bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2217: Analysis of mitochondrial variations in non-small cell lung cancer (NSCLC) by microarray-based genome resequencing
Authors: Wang Zhaoxi , Choi Sojung , Huang Yen-Tsung , Chen Feng , Lin Xihong , Neuberg Donna , Kim Jhin , Christiani David C.
Published: February 2011

The human mitochondrial genome is a circular double-stranded DNA molecule (mtDNA) of 16,569 base pairs, encoding 13 proteins that are essential in oxidative phosphorylation, two ribosomal genes, and 22 tRNAs. The mtDNA genome has a high copy number per cell, replicates independently of the nuclear genome, and has a maternal-inherited pattern. Mutations in mtDNA genome have long been suspected to play an important role in the development of cancer. Although most cancer cells harbor mtDNA mutations, the question of whether such mutations are associated with clinical prognosis of cancer remains unclear. In this study, we resequenced the entire mitochondrial genomes of tumor tissue from a population of 249 Korean non-small cell lung cancer (NSCLC) patients using the Affymetrix GeneChips Human Mitochondrial Resequencing Array 2.0 (Santa Clara, CA). A total of 892 single nucleotide polymorphisms (SNPs) were identified, including 269 (30.2%) synonymous SNPs, 142 (15.9%) non-synonymous SNPs, 78 (8.7%) SNPs in non-coding region, 188 SNPs (21.1%) in control region, as well as 215 (24.1%) previously unreported SNPs. Each tumor sample contained at least 6 mitochondrial SNPs, with maximum 41 SNPs. We selected 189 (21.2%) SNPs with frequency >2.5% for subsequent analyses. Single SNP analysis using Cox proportional hazards models identified 19 SNPs with p value < 0.05, but none of them remained significant after adjustment for multiple comparisons. Unsupervised clustering analysis revealed two major SNP signature panels, including Signature Panel 1 (8 SNPs) and Signature Panel 2 (5 SNPs). Accordingly, the patient population was divided into three groups: 68 (27.3%) subjects with both signature panels, 73 (29.3%) subjects with only the Signature Panel 1 but not the Signature Panel 2, and 108 (43.4%) subjects without signature panel. In early stage (stage I/ II) NSCLC, patients with the Signature Panel 2 had the worst clinical prognosis, and patients without any signature panel performed best in the survival analyses (adjust hazard ratio=1.83; 95% confidence interval, 1.25-2.66; p=0.003). Further mitochondrial haplogroup analysis identified that the Signature Panel 1 had most defining mutations for haplogroup M and N, and the Signature Panel 2 contained all defining mutations of haplogroup D4a. Interestingly, haplogroup D4a was previous reported as a marker for extreme longevity in Japanese. We conclude that mtDNA mutations may play an important role in survival among early stage lung cancer patients.

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 2217.
Abstract 2220: Analyzing cancer with single cell resolution using droplet technology
Authors: Samuels Michael L. , Kennedy Scott
Published: February 2011

Cancers develop and grow in the complex environments present in normal tissue, and tumor cells are composed of heterogeneous mixtures of both clonal transformed cells and other cell types. We demonstrate the use of microfluidic-generated single cell microdroplets in workflows that capture both phenotypic and genomic information from individual cells, enabling analysis of tumors with single cell resolution. This transformative new tool will allow for more thorough examinations of the variations that influence the predisposition and composition of cancers, and the cellular responses to therapeutic and prevention agents. Uniform picoliter-nanoliter volume aqueous droplets suspended in an immiscible oil provide unique opportunities for fluorescence-based analytical techniques, including sorting relevant sub-populations at rates of several hundred cells per second, coupled directly to single cell genomic characterization. We use a defined set of control samples to perform low-bias single cell whole genome amplification (WGA) within the sorted microdroplets, and show how following this with droplet PCR-based targeted amplification (of many thousands of selected genomic loci using a cancer-focused droplet primer library) provides an automated sample preparation method for any Next Gen sequencing platform. Single cell microdroplet technology results in a cost-effective method to gain sequence information from phenotypically selected individual cells, addressing cancer gene discovery and validation by powering the larger studies required to classify driver or passenger mutations and analyzing how epigenomic features (such as cytosine methylation) regulate transcriptional changes that can underlie cancer 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 2220.
Abstract 2224: Next-generation sequencing and bioinformatics analysis identified up-regulation of TGFBI and SOX4 in human glioblastoma
Authors: Lin Biaoyang , Madan Anup , Fang Xuefeng , Yoon Jae-Geun , Foltz Gregory
Published: February 2011

A comprehensive network-based understanding of molecular pathways abnormally altered in glioblastoma multiforme (GBM) is essential for developing effective therapeutic approaches for this deadly disease. Applying a next generation sequencing technology, massively parallel signature sequencing (MPSS), we identified a total of 4535 genes that are differentially expressed between normal brain and GBM tissue. The expression changes of three up-regulated genes, CHI3L1, CHI3L2, and FOXM1, and two down-regulated genes, neurogranin and L1CAM, were confirmed by quantitative PCR. Of note, pathway analysis revealed that TGF- β was significantly up-regulated in GBM tumor samples. An integrative pathway analysis of the TGF β signaling network identified two novel TGF—β signaling pathways mediated by SOX4 (sex determining region Y-box 4) and TGFBI (transforming growth factor beta 1 induced). Quantitative RT-PCR and immunohistochemistry staining showed that their expressions are elevated in GBM tissues compared with normal brain tissues at RNA and protein levels. In vitro functional studies confirmed that TGFBI and SOX4 expression is increased by TGF- β stimulation and decreased by a specific inhibitor of TGF- β receptor 1 kinase. The comprehensive identification of genes involved in TGF- β signaling provides further insight into GBM biology and novel therapeutic targets for further investigation.

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 2224.
Abstract 2234: Circulating tumor cells are rare in patients with curable oligometastatic cancer
Authors: Boffa Daniel J. , Guo Xiaojie , Molinaro Annette , Finan Betsy , Detterbeck Frank , Rimm David L.
Published: February 2011

Background: In the majority of patients with solid organ malignancies, tumor metastasis marks a transition from curable to terminal. However, a subset of metastatic cancer patients develops only a very limited number of metastases (known as oligometastatic). Survival in this cohort is considerably better than patients with widely disseminated cancer. In fact, in highly select subgroups, as many as 30% of oligometastatic patients are actually cured of cancer.

The mechanism that distinguishes limited (or oligometastatic) cancer, from the more lethal, widely disseminated, is unknown. Tumor cell circulation is a critical component of metastasis formation, with circulating tumor cells being detectable in as many as 40-70% of patients with metastatic cancer. We hypothesize that patients with limited metastasis formation, have limited tumor cell circulation. To test this hypothesis, we quantified circulating tumor cells in patients with oligometastatic disease.

Methods: Circulating tumor cells (CTCs) were quantified in patients with metastatic epithelial malignancies using the Veridex assay. Patients with > 2 CTCs in a 7.5 ml specimen were considered positive, as a single CTC has previously been shown to carry a substantially higher false positive rate (6.4%) than >2 CTCs (.3%).

Results: From April 2008 to December 2009, a total of 20 patients with oligometastatic cancer were enrolled. In all patients the primary tumor had been previously removed and each was being evaluated for curative surgery to remove the oligometastatic cancer. The origin of the primary tumor was colon (17 patients), head and neck (1 patient), ovarian (1 patient), and pancreas (1 patient). The site of oligometastatic cancer was the liver (12 patients), lungs (4 patients) or both liver and lungs (4 patients). Only 1 of the 20 patients with oligometastatic cancer (5%) was found to have >2 circulating tumor cells. During the same time frame, CTCs were more common in patients with disseminated cancer as 4 of 7 patients (57%) with incurable, widely disseminated metastatic cancer had > 2 CTCs (two tailed P value = .0089).

Conclusions: Circulating tumor cells are rare in patients with oligometastatic cancer.

One mechanism by which oligometastatic cancer patients avoid more widely disseminated cancer may relate to the frequency and duration of tumor cell circulation.

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 2234.
Abstract 2246: Identification of genomic alterations associated with metastasis and poor prognosis in clear cell renal cell carcinoma
Authors: Sanjmyatav Jimsgene , Schwaenen Carsten , Wessendorf Sven , Kreuz Marcus , Steiner Thomas , Wunderlich Heiko , Fuessel Susanne , Junker Kerstin
Published: February 2011

Introduction: Clear cell renal cell carcinoma (ccRCC) is the most frequently encountered renal malignancy with a high metastatic potential and the prognosis of patients in this subgroup is very poor. Prognostic parameters which can be used for an individual prognostic evaluation are still not available, So the aim of our study was high resolution screening of genomic imbalances of metastasized and non-metastasized primary ccRCC by array CGH in order to identify regions of DNA copy number changes significantly associated with metastasis and clinical outcome of patients.

Methods: We conducted genome-wide copy number profiles in 56 primary ccRCC including 32 metastasized and 24 non-metastasized tumours by array CGH with a median resolution of 2Mbp. The median follow up of the patients was 47 months.

Results: Array CGH identified 6 recurrent chromosomal aberrations: gains of 1q21.3, 12q13.11, 12q13.2 and 20q11.21q13.2 and losses of 8p11.23p12 and 9p21.3p24.1, which were significantly associated with metastasis occurrence. In multivariate analysis including these aberrations gains of 1q21.3 and of 20q11.21q13.2 and loss of 9p21.3p24.1 attained as independent predictors for metastasis in our cohort with 81.5% sensitivity and 95.8% specificity of 81.5% and 95.8%, respectively. Kaplan-Meier survival analysis showed that gains on chromosomes 7, 12, 16 and 20q and losses on chromosome 9 have a significant correlation with disease specific survival. Multivariate Cox-regression analysis revealed gains of 7q36.3 and of 20q11.21q13.2 and a loss of 9p21.3p24.1 as independent prognostic factors for patients' outcome of patients.

Conclusions: Our data suggest that specific common copy number alterations in ccRCC could serve as independent predictors for metastasis occurrence and cancer specific survival in patients with ccRCC.

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 2246.
Abstract 225: Cytotoxic extracts of ethnomedicinal plants Podophyllum peltatum (Mayapple) and Echinacea angustifolia
Authors: McDowell Olena T. , Myles Elbert L.
Published: February 2011

Phytochemicals and other natural compounds are historically established as cures for many ailments in rural America and developing countries. Our lab is testing plants that show potential for inhibiting the growth of cancer cells. The family Asteraceae and the family Berberidacae are very well known for there values as medicinal plants. More specifically, the genus species Echinacea angustifolia and genus species Podophyllum peltatum commonly known as the Mayapple, both are noted for there unique medicinal purposes. The crude methanolic extracts of these plants were tested against human metastatic cell lines representing breast (MCF-7),(BT-549), and (BT-20) tissues. Trypan blue exclusion is used to determine viable cell counts. Alamar BlueTM and florescent analysis were used to evaluate the cytoxicity of the extracts. Analysis of the crude extracts of Echinacea angustifolia has shown on MCF-7, BT-549 and BT-20 that most of them exhibit anticancer activity against the cell lines at the maximum concentration of 200 g /mL. In particular, less diluted concentrations have demonstrated exceptional inhibitory effects with IC50 values ranging between 0.10-15 g /mL, respectively. On the other hand, analysis of the crude extracts of Podophyllum peltatum (Mayapple) has shown on MCF-7, and BT-549 some anticancer activity against cell lines at the maximum concentration of 200 g /mL. More specifically, the more diluted concentrations have demonstrated exceptional growth inhibition with IC50 values ranging between 2-8g /mL

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 225.
Abstract 2258: Targeting MACC1: In vivo imaging of distant metastases reduction by MACC1-shRNA in human colon cancer xenografts
Authors: Stein Ulrike S. , Sack Ulrike , Kobelt Dennis , Kelch Inken , Fichtner Iduna , Walther Wolfgang , Schlag Peter M.
Published: February 2011

We identified the previously undescribed gene MACC1 by a genome-wide search for differentially expressed genes in human colon cancer tissues, metastases and normal tissues. MACC1 plays an important role for metastasis prediction, in particular for early-stage patients. Based on MACC1 mRNA expression in not (yet) metastasized primary colon cancers of stages I, II and III, our negative and positive prediction of the development of metachronous distant metastases was correct to 80% and 74%, respectively. The 5-year-survival for subjects suffering from colon cancer was 80% for MACC1 low expressors, but 15% for individuals with high MACC1 expression in their primary tumors. Moreover, we found MACC1 to act as an inducer of migration, invasion and proliferation in cell culture, and of distant metastases in several xenograft models. MACC1 transcriptionally regulates the receptor tyrosine kinase Met, that activates the HGF/Met signaling pathway resulting in enhanced cell motility, invasion and metastasis. Here we report the reduction of metastasis by using shRNA targeting the metastasis-inducer MACC1 in vivo. MACC1 high expressing human colon cancer cells were transfected with MACC1-shRNA, Met-shRNA, or control-shRNA harboring plasmids, together with luciferase-containing constructs. Cell clones were intrasplenically transplanted into mice. Tumor growth and metastases formation was demonstrated by in vivo imaging. Signals in MACC1-shRNA/luc mice and in Met-shRNA/luc mice originated exclusively from spleen tumors, not from liver metastases, as demonstrated with the isolated organs by luminescence. By contrast, signals in control-shRNA/luc mice originated from spleen tumors and liver metastases. These findings were confirmed by brightfield images, demonstrating reduced tumor growth and metastases development in MACC1-shRNA/luc and Met-shRNA/luc mice compared to control-shRNA/luc mice. In summary, knock down of MACC1 by MACC1-shRNA led to reduction of tumor growth and metastasis in mice, making MACC1-shRNA a useful tool for metastasis intervention of colon cancer.

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 2258.
Abstract 2259: Blockade of CXC chemokines/CXCR2 axis inhibits pancreatic ductal adenocarcinoma progression through anti-angiogenesis
Authors: Ijichi Hideaki , Tada Motohisa , Miyabayashi Koji , Imai Mitsuho , Mohri Dai , Asaoka Yoshinari , Ikenoue Tsuneo , Tateishi Keisuke , Omata Masao , Moses Harold L.
Published: February 2011

Pancreatic ductal adenocarcinoma (PDAC), one of the most lethal neoplasms, is characterized histologically by expanded stromal components with marked fibrosis, which suggests that tumor-stromal interaction might play an important role in PDAC progression.

We have established pancreas epithelium-specific transforming growth factor-ß receptor type II (Tgfbr2) knockout mice in the context of active Kras (KrasG12D) expression (Kras+Tgfbr2KO) (Ijichi et al., Genes Dev. 2006), which developed differentiated PDAC with 100% penetrance and recapitulated histology of human PDAC described above.

In the AACR annual meeting 2009, we reported that tumor-stromal interaction in the PDAC model promoted tumor progression and blockade of CXC chemokines/CXCR2 axis inhibited tumor growth and extended survival of the mice.

We examined the underlying mechanisms of anti-tumor effects. Treating the Kras+Tgfbr2KO PDAC mice with the CXCR2 inhibitor decreased CTGF expression and macrophage infiltration in vivo, resulting in decrease of microvessel density and inhibiton of tumor size. The CXC chemokines released from the PDAC cells, CTGF, macrophage, are all considered to promote angiogesis. Taken together, treatment of the mice with the CXCR2 inhibitor achieved anti-tumor effects through anti-angiogenesis via modulating network of an array of components in the tumor microenvironment. On the other hand, treating the mice with gemcitabine, a standard chemoreagent for PDAC, also showed anti-tumor effects, but it was not through anti-angiogenesis.

PDAC is well known as a hypovascular tumor, however, angiogenesis might be important for tumor progression and could be a potent therapeutic target. Pursuing optimum combination of anti-angiogenesis and a conventional chemotherapy would provide us a more effective therapeutic strategy for PDAC.

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 2259.
Abstract 2261: Antitumor and anti-inflammatory effect of a novel nutrient mixture on human lymphoma cell line U-937
Authors: Roomi M. Waheed , Roomi Nusrath M. , Rath Matthias , Niedzwiecki Aleksandra
Published: February 2011

Introduction: Phytochemicals and dietary antioxidants are known to decrease the risk of inflammation and prevent cancer development. We have developed strategies to inhibit cancer progression and its spread by natural products. A nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract has exhibited anticancer activity in vitro and in vivo in a number of cancer cell lines.

Objective: We investigated the effect of NM on human lymphoma cell line U-937 in vitro by measuring: cell proliferation, MMP expression, invasion, apoptosis, and Cox-2 and Cox-1 protein expression.

Materials and Methods: Human lymphoma cell line U-937 (ATCC) was cultured in RPMI medium supplemented with fetal bovine serum and antibiotics. U-937 cells were seeded on 24-well tissue culture plates with 40nM phorbol 12-myristate 13-acetate (PMA). After 24 hrs, the cells were treated with NM at 0, 50, 100, 250, 500 and 1000 mcg/ml, in triplicate at each dose. Cell proliferation was evaluated by MTT assay, MMP expression by gelatinase zymography, invasion through Matrigel, apoptosis by using live green caspase detection kit (Molecular Probe), and COX-2 and COX-1 expression by Western blot.

Results: NM was not toxic to U-937 cells at a concentration of 250 mcg/ml and exhibited an antiproliferative effect at 500 mcg/ml concentration. Zymography demonstrated only one band corresponding to MMP-9, which was inhibited by NM in a dose-dependent manner. Matrigel invasion was significantly reduced (by 95%) at 250 mcg/ml NM and completely blocked at 500 mcg/ml NM. NM induced slight apoptosis at 100 mcg/ml and moderate at 500 and 1000 mcg/ml concentration. NM inhibited Cox-2 expression in a dose-dependent fashion and had no effect on Cox-1 expression.

Conclusions: Our results suggest that NM is an excellent candidate for therapeutic use in the treatment of hematological malignancies.

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 2261.
Abstract 2265: Expression of hyaluronan in human cancer tissue: A predictive biomarker for PEGPH20-mediated therapy
Authors: Jiang Ping , Kadhim Salam , Li Xaioming , Thompson Curtis , McMannus Craig , Osgood Ryan , Anderson Jennifer , Symons Rebecca , Fieser Gail , Bahn Jesse , Tammi Raija , Shepard Michael , Frost Gregory
Published: February 2011

Background: Hyaluronan (HA), a ubiquitous, high-molecular-mass polysaccharide in the precellular and extracellular matrix, is frequently elevated in malignancy, and associated with aggressive disease. Interaction of HA and tumor cells may activate receptor tyrosine kinases, enhance drug resistance, and facilitate tumor progression. A pegylated recombinant human PH20 hyaluronidase (PEGPH20) has been developed to systemically target tumors that accumulated with HA. Systemic PEGPH20 administration in mice bearing PC3 tumors induced a decrease in tumor interstitial fluid pressure (IFP), a decrease in tumor water content, and an increase in tumor vascular volume by enzymatically depleting the HA from the extracellular tumor microenvironment and resulted in significant tumor growth inhibition. PEGPH20 therefore represents an innovative potential treatment approach that may provide improved therapeutic outcomes for cancer patients.

Methods: The present study investigated the expression of HA in paraffin embedded human tissue from 9 tumor types as well as corresponding normal tissue by immunohistochemical staining of BioMax tissue arrays with biotinylated HA binding protein (HABP) as a probe. Staining intensity was quantified using Aperio Spectrum plus software. The correlation between HA expression and PEGPH20 response has also been explored using established preclinical models.

Results: Strong HA expression was found in 87% of pancreatic, 64% of breast, 50% of multiple myeloma, 46% of NSCLC, 39% of gastric, and 22% of prostate cancer tissues, when compared with corresponding normal tissue (p<0.05). BxPc3 (HA high), MIA PaCa-2 (HA low) and AsPC-1 (HAlow) pancreatic tumor xenografts were evaluated for tumor growth inhibition (TGI) after systemic PEGPH20 treatment and TGI's of 25%, 9% and 15% were observed respectively. In addition, 3 prostate cancers, PC3 (HA high), MatLylu (HA high) and Du145 (HA low) were also evaluated in xenografts with significant growth inhibition shown for PC3 (72%) and MatLylu (34%). Systemic PEGPH20 treatment also induced tumor growth inhibition (60%) for 4T1 (HA high) breast cancer.

Conclusion: Accumulation of HA compared to normal tissue was commonly observed and most frequent in pancreatic adenocarcinomas (87%). Furthermore, accumulation of HA was shown to predict response to PEGPH20 therapy in xenograft models (R2=0.805, p<0.01). This work provides strong rationale for exploring efficacy of PEGPH20 in clinical trials.

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 2265.
Abstract 2269: M402 - A novel heparan sulfate proteoglycan mimetic targeting tumor-host interactions
Authors: Zhou He , Roy Sucharita , Cochran Edward , Zouaoui Radouane , Karlgren Juliane , Dussault Nancy , Kwan Rain YQ , Duffner Jay , Zhao Ganlin , Smith Sean , Galcheva-Gargova Zoya , Moy Erick , Barnes Marishka , Honan Chris , Chu Chia Lin , Qi Yi Wei , Ganguly Tanmoy , Schultes Birgit , Venkataraman Ganesh , Kishimoto Takashi Kei
Published: February 2011

Heparan sulfate proteoglycans (HSPGs) play important roles in tumorigenesis by mediating tumor-stromal interactions through the presentation of growth factors, cytokines, and chemokines critical for tumor progression, survival and metastasis. M402 is a rationally engineered, non-cytotoxic HSPG mimetic, designed to disrupt tumor-host interactions. M402 binds and inhibits multiple factors including VEGF, FGF2, SDF-1α, and P-selectin. A single 20 mg/kg subcutaneous (s.c.) dose of M402 effectively reduced seeding of B16F10 murine melanoma cells to the lung in a syngeneic experimental metastasis model. Chronic administration of M402, alone or in combination with cisplatin or docetaxel, inhibited spontaneous metastasis of orthotopically implanted 4T1 murine mammary carcinoma in this model. M402 treatment also normalized circulating levels of GR1+ immature myeloid cells and platelet counts in 4T1 metastatic tumor bearing mice. Fluorescently-labeled M402 exhibited selective accumulation in the primary tumor. Immunohistological analyses of primary tumor presented a decrease in microvessel density in M402-treated animals, suggesting anti-angiogenesis may be one of the mechanisms involved in vivo. Importantly, M402, as monotherapy or in combination with chemotherapeutics, also revealed significant survival benefits in this aggressive tumor model. These data demonstrate that targeting HSPG biology may provide a useful approach to attenuate multiple pathways involved in tumor progression and metastasis.

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 2269.
Abstract 227: Antitumor activity of glyceollins through induction of apoptosis
Authors: Kim Hyo Jung , Seo Jiyeon , Lim Ji Sun , Park Jia , Nam Dae Hwan , Garcia Annalene , Kim Jong-Sang
Published: February 2011

Glyceollins, which are synthesized from daidzein in soybean infected with fungi, have been shown to have anti-fungal effects and cancer preventive properties. However, the effects of glyceollins on apoptosis of cancer cell are not well known. We have investigated the antitumor effects of glyceollins on murine hepatoma cell lines. Cell viability was tested with a MTT assay. In Hepa1C1C7, glyceollins markedly reduced cell viability which was due to apoptosis. Glyceollins were observed to induce apoptosis of Hepa1C1C7 cells as evidenced by cell-morphological changes, an early redistribution of plasma membrane phosphatidylserine, increases in membrane leakage (propidium iodide) staining), Annexin-V binding, sub G1 phase in the cell cycle, and DNA laddering. Western blot analysis showed that upon treatment of tumor cells with the glyceollins, cleavage of pro-caspases-3 and significant release of mitochondrial cytochrome c into the cytosol were readily observed. This study demonstrated that glyceollins caused cell death via an apoptotic pathway, which might be exploited for prevention of tumor 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 227.
Abstract 2270: Design, synthesis, and functional characterization of new cycle-peptides inhibitors for C-X-C chemokine receptor-4 (CXCR4)
Authors: Scala Stefania , Portella Luigi , Napolitano Maria , Consales Claudia , Polimeno Maria Neve , D'Alterio Crescenzo , Palma Giuseppe , Arra Claudio , Vitale Rosamaria , Amodeo Pietro , Luca Stefania De , Monfrecola Luca , Castello Giuseppe
Published: February 2011

Modulation of the axis CXCR4/CXCL12 affected primary tumor growth and secondary lesions development. Moreover, CXCR4 inhibition increases hematopoietic stem cell mobility in patients undergoing autologous transplantation for myeloma and non-Hodgkin lymphoma. Although several CXCR4 inhibitors were described neither one reached satisfactory efficacy, bioavailability and acceptable toxicity. To develop new CXCR4 inhibitors through a rationale design approach, comparative studies were undertaken evaluating the N-terminal structure of the ligand CXCL12 and v-MIP II, an inhibitory chemokine like produced by the herpes virus 8-Kaposi associated. A common amino acidic motif (RFF) was identified in both structures although in inverted orientation. Having as a core this small domain, a new library (20 units) of cycle-peptide molecules was generated. 20 units of cycle-peptide molecules were synthesized that consists of 5 and 7 amino-acid residues cycled by a S-S bridge designed to interact with the receptor CXCR4. The peptides were characterized for functionally CXCR4 interaction through: 1. inhibition of CXCR4 binding; 2. inhibition of migration CXCL12-induced; 3. inhibition of calcium release CXCL12-induced; 4. Inhibition of P-Erk CXCL12-induced. Four peptides were identified as CXCR4 inhibitors. These four CXCR4 inhibitory peptides were further evaluated for the in vivo efficacy through inhibition of metastasis formation. B16 mouse melanoma cells transfected with CXCR4 were injected in the vein tail in C57 Bl mice and peptide treatment followed for 10 days. Dramatic reduction in number and size of lung metastases were registered in mice peptides-treated. Moreover, xenograft of human renal cancer cells SN12C-EGFP were subcutaneously injected and systemic peptides treatment followed for 10 days. Statistically significant decrease in tumor growth was showed in peptides treated mice. Taken together the data define four new CXCR4 inhibitory peptides, designed through innovative strategy, effective in “in vivo” systems deserving further development.

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 2270.
Abstract 2277: Anti-tumor effect of an apoptosis-targeting peptide-conjugated heparin derivative in breast cancer xenografts
Authors: Bae Sang Mun
Published: February 2011

Anti-tumor effect of an apoptosis-targeting peptide-conjugated heparin derivative in breast cancer xenografts.

Sang Mun Bae1, Jong-Ho Kim1, Kai Wang1, Sungwoo Chung2, Youngro Byun2, Byung-Heon Lee1, In-San Kim1 and Rang-Woon Park1

1Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea

2College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea

Abstract

Angiogenesis, the development of new blood vessels, is an essential process during tumor growth and metastasis. In the process of tumor angiogenesis, tumor cells secret various angiogenic growth factors such as vascular endothelial growth factor (VEGF), fibroblast growth factor basic (bFGF), platelet-derived endothelial cell growth factor (PDGF), insulin-like growth factors (IGF-1 and IGF-2) and scatter factors, which are potential targets for development of angiogenesis inhibitor and depend on heparin sulfate for their biological functions. Angiogenic endothelial cells are a central challenge for therapeutic intervention since its easy accessibility. Recently, researchers have sought to develop angiogenesis inhibitor for tumor therapy since tumor cannot grow without blood-supplied oxygen and nutrients. Heparan sulfate is located in cell surface and in the extracellular matrix and plays pivotal regulatory roles in growth factor. For this reason, heparin has been highlighted as a growth factor inhibitory therapeutic agent beyond its anticoagulant indications. In a previous study, we reported new angiogenesis inhibitor, low molecular weight heparin derivative (heparin-taurocholic acid conjugate; HT), showed delay of in vivo tumor growth in MDA-MB231 human breast cancer. In this study, we sought to design a new angiogenesis inhibitor targeting tumor tissue by introducing the actively targeting moiety, tumor homing peptide, to previously developed HT. Here, we evaluate the inhibitory effect of HT-Apopep by in vivo antitumor efficacy and show the body distribution image of HT-Apopep in tumor-bearing mice.

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 2277.
Abstract 2284: Dissecting signaling networks involved in the tumor-microenvironment crosstalk in chronic lymphocytic leukemia (CLL)
Authors: Bhattacharya Nupur , Caudron-Herger Maiwen , Busch Hauke , Rippe Karsten , Doehner Hartmut , Stilgenbauer Stephan , Mertens Daniel
Published: February 2011

Crosstalk between tumor cells and the microenvironment is often a prerequisite for carcinogenesis. CLL, the most common leukemia in the western world, is an excellent model system to investigate this microenvironmental crosstalk. The dependence of the CLL cells for pro-survival support from the microenvironment is apparent from the induction of rapid spontaneous apoptosis in CLL cells in vitro unless they are cultured on feeder cells or treated with microenvironmental cytokines. CLL cells are also capable of inducing other non-malignant microenvironmental cells to form a pro-survival niche.

We have used a tripartite strategy to characterize the interaction between CLL cells and their microenvironment. Firstly, we have quantified the interaction of pro-survival ligand-receptor pairs and their dose-response curves in primary CLL cells and non-malignant B cells. Of the ligand-receptor pairs tested, IL4 and CD40L were found to rescue primary CLL cells most effectively from apoptosis. The effector dependence of IL4 and CD40L displayed different saturation curves in CLL versus non-malignant B cells, pointing towards distinct differences in ligand-receptor interactions. Secondly, we have used a systems biology approach to identify central nodes in the survival network induced by the microenvironment. The model is generated from the kinetics of the transcriptome acquired from gene expression profiling CLL cells after microenvironmental stimulation. Via a procedure that involves gene-ranking, significance filtering and kinetic classification, deregulated genes have been identified. Model construction using these genes will yield central nodes which will be functionally validated on a cohort of patients. As a third approach, the pro-survival niche inducing property of CLL cells on other microenvironmental cells is studied by gene expression profiling differentiated monocytes, which arise when monocytes are cocultured with CLL cells (which support CLL survival) versus the differentiated monocytes which arise when cultured with non-malignant B cells. Analysis of the gene expression profiles revealed genes specifically deregulated due to the effect of CLL cells.

This study will give us a deep insight into the signaling networks operating between the tumor and its microenvironment. This is certainly important for the identification of drug targets which can abrogate this interaction and thereby reduce the viability of the CLL cells in vivo.

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 2284.
Abstract 2287: Tumors induce complex DNA damage in distant proliferative tissues in vivo
Authors: Redon Christophe E. , Dickey Jennifer S. , Nakamura Asako J. , Kareva Irina G. , Naf Dieter , Nowsheen Somaira , Kalogerinis Peter K. , Bonner William M. , Georgakilas Alexandros G. , Sedelnikova Olga A.
Published: February 2011

The influence of a tumor on its surrounding tissues has been extensively investigated, but whether a tumor can influence distant tissues in an organism is unknown. This question is important in understanding the relationship between cancer and overall patient health. To explore this issue we chose to investigate the DNA damage endpoint, prompted by its utility in defining the “bystander effect” which denotes the phenomenon that cells exposed to ionizing radiation (or other stresses) communicate with their neighbors resulting in the appearance of DNA damage in unstressed cells. We measured DNA damage in tissues proximal and distal to the tumor site in C57BL/6 and BALB/c mice with early-stage subcutaneous syngeneic grafts, namely, B16 melanoma, MO5076 sarcoma, and COLON26 carcinoma. When the tumors approached 100 mg, the animals were sacrificed and various organs examined. Samples of duodenum, colon, rectum, stomach, liver, kidney, lung, ovary, skin, splenic and blood lymphocytes were taken for analysis. We report that levels of two types of DNA damage, double-strand breaks (DSBs) measured by γ-H2AX focus formation and oxidatively-induced non-DSB clustered DNA lesions (OCDLs), were elevated in tissues distant from the tumor site in the tumor-bearing mice compared to their age- and sex-matched controls. The most affected tissues were crypts in the gastrointestinal tract organs and the skin, particularly the hair follicles. These tissues have high rates of proliferation, indicating that S-phase cells were the most vulnerable to influence from the distant tumor. This study is the first direct demonstration of the tumor-induced bystander effect in vivo, i.e. that the presence of a tumor impacts distant tissues, and suggests that tumors may constitutively promote complex oxidative DNA damage in highly proliferating tissues throughout an organism. These findings may have important implications for the study of genomic instability resulting from a primary tumor as well as for understanding second cancers which are a growing concern as long-term survival after a first cancer increases. This study also suggests a role for DNA damage markers as general cancer-related stress biomarkers.

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 2287.
Abstract 2297: Transcriptome profiling of invasion phenotype associated with epithelial-mesenchymal transition in esophageal cancer
Authors: Li Hsiao-Fang , Peng Talun , Cheng Ann-Joy
Published: February 2011

The esophageal cancer (ESC) is one of the ten-leading cancers in Taiwan with highly metastatic potential. The aim of this study was to clarify relevant alterations of gene expression associated with the invasive phenotype of ESC. To reduce heterogeneity and to obtain data on genes specifically involved in invasive mechanism, we established a highly invasive ESC subline (CE48) through 10 passages of in vitro Matrigel selections. This subline exhibited increases of invasion and migration abilities with the ascending generations, in accompany of morphological alteration from epithelial type to mesenchymal type, suggesting an epithelial- mesenchymal transition (EMT) was occurred during this Matrigel selection. In support of this finding, five molecular EMT markers were also changed accordingly. Affymetrix cDNA microarray was used to compare the transcriptomes between the parental and the EMT-associated invasion subline. Heretical cluster and algorithmic analyses of the microarray dataset revealed that several functional pathways associated with these phenotypic alterations. On the other hand, 10 genes were examined for the potential differential expressions between parental and invasion subline by RT-PCR or western blot analysis. In which, TKT, FLNA, FN1, Gp96, Slug and IQGAP1 were found over-expressed in the invasion subline, suggesting a positive association in the regulation of EMT-associated invasion. Knockdown of TKT by siRNA further demonstrated suppression of cell invasion. Therefore, the EMT-associated invasion sublin and transcriptome profiling of the cells were determined. Results of this study provide molecular knowledge in the EMT-associated metastasis in ESC.

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 2297.
Abstract 2298: BP1 upregulates Twist, a trigger of EMT and cancer metastasis
Authors: Berg Patricia E. , Hwang Bin-Jin , Rheey Jinguen
Published: February 2011

BP1 Upregulates Twist, a Trigger of EMT and Cancer Metastasis

Bin-Jin Hwang, Jinguen Rheey, and Patricia E. Berg

George Washington University, Department of Biochemistry and Molecular Biology,

Washington, D.C.

Background. We have cloned a gene, BP1, which is a member of the homeobox gene family of transcription factors. Increasing evidence suggests that homeobox genes are important in malignant transformation and tumorigenesis, and our recent studies have shown that BP1 may play a role in breast cancer cell survival, aggressiveness and metastasis. BP1 protein (pBP1) is expressed in 80% of invasive ductal breast tumors, and is associated with breast cancer aggressiveness: 100% of estrogen receptor negative tumors and 89% of the tumors of African American women are BP1 positive. Now we have evidence that BP1 may regulate the epithelial to mesenchymal transition (EMT). EMT, a process in which cancer cells lose their epithelial features and gain mesenchymal markers, enables tumor cells to become more invasive and migratory, and can lead to metastasis. BP1 upregulates Twist, a basic helix-loop-helix (bHLH) transcription factor which triggers EMT. Our objective is to determine whether BP1 plays a role in induction of EMT. Materials and Methods. The expression levels of Twist, E-cadherin and vimentin were observed on microarrays after probing with RNA from MCF-7 cell lines overexpressing BP1. RNA levels were verified by real-time PCR and protein levels by Western blotting and, in some cases, by confocal microscopy. Summary. Twist mRNA expression on microarrays is higher in MCF-7 cells overexpressing BP1 compared with cells containing an empty vector. Real-time PCR confirmed that Twist mRNA is increased five-fold in cells with high levels of BP1, and Western blots demonstrated that Twist protein is also elevated in the same cells. We have discovered a potential DNA binding site for pBP1 in the Twist promoter, suggesting that BP1 may directly regulate Twist. This will be verified by ChIP. Expression of two genes regulated by Twist was also examined: Twist activates vimentin and represses E-cadherin. Cells overexpressing pBP1 also show increased vimentin and decreased E-cadherin. The effect of increased pBP1 on cell migration is under investigation. Conclusions: BP1 stimulates Twist expression in MCF-7 breast cancer cells, resulting in increased vimentin and decreased E-cadherin levels. We therefore hypothesize that BP1 is upstream of Twist and induces EMT. If BP1 is required for EMT, it may be a good target for therapy of metastatic tumors.

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 2298.
Abstract 2301: Cross-species expression profiling identifies novel genes differentially modulating epithelial-mesenchymal transition program and cell migration/invasion capability
Authors: Zhang Haijun , Wu Sherwin , Wu Feng , Ethier Stephen P. , Miller Fred , Wu Guojun
Published: February 2011

Despite remarkable advances in research and clinical management, metastatic breast cancer remains incurable and causes most deaths in human breast cancer patients due to the unavailability of specific, targeted therapy. Therefore, identification of functional metastasis genes and further unveiling the molecular mechanisms underlying metastasis process remains the top priority in cancer research field. In recent years, much evidence supports the notion that epithelial-mesenchymal transition (EMT), a developmental program involved in a variety of tissue remodeling events, contributes to cancer cell plasticity and to the invasive potential of transformed epithelial cells. However, the legitimacy of EMT is still challenged by disagreements among pathologists who dismiss EMT as a fallacy.

Utilizing a cross-species expression profiling strategy which combined microarray technique and model cell lines, we identified 22 up-regulated and 12 down-regulated genes as an essential genetic program in both human and mouse highly-metastatic model cell lines. About half of the gene candidates are novel and their roles in metastasis have not been reported. We randomly selected four genes and investigated their roles in metastatic characteristics including EMT program and cell migration and invasion. Overexpression of GCNT2 gene (an up-regulated gene candidate) not only led to a marked increase of cell migration and invasion, but also led to marked alterations in epithelial-mesenchymal transition (EMT) program. Ectopic expression of MAPK12 (another upregulated gene) only changed the EMT program but has no effect on cell migration and invasion. Overexpression of IRX5 (a down-regulated gene) not only led to a marked decrease of cell migration and invasion in vitro, but also a modulation of EMT program. However, expression of PHLDA3 (another down-regulated gene) only affect the cell migration and invasion, but has no effect on EMT program. These results highlight the feasibility of the cross-species expression profiling approach as a strategy to identify gene targets that are involved in breast cancer metastasis. The results of the functional studies for randomly selected genes further suggest that this multigenic program is consisted of different genes that play various roles in controlling EMT and/or Cell migration and invasion ability. Further studies are needed to investigate the effect of these novel gene candidates on metastasis in vivo and as therapeutic targets for treatment of breast cancer metastasis.

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 2301.