bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1580: Molecular mechanisms of OSM-induced VEGF in breast cancer
Authors: Nandakumar Madhuri , Fox Ryan , Jorcyk Cheryl
Published: June 2012

Oncostatin M (OSM) is an interleukin-6 (IL-6) family cytokine that has been shown to induce expression of vascular endothelial growth factor (VEGF) in astroglioma and in hepatocellular and breast carcinoma. It is also known to induce VEGF in non-cancerous cells such as cardiac myocytes, hepatocytes, and smooth muscle cells. Under hypoxic conditions, VEGF expression is known to be dependent on the transcription factor hypoxia-inducible factor 1 alpha (HIF1α). Our current studies were designed to determine whether VEGF expression induced by OSM and other IL-6 family members, including IL-6 and leukemia inhibitory factor (LIF), is likewise dependent on HIF1α in human breast cancer cells. To analyze VEGF expression levels, MDA-MB-231 and T47D human breast cancer cells were cultured in the presence or absence of OSM, IL-6, and LIF, with or without a HIF1α siRNA, for 12 to 72 hrs. All three cytokines significantly induced HIF1α expression by Western blot analysis at each time point in both MDA-MD-231 and T47D cells. Conditioned media from these experiments was also analyzed using ELISA to determine HIF1α-induced VEGF expression. Treatment with OSM resulted in a two- to three-fold induction of secreted VEGF, while treatment with IL-6 and LIF showed no significant induction. In addition, OSM-induced VEGF expression was independent of HIF1α in MDA-MB-231 cells. This was indicated by unchanged secreted VEGF levels despite the reduction of HIF1α expression in siRNA treated groups. In T47D cells however, HIF1α siRNA-treatment for 72 hrs reduced VEGF levels by 40% suggesting that OSM-induced HIF1α may be partially responsible for VEGF expression in this cell line. To summarize, it is important to note that IL-6 and LIF did not induce VEGF expression in two commonly used human breast cancer cell lines, MDA-MB-231 and T47D. Furthermore, OSM-induced VEGF was not dependent on HIF1α in MDA-MB-231 cells, suggesting regulation of VEGF by an alternative molecular mechanism. Investigations are in progress to better understand mechanisms utilized by breast cancer cells for OSM-induced VEGF expression. Future results could lead to the potential design of OSM-targeted therapeutics. Funded by NIH R15CA137510, ACS RSG-09-276-01-CSM, Susan G. Komen for the Cure KG100513, and NIH/NCRR P20RR016454.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1580. doi:10.1158/1538-7445.AM2011-1580
Abstract 1584: Characterization of 28 patient-derived colorectal cancer xenografts and analysis for predictive response markers to current standard treatment
Authors: Becker Michael , Rivera-Markelova Maria , Merk Johannes , Sers Christine T. , Slisow Wassilij , Rosenthal Andre , Hinzmann Bernd , Schlag Peter M. , Hoffmann Jens , Fichtner Iduna
Published: June 2012

Improved methods for the preclinical characterization of anti-cancer drugs in larger panels of primary lung cancer xenograft models have recently described (Fichtner et al. Clin Cancer Res. 2008;14(20):6456-68). These models consider more thoroughly the heterogeneity and genetic instability of tumors in a way ultimately enabeling treatment decisions on an individualized basis.

The approach is based on large scale screening for activity in new in vivo models combined with extensive characterization of these models (gene expression and mutations). Bioinformatic methods are then used to further evaluate the mechanism of action of new drugs, to investigate possible resistance mechanisms and provide a rationale for combination therapies.

Here we describe the development of a panel of primary patient derived colorectal cancer models. Primary colon carcinoma tissue samples were collected using a standardised procedure. Tumour pieces were transplanted onto immunodeficient mice immediately after surgery. A panel of 28 stably passagable colon cancer xenografts could be established as permanent tumour models. The results obtained so far, show that these patient-derived colon cancer models feature a high coincidence with the original tumour regarding histology and gene expression profiling. Response analysis according to clinical trial criteria revealed that the standard drug Oxaliplatin induce a overall response in 8 of the 28 xenograft models.

Gene expression analyses were performed to identify predictive profiles for Oxaliplatin sensitivity. As one example, the effectiveness of Oxaliplatin seems to be related to the differential expression of genes which products are involved in the wnt pathway.

The analysis of drug sensitivity in this set of models together with bioinformatics coorelations will provide hypotheses for further studies to identify patients that will more likely benefit from a drug as well as provide a rationale for combinations to be tested in clinical trials.

These new approaches may help to better integrate research results in clinical development and may increase the success of new anti-cancer drugs in clinical studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1584. doi:10.1158/1538-7445.AM2011-1584
Abstract 1588: Transformation of primary human fallopian tube secretory epithelial cells: A cell of origin for high-grade serous ovarian carcinoma
Authors: Karst Alison M. , Levanon Keren , Drapkin Ronny
Published: June 2012

High-grade serous ovarian carcinoma (HGSOC) is a lethal disease for which improved screening and treatment strategies are urgently required. Progress in these areas has been impeded by our poor understanding of HGSOC pathogenesis. The vast majority of ovarian cancer research is been based on the hypothesis that HGSOC develops from ovarian surface epithelial cells. However, recent studies suggest that > 50% of high-grade serous carcinomas involving the ovary likely arose from secretory cells of the fallopian tube epithelium. Consequently, current ovarian-based experimental models of HGSOC are inadequate in representing the full spectrum of the disease. The aim of our study was to develop the first experimental model of fallopian tube secretory epithelial cell (FTSEC) transformation and to use this model to answer three basic questions: 1) Can FTSECs be transformed in vitro?, 2) What genetic alterations are capable of transforming FTSECs?, and 3) Do transformed FTSECs give rise to high-grade serous carcinomas? To this end, we isolated primary human FTSECs from fresh samples of normal fallopian tube fimbriae and immortalized them via transduction with hTERT and SV40 TAg. Immortal FTSECs were transformed by either H-RasV12 or c-Myc expression, resulting in accelerated proliferation, enhanced clonogenicity, anchorage-independent growth, and tumor formation in immunodeficient mice. When SV40 TAg was replaced with equivalent non-viral genetic alterations (sh-p53, CDK4R24C, and sh-PP2A-B56γ), FTSECs remained susceptible to c-Myc-induced transformation and tumorigenesis. These results indicate that hTERT expression plus combined dysfunction of the p53, Rb, and PP2A tumor suppressor pathways is sufficient to support c-Myc-mediated transformation. Importantly, all of the transformed FTSECs in our study produced high-grade serous tumors with Müllerian and epithelial characteristics. Moreover, these tumors were grossly, histologically, and immunophenotypically consistent with human HGSOC. The genetic elements employed by our model represent the most common genomic alterations in human HGSOC – namely, TP53 mutation, RB1 dysfunction, and c-MYC amplification – and are therefore appropriate for modeling this disease. In summary, we have presented the first proof-of-principle evidence that FTSECs can be transformed into high-grade serous carcinomas resembling HGSOC. Our FTSEC transformation model provides a novel framework for assessing the transformative effects of specific genetic alterations on primary FTSECs and thus determining what role they may play in the pathogenesis of HGSOC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1588. doi:10.1158/1538-7445.AM2011-1588
Abstract 159: Prognostic and diagnostic significance of microRNA in bladder cancer
Authors: Boyd Sutton , Bissada Nabil K. , Ghoneim Mohamed A. , Keane Thomas E. , Fowler Elizabeth , Sobolesky Philip , Wolff Daynna J. , Watson Dennis K. , Moussa Omar
Published: June 2012

Purpose: Recently the role of microRNA (miRNA) in the development and progression of cancer including genitourinary tumors, and lymphomas to name a few have illustrated specific miRNA's to be dysregulated. Consequently miRNA's are currently being investigated as potential diagnostic and prognostic markers in neoplastic diseases. Because miRNA's are highly conserved they are promising as a potential biomarker that may be acquired by non-invasive means. This study investigated the presence of miR-21, miR-17-5p, miR-155, miR-26, miR-126, and miR-182 in bladder cancer tumor and tumor normal tissue specimens. We further investigated the presence of miR-21, miR-17-5p and miR-155 in urine sediment of bladder cancer patients.

Materials and Methods: quantitative Real time (RT-PCR) analysis was utilized for the detection of the precursors of miR-21, miR-17-5p, miR-155, miR-26, miR-126, and miR-182 in 23 RNA samples obtained from with bladder cancer tissue specimens. The relative expression values were compared to pooled matched non-tumor tissue. The presence of precursors of miRNA miR-21, miR-17-5p and miR-155 were also tested in 104 urine specimens from patients with active disease following positive cystoscopy (n=57) and patients previously diagnosed and treated for bladder cancer (n=34) with negative cystoscopic examination, patients with non-bladder cancer urological disease (n=6) and 7 healthy volunteers.

Results: The precursors of the oncogenic miRNA miR-21, miR-155, and miR-26 were up-regulated in the early grades and stages of the disease and the levels of expression are maintained in high grades and late disease stage. Interestingly, high levels of miR-17-5p were detected in higher grades and stages of bladder cancer and correlated with poor long term survival. Our data also indicated that the miR-126 is down-regulated in bladder cancer specimens compared to the non-tumor tissues. Our preliminary data have also shown that high levels of the precursors of miR-21, miR-17-5p and miR-155 can be detected in urine sediment collected from patients with active disease in higher levels compared to cytoscopy negative patients, patients with non-malignant urological diseases and healthy controls.

Conclusion: Our data have shown that miRNA detection in tissue and urine specimens in patients with bladder cancer has a promising prognostic and diagnostic significance and this preliminary data warranting further investigation to look at miRNA's as potential biomarkers. Illustrating the presence of miRNA in urine sediment of bladder cancer patients may prove useful in noninvasive evaluation of bladder cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 159. doi:10.1158/1538-7445.AM2011-159
Abstract 1597: A novel preclinical model of prostate cancer bone metastasis derived from a human primary tumor
Authors: Ainakouzi Nader , Bawa Olivia , Opolon Paule , Lerondel Stephanie , Lepape Alain , Gaudin Catherine , Massard Christophe , Loriot Yohann , Fizazi Karim , Chauchereau Anne
Published: June 2012

Background: Prostate cancer is frequently associated with bone metastases which are the main cause of morbidity and mortality for this tumor. Animal models are important tools to investigate the pathogenesis and develop treatment strategies for bone metastases in humans. However there are few animal models that recapitulate spontaneous clinical bone metastases. We recently obtained the new prostate cell line IGR-CaP1 derived from primary epithelial prostate cancer that has been characterize in vitro.

Methods: The IGR-CaP1 cell line was stably transduced with a luciferase expressing viral vector to monitor tumor growth by bioluminescence imaging (BLI) in male nude mice. Bone lesions were observed after direct intratibial injections of 5×105 luciferase-expressing IGR-CaP1 cells and after injection of 106 cells into the left cardiac ventricle of mice. Bone metastases were detected by bioluminescence imaging and were evaluated by both Computerized Tomography (CT) and micro Single-Photon Emission Computed Tomography (SPECT) measuring the bone fixation of 99Tc methylenediphosphonate (m99Tc-MDP).

Results: In contrast with the widely used PC3 preclinical model showing only osteolytic activity after bone injection, high resolution CT scan showed both lytic lesions in the cortical bone and osteoblastic lesions in the trabecular bone in the IGR-CaP1-injected mice 5 weeks after injection. The mixed nature of the lesions and the intensive bone remodeling were confirmed by bone scintigraphy with a ∼5 fold increase in m99Tc-MDP fixation in injected tibia versus control tibia. Intracardiac injection led to bone metastasis mainly localized in the spinal column, fore and hind limbs and in the mandible. Immunohistochemistry confirmed the presence of these bone metastases and showed high expression of vimentin in metastatic tumor cells. In this model, we are currently analyzing by RT-PCR, the expression of a 25-gene signature that corresponds to genes highly expressed in human bone metastases.

Conclusion: IGR-CaP1 is a unique model issued from a primary tumor able to reconstitute human prostate adenocarcinoma in animals and provide bone metastases. This model provides a novel means for identifying and understanding mechanisms that contribute to bone metastasis and allows for preclinical testing of new anticancer therapies, specifically bone-targeting therapies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1597. doi:10.1158/1538-7445.AM2011-1597
Abstract 16: Involvement of BH4 domain of bcl-2 in the regulation of HIF-1-mediated VEGF expression in hypoxic tumor cells
Authors: Gabellini Chiara , Trisciuoglio Daniela , desideri Marianna , Ragazzoni Ylenia , De Luca Teresa , Del Bufalo Donatella
Published: June 2012

In addition to act as an antiapoptotic protein, bcl-2 can also promote tumor angiogenesis. In this context, we have previously demonstrated that under hypoxia bcl-2 promotes hypoxia inducible factor 1 (HIF-1) -mediated vascular endothelial growth factor (VEGF) expression in melanoma and breast carcinoma. Here, we report on the role of the BH4 domain in bcl-2 functions, by showing that removal of or mutations at the BH4 domain abrogates the ability of bcl-2 to induce VEGF protein expression and transcriptional activity under hypoxia in human melanoma cells. We have also extended this observation to other human tumor histotypes such as colon, ovarian and lung carcinomas. The involvement of BH4 on HIF-1α protein expression, stability, ubiquitination and HIF-1 transcriptional activity was also demonstrated in melanoma experimental model. Moreover, we validated the role of the BH4 domain of bcl-2 in the regulation of HIF-1/VEGF axis, demonstrating that BH4 peptide is sufficient to increase HIF-1α protein half-life impairing HIF-1α protein ubiquitination, and to enhance VEGF secretion in melanoma cells exposed to hypoxia. Finally we found that the mechanism by which bcl-2 regulates HIF-1-mediated VEGF expression does not require BH1 and BH2 domains, and it is independent of antiapoptotic and prosurvival function of bcl-2.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 16. doi:10.1158/1538-7445.AM2011-16
Abstract 1600: A xenograft model of spontaneous metastases in NOD SCID mice of human non-small cell lung cancer
Authors: Harris James E. , Shin James , Lee Beverly , Pelosky Kristen , Zahnow Cynthia , Brayton Cory , Yang Stephen , Baylin Stephen , Brock Malcolm V.
Published: June 2012

Introduction: Xenograft models of human non small cell lung cancer cell lines have been unreliable in studying distal organ metastases. Although the flank, as a primary xenograft site, is typically used for the convenience of measurement, rarely do even the most aggressive cell lines develop metastases.

Methods: Using 15 anesthetized NOD.CB17-Prkdcscid/NcrCrl (NOD/SCID) mice, we xenografted 10,000 A549 cells into the liver under direct visualization and allowed the tumors to grow for 7 weeks. The mice were then euthanized, perfused with formalin, and grossly dissected. The liver, lung and spleen were processed, embedded in paraffin, sectioned and stained. A second group of 7 NOD/SCID mice were subsequently xenografted in the liver with A549, and a mouse each week was euthanized for 7 weeks and evaluated.

Results: At 7 weeks, all 15 mice had gross primary tumor in the liver as well as grossly evident splenic (10/15) and mediastinal metastases (5/15). There were no gross pulmonary metastases observed (0/15). Histologic review, however, confirmed multiple microscopic foci of metastatic disease in all mice (15/15) throughout the lungs bilaterally, the mediastinal lymph nodes and spleen. Serially euthanized mice revealed histological evidence of a gradual spontaneous metastatic model system. Histologically evident primary tumor was apparent by week 1 with gross primary tumor development in the liver by week 2. Weeks 3 and 4 showed gross and histologic progression of primary tumor size with no apparent evidence of microscopic distal metastases. By week 5, the first histological findings of micrometastases appeared in the lungs which became wide spread in all the pulmonary parenchyma by week 7. Splenic and mediastinal lymph node involvement were observed histologically by week 6.

Conclusions: Liver xenografting of A549 into NOD/SCID mice is a reliable way of developing widespread micrometastases to the lungs, mediastinal lymph nodes, and spleen. This model is clinically relevant since, unlike tail vein injection, it allows the study of the biology and treatment of a gradually developing primary lung tumor with subsequent metastatic spread.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1600. doi:10.1158/1538-7445.AM2011-1600
Abstract 1601: Hsp90 inhibition results in a significant delay in tumor progression in a model of emerging EGFR TKI resistance in non-small cell lung cancer
Authors: Slocum Kelly L. , Sheikh Hana , West Kip , Normant Emmanuel , Robison Keith , MacDougall John , Fritz Christian
Published: June 2012

Heat shock protein 90 (Hsp90) plays a role in regulating the stability of key cancer-causing proteins through its role as a protein chaperone. Proteins chaperoned by Hsp90, known as client proteins, include cancer-causing forms of ALK, BCR-ABL, EGFR, FLT3 and HER2. Infinity is developing two drug candidates in its Hsp90 chaperone inhibitor program: IPI-504 (retaspimycin hydrochloride), an intravenously-administered small molecule, and IPI-493, which is administered orally.

EGFR tyrosine kinase inhibitors (TKIs) are an effective treatment for lung cancer patients with activating mutations in EGFR. After a dramatic initial response, however, most patients become resistant to drug treatment and progress. In about half of these cases, resistance is due to a second point mutation in EGFR (T790M). It is believed that in at least some of these cases, the TKI resistance mutations are pre-existing and that treatment with TKIs selects for the resistant cells.

In an effort to model the emergence of resistance to TKIs from pre-existing mutations, we developed a novel in vivo model, where gefitinib treatment initially leads to tumor regression followed by rebound of tumor growth and outgrowth of drug resistant clones containing the T790M mutation. We show that in this model, treatment with IPI-493 alone and IPI-493 following gefitinib resulted in tumor growth inhibition of 61 and 77%, respectively, when compared with gefitinib treatment alone. Treatment with IPI-493 alone also resulted in a significant delay in time to tumor progression with ∼40% of animals still on study 45 days following tumor implant; all animals treated with either vehicle or gefitinib had been removed due to tumor progression. Interestingly, treatment with IPI-493 following gefitinib resulted in an even more impressive delay in time to progression, with >50% of animals still on study on day 65 post-implant.

These results suggest that further studies with Hsp90 inhibitors in EGFR mutant NSCLC patients who have been pre-treated with a TKI may be warranted.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1601. doi:10.1158/1538-7445.AM2011-1601
Abstract 1603: Primary breast cancer xenografts maintain the molecular profiles of corresponding patients’ tumors and recapitulate a part of stroma microenvironment
Authors: Reyal Fabien , Guyader Charlotte , Decraene Charles , Lucchesi Carlo , Auger Nathalie , Assayag Franck , De Plater Ludmilla , Gentien David , Poupon Marie-France , Cottu Paul , De Cremoux Patricia , Vincent-Salomon Anne , Roman-Roman Sergio , Delattre Olivier , Decaudin Didier , Marangoni Elisabetta
Published: June 2012

Identifying new therapeutic agents for breast cancer (BC) treatment requires preclinical models that recapitulate the molecular characteristics of their respective clinical tumors. In this work, we analyzed the genomic and expression profiles of human BC xenografts and their corresponding patient's tumors. Eighteen BC xenografts were obtained by grafting tumor fragments from patients into Swiss nude mice. The molecular characterization of patient's and xenograft's tumors (early and late in vivo passages) was performed by DNA copy number analysis (aCGH) and gene expression analysis using Affymetrix Microarrays. Comparison of genomic profiles of BC xenografts with those of respective patient's tumors, showed a correlation coefficient higher than 0.50 in 14/18 paired tumors. Unsupervised hierarchical clustering analysis showed that 16/18 pairs segregated together further indicating the striking similarity between pairs. At the level of gene expression profile few variations were found between patient's tumor and their respective xenografts, i.e. 371 genes that were differentially expressed in more than 84% of paired tumors and that largely represent human stromal compartment related genes. Furthermore, analysis of gene expression by murine stroma revealed a partial but significant recapitulation of the primitive human stroma.

Finally, we found that sequential mouse to mouse tumor passages did not affect neither genomic rearrangements nor gene expression profiles, suggesting a striking genetic stability of these models across the time. In conclusions, this panel of human BC xenografts accurately maintains the genomic and the expression profiles of their corresponding patient's tumors and are stable across sequential in vivo passages. Consequently, these xenografts represent a validated model for preclinical investigation of new therapeutic agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1603. doi:10.1158/1538-7445.AM2011-1603
Abstract 1609: In vivo and ex vivo comparison of two human AML bone marrow engraftment mouse models using fire fly luciferase expressing MOLM-13 and MV4-11 cells
Authors: Lingnau Andreas , Feger Daniel , Hoffmann Steffen , Umber Sarah , Kubbutat Michael HG
Published: June 2012

Acute myeloid leukemia (AML) is the most common leukemia affecting adults. In 70-90% of all AML patients FLT3, a receptor tyrosine kinase which plays an important role in normal hematopoiesis, is highly expressed. Furthermore, in about 30% of all AML patients activating internal tandem duplications (ITDs) or point mutations within the FLT3 gene have been identified. FLT3, therefore, is one of the most promising drug targets for the treatment of AML.

Most often novel FLT3 protein kinase inhibitors are tested in subcutaneous xenograft mouse models using various human AML cell lines. To generate more relevant, orthotopic AML in vivo models, we stably transduced the human AML cell lines MOLM-13 and MV4-11 with fire fly luciferase (MOLM-13-Luci and MV4-11-Luci) suitable for in vivo bioluminescence imaging. MOLM-13 carries a heterozygous and MV4-11 a homozygous FLT3 internal tandem duplication (ITD) mutation. In vivo growth of MOLM-13-Luci and MV4-11-Luci cells after subcutaneous implantation in NMRI nude mice proofed that stable expression of luciferase did not alter growth characteristics of these cells.

To initiate orthotopic growth, both AML cell lines were implanted intravenously via the tail vein into NOD SCID mice after Cyclophosphamide treatment. Cyclophosphamide facilitates engraftment of the human cells by reducing the endogenous bone marrow cell population.

Approximately two (MOLM-13-Luci), or three (MV4-11-Luci) weeks after implantation, luciferase activity was detectable in the pelvic region of the mice indicating growth of both tumor cell lines. Within the next weeks luciferase activity was also seen in additional parts of the mouse at different levels.

After necropsy a set of different organs and tissues was analysed ex vivo for luciferase activity. The pattern of metastasising tumor cells was compared in both models. Furthermore, detection of human Ki-67 protein using appropriate human specific antibodies allowed verification of growth of MOLM-13-Luci and MV4-11-Luci cells in different tissues.

Sutent, a small-molecule kinase inhibitor, totally abolished tumor growth of both orthotopically growing tumors, indicating that both in vivo models are highly suitable for the characterisation of novel drugs directed against AML.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1609. doi:10.1158/1538-7445.AM2011-1609
Abstract 161: Plasma-based biomarkers for breast cancer diagnosis
Authors: Hu Zhibin , Shen Hongbing , Dong Jing , Ma Hongxia , Dai Juncheng , Wei Qingyi , Wang Li-E
Published: June 2012

Introduction It has been demonstrated that there are abundant, stable microRNAs (miRNAs) in plasma/serum, which can be detected and is potentially disease-specific. In this study, we aimed to assess whether serum/plasma miRNAs may be used as potential biomarkers for breast cancer diagnosis.

Methods We used systematically Solexa sequencing as well as TaqMan Low Density Array followed by individual quantitative reverse transcription polymerase chain reaction assays to screen plasma miRNAs in 48 breast cancer patients and 48 age-matched controls of Chinese origin for the discovery stage and validated in an internal validation set (50 patients and 50 controls in Chinese) and in an external validation set (24 patients and 24 controls in Caucasians).

Results Four miRNAs (miR-16, miR-25, miR-222 and miR-324-3p) were differentially expressed between breast cancer cases and controls and subsequently validated in the internal and external validation stage samples. Taken together, Chinese patients with high plasma miRNA expression levels had a significantly increased risk of breast cancer for all the four miRNAs. The Area Under the Receiver Operating Characteristic Curve was 0.917 for the four-miRNA signature in the discovery stage (sensitivity=0.917 and specificity=0.917), 0.930 in the internal validation stage (sensitivity=0.920 and specificity=0.940).

Conclusions The four-miRNA signature from plasma may serve as a noninvasive diagnostic biomarker for breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 161. doi:10.1158/1538-7445.AM2011-161
Abstract 1612: Investigating NF-κB pathway as a novel therapeutic target in preclinical lung cancer mouse models
Authors: Xue Wen , Meylan Etienne , Oliver Trudy , Jacks Tyler
Published: June 2012

Lung cancer represents the most frequent cancer type and the leading cause of cancer death (1.3 million worldwide). Non-small cell lung cancer (NSCLC) accounts for 80% of all lung cancer cases. NSCLC development is associated with frequent mutations in well-defined oncogenes and tumor suppressor genes such as Kras (30%) and p53 (50-70%). We recently demonstrated that hyper-activated NF-kB pathway in NSCLC is required for the initiation and maintenance of lung cancer, indicating this pathway as a promising therapeutic target. In this study, we tested therapeutic delivery of proteasome inhibitor Bortezomib (Velcade) to inhibit NF-κB pathway in NSCLC mouse models. In mouse lung cancer cell lines harboring KrasG12D and p53 loss (KP cells), Bortezomib efficiently reduced nuclear p65, repressed NF-kB target genes such as Bcl2 and rapidly induced apoptosis. The cell lines with high NF-kB activity are especially sensitive to Bortezomib. Using autochothonous mouse models and microCT imaging, we showed that Bortezomib treatment at a well-tolerated dose leads to lung tumor regression in mice and prolonged survival. Our results demonstrated using preclinical mouse models to study cancer treatment response and supported NF-kB pathway as an effective therapeutic target in lung cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1612. doi:10.1158/1538-7445.AM2011-1612
Abstract 1619: Discovering a new physiological substrate of Aurora-A kinase
Authors: Toughiri Raheleh , Padmanabhan Achuth , Li Xiang , Bieberich Charles
Published: June 2012

Aurora-A is a serine/threonine kinase that has oncogenic properties in vivo. The expression and kinase activity of Aurora-A are up-regulated in many cancers including prostate cancer, which is the most frequently diagnosed malignancy in men and second leading cause of cancer death in the United States. Aurora-A is one of the key regulators of mitosis that localizes to the centrosome from G2 phase through mitotic exit, where it regulates mitotic spindle formation and centrosome separation. Although Aurora-A is known to be an oncogene, few Aurora-A substrates have been identified. It is essential to elucidate Aurora-A substrates to develop biomarkers for Aurora-A inhibitors, and to identify new targets downstream of Aurora-A for cancer therapy.

To identify new Aurora-A substrates, an Aurora-A Reverse In-gel Kinase Assay was developed. The basis of this method is co-polymerization of a denatured protein kinase throughout a polyacrylamide gel followed by protein refolding. Subsequently, an in-gel kinase reaction in the presence of γ-32P-ATP allows phosphorylation of potential kinase substrates that can be located by autoradiography. Phosphorylated proteins can be extracted from the gel and identified by liquid chromatography mass spectrometry. Using this method, we were able to identify Nuclear Mitotic Apparatus (NuMA) as an Aurora-A kinase substrate. Preliminary in vitro kinase experiments verify phosphorylation of recombinant NuMA C-terminus by Aurora-A. The identification of Aurora-A phosphorylation site/s will permit a better understanding of Aurora-A function in normal and cancer cells. The location of Aurora-A phosphorylation of NuMA and consequences of their mutation will be discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1619. doi:10.1158/1538-7445.AM2011-1619
Abstract 1629: A protein-protein interaction network for RalB-TBK1 signaling pathway in KRAS mutant non small cell lung cancer
Authors: Kim Jae-Young , Fang Bin , Koomen John , Zhang Guolin , Haura Eric B.
Published: June 2012

KRAS mutation accounts for development of approximately 30% of Non Small Cell Lung Cancer (NSCLC) however therapeutic targets for KRAS mutant NSCLC have poorly been identified. A better understanding of the KRAS signaling network could uncover novel therapeutic targets or therapeutic strategies. RalB and TBK1 are important proteins used by KRAS to maintain tumor cell survival. To better understand oncogenic properties of RalB-TBK1 signaling for KRAS mutant lung cancer development, we are currently developing a protein-protein interaction network for RalB and TBK1 using Tandem Affinity Purification (TAP) coupled with Liquid Chromatography-Mass Spectrometry (LC-MS/MS). By interrelating protein-protein interactions and data from a RNA interference screen, a map of signaling will be constructed to highlight important components vulnerable to attack. Data from this mapping will be used to interrogate novel targets in a larger collection of lung cancer cell lines. We have expressed Strep-HA tagged versions of both RalB and TBK1 in KRAS mutant lung cancer cells using retroviruses. A two step purification coupled to LC-MS/MS is used to purify and identify protein complexes. Preliminary studies have identified the bait (TBK1) in the MS results along with HSP90 isoforms. Based on these results, we determined if HSP90 inhibition can modulate TBK1 levels. We found that 17-DMAG and AUY922, two HSP90 inhibitors, lead to reduced levels of TBK1 proteins. AUY922 also had potent effects on cell viability in vitro. Suppression of TBK1 expression using shRNA significantly induced cell death in KRAS mutant NSCLC cells, confirming that TBK1 signaling is essential for survival of those cancer cells, and loss of TBK1 by HSP90 inhibitors could in part lead to their cytotoxic effect. Collectively, we expect this network approach to KRAS-RalB-TBK1 signaling pathways will provide better insight into molecular mechanism by which TBK1 mediates KRAS mutant lung cancer cell survival and novel therapeutic targets for this cancer subtype. Updated work will be presented. Supported by the SPORE in Lung Cancer (P50-CA119997).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1629. doi:10.1158/1538-7445.AM2011-1629
Abstract 1634: Targeting L1 cell adhesion molecule using lentivirus-mediated short hairpin RNA interference reverses aggressiveness of oral squamous cell carcinoma
Authors: I-Hui Wu , Hung Shiao-Chen , Hsue Shui-Sang , Liao Chia-Hui , Wang Hsien-Chi , Chuang Pei-Hsin , Sung Shian-Ying , Hsieh Chia-Ling
Published: June 2012

Oral cancer consistently ranks as one of the ten most frequently diagnosed cancers in the world. Despite multidisciplinary treatment with surgery, chemotherapy, and radiation, more than 50% of patients with oral squamous cell carcinoma (OSCC) receiving therapy will eventually develop recurrent and metastatic disease, which carries a particularly poor prognosis. Characterization of the specific molecular alternations associated with OSCC cell growth, invasion, and metastasis can advance our understanding of the molecular mechanisms underlying cancer induction and progression as well provide a basis for the development of new and effective therapeutic treatments for cancer patients. The L1 cell adhesion molecule (L1CAM) has been implicated in tumor progression of many types of cancers, but its role in OSCC has not been investigated. In the present study, we demonstrated overexpression of L1CAM in OSCC cells, but not in normal keratinocytes, using both clinical specimens and cell lines. This overexpression demonstrated a strong correlation with less differentiation and a higher invasion potential of cancer cells, supporting the significance of L1CAM in human OSCC tumor progression. Targeting L1CAM gene expression in SCC4 cells overexpressing L1CAM using a lentivirusmediated small hairpin RNA (shRNA) led to a significant reduction in cell proliferation in vitro via retardation of cell cycle at the G1 phase. In addition, shRNA knockdown of L1CAM strongly attenuated the migration and invasion of SCC4 cells, and this was also observed to parallel increased E-cadherin levels and decreased levels vimentin, fibronectin, and Snail-family transcription factors, indicating that L1CAM expression was related to the epithelial-mesenchymal transition. Furthermore, while mice receiving orthotopically placed control SCC4 cells died within 40 days due to invasive tumor growth and regional lymph node metastasis, prolonged animal survival and complete suppression of tumor progression was observed in mice implanted with L1CAM-deficent SCC4 cells, further substantiating the fundamental importance of L1CAM in OSCC pathophysiology. Our findings suggested that L1CAM is a critical mediator of tumor progression in OSCC, and targeting L1CAM using lentivirus-mediated shRNA may be a useful molecular pharmaceutical approach for the treatment of advanced OSCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1634. doi:10.1158/1538-7445.AM2011-1634
Abstract 1639: Targeting the p38 MAPK pathway inhibits irinotecan resistance in colon adenocarcinoma
Authors: Paillas Salomé , Boissiere Florence , Bibeau Frederic , Mollevi Caroline , Causse Annick , Denis Vincent , Vezzio-Vié Nadia , Ait-Arsa Imade , Pourquier Philippe , Martineau Pierre , Del Rio Maguy , Gongora Céline
Published: June 2012

Despite recent advances in the treatment of colon cancer, tumor resistance is a frequent cause of chemotherapy failure. To better elucidate the molecular mechanisms involved in resistance to irinotecan (and its active metabolite SN38), we established SN38-resistant clones derived from the colon adenocarcinoma HCT-116 and SW48 cell lines. These clones show various levels (6 to 60 fold) of resistance to SN-38 and display enhanced levels of activated MAPK p38 as compared to the corresponding parental cells. Because four different isoforms of p38 have been described, we then studied the effect of p38 overexpression or downregulation of each isoform on cell sensivity to SN38 and found that both alpha and beta isoforms are involved in the development of resistance to SN38. In this line, we show that cell treatment with SB202190, which specifically inhibits p38 alpha and beta, enhanced the cytotoxic activity of SN38, but not of 5-FU or oxaliplatin. Moreover, p38 inhibition sensitized tumor cells derived from both SN38-sensitive and -resistant HCT116 cells to irinotecan treatment in xenograft models. Finally, we detected less phosphorylated p38 in primary colon cancer of patients sensitive to irinotecan-based treatment, compared to non-responder patients. This indicates that enhanced level of expression of phosphorylated p38 could predict the absence of clinical response to irinotecan. Altogether, our results show for the first time that the p38 MAPK pathway is involved in irinotecan sensitivity and suggest that phosphorylated p38 expression level could be used as a marker of clinical resistance to irinotecan. They further suggest that targeting the p38 pathway may be a potential strategy to overcome resistance to irinotecan-based chemotherapies in colorectal cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1639. doi:10.1158/1538-7445.AM2011-1639
Abstract 1646: In vivo target validation in orthotopic hepatocellular carcinoma model using formulated siRNA
Authors: Liao Baisong , Kellner Debra , Wang Chunrong , Wu Shilu , Molina Marion , Zhu De-Min , Gollerkeri Ashwin , Pachuk Catherine
Published: June 2012

Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and its incidence continues to increase. Although there are approved therapies for HCC, the prognosis remains poor. The 5-year survival rate is < 5%. Nucleic acid-based therapeutics such as RNAi may improve patient outcome. Nevertheless, target identification/validation for HCC and delivery of siRNA are major challenges facing the development of HCC-specific RNAi based therapeutics. To enable target validation in vivo using RNAi, we have developed nanolipid siRNA formulations for the in vivo delivery of siRNA to orthotopic tumors. PK/PD studies in model systems demonstrate that up to 80% knockdown of target protein is achievable within tumors and that significant knockdown persists for at least 10 days following a single dose of formulated siRNA. Efficacy studies using similar formulations of siRNAs against potential HCC tumor targets, including mTOR, will be presented.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1646. doi:10.1158/1538-7445.AM2011-1646
Abstract 165: MicroRNA-29c expression as a new prognostic marker for esophageal squamous cell carcinoma
Authors: Kinoshita Koichi , Watanabe Masayuki , Kamohara Hidenobu , Tanaka Yohei , Kurashige Junji , Saito Seiya , Karashima Ryuichi , Hirashima Kotaro , Imamura Yu , Nagai Yohei , Iwagami Shiro , Iwatsuki Masaaki , Hayashi Naoko , Baba Hideo
Published: June 2012

Introduction and Objective: MicroRNAs (miRNAs) are small non-cording RNAs that act at the posttranscriptional level to control gene expression in virtually every biological process, including oncogenesis. Lately, the discovery of miRNAs in cancer patients opened up the exciting prospective of using miRNAs as powerful cancer biomarkers. We screened miRNAs which would become the prognostic factor in esophageal squamous cell carcinoma (ESCC) patients and performed validation about miRNAs identified by screening.

Matherial and Method: Tissues of ESCC and the matched normal counterparts were obtained from surgical specimens after resection from patients undergoing primary surgical treatment of ESCC. Total RNAs were obtained from the archive of formalin-fixed paraffin-embedded (FFPE) tissues using the RecoverAll Total Nucleic Acid Isolation kit for FFPE (Ambion) according to the manufacturer's instructions. Here, we investigated 45 miRNAs expression levels and prognosis in 20 patients with ESCC and assessed by reverse transcription-PCR analysis of the endogenous control RNU6B. About miRNAs which was significant difference for prognosis, we examined a correlation with the clinicopathologic factors including gender, age, TNM stage, vessel invasion and presence of recurrence with 65 ESCC patients.

Result: MiR-29c was identified to have an influence on prognosis. We re-analyzed the miR-29c expression level in 65 ESCC samples and investigated the various clinicopathological factors associated with miR-29c expression. The venous invasion and presence of recurrence were significantly correlated with high expression of miR-29c(p<0.05). The Kaplan-Meier analyze showed that, high expression level of miR-29c was found to be correlated with a low overall survival rate(p<0.001).

Conclusion: Our results suggest that the miR-29c overexpression might be a poor prognotic factor and miR-29c acts as an oncomir associated with the development and progression of ESCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 165. doi:10.1158/1538-7445.AM2011-165
Abstract 1652: MRGBP (C20orf20) contributes to development of colorectal cancer
Authors: Yamaguchi Kiyoshi , Sakai Michihiro , Kim JooHun , Yamada Yoshinao , Akiyama Yoshiyuki , Nakamura Yusuke , Furukawa Yoichi
Published: June 2012

MRGBP (MRG-binding protein, also known as C20orf20) encodes a subunit of TRRAP/TIP60-containing histone acetyltransferase complex. In our earlier study, a genome-wide expression analysis found that MRGBP was up-regulated in the majority of colorectal tumors. Importantly, the enhanced expression was associated with cell proliferation. In the present study, we extensively investigated MRGBP expression in 22 adenomas and 47 carcinomas of the colon and rectum by immunohistochemical analysis. Each stain result was sub-divided into three groups according to the percentage of positive stains in tumor cells (negative, <10%; low, 10 – 50%; high, >50%). As a result, carcinomas with negative, low, and high expression were comprised of 15, 11, and 21 tumors, respectively. On the other hand, adenomas with negative, low, and high expression were 12, 9, and 1, respectively. Statistical analysis has revealed that MRGBP expression in carcinomas was significantly increased compared with that in adenomas. No clinicopathological factors were associated with the expression of MRGBP in colorectal cancer. We further investigated genes regulated by MRGBP using DNA microarray. Comparison of gene expression profiles between MRGBP-knocked down cells and control cells identified a total of 41 genes whose expressions were clearly suppressed. The 41 genes included MCM2, MCM5, CDT1, and CDCA7, implying that MRGBP plays a role in DNA replication, minichromosome maintenance, and cell division. These data suggest that MRGBP may contribute to the development of colorectal cancer through rendering advantages in cell proliferation to cancer cells. Our findings may be helpful for the identification of biomarker and development of therapeutic approach to colorectal tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1652. doi:10.1158/1538-7445.AM2011-1652