bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3397: The role of Pdef in mammary development, tumorigenesis and metastasis
Authors: Findlay Victoria J. , Watson Patricia M. , Feldman Ron J. , Sigmon Stacey A. , Gong Yong Z. , Moussa Omar , Sypropoulos Demetri D. , Turner David P. , Watson Dennis K.
Published: February 2011

ETS proteins represent one of the largest families of transcription factors with diverse functions that activate or repress the expression of genes that are involved in various biological processes, including cellular proliferation, differentiation, development, transformation and apoptosis. The ETS family gene, PDEF (prostate derived ets factor), is expressed in normal epithelial tissues including prostate, breast, colon and bladder. Significantly, PDEF protein is present in non-invasive cancers but reduced or absent in most invasive cancers. By re-expression and knock-down of PDEF in cancer cell lines, we and others have shown that PDEF target genes control aspects of the metastatic process, including cell growth, migration and invasion. We hypothesize that PDEF is important for normal developmental processes and that loss of PDEF regulatory networks is a key event in the development of invasive cancer. To study the function of Pdef in vivo, we generated mice with constitutive Pdef knockout (Pdef −/−). Specific stages of mammary development were examined, including early-pubescent (5 week), virgin adult (8 week), pregnancy (days 7.5 and 13.5), lactation (days 1 and 7) and involution (day 7). Inhibition of the development of the ductal tree and an increase in ductal side branching and terminal end bud proliferation was observed in 5-8 week old virgin Pdef −/− mice. TGFβ1 and ERα are required for development of the ductal tree during puberty and mRNA levels for both are reduced in the

Pdef −/− mice at 5 and 8 weeks of age. Incomplete lobuloalveoli development was observed in Pdef −/− pregnant and lactating mice, although no phenotypic effects were observed in the resultant pups. To explore the role of Pdef in mammary tumor progression and metastasis, we generated mice expressing the MMTV-PyMT or MMTV-Neu oncogene in wild type and Pdef −/− backgrounds. No impact of Pdef dosage upon tumor incidence or growth was observed in the PyMT model; however, initial tumor formation was delayed by 4 weeks (8.9 vs. 4.8) and time from tumor initiation to sacrifice was 3 weeks longer (29.4 vs 26.4) in the Pdef −/− Neu model. Significantly, an increase in lung metastasis was observed in the Pdef −/− mice in both the PyMT and Neu models. Thus, loss of Pdef in vivo is associated with defects in mammary development and increased metastasis in mouse models of breast cancer, suggesting Pdef is a metastasis suppressor.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3397.
Abstract 3398: Double-faced Sema3E-PlexinD1 signaling inhibits tumor growth but promotes metastatic spreading
Authors: Casazza Andrea , Capparuccia Lorena , Finisguerra Veronica , Mazzone Massimiliano , Comoglio Paolo M. , Tamagnone Luca
Published: February 2011

Semaphorins comprise a wide family of secreted and membrane-bound signals, originally found in neural development, and then implicated in a range of functions: from angiogenesis, to the immune response, to cancer progression. We have previously shown that high-affinity semaphorin receptors are found in the families of the plexins and of the neuropilins. Moreover, additional molecules interact with semaphorin receptors in cell-context dependent manner, featuring a complex scenario of multiple potential signaling pathways. The role of semaphorin signaling in tumor progression is currently under close scrutiny; it is known that cancer cells release semaphorins regulating their own behavior as well as that of cells in the tumor microenvironment, such as endothelial cells and recruited leukocytes.

In this work, we focused on the function of Semaphorin 3E (Sema3E) and its high affinity receptor PlexinD1, and on their potential role to regulate cancer progression. Here we found that Sema3E and PlexinD1 are highly expressed in human metastatic tumors. Moreover, we provided experimental evidences that Sema3E released by tumor cells is a double-faced signal, acting via two distinctive signaling pathways. In paracrine manner, Sema3E-PlexinD1 signaling can elicit endothelial cell repulsion, thereby reducing vessel density and tumor growth. Conversely, Sema3E autocrine signaling in tumor cells thrusts their invasiveness, trans-endothelial migration, and metastatic spreading, through the trans-activation of PlexinD1-associated oncogenic kinases. Notably, by silencing the endogenous expression of either Sema3E or PlexinD1 the metastatic potential of cancer cells is significantly reduced, identifying this pathway as a major driver of the metastatic spreading. In sum, Sema3E autocrine signaling in cancer cells is crucially implicated to induce a metastatic behavior, and it appears as a promising target for strategies aimed at blocking tumor metastasis.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3398.
Abstract 3410: Discovery of prognostic factors in Ewing's sarcoma family of tumors by use of Intercohort co-analysis of public gene expression datasets
Authors: Bennani-Baiti Idriss M. , Lawlor Elizabeth R. , Cooper Aaron , Kauer Maximilian , Ban Jozef , Aryee Dave N.T , Kovar Heinrich
Published: February 2011

Cancer genomics can help uncover genes that may serve as indicators of tumor susceptibility, progression, or treatment outcome, as exemplified by studies of adult tumors for which large cohorts are readily available. Childhood cancers comprise a large number of low-incidence histogenetically and molecularly distinct neoplasms, rendering population-based genomic studies unrealistic for these diseases. In recent years, several small-size cohort childhood cancer genomic studies have been published. We report here a novel analytical approach, named intercohort co-analysis or Ican, whereby gene expression significance is inferred from gene expression levels in biologically relevant normal tissues, and gene-disease association significance is assessed in each individual cohort. Next, only genes found to be consistently significant in all cohorts are considered as putative markers of specific disease states. We tested Ican in Ewing's sarcoma (ESFT), a family of tumors that lacks a reliable molecular prognostic or metastasis indicator. We report here that this approach revealed a high intercohort gene expression significance variability, and identified CXCR4 and CXCR7, two chemokine receptors known to correlate to metastasis and/or to poor prognosis mostly in adult cancers, as significantly and consistently associated with ESFT subsets. Accordingly, we show for the first time that CXCR4 expression marks metastatic ESFT and that of CXCR7 serves as a predictor of adverse patient survival. We submit these findings as a proof-of-principle that Ican of publicly available gene expression profiles can yield genes with predictive value for the progression or outcome of low-incidence diseases. We further submit that this approach is poised to gain momentum as many journals now request the mandatory contribution of gene expression profiles to public repositories. These policies, along with the growing number of cancer gene expression profiling investigations, we predict, will swiftly drive upward the number of publicly available low-incidence disease expression datasets, and consequently, of Ican applications.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3410.
Abstract 3419: NOS1 imparts a survival benefit to PAX-FKHR expressing alveolar rhabdomyosarcomas
Authors: Bajaj Sheetal V. , Shahbazian Violette , Anderson Michael J. , Triche Timothy J.
Published: February 2011

BACKGROUND: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood and adolescence. Based on histology, two major forms are identified: embryonal and alveolar. Molecularly, these tumors are classified as translocation-positive alveolar RMS (ARMS, characterized by PAX3-FKHR or PAX7-FKHR chimeric fusions), translocation-negative alveolar RMS (nARMS), and embryonal RMS (ERMS). The PAX-FKHR oncogenes have been shown to be responsible, in part, for the aggressive clinical behavior and poor outcome of ARMS. It is thus important to identify biomarkers in this subgroup that can be used as potential therapeutic targets.

METHODS: Using Affymetrix Human Exon 1.0 ST (HuEx) microarrays, genome-wide expression profiles were generated on primary tumors from RMS patients accrued on COG treatment protocols. Data analysis was performed using institutional software (Genetrix) by standard classification methods. Expression of neuronal nitric oxide synthase (NOS1) was validated by qPCR, immunoblot and immunohistochemistry. A functional relationship, if any, to PAX-FKHR was determined by qPCR after siRNA-mediated knockdown of PAX-FKHR in ARMS cell lines and stable transduction of PAX-FKHR into an ERMS cell line. Further, shRNA targeting NOS1 using a lentiviral delivery system was used for functional NOS1 studies in ARMS cell lines.

RESULTS: Expression analysis revealed that NOS1 is highly expressed in ARMS with a PAX3 or PAX7 translocation. Little expression was found in nARMS, ERMS or normal muscle. This was confirmed by qPCR and immunoblotting in two independent groups of primary tumors and RMS cell lines. PAX-FKHR knockdown resulted in decreased expression of NOS1 and its corresponding protein in ARMS cell lines. PAX-FKHR expression in an ERMS cell line induced the expression of NOS1 and its corresponding protein where the native cell line showed no corresponding expression. Further, targeting NOS1 using shRNA in ARMS cell line caused decreased NOS1 expression leading to poor cell survival and cell death.

CONCLUSIONS: NOS1 is over-expressed in the more aggressive PAX3 or PAX7 ARMS when compared to translocation-negative RMS. Its expression appears to be regulated by PAX-FKHR, a tumor specific chimeric gene that has been implicated in regulating the malignant phenotype of ARMS. It seems to play an important role in the survival of these tumor cells. Taken together, these observations suggest that NOS1 is a downstream effecter of PAX-FKHR mediated oncogenesis and an essential factor for tumor cell survival that could be a therapeutic target, using suitable small molecule therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3419.
Abstract 3454: Molecular markers for breast carcinogenesis models induced by environmental substances and estrogen
Authors: Calaf Gloria M. , Balajee Adayabalam S. , Roy Debasish
Published: February 2011

Breast cancer is the most common cancer in women throughout the world. Exogenous and endogenous agents such as environmental carcinogens and female hormones seem to be involved in the etiology of breast cancer. It is a multistage process involving a series of genetic alterations, identification of potential genes responsible for breast carcinogenesis is critical for timely diagnosis and prevention of breast cancer. To gain insights into the mechanisms for breast cancer initiation and development processes by exogenous environmental agents, we have developed many experimental breast cancer model systems: In vitro model with the immortalized human breast epithelial cell line, MCF-10F exposed to (I) low doses of high LET (linear energy transfer) alpha particles (150 keV/μm) (Carcinogenesis 21: 769, 2000) and b), (II) organophosphorous pesticides, either with Parathion (P) or Malathion (M) and (III) combination of either P or M in the presence of estrogen (E). Results showed that MCF-10F cells treated either with double dose of 60 cGy alpha particles in the presence of E or pesticides induced malignant transformation of MCF-10F. The malignant transformation was determined by multiple biological assays: increased cell proliferation, anchorage independency, invasive capabilities and tumor formation in nude mice, microsatellite instability and loss of heterozygosity in chromosomes 17, 11, 6, 8. Gene expression analysis using cancer pathway specific and affymetrix arrays detected alterations in the expression levels of p53, ErbB2, BRCA1, c-Ha-ras, Rho-A, PTEN, RB, c-Ha-ras, transforming protein Rho-A, F, GDP, TGF alpha, beta receptor, integrin B6, Notch3 and cathepsin. In addition to the in vitro human model, in vivo rat mammary gland model was also generated: (I) control (II) either P or M (III) E and (IV) P or M and E. Animals were treated for 5 days. These combined treatments induced significant progressive morphological and molecular changes. Alterations in the expression levels of RNA and protein were observed for c-fos, c-myc, mutant p53, ErbB2, BRCA1, c-Ha-ras, Rho-A, CYP1A1 gene and protein expression in the rat mammary gland after 240 days of treatment in comparison to control. Such stimulation led to mammary tumor formation. Collectively, our study shows the molecular signature of oncogenic deregulation in breast cancer progression induced by the combination of environmental substances and estrogen. Thus, aberrant expression of multiple genes involved in key signaling pathways renders these models as important tools for monitoring carcinogenic progression and chemo-intervention. Grant FONDECYT # 1080482 (GMC).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3454.
Abstract 3464: Epigenetic mechanism is involved in depleted uranium-induced transformation in human lung epithelial cells
Authors: Xie Hong , LaCerte Carolyne , Wise John P.
Published: February 2011

Depleted uranium (DU) is commonly used in military applications and is also used in civilian industry and thus exposure of soldiers and others is frequent and widespread. There is limited research information on the potential health hazards of DU exposure. In our study, we used the human bronchial epithelial cell line (BEP2D) to study the potential carcinogenic effects of DU. DNA strand breaks were observed at 25 ug/cm2 DU treatment. We found that DU induces cell transformation including cell contact inhibition and anchorage independent phenotype. Doses of 0.25, 2.5 and 25 ug/cm2 uranium trioxide induces 23, 24 and 17 growth foci in 30 dishes, respectively. These foci were cloned and were found to exhibit anchorage independent growth and an aneuploid phenotype. We next investigated DU-induced epigenetic modifications. We found that DU decreases trimethylated H3K9 (H3K9me3), along with inhibition of the DNA damage repair protein ATM. Our study demonstrates that DU-induced both genotoxicity and aberrant histone methylation which may contribute to its carcinogenesis. This work was supported by ARO grants # W911NF-04-1-0240 and W911NF-08-0333 (J.P.W.).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3464.
Abstract 3465: DNA adducts of tobacco-specific nitrosamines in human pancreatic tissue resulting from pyridylhydroxybutylation and pyridyloxobutylation
Authors: Eggert Heike , Heppel Christopher W. , Peschel Oliver , Kunz Sebastian , Giebe Birkhild , Richter Elmar
Published: February 2011

Pancreatic cancer is the fourth leading cause of cancer-related deaths in the United States. Whereas cigarette smoking is a well established important risk factor for pancreatic cancer, the contribution of smokeless tobacco consumption is still a matter of debate. The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is the only known compound present in tobacco and tobacco smoke which induces pancreatic tumors in animal models. In rats, the major NNK metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), proved to be an even more potent pancreatic carcinogen. NNK and NNAL have been detected in pancreatic juice. However, activation of NNK and NNAL leading to pyridyloxobutyl (POB) and pyridylhydroxybutyl (PHB) DNA adducts, respectively, has not been proven in human pancreas. Determination of POB DNA adducts was achieved by acid hydrolysis of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB) from DNA after addition of [3,3,4,4-D4]-HPB as internal standard, derivatization with pentafluorobenzoyl chloride (PFBC) and quantification by gas chromatography/high resolution mass spectrometry with negative chemical ionization according to Heppel et al. (Anal Bioanal Chem 393:1525-30, 2009). POB DNA adducts could be easily detected in 8 of 11 pancreatic samples of sudden death victims from Germany at 2.95±1.08 fmol/µg DNA (mean±standard error; values below detection limit set at zero). Using [3,3,4,4-D4]-diol as internal standard, this analytical method also revealed the presence of PHB adducts in pancreatic DNA releasing 4-hydroxy-1-(3-pyridyl)-1-butanol (diol). However, because of massive peak broadening the PFBC ester of diol could not be quantified reliably. To improve the chromatographic resolution, the PFBC ester of diol was further derivatized according to Kao and Giese (Chem Res Toxicol 18:70-5, 2005) with pivalic anhydride to cap the secondary hydroxy group. The resulting tert-butyl esters of the PFBC esters of diol and [3,3,4,4-D4]-diol show excellent performance in capillary gas chromatography and allow quantification by single ion monitoring at the molecular masses of m/z 445.13 and m/z 449.16, respectively. In summary, the presence of POB and PHB DNA adducts in human pancreatic tissue has been clearly demonstrated for the first time. Further studies will show the specificity of these pancreatic DNA adducts for smoking and smokeless tobacco consumption and the correlation with other tobacco-specific biomarkers. Supported by a grant of the Institute for Science and Health, St. Louis, MO.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3465.
Abstract 3470: Effects of combined phytochemicals on skin tumorgenesis in SENCAR mice
Authors: Kowalczyk Magdalena C. , Junco Jacob , Kowalczyk Piotr , Tolstykh Olga , Hanausek Margaret , Slaga Thomas J. , Walaszek Zbigniew
Published: February 2011

The purpose of our study was to determine the effect of combined action of phytochemicals on early stages of skin tumorgenesis, i.e., initiation and promotion. We tested calcium D-glucarate (CG) given in the diet, while resveratrol (RES) and ursolic acid (URA) were applied topically. The 7,12-dimethylbenz[a]anthracene (DMBA)-initiated, 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted multistage skin carcinogenesis model in SENCAR mice was used. Mice (6-7 weeks old) with shaved backs received one topical dose of DMBA (20 nmol), then after one month, two weekly doses (2 µg each) of TPA (tumor promoter) for 14 weeks until sacrifice. The phytochemicals RES (2.5 μmol) or URA (1 μmol) were applied topically 20 min prior to DMBA or TPA treatment and 2% dietary CG was given in the AIN-93G diet from 2 weeks prior to 2 weeks after the DMBA dose or continually beginning 2 weeks prior to the first dose of TPA. We determined the tumor incidence (% of mice with tumors) and tumor multiplicity (number of tumors per mouse) and using RT-PCR and immunochistochemical analyses, we checked the expression of some cell proliferation and inflammation related genes. URA applied alone and in combination with CG during the promotion stage was a strong inhibitor of tumor multiplicity and tumor incidence. Combinations of URA with dietary CG appeared to show some synergistic effects. All compounds and their combinations reduced epidermal proliferation, but only URA and a combination CG/URA applied during promotion significantly reduced hyperplasia, as determined by epidermal thickness measurements taken at 14 weeks of TPA treatment. DMBA/TPA treatments resulted in epidermal hyperplasia, characterized by significant increase in transcription factor AP-1 components and nuclear factor kappa B (NFkB) expression levels with a simultaneously increased expression of the inflammation markers COX-2 and IL-6, compared to untreated controls. All anti-promotion treatments (either single compounds or their combinations) caused a marked decrease of inflammation related genes levels compared to the DMBA/TPA control. On the other hand, only URA and URA with dietary CG diminished the induced expression of proto-oncogenes c-jun and c-fos (components of AP-1) and the expression of NFkB protein p50. These results show that URA is a potent inhibitor of skin tumor promotion and it may be useful in the prevention of skin cancer and other epithelial cancers in humans. Supported by NIH grants CA 102747 and P30 CA 54174-16S1.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3470.
Abstract 348: Prognostic value of mTOR activation and therapeutic effects of everolimus in esophageal squamous cell carcinoma
Authors: Hirashima Kotaro , Baba Yoshifumi , Saito Seiya , Kinoshita Kouichi , Satou Nobutaka , Kurashige Junji , Karashima Ryu-ichi , Imamura Yu , Ikeda Osamu , Hayashi Naoko , Watanabe Masayuki , Iyama Ken-ichi , Baba Hideo
Published: February 2011

Background: The mammalian target of rapamycin (mTOR) plays central roles in cell growth and homeostasis. Aberrant activation of mTOR in relation to clinical outcome has been reported in several types of cancers. mTOR inhibitor, particularly everolimus (RAD001), has been attracting attentions as a promising anticancer drug. Nonetheless, no study has shown the prognostic value of mTOR activation in oesophageal squamous cell carcinoma (ESCC) and the therapeutic effect of everolimus on human ESCC cell lines. The aims of this study are [1] to evaluate phosphorylated-mTOR (i.e., mTOR activation) expression in ESCC and to clarify its clinical, pathologic, and prognostic features and [2] to examine the therapeutic effect of everolimus on ESCC cell lines.

Methods: We examined p-mTOR expression (i.e., mTOR activation) by immunohistochemistry using 143 ESCC specimens. The present study involved 143 consecutive patients with ESCC who underwent the curative surgical resection at Kumamoto University Hospital from January 1996 to December 2006. We determined its clinical, pathologic, and prognostic features. Prognostic significance of p-mTOR expression was examined by Cox regression and Kaplan-Meier analysis. Subsequently. the effects of everolimus on ESCC cell lines were evaluated by in vitro assays (proliferation, cell cycle, apoptosis, invasion).

Results: p-mTOR expression was detected in 71 (49.7 %) tumors and was independently associated with higher overall mortality. Compared to p-mTOR negative patients, p-mTOR positive patients experienced high overall mortality [HR 2.44; 95% confidence interval (CI), 1.24-4.83, P = 0.01], which persisted in multivariate analysis (adjusted HR 2.92; 95% CI, 1.48-5.78, P = 0.002). p-mTOR expression was not related with clinical or pathologic variables including age at diagnosis, sex, tumor location, histological grading, T classification (tumor invasion) or N classification (lymph-node metastasis). In vitro, in both TE4 (p-mTOR expression-high in vitro) and TE11 (p-mTOR expression-low in vitro) squamous cell carcinoma cell lines, everolimus suppressed the phosphorylation of downstream p70S6 kinase and 4E-BP1in a dose-dependent manner. Everolimus significantly inhibited cancer cells proliferation and invasion, and induced cells apoptosis in vitro. Conclusion: p-mTOR expression in ESCC was independently associated with poor prognosis, supporting the potential for mTOR as a therapeutic target for ESCC. Everolimus may be useful as an anticancer drug for ESCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 348.
Abstract 3495: Targeting gliomas with a dual kinase inhibitor and stem-cell delivered TRAIL
Authors: Bagci-Onder Tugba , Wakimoto Hiroaki , Anderegg Maarten , Shah Khalid
Published: February 2011

Gliomas are difficult to eradicate due to several factors: a) the lack of efficient delivery of anti-tumor agents to the tumors without affecting the normal brain tissue; b) the inherent or acquired resistance of tumor cells to anti-tumor agents; and c) the highly invasive nature of tumor cells that escape the primary tumor mass and subsequently cause recurrence. We previously established that a secreted form of tumor necrosis factor-related apoptosis inducing ligand (S-TRAIL) specifically eradicates glioma cells when delivered by various stem cell types in culture and in orthotopic glioma models. However, recent evidence has shown that a number of established and primary glioma lines have varying degrees of resistance to TRAIL, due to the heterogeneity of genetic alterations and possibly due to the over-activation of phosphatidylinositol 3 kinase (PI3K) pathway. In this study, we report the effect of a novel PI3K inhibitor, PI-103, alone or in combination with neural stem cell (NSC)-delivered S-TRAIL in mouse models of malignant and invasive gliomas. Using established and primary CD133+ glioma initiating cells engineered to express fluorescent and bioluminescent markers, we show that PI-103 alone inhibits proliferation and invasion, and causes G0-G1 arrest in cell cycle of most of the glioma cells tested. Furthermore, PI-103 augments the response of glioma cells to stem cell delivered S-TRAIL in co-cultures of NSCs and glioma cells suggesting that it can synergize with S-TRAIL-induced apoptosis. Using orthotopic malignant glioma models and bioluminescence imaging, PI-103 treatment in combination with NSC-derived S-TRAIL was shown to result in significant reduction in tumor volumes compared to either treatment alone in vivo. Our results reveal that combination of the novel PI3K inhibitor, PI-103, and stem cell delivered TRAIL might represent a novel therapy for malignant gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3495.
Abstract 3516: Targeting cancer by cisplatin-containing nanovectors
Authors: Sengupta Poulomi , Soni Shivani , Basu Sudipta , Sengupta Shiladitya
Published: February 2011

Cisplatin [cis-dichlorodiammineplatinum(II)] is a first-line chemotherapeutic agent, and is widely used for the treatment of many malignancies including testicular, ovarian, cervical, head and neck, and non-small cell lung cancer. Its use is dose-limited mainly because of nephrotoxicity. To address this limitation, we have engineered novel nanovector to deliver the drug directly to the tumor site with reduced toxicity.

In this study, we have functionalized cholesterol with succinic acid and ethyelene diammine to complex platinum through a monocarboxylato and a coordinate bond. The Chol-Pt complex was mixed with phosphatidylcholine and DSPE-PEG at 5:10:1 weight ratio and cisplatin containing nanoparticles were synthesized through evaporation-hydration-extrusion technique. The nanoparticles were found to be in the size range of 100-150 nm and spherical in morphology (characterized by DLS and TEM respectively). The physicochemical release kinetics of cisplatin from the nanoparticles in different hypoxic cell lysates showed a sustained release of the drug.

Our in vitro studies of the cisplatin liposomes indicate that cisplatin liposomes are highly effective in 4T1 breast cancer and LLC (Lewis Lung Carcinoma) cell lines in 48 hours, as compared with free cisplatin. Most interestingly, our cisplatin containing liposomes demonstrated efficacy in cisplatin resistant CP20 (liver adenocarcinoma) cell line at 10 μM concentration. Hence, our study showed that our novel cisplatin nanovector delivery device could potentially be harnessed for lung, breast and cisplatin resistant cancer chemotherapy with increased therapeutic index compared to cisplatin.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3516.
Abstract 3517: Complete remission of xenograft tumor in nude mice induced by N-mustard-quinoline conjugates bearing hydrazinecarboxamide linker
Authors: Su Tsann-Long , Kakadiya Rajesh , Dong Huajin , Kumar Amit , Lee Te-Chang , Tsai Tung-Hu , Chou Ting-Chao
Published: February 2011

Designing DNA-directed alkylating agents is a promising strategy to overcome the drawbacks of alkylating agents by enhancing DNA specific cross-linking and reducing the chemical reactivity of N-mustard. These compounds are generally synthesized by linking the alkylating warhead (such as alkyl or phenyl N-mustard pharmacophore) to the DNA-affinic carrier (such as DNA intercalating or groove binding agents). Recently, we reported that N-mustard-9-anilinoacridine conjugates via a urea, carbamate, or carbonate linker exhibited potent cytotoxicity and therapeutic efficacy against various human tumor xenografts indicating that these linkers are able to stabilize the reactive alkylating war head. Thus, these derivatives have a long half-life in plasma. Some of these conjugates are able to induce complete tumor remission (CR) in inhibiting human tumor xenograft model with low toxicity. To continue our research in finding potent DNA-directed alkylating agents, we have recently synthesized a series of N-mustard-quinoline conjugates bearing a hydrazinecarboxamide linker. These conjugates were prepared in good yield by the condensation of appropriate 4-hydrazinoquinolines with 4-bis(2-chloroethyl)- aminophenyl isocynate. The results showed that these conjugates possessed significant cytotoxicity against human lymphoblastic leukemia (CCRF-CEM) and various solid tumor xenografts (such as MX-1, HCT-116, H1299, CL 1-0, CL 1-5, PC3 and MCF-7) cell growth in vitro. Among these agents, BO-1392 and BO-1393 were able to induce CR against human breast carcinoma MX-1 xenograft and significantly suppress against human ovarian SK-OV3 xenograft in animal model with relative low toxicity. Interestingly, we found that BO-1393 (iv. injection) can significantly prolong the life span of nude mice bearing human glioma SK-NA-S xenograft (orthotropic implanted in brain). The experiment was compared with Temozolomide (oral administration). The modified comet assay and alkaline agarose gel shift assay showed that these conjugates are capable of inducing DNA interstrand cross-linking in human non-small lung cancer H1299 cells. In conclusion, the present studies demonstrated that the newly synthesized N-mustard-quinoline conjugates having a hydrazinecarboxamide linker exhibited a broad spectrum of antitumor activity and are chemically stable. The remarkable therapeutic efficacy of BO-1393 demonstrated that this agent has high potential for clinical development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3517.
Abstract 3539: Overexpression of the ATP Binding Cassette Gene ABCA1 determines resistance to curcumin in M14 melanoma cells
Authors: Kronski Emanuel , Iancu Cristina M. , Killian Peter , Mirisola Valentina , Jochum Marianne , Nerlich Andreas , Pfeffer Ulrich , Bachmeier Beatrice
Published: February 2011

Background: Curcumin induces apoptosis in many cancer cells and it reduces xenograft growth and the formation of lung metastases in nude mice. Moreover, the plant derived polyphenol has been reported to be able to overcome drug resistance to classical chemotherapy. These features render the drug a promising candidate for tumor therapy especially for cancers known for their high rates concerning therapy resistance like melanoma.

Results: We show here that the melanoma cell line M14 is resistant to Curcumin induced apoptosis, which correlates with the absence of any effect on NFkappaB signaling. We show that CXCL1 a chemokine that is down regulated in breast cancer cells by Curcumin in an NFkappaB dependant manner is expressed at variable levels in human melanomas. Yet in M14 cells, CXCL1 expression did not change upon Curcumin treatment. Following the hypothesis that Curcumin is rapidly removed from the resistant cells, we analyzed expression of known multi drug resistance genes and cellular transporters in M14 melanoma cells and in the Curcumin sensitive breast cancer cell line MDA-MB-231. ATP-binding cassette transporter ABCA1, a gene involved in the cellular lipid removal pathway is over-expressed in resistant M14 melanoma as compared to the sensitive MDA-MB-231 breast cancer cells. Gene silencing of ABCA1 by siRNA sensitizes M14 cells to the apoptotic effect of Curcumin most likely as a result of reduced basal levels of active NFkappaB. Moreover, ABCA1 silencing alone also induces apoptosis and reduces p65 expression. Conclusion: Resistance to Curcumin thus follows classical pathways and ABCA1 expression should be considered as response marker.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3539.
Abstract 3540: Down-regulation of solute carrier family 17 member 1(SLC17A1) expression contributes to chemotherapy resistance in acute myeloid leukemia
Authors: Lee Chansu , Park Juwon , Oh Jeong In , Kim Byung-Su , Kwon Jung Hye , Kim Byoung Kook , Ahn Kwang-Sung , Yoon Sung-Soo
Published: February 2011

Cytosine arabinoside (Ara-C), an structural analogue of deoxycytidine, is generally considered to be the major drug in the treatment of AML. In acute myeloblastic leukemia (AML) treatment, resistance to cytotoxic nucleoside analogues is a major problem. To investigate novel genes associating with becoming Ara-C resistant cells, whole genome SNP chip analysis was performed. However, there was no significant in Ara-C metabolite genes. In that analysis, 12 region of loss of heterozygote (LOH) was found in non-CR group. SLC17 family gene and HIST1H family gene located in major of LOH in non-CR group. Since SLC29A1 expression levels directly affected the Ara-C mediated Apoptosis, we selected SLC17A1 among them and functional analysis of SLC17A1 was performed. Its expression levels were examined in four different AML cell lines using RT-PCR. Its expression level was a little different but that gene seemed to be constitutively expressed. To detect the pattern change of SLC17A1 in survival AML cells against Ara-C treatments, four AML cell-lines underwent 10−8 M of Ara-C addition/ depletion for 2 and 4 cycles. The results showed that expression of SLC17A1 was decreased more than SLC29A1 expression in the same treated condition. To examine whether the expression levels of SLC17A1 was associated with Ara-C resistance, Ara-C resistant HEL cell was established in vitro and in vivo (HEL-R1 and HEL-R1-vivo). Our results indicated that SLC17A1 mRNA expression of both resistant cells is decreased or less increased than HEL cells in Ara-C treatment. And the reduction of SLC17A1 mRNA by shRNA of SLC17A1 effectively suppressed Ara-C mediated apoptosis. Our results suggested that the function of SLC17A1 gene was similar to that of SLC29A1. Taken together, the expression levels of SLC17A1 also might involve the cytotoxic effects of Ara-C treatment. SLC17A1 could be a novel gene controlling Ara-C mediated apoptosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3540.
Abstract 3547: The differences in intestinal absorptions between CZ48 and its parental Camptothecin
Authors: Liu Xing , DeJesus Albert , Cao Zhisong , Giovanella Beppino
Published: February 2011

The determination of oral bioavailability is a critical step in early phase drug development. In part, the ability to predict whether a drug will be effectively transported across the gastrointestinal mucosa can be estimated from the physicochemical properties of the compound. Currently, the Caco-2 human colon carcinoma cell line is often used by the pharmaceutical industry to evaluate intestinal absorption of drugs. The purpose of the present study was to investigate the membrane transport mechanism of CZ48, a hydrated C20-propionate ester of Camptothecin (CPT), in the Caco-2 cell culture model, in order to understand the possible role of membrane transporters on its oral bioavailability and disposition. Caco-2 cells were grown onto Millicell™ to form monolayers. Directional transport of CZ48 and CPT was determined. The results indicated that apical (AP) to basolateral (BL) transport of CZ48 and CPT was distinctively different from BL to AP transport, with AP to BL transport surpassing the BL to AP transport. The ratio of BL-AP/AP-BL transport rate was 0.36 and 3.0 for CZ48 and CPT at the concentration of 5µM, respectively. The results also showed that AP to BL permeability of CZ48 was ten times lower than CPT, but the accumulation of CZ48 in intact cells was higher than CPT (0.67 and 0.49 nmol, respectively). In conclusion, the p-glycoprotein may be involved in the transepithelial transport of CPT. An additional carrier mechanism may be involved in the AP uptake of CZ48, since Apical transport of CZ48 in the Caco-2 cells is mediated, at least in part, by uptake of a nucleic bases into cells via an nucleobase transporter.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3547.
Abstract 3673: Identifying the target of an orphan compound: CB30865 is a Nampt inhibitor
Authors: Fleischer Tracey C. , Flick Jeff S. , Terry-Lorenzo Ryan T. , Gao Zhong-Hua , Murphy Brett R. , Willardson J Adam , Boniface J Jay
Published: February 2011

Background: Drug discovery based on cellular phenotypes is impeded by the challenge of identifying the molecular target. To alleviate this problem, Myriad Pharmaceuticals has developed a chemical proteomics process to identify cellular proteins that bind to small molecules. CB30865 is a potent and selective cytotoxic compound of previously unknown mechanism of action (1). Using our technology, we have unequivocally demonstrated that CB30865 cytotoxicity is due to inhibition of nicotinamide phosphoribosyltransferase (Nampt). Nampt catalyzes the first step in the synthesis of NAD from nicotinamide. Cancer cells develop dependence on Nampt due to increased energy requirements and the elevated activity of NAD consuming enzymes such as sirtuins and mono and poly(ADP-ribose) polymerases.

Materials and Methods: Direct target affinity purification (DTAP) was performed using analogs of CB30865 (MPI-0479883, MPI-0482594) coupled to beads. Purified proteins were identified by nano-flow reversed-phase liquid chromatography and tandem mass spectrometry using an LTQ-Orbitrap. In vitro Nampt activity was measured in a coupled biochemical assay based on the production of fluorescent resorufin. Nampt cellular activity was assayed by measuring levels of NAD and NAD-dependent polyADP-ribosylation.

Results: As reported (1), 3-pyridyl, but not 2-pyridyl, analogs of CB30865 were potently cytotoxic. A 55 kD protein was purified efficiently using DTAP with MPI-0479883 (a 3-pyridyl CB30856 analog) but not MPI-0482594 (a 2-pyridyl analog). Preincubation of cellular lysates with uncoupled 3-pyridyl but not 2-pyridyl analogs potently blocked binding of the 55 kD protein. Based on LC-MS/MS analyses, the 55 kD protein was identified as Nampt. Immunoblotting of DTAPs using a Nampt specific antibody confirmed the MS identification. Furthermore, biochemical and cellular assays of Nampt activity demonstrated pyridyl-isomer specific enzyme inhibition. Finally, cellular Nampt inhibition and cytotoxicity were completely rescued by treating cells with 10 µM nicotinic acid as an alternate NAD source.

Conclusions: The chemical proteomics DTAP approach is a powerful tool to identify molecular targets of orphan compounds. We demonstrate this here through identification of Nampt as the molecular target of CB30865 and further implicate NAD metabolism as a functional target for cancer.

(1) Skelton LA, Ormerod MG, Titley J, Kimbell R, Brunton LA, Jackman AL. A novel class of lipophilic quinazoline-based folic acid analogues: cytotoxic agents with a folate-independent locus. Br J Cancer. 1999 Apr;79(11-12):1692-701.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3673.
Abstract 3700: Effect of gender on pharmacokinetic disposition of pegylated liposomal CKD-602 (S-CKD602) and optisomal topotecan (TLI) in rats
Authors: Song Gina , Wu Huali , La-Beck Ninh M. , Zamboni Beth A. , Srychor Sandra , Santory Melissa M. , Deither Steven R. , Zamboni William C.
Published: February 2011

Introduction: S-CKD602 is a pegylated liposomal formulation of CKD-602, a camptothecin analogue. Optisomal Topotecan (TLI) is a sphingomyelin stabilized liposomal formulation of topotecan. The cytotoxicity of CKD-602, topotecan and other camptothecin analogues is related to the duration of exposure. Pegylated and sphingomyelin liposomal formulations have been designed to prolong drug circulation time, increase tumor delivery, and improve the therapeutic index. The pharmacokinetics (PK) of liposomal agents is highly variable in patients. Moreover, the factors associated with this variability are unknown. Thus, we evaluated the plasma PK disposition of TLI and S-CKD602 in male and female rats.

Methods: TLI was administered at 0.93mg/kg IV x1 and S-CKD602 at 0.6 mg/kg IV x1 to male and female Sprague-Dawley rats via a bilateral jugular vein cannula. Plasma samples for TLI were obtained in 6 cohorts (n= 3 rats per cohort, total = 18 rats per gender): 1) pre, 1, and 3 min; 2) 5, 10, and 20 min; 3) 0.5, 1, and 2 h; 4) 4, 8, and 12 h; 5) 24, 36, and 48 h; 6) 60 and 72 h post dose. Plasma samples (n= 3 rats per time point per gender) for S-CKD602 were obtained prior to the end of the infusion, and at 4, 8, 24, 48, and 72 h after administration. Total (lactone and hydroxyl acid) form of sum total (encapsulated and released) CKD-602 and topotecan were measured via LC/MS. The clearance (CL) was estimated using noncompartmental PK analysis.

Results: For TLI, the CL in male and female rats were 0.026 ± 0.0038 and 0.021 ± 0.0015 L/h/kg, respectively. For S-CKD602, the CL in male and female rats were 0.0037 ± 0.0004 and 0.0027 ± 0.0001 L/h/kg, respectively. For TLI and S-CKD602, CL was 1.2-fold (p=0.14) and 1.4-fold (p=0.009) lower in female rats compared with male rats, respectively.

Conclusions: The CL of TLI and S-CKD602 was lower in female rats as compared with male rats. These studies suggest that gender may affect the disposition of liposomal formulations of drugs and may play a role in the high PK variability reported in patients treated with liposomal formulations of anticancer agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3700.
Abstract 3774: Congenital abnormalities and hepatoblastoma: A report from the Children's Oncology Group (COG)
Authors: Spector Logan G. , Puumala Susan E. , Tomlinson Gail E. , Georgieff Michael K. , Krailo Mark , Ross Julie A.
Published: February 2011

Hepatoblastoma (HB) is a rare pediatric tumor which comprises the majority of liver cancer in children. Previous research has established that Beckwith-Wiedemann Syndrome (BWS) and Familial Adenomatous Polyposis (FAP) predispose to HB, but information on its association with isolated congenital abnormalities is sparse. We examined data from a case-control study of HB conducted through the Children's Oncology Group (COG). Cases were diagnosed by COG institutions during 2000-2008 and controls were recruited from state birth registries, frequency matched for sex, region, year of birth, and low birth weight (a strong risk factor for HB). Data on congenital abnormalities among subjects and covariates were obtained by maternal telephone interview. There were 359 cases and 372 controls included in this analysis, which comprises approximately 90% of the final sample. Odds ratios (OR) and 95% confidence intervals (CI) describing the association between congenital abnormalities with HB, adjusted for birth weight and sex, were calculated using unconditional logistic regression. Twelve cases were reported to have BWS and 4 to have FAP, while no controls had either condition; further analyses excluded abnormalities among children with these syndromes to better examine isolated defects. There was a significant association of HB with kidney, bladder, or sex organ abnormalities (OR = 4.22; 95% CI: 1.57-11.29) which appeared to be specific to kidney/bladder defects (OR = 5.24; 95% CI: 1.40-19.59) but not those of sex organs (OR = 1.58; 95% CI: 0.45-5.55). Elevated but non-significant ORs were found for spina bifida or other spinal defects (OR = 2.39; 95% CI: 0.40-14.07), large or multiple birthmarks (OR = 1.63; 95% CI: 0.97-2.76). No significant associations of cleft lip or palate, Down syndrome or other numerical chromosomal anomalies, microcephaly, or rib abnormalities, although <4 subjects were reported to have each of these defects, precluding precise estimation of ORs. These results confirm the previously noted associations of HB with BWS and FAP, and further suggest a novel association with kidney/bladder abnormalities. Supported by NIH CA111355, and the Children's Cancer Research Fund, Minneapolis, MN.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3774.