant reduction in cell proliferation assessed by Ki-67 and Phospho-histone H3 (pHH3). While cancer protection by methyl donor deficiency is partially dependent on reduced caloric intake, additional mechanisms dependent on altered cell turnover must be invoked to explain these results.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-182. doi:1538-7445.AM2012-LB-182
Abstract LB-183: Identification of anti-cancer activity in structural components of ancestral wheat
Authors: Matthews Shawna B. , Santra Meenakshi , Jiang Weiqin , Zhu Zongjian , Thompson Henry J.
Published: August 2012

In 2009 the average American consumed 134.9 lbs of wheat, making this staple crop a primary source of calories, fiber, and nutrients in the human diet; however little is known about the anti-cancer activity of small molecules present in wheat. In this study, more than 75 wild and less domesticated wheat varieties were extracted using a modified Bligh Dyer technique for in vitro screening. Dried extracts were dissolved in DMSO, diluted with cell culture medium, and evaluated for anticancer activity against MDA-MB-468 (ER- & PR-) and T47D (ER+ & PR+) human breast cancer cell lines in 96-well plate format as assessed by crystal violet staining. The results showed that diploid species of Middle Eastern origin, where human cultivation of wheat began 9000 years ago, have a remarkable ability to inhibit the proliferation of breast cancer cells compared to the domesticated species of tetraploid and hexaploid wheat, which have little or no effect. Furthermore, this anti-cancer activity resides exclusively in the straw-like fibrous hull surrounding the wheat seed, which is not consumed by humans. Principal components analysis of wheat metabolites, analyzed via UPLC-TOF-MS, demonstrated that wheat varieties capable of inhibiting the growth of human breast cancer cells in vitro are chemically distinct from the varieties that do not have inhibitory capability, suggesting that small molecules may be associated with the inhibitory effect. Taken together, these data suggest that ancestral diploid wheat, which is now rarely grown and even more rarely consumed, may possess significant anti-cancer potential in the hulls, and that this anti-cancer potential is an opportunity for advancement of breast cancer prevention through identification of bioactive components and wheat varieties. This work was supported by a grant from the Colorado Wheat Research Foundation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-183. doi:1538-7445.AM2012-LB-183
Abstract LB-184: Differential effects of sulforaphane and related isothiocyanates on HDAC turnover and the DNA damage response in colon cancer cells
Authors: Rajendran Praveen , kidane Ariam I. , Yu Tian-Wei , Dashwood Wan-Mohaiza , Ho Emily , Williams David E. , Dashwood Roderick H.
Published: August 2012

Protein acetylation is mediated by histone deacetylases (HDACs) and acetyltransferases, which influence chromatin dynamics, protein turnover, and the DNA damage response. HDACs overexpressed in cancer cells have been implicated in protecting against genotoxic insults, whereas HDAC inhibitors circumvent this protection (Rajendran et al. Clin Epigenetics 2011,3:4). Here, we show in human colon cancer cells that sulforaphane and related isothiocyanates (ITCs) inhibited HDAC activity and increased HDAC protein turnover, with the potency directly proportional to alkyl chain length. Under these conditions, DNA damage signaling was triggered by ATR kinases, leading to increased double-strand breaks and histone (H2AX) phosphorylation. Activation of checkpoint kinase-2 was followed by growth arrest and cell death. HDAC inhibition by ITCs enhanced the acetylation of repair proteins, like CtIP, leading to their degradation. Notably, cancer cells were more susceptible than normal cells, the latter exhibiting efficient double-strand-break processing and repair. Thus, dietary ITCs preferentially exploit the HDAC turnover pathways, faulty DNA repair mechanisms, and genomic instability in cancer cells. Supported by NIH grants CA090890, CA65525, CA122906, CA122959, CA80176, and ES007060.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-184. doi:1538-7445.AM2012-LB-184
Abstract LB-185: Cytokinesis-blocked micronuclei Assay (CBMN) as a biological cancer risk assessment tool
Authors: Lloyd Stacy M. , Lopez M , El-Zein Randa
Published: August 2012

Lung cancer is the leading cause of all cancer-related deaths in the US. Given the lack of effective of screening modalities, the need to develop more accurate cancer risk assessment tools is imperative to improve the ability to identify individuals that are at greatest risk of developing this disease. The Cytokinesis-Blocked Micronuclei Assay (CBMN) presents a sensitive, specific, and rapid method of assessing DNA damage, a hallmark of cancer. We have previously reported that this assay is extremely sensitive to genetic damage caused by the tobacco specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and that the binucleated cells with micronuclei, nucleoplasmic bridges, and nuclear buds in lymphocytes (chromosome damage endpoints measured by the assay) are strong predictors of lung cancer risk. To further identify the specific chromosomes involved in lung carcinogenesis, we conducted Spectral Karyotyping (SKY) on a population of n=116 cases and n=126 controls on whom CBMN data are available. SKY was performed on both baseline and NNK-treated blood lymphocytes. After adjusting for age, gender, race/ethnicity, smoking status, pack years (intensity), and smoke years (duration), we observed consistent significant results for damage on chromosomes 1, 3, 4, 9, 13, 16, 17, 19, 22, and X (all p's ≤ 0.05) and lung cancer risk. Several of these identified chromosomes are well known to harbor critical genes involved in lung carcinogenesis, such as the FHIT gene (chromosome 3), p16/CDKN2A (chromosome 9), PADPRP (chromosome 13) and TP53 (chromosome 17). Our results support the utilization of CBMN as a cancer risk assessment tool, and when used in conjunction with other cytogenetic methodologies can increase our ability to identify specific regions of DNA damage, thereby improving our understanding of the underlying mechanisms involved in individual cancer predisposition. This work was supported by the following: Kellogg Health Scholars Program (W.K. Kellogg Foundation P0117943) NIMHS P60 MD000503 CA129050 NIHS P30 ES007784

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-185. doi:1538-7445.AM2012-LB-185
Abstract LB-187: Modulation of redox status: An alternative approach for prevention and treatment of prostate cancer
Authors: Das Trinath P. , Suman Suman , Gulappa Thippeswamy , Kurisetty Vittal , Vadodkar Aditi Shirish , Lakshmanaswamy Rajkumar , Damodaran Chendil
Published: August 2012

Radical prostectomy, radiation treatment, and/or hormone ablation therapy are the first lines of treatment for prostate cancer (CaP). Despite initial sensitivity to these treatments, CaP eventually progresses to castration-resistant CaP (CRPC), which is both aggressive and refractory to current therapeutic options. Accordingly, there is a tremendous need for a novel agent that can either retard or prevent the emergence of CRPC. Changes in reactive oxygen species (ROS) play a pivotal role in these processes. For example, a moderate increase in ROS promotes cell proliferation, but excessive amounts of ROS can induce apoptosis in cancer cells. Hence, it is possible to manipulate the redox (oxidative stress) system to selectively kill cancer cells. Psoralidin (Pso), a natural bioactive compound derived from Psoralea Corylifolia which is extensively used in Asian and African medicines, is an orally bioavailable dietary agent. We are the first group to demonstrate that Pso is non-toxic to normal prostate epithelial cells but specifically targets CaP cell lines (PC-3, DU-145, LnCaP and C4-2B) by inhibiting Akt and its downstream pro-survival signaling pathways. While dissecting the upstream events of Akt, we found that Pso generates ROS production and inhibits Akt mediated pro-survival signaling in both cell cultures and animal models of CRPC. Importantly, over expression of antioxidants, genetically (SOD/Catalase) or pharmacologically N-acetylcysteine (NAC) or treatment with ROS inhibitors Rotenone (mitochondrial) or Diphenylenel iodonium DPI (extra mitochondrial) inhibited Pso induced oxidative stress (both Intracellular and NADPH oxidase mediated) in CaP cells. Further, Pso in combination with anti-oxidants/ROS inhibitors significantly decreased the migratory and invasive property of CaP cell lines. Currently, we are delineating the role of Pso induced non-mitochondrial ROS generation, by analyzing expression patterns of NADPH oxidase and their subunits. We will apply genetic and/or pharmacological approaches to determine the key Pso target in CaP cell lines. Further, our chemopreventive and chemotherapeutic studies of Pso in both xenograft models and pilot experiments in transgenic adenocarcinoma of mouse prostate (TRAMP) model showed that Pso inhibits prostate tumor growth. These studies may suggest modulation of redox status could be a novel and powerful approach to chemoprevention and/or treatment of CaP.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-187. doi:1538-7445.AM2012-LB-187
Abstract LB-198: Ryl hydrocarbon receptor agonist Aminoflavone disrupts mammosphere formation via alpha 6 integrin inhibition
Authors: Brantley Eileen J. , Yee Michael , Quitugua Ashley , Zylstra Dain , Soto Ubaldo
Published: August 2012

Introduction: Emerging evidence has suggested that cancer stem cells (CSCs) play a prominent role in the initiation and dissemination of a myriad of malignancies including breast cancer. CSCs have the capacity to self-renew within heterogeneous tumor cells. Currently available therapeutic options for breast cancer are incapable of suppressing CSC growth. Alpha 6 integrin contributes to CSC growth while aryl hydrocarbon receptor (AhR) activation impedes CSC proliferation. AhR agonist Aminoflavone, a synthetic flavonoid, is currently undergoing clinical evaluation for the treatment of breast cancer. We hypothesize that Aminoflavone alters the integrity and formation of cell suspension clusters initiated by breast cancer stem cells (mammospheres) by modulating alpha 6 integrin expression in an AhR-dependent manner Experimental Procedures: Mammospheres derived from MCF-7 breast cancer cells were exposed to Aminoflavone at varying times. We also treated mammospheres with Tamoxifen an anticancer agent frequently used in the treatment of breast cancer. Following Aminoflavone treatment, cells were analyzed microscopically to evaluate alterations in mammosphere structure. In addition, the expression of molecular targets associated with CSC growth in addition to alpha 6 integrin were evaluated by Western blotting and quantitative RT-PCR analysis. Results: We observed that Aminoflavone, but not Tamoxifen, disaggregates MCF-7 mammospheres. In contrast, Aminoflavone did not affect mammospheres produced from AhR defective MCF-7 (AHR100) cells. Aminoflavone abrogated the ability of secondary mammospheres to form. In addition, alpha-6-integrin expression was abolished following Aminoflavone treatment in MCF-7 but not AHR100 cells. The expression of several genes whose products regulate alpha-6 integrin expression were also modulated following Aminoflavone treatment in MCF-7 cells. Conclusion: These data suggest that Aminoflavone disrupts mammosphere formation via AhR signaling activation that results in alpha 6 integrin inhibition. These findings support continued efforts to develop novel AhR ligands with efficacy against breast CSCs as a viable therapeutic strategy for breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-198. doi:1538-7445.AM2012-LB-198