bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2764: High-resolution HLA typing by next-generation sequencing and risk of follicular lymphoma
Authors: Akers Nicholas K. , Conde Lucia , Smith Martyn T. , Bracci Paige M. , Skibola Christine F.
Published: June 2012

The molecular basis of follicular lymphoma (FL) has been fairly well characterized, although its root causes remain less clear. Recently, in two genome-wide association studies of European-Americans, we identified novel genetic susceptibility loci for FL, narrowed down to two independent regions on chromosome 6p21.3 in the major histocompatibility complex (MHC) (Skibola et al., Nat Gen 41(8):873-5., 2009; Conde et al., Nat Gen 42(8):661-4, 2010). The first susceptibility locus, rs6457327 (combined allelic p-value=4.7 × 10-11 in 645 patients and 3,377 controls), is part of a 26-kb region of high linkage disequilibrium (LD) in the MHC Class 1 region at 6p21.33 in close proximity to the psoriasis susceptibility region 1 (PSORS1). A second locus at 6p21.32, in the MHC Class II region upstream of HLA-DQB1, two SNPs in high LD, rs10484561 and rs7755224, were associated with 2-fold increased risks for FL in 1,465 patients and 6,958 controls (combined p-values=1.12×10-29, 2.00×10-19). Using tag SNPs as surrogates for HLA allelotypes, the latter loci were found to be part of an extended haplotype that includes HLA-DRB1*0101-HLA-DQA1*0101-HLA-DQB1*0501 and that was associated with increased FL risk [odds ratio (OR) = 2.07, 95% confidence interval (CI) = 1.40-3.06]. These findings were further validated by direct typing of HLA-DQB1 alleles in a subset of 265 FL cases and 757 controls using multiplexed, ligation-dependent probe amplification. We verified the positive association between FL and the HLA-DQB1*05 allele group (OR=1.56, 95% CI 1.22-1.99; adjusted p-value=7.7×10-3), and found an inverse association between the HLA-DQB1*06 allele group and FL risk (OR=0.53, 95% CI 0.40-0.69; adjusted p-value=1.98×10-5).

To gain further insight into the role of HLA alleles in FL susceptibility, high-resolution HLA typing by next-generation sequencing of the HLA class I (A, B, C) and class II (DRB1/3/4/5, DQA1, DQB1, DPB1) genes was conducted in 222 FL patients and 220 sex- and age-matched controls. HLA typing was performed by sequencing exon 2 for the class II genes and exons 2-4 for the class I genes, as described previously (Bentley, G et al. Tissue Antigens. 74:393-403, 2009), and data analysis is underway. To our knowledge, this study will present the highest resolution (up to 8-digit) HLA typing reported in a sample of European-Americans, and is an important next step to determine the role of MHC genetic variation in the pathogenesis of FL.

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 2764. doi:10.1158/1538-7445.AM2011-2764
Abstract 2769: Evaluation of PPP2R2A as a prostate cancer susceptibility gene: Comprehensive germline and somatic study
Authors: Cheng Yu , Liu Wennuan , Sun Jishan , Kim Seong-Tae , Lu Lingyi , Sun Jielin , Zheng Siqun , Isaacs William B. , Xu Jianfeng
Published: June 2012

PPP2R2A, a gene mapped to 8p21.2, encodes α isoform of the regulatory B55 subfamily of the protein phosphatase 2 (PP2A). PP2A is one of the four major Ser/Thr phosphatases, and is implicated in the negative control of cell growth and division. Because of its known functions and its location within a chromosomal region where evidence for linkage and somatic loss of heterozygosity were found, we hypothesized that either germline sequence variants or somatic copy number changes in PPP2R2A may increase prostate cancer (PCa) risk. To address this, we first sequenced PPP2R2A in 96 probands of hereditary prostate cancer (HPC) families recruited at John Hopkins Hospital; each family had at least three first-degree relatives affected with prostate cancer. Twenty-five germ-line variants were identified in this analysis, including 15 known SNPs and 10 new sequence variants. Most new sequence variants were rare, except 2 new sequence variants with minor allele frequency (MAF) ≥ 0.05 (−482T/C MAF=0.072 and -78C/G MAF=0.058). Thereafter, it was less likely that these sequence variants of PPP2R2A would be co-segregated with PCa. Second, to see if there are any genetic variants in PPP2R2A associated with sporadic PCa, we analyzed 10 SNPs of 15 known SNPs that span the complete PPP2R2A gene in Cancer Genetic Markers of Susceptibility (CGEMS, 737 cases and 1,105 controls) and in Cancer of the Prostate in Sweden (CAPS, 498 cases and 494 controls) from a Sweden population, as genotypes of these 10 SNPs had already been available in both studies. No consistent significant differences in the allele and genotype frequencies were observed among these sporadic PCa cases and controls. Finally, we examined the deletion status in 141 clinical PCa samples from John Hopkins using Affymetrix 6.0 SNP arrays. With the allele-specific analysis of intensities from all paired tumor and normal tissue samples, it was found that PPP2R2A was commonly (67.1%) deleted in these tumor samples including a homozygous deletion in 3 tumors (2.1%). These results suggest that somatic deletion rather than germline sequence variants of PPP2R2A may play a more important role in prostate cancer susceptibility.

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 2769. doi:10.1158/1538-7445.AM2011-2769
Abstract 2775: Meat intake, genetic variation in carcinogen metabolic enzymes, and prostate cancer risk in a multiethnic case-control study of non-Hispanic Whites, African Americans and Hispanics
Authors: Joshi Amit D. , Corral Roman , Catsburg Chelsea , Koo Jocelyn , John Esther M. , Ingles Sue A. , Stern Mariana C.
Published: June 2012

Background: The role of meat and meat-induced carcinogens in prostate cancer (PCa) is still inconclusive. The process of cooking meat increases the content of chemical carcinogens such as heterocyclic amines (HCAs) and polycyclic aromatic hydrocarbons (PAHs), which are metabolized to active carcinogens, or detoxified by metabolic enzymes in the liver and the prostate.

Study objectives: We tested whether diets high in red or white meats are associated with PCa risk, taking into account cooking practices, and whether polymorphisms in carcinogen metabolism genes modify these associations.

Methods: We used data and DNA samples from the California Collaborative Prostate Cancer Study, a multiethnic, population-based study of non-Hispanic White, African American and Hispanic men (717 localized PCa cases, 1,140 advanced PCa cases, and 1,096 controls). We considered nutrient density (g/ 1000 kcal / day) adjusted variables that captured intake of red meat, poultry, and processed meat, taking into account cooking methods, levels of doneness, and estimated meat mutagens, obtained using the CHARRED database. We considered the following polymorphisms: CYP1A2 -154 A>C, CYP1B1 L432V, CYP2E1 -1054 C>T, COX2 -765 G>C, EPHX1 Y113H, GSTP1 I105V, UGT1A6 T181A and the NAT2 predicted phenotype. Conditional logistic regression was used to estimate odds ratios for PCa risk adjusting for age, body mass index, total calorie intake, family history of PCa, saturated fat intake, pack-years of cigarette smoking, total fruit consumption and total vegetable consumption.

Results: Diets high in baked poultry, were inversely associated (Ptrend = 0.028), whereas diets high in pan-fried red meat (Ptrend = 0.040) and well done red meat (Ptrend = 0.016) were positively associated with advanced PCa risk. Estimated meat mutagens (PhIP, MeIQX) were positively associated with advanced PCa risk only among Hispanic men (Ptrend = 0.022 and 0.04, respectively). The UGT1A6 T181A polymorphism modified the associations of localized PCa risk with consumption of high temperature cooked, well done, or oven broiled poultry and with estimated PhIP, suggesting positive associations between these exposures and localized PCa risk only among carriers of 1 or 2 copies of the Alanine allele.

Conclusions: Diets high in red or white meat cooked with high temperature methods may be modifiable risk factors for advanced PCa, and risk factors for localized disease among carriers of selected metabolic enzyme variants.

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 2775. doi:10.1158/1538-7445.AM2011-2775
Abstract 278: Role of AF4 and Dot1 in MLL-AF9 mediated leukemogenesis
Authors: Lumba Bhavna , Hemenway Charles S.
Published: June 2012

Mixed lineage leukemia (MLL) is characterized by rearrangement of the MLL gene at chromosome band 11q23. MLL gene rearrangements give rise to chimeric proteins that contain the N-terminal portion of MLL fused to the C-terminal portion of over 50 different fusion partners. MLL chimeras do not retain the histone methyltransferase activity of wild type MLL and it is unclear how they deregulate the expression of MLL target genes. Our laboratory studies the two most common MLL fusion partners encountered in patients with leukemia, AF4 and AF9. We have previously shown that AF4 and AF9 heterodimerize and peptides that block this interaction are toxic to MLL leukemia cell lines. This supports the hypothesis that the interaction of MLL-AF9 with AF4 is required for leukemogenesis. However, we have also shown that AF4 and H3K79 methyltransferase, Dot1 have similar binding sites on AF9. Furthermore, we have found that Dot1 is required for immortalization of hematopoietic progenitor cells by MLL fusion genes. The work of Yokoyama and others demonstrated that AF4, ENL (a close homologue of AF9) and Dot1L do not exist in the same multimeric complex. To understand the role of AF4 and Dot1 in MLL-AF9 mediated leukemogenesis, we made point mutants of AF9 with differential binding for AF4 and Dot1. Based on our preliminary data and recent studies, we hypothesize that the binding of AF4 and not Dot1 to MLL-AF9 is required for MLL-AF9 mediated leukemogenesis. Nevertheless, Dot1 remains a critical factor in MLL-AF9 leukemia. We hypothesize that Dot1 recruitment is mediated by wild type AF9 rather than MLL-AF9.

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 278. doi:10.1158/1538-7445.AM2011-278
Abstract 2791: SINE retrotransposons cause epigenetic reprogramming of adjacent gene promoters
Authors: Estecio Marcos R.H , Dekmezian Mhair , Gallegos Juan , Liang Shoudan , Issa Jean-Pierre
Published: June 2012

Retrotransposons of SINE and LINE classes have been extremely successful in colonizing mammalian genomes. Almost half of the human genome and as much as 40% of the mouse genome is composed of repetitive DNA sequences. The comparison of human and mouse genomes revealed a spatial concordance in positioning of SINE but not LINE repeats, indicating evolutionary pressure to maintain and/or exclude these repeats from orthologous regions. We hypothesized that this phenomenon is due to a negative influence of SINE repeats on gene expression and counter selection over evolution, resulting in exclusion of these repeats from vulnerable genomic environments. To test the capacity of SINE B1 elements in promoting gene silencing, we generated a system where the luciferase gene is under the control of three different mouse gene promoters (cdkn2a, p14arf and mlh1), and we inserted two and four copies of SINE B1 elements upstream to these promoters. Using this in vitro system, we found that SINE B1 elements can influence the activity of downstream gene promoters, with acquisition of DNA methylation and loss of activating histone marks, thus resulting in a repressed state. Interestingly, SINE sequences themselves do not immediately acquired DNA methylation, but became marked by H3K9me2 and H3K27me3. Moreover, gain of DNA methylation depended on cryptic genomic methylation centers rather than spreading from SINE B1 repeats. Since mouse SINE B1 elements caused transcriptional repression, we sought to investigate whether human SINE Alu repeats could have the same effect. The human E-cadherin gene has three SINE Alus adjacent to the 3’ region of its CpG island, and in reporter assays the removal of these repeats resulted in higher, stable promoter activity. Genome-wide analysis of SINE repeats distribution in the human and mouse genomes showed that they are excluded from proximal promoters and their enrichment is directly correlated with the presence of USF1, USF2 and CTCF binding, proteins with insulator function. In conclusion, our work supports the concept that SINE repeats interfere negatively with gene expression and that their presence near gene promoters is dependent on, and co-evolved with the presence of insulator elements.

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 2791. doi:10.1158/1538-7445.AM2011-2791
Abstract 2800: BAY 94-9392 – A new derivative of L-glutamate for tumor specific PET imaging
Authors: Koglin Norman , Mueller Andre , Friebe Matthias , Berndt Mathias , Schmitt-Willich Heribert , Gekeler Volker , Dinkelborg Ludger M.
Published: June 2012

There is a high demand for a tumor specific imaging, staging and therapy monitoring by PET. Currently, FDG (2-[F-18]fluoro-2-deoxyglucose) is the commonly used agent for tumor detection and staging. However, this method cannot differentiate between tumor and inflammatory lesions. In addition, FDG does not accumulate in tumors with low glycolytic activities, such as prostate cancer. It is not optimal for specific detection of brain tumors and metastases due to a high background signal resulting from healthy brain metabolism. The amino acids L-glutamine and L-glutamate as well as their corresponding transporters play a key role in the adapted intermediary metabolism of tumors. We synthesized and evaluated a new PET imaging agent BAY 94-9392, a derivative of L-glutamate. This agent was successfully used for tumor detection with high sensitivity and specificity as demonstrated in various preclinical models.

Methods: The radiosynthesis of (4S)-4-(3-[F-18]fluoropropyl)-L-glutamate (BAY 94-9392) was established on a commercial synthesis module. BAY 94-9392 was investigated in a variety of tumor cell lines as well as tumor models for uptake, biodistribution, mode of action, and its potential to differentiate between tumor versus inflammation.

Results: BAY 94-9392 is specifically transported by the cystine/glutamate exchanger (xCT, SLC7A11) and it is not metabolized. Compared to FDG, the agent displayed a high accumulation rate in a panel of tumor cells. PET imaging with excellent tumor visualization and high tumor to background ratios was achieved in a variety of preclinical tumor models. BAY 94-9392 did not accumulate in inflammatory lesions, whereas FDG shows a strong uptake in this turpentine oil induced and histopathologically confirmed rat model.

Conclusions: BAY 94-9392 is a new, promising tumor specific PET tracer. Both preclinical and clinical studies are in progress for further characterization.

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 2800. doi:10.1158/1538-7445.AM2011-2800
Abstract 2803: The role of the sarcosine pathway in prostate cancer progression
Authors: Khan Amjad P. , Rajendiran Thekkelnaycke M. , Ateeq Bushra , Asangani Irfan , Yocum Anastasia K. , Mehra Rohit , Sreekumar Arun , Chinnaiyan Arul M.
Published: June 2012

Differential metabolomic alterations occur during prostate cancer progression. Exploring the metabolome of prostate cancer progression, may lead to the identification of critical biomarkers for cancer invasion and disease aggressiveness. Recently, we have identified sarcosine, an N-methyl derivative of the amino acid glycine, as a key metabolite increased most robustly in metastatic prostate cancer and detectable in the urine of men with organ-confined disease (Sreekumar et al., Nature, 2009). Here, we examined if the proximal regulatory enzymes of sarcosine, glycine-N-methyl transferase (GNMT), sarcosine dehydrogenase (SARDH) and pipecolic acid oxidase (PIPOX), play a functional role in prostate cancer progression. By tissue microarray analysis, we observed that GNMT protein levels are strongly associated with prostate cancer aggressiveness. Here, we demonstrate that stable knockdown of GNMT inhibits cell proliferation, induces apoptosis, attenuates cell invasion in Matrigel coated transwells, and blocks the anchorage-independent growth of the cancerous DU145 cells. Overexpression of GNMT in benign RWPE cells showed significantly increased cell invasion and increased sarcosine level. Importantly, knockdown of GNMT in DU145 cells also inhibited intravasation using chick chorioallantoic membrane (CAM) assay and tumor growth in xenograft assays. In contrast, we also showed that while the knockdown of sarcosine degrading enzymes, SARDH and PIPOX, induced invasion and increased sarcosine level in RWPE cells, the overexpression of these enzymes attenuated invasion and decreased sarcosine level. Further, overexpression of SARDH inhibited cell proliferation, blocked the anchorage-independent growth and also inhibited intravasation using CAM assay. Taken together, this study shows that the components of the sarcosine pathway may have potential as biomarkers of prostate cancer progression and serve as new avenues for therapeutic intervention.

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 2803. doi:10.1158/1538-7445.AM2011-2803
Abstract 2809: Pathway dependence on the tyrosine kinase TYK2 in T-cell acute lymphoblastic leukemia
Authors: Sanda Takaomi , Tyner Jeffrey W. , Gutierrez Alejandro , Ngo Vu N. , Moriggl Richard , Ahn Yebin , Glover Jason , Chang Bill H. , Willis Stephanie G. , Zhou Wenjun , Gray Nathanael S. , Staudt Louis M. , Druker Brian J. , Look A Thomas
Published: June 2012

To identify novel oncogenic pathways that are dysregulated in T-cell acute lymphoblastic leukemia (T-ALL), we carried out an RNA interference (RNAi) screen using a retroviral library of inducible short-hairpin RNAs (shRNAs) in T-ALL cell lines. We found that loss of TYK2, a JAK family tyrosine kinase, was lethal in each of three T-ALL cell lines that we tested in this screen. An independent RNAi screen using a small-interference RNA (siRNA) library that individually silences each member of the tyrosine kinase gene family in primary T-ALL cells from a pediatric T-ALL specimens also demonstrated a dependence on TYK2 for cell viability. We confirmed by targeted knock-down analysis with multiple independent shRNAs that the loss of TYK2 induces apoptosis in T-ALL cells, whereas knock-down of other JAK proteins (JAK1, JAK2 or JAK3) had no effect. The TYK2 protein is constitutively phosphorylated on tyrosine residues in many T-ALL cell lines, and sequence analysis of the TYK2 gene in T-ALL cell lines and patient samples revealed a diversity of TYK2 point mutations in the FERM domain (V15A), pseudo-kinase domain (V731I) and kinase domain (E957D and R1027H). TYK2 cDNAs harboring each of these mutations, but not wild-type TYK2 cDNA, could transform Ba/F3 cells to factor-independent growth, and TYK2 was heavily phosphorylated in the Ba/F3 cell lines transformed with mutant cDNAs, indicating that these mutations activated the TYK2 kinase. TYK2-dependent T-ALL cell lines and the transformed Ba/F3 cells were sensitive to small-molecule JAK/TYK2 inhibitors. Moreover, we identified that inhibition of TYK2 induces downregulation of STAT1 phosphorylation, and that loss of STAT1 inhibits the growth of TYK2-dependent T-ALL cells, indicating that this transcription factor is involved in the TYK2 pathway and required for cell survival. Gene expression analysis after TYK2 or STAT1 knock-down in TYK2-dependent T-ALL cell lines revealed that BCL2 is upregulated by this pathway. In conclusion, our results establish a requirement for the TYK2-STAT1 pathway to promote T-ALL cell the survival, and establish TYK2 as a novel therapeutic target for this disease.

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 2809. doi:10.1158/1538-7445.AM2011-2809
Abstract 2813: Serum biomarker panel detects lung cancer in never smokers
Authors: Birse Charlie , Laiger Robert , Bruce Robert , Tomic Jennifer , Ruben Steve , Lenk Thomas
Published: June 2012

Introduction: Lung cancer is the leading cause of cancer mortality worldwide with more than 1 million deaths each year. Although most lung cancers are attributable to smoking tobacco, it is estimated that as many as 25% of all lung cancer cases are in subjects who have never smoked. A number of clinical, epidemiological, and molecular variations suggest that lung cancers that arise in smokers and non-smokers are significantly different. Biomarker panels may have considerable value when combined with imaging protocols in detecting and diagnosing lung cancer. We previously employed a novel mass spectrometry-based approach to identify serum biomarkers which we have previously shown to detect non-small cell lung cancer (NSCLC) in a smoking population representing all 4 stages of disease. In this study we extend these findings to a cohort of lung cancer subjects who have never smoked.

Methodology: Initially 9 biomarkers were assayed in serum collected from smoking subjects with NSCLC and appropriate controls. More than 600 specimens collected from 4 independent sites were employed in the study. Samples were randomly divided into a training set (NSCLC n=128, controls n = 191) and a testing set (NSCLC n=141, controls n=175) and used to develop a regression-based algorithm for lung cancer detection. Subsequently, an independent validation study was undertaken in cohort of lung cancer subjects who had never smoked (interview questionnaire). All stages of cancer (stage I n = 8, stage II n = 4, stage III n = 17, stage IV n = 11) and all major histological cell types (adenocarcinoma n = 21, squamous n = 7, bronchioloalveolar = 8, others n=4) were included. Controls were matched on age/gender (n=40).

Results: A global 6-marker regression model identified smoking associated cancer cases with good performance (Training AUC=0.877; Testing AUC=0.868). All stages of cancer were distinguished as well as all of the major histological cell types. Fitting of the model to data from the never smoker cohort revealed that the algorithm again discriminated the malignant cases with strong performance AUC=0.906 (sensitivity = 83% at specificity = 83%).

Conclusion: Lung cancer biomarkers identified initially through proteomic analysis have shown robust performance in a study cohort of lung cancer subjects who have never smoked. The findings from these studies suggest that these biomarkers may provide suitable performance across all lung cancer populations to design tests for a variety of diagnostic applications. For example, it is possible that these biomarkers could be employed to enhance the discrimination of malignant nodules identified by radiologic imaging.

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 2813. doi:10.1158/1538-7445.AM2011-2813
Abstract 2832: Development of AGS-22M6E, a novel antibody drug conjugate (ADC) targeting Nectin-4 for the treatment of solid tumors
Authors: Satpayev Daulet , Torgov Michael , Yang Peng , Morrison Karen , Shostak Yuriy , Raitano Art , Liu Wendy , Lortie Dawn , An Zili , Capo Linnette , Leavitt Monica , Perez Myra , Verlinsky Alla , Shirasuna Kenna , Avina Hector , Guevara Claudia , Morrison Kendall , Challita-Eid Pia , Jia Xiao-Chi , Gudas Jean , Stover David
Published: June 2012

Nectin-4 is a type I transmembrane antigen identified by Agensys to be highly expressed at the RNA level in a significant number of bladder and breast cancer specimens. Analysis of protein expression carried out in an extensive panel of tumor specimens by immunohistochemistry confirmed strong to moderate expression levels of Nectin-4 in bladder, breast and pancreatic cancers (40-60%). A subset of lung and ovarian cancers (up to 30%) also showed significant expression levels of this protein. We have generated an antibody-drug conjugate (ADC) targeting Nectin-4, AGS-22M6E. The highly potent microtubule disrupting drug monomethyl auristatin E (MMAE) was conjugated via valine-citrulline cleavable linker to a fully human IgG1k antibody specific for Nectin-4. AGS-22M6E was characterized for binding affinity, species crossreactivity and cytotoxicity in vitro. AGS-22M6E binds to cell surface Nectin-4 with high affinity and crossreacts with Nectin-4 orthologs of cynomolgus monkey, rat and murine origin. Upon binding to its target on the cell surface AGS-22M6E is internalized and induces cell death in vitro in a dose dependent manner. In vivo efficacy of AGS-22M6E was tested in SCID mice on xenograft models of breast, bladder, pancreatic and lung cancers in subcutaneous and orthotopic modalities. Treatment of well established tumor xenografts with the ADC resulted in significant growth inhibition or complete eradication of tumors in various models covering multiple cancer indications. Pharmacokinetics of AGS-22M6E in non-tumor bearing mice was also evaluated and will be discussed. Overall, these data validate AGS-22M6E as an attractive therapeutic drug candidate for multiple solid tumor indications and support further clinical investigation.

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 2832. doi:10.1158/1538-7445.AM2011-2832
Abstract 2840: Efficacy of radiation therapy in mammary carcinomas is regulated by macrophages
Authors: Shiao Stephen L. , DeNardo David G. , Fujikawa Kerri A. , Faddegon Bruce A. , Park Catherine C. , Coussens Lisa M.
Published: June 2012

In patients with locally advanced breast cancer, radiation therapy (RT) and chemotherapy following surgery has been a mainstay of treatment with demonstrated survival advantage in numerous randomized trials. However, despite recent advances in treatment, many women still ultimately succumb to disease highlighting the need to improve therapeutic strategies. Results from several groups have recently revealed that the therapeutic effects of RT are in part dependent on activation of immune-mediated mechanism(s); however, little is known regarding the mechanistic details of this activation. To address this, we examined changes in the immune microenvironment of mammary carcinomas following RT. We found that following RT (5 Gy), the density of infiltrating macrophages was significantly increased (20 vs 35%, p = 0.02) as compared to mammary tumors in mice not receiving RT. When macrophages were depleted following RT, via use of a CSF-1 antagonist, a significant delay in tumor growth, as compared to tumors treated with RT alone was observed. Since this data indicated that macrophage infiltration blunted RT responsiveness, we hypothesized that reprogramming tumor-infiltrating macrophages more towards a classical cytotoxic phenotype would result in enhanced RT response and delayed tumor re-growth. Thus, we treated tumor-bearing mice with anti-IL-4 neutralizing mABs in combination with RT, and evaluated tumor re-growth, as compared to tumor-bearing mice treated with either RT or anti-IL4 mAB alone, and found that indeed, reprogramming tumor-infiltrating macrophages to favor classical (M1)-type activity significantly enhanced RT response and delayed tumor re-growth. Together these studies indicate that: 1) RT induces macrophage infiltration into mammary tumors, 2) protumor-type macrophages infiltrating mammary carcinomas blunt response to RT, and 3) reprogramming of tumor-infiltrating macrophages to favor a cytotoxic phenotype improves efficacy of RT.

This work was supported by grants from the NIH/NCI, and Era of Hope (W81XWH-06-1-0416), and Program in Mesothelioma (PR080717) grants from the Department of Defense to LMC.

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 2840. doi:10.1158/1538-7445.AM2011-2840
Abstract 2848: SHLP6: A naturally occurring mitochondrial-derived peptide has therapeutic potential in prostate cancer
Authors: Cobb Laura J. , Nakamura Hiromi , Cohen Pinchas
Published: June 2012

Mitochondrial DNA (mtDNA) mutations have been demonstrated in prostate cancer, particularly at the 16S rRNA region; and abnormal mRNA transcripts from that locus have been described. We and others showed that a novel open reading frame (ORF) within the mitochondrial 16S rRNA region, encoding for a 24 AA peptide called humanin, is a potent cell-survival and metabolic factor.

Hypothesizing that additional bioactive peptides may be encoded from this region we identified an additional six ORFs that encode peptides transcribed from within the 16s rRNA gene in the mtDNA, which we have named small humanin-like peptides, or SHLPs. SHLP1-5 are functionally analogous to humanin, and act as potent survival factors.

Intriguingly an additional peptide, SHLP6, functions in a completely opposing manner. We demonstrate that endogenous SHLP6 is expressed in serum and in multiple tissues, including the prostate, and that its expression declines with age. In addition, the addition of exogenous SHLP6 promotes apoptosis in prostate cancer cell lines including 22RV1, LNCaP and DU145. To assess the in vivo potency of SHLP6, we treated 22RV1 xenografts in SCID mice with SHLP6 for 1 week, and observed a potent inhibition of CaP xenograft growth and angiogenesis in vivo. In addition, we show that SHLP6 expression is decreased in preliminary staining of human prostate cancer, compared with normal prostate tissue.

Analysis of gene and protein expression of SHLP6 treated cells reveals modulation of cell cycle and apoptosis genes and a dramatic inhibition of VEGF expression. Importantly, treatment of prostate cells with siRNA against SHLP6 leads to a dramatic increase in VEGF expression, suggesting that endogenous SHLP6 plays a role in the regulation of VEGF levels. We therefore propose the revolutionary concept that a previously unrecognized family of peptides is produced from the mitochondria and that one of these, SHLP6, plays a key role in regulating angiogenesis, is altered in prostate cancer, and may serve as a novel therapeutic and diagnostic agent in this disease.

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 2848. doi:10.1158/1538-7445.AM2011-2848
Abstract 2864: Targeting cathepsin S induces tumor cell autophagy via the EGFR-ERK signaling pathway
Authors: Chen Kuo-Li , Chang Wun-Shaing Wayne , Lin Chun-Cheng , Cheung Chun-Hei Antonio , Kuo Chang-Po , Kuo Ching-Chuan , Chang Yi-Hsun , Liu Ko-Jiunn , Chang Jang-Yang
Published: June 2012

Cathepsin S is a vital cellular cysteine protease that is frequently amplified and over-expressed in various human cancers. In this study, we report that targeting cathpesin S could induce tumor cell autophagy via the EGFR-ERK signaling pathway. Cancer cells treated with cathepsin S inhibitors [α-ketoamide inhibitor (6r), Z-FL-COCHO (ZFL)] and cathepsin S-specific siRNA induced autophagy as indicated by an increase in the cleavage of the microtubule-associated protein light chain 3B (LC3B) and the formation of membrane-bound autophagic vacuoles (AVOs). In addition, co-treatment of a specific inhibitor of autophagy, 3-methyladenine (3MA), inhibited the 6r-induced autophagy in cancer cells. Further Western blot analysis revealed that targeting cathepsin S induced cancer cell autophagy through the activation of EGF receptor and its downstream MAPK-related signaling pathways. The induction of autophagy by targeting cathepsin S subsequently leads to the activation of apoptosis as indicated by both the up-regulation of caspase-9/3 activity, the down-regulation of Bcl-2, Bcl-XL and the altered mitochondrial membrane retention potential in cells. The application of the autophagy inhibitor, 3MA, was able to inhibit the process of apoptosis induced by the cathepsin S inhibitor 6r in cancer cells. In conclusion, the current study reveals that cathepsin S inhibitor is able to induce cancer cell autophagy through the EGF receptor signaling pathways, which may provide therapeutic benefit in cancer patients who are less sensitive to apoptosis-inducing 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 2864. doi:10.1158/1538-7445.AM2011-2864
Abstract 2867: Docosahexaenoic acid induces autophagy through p53/AMPK/mTOR signaling in human cancer cells
Authors: Jing Kiapeng , Song Kyoung-Sub , Shin Soyeon , Kim Kim Nayeong , Jeong Soyeon , Heo Jun-Young , Park Ji-Hoon , Seo Kang-Sik , Kwon Hyun-Joo , Park Hae-Duck , Han Chang , Wu Tong , Park Jong-Il , Park Seung-Kiel , Kweon Gi-Ryang , Yoon Wan-Hee , Hwang Byung-Doo , Lim Kyu
Published: June 2012

Although Omega 3-polyunsatuarated fatty acids (Omega 3-PUFAs) induce cytotoxicity in several cancer cell lines, the exact mechanisms are not identified yet. In this study, we showed that autophagy is involved in the Omega 3-PUFAs-induced cytotoxicity in cancer cells. Autophagy is characterized by the sequestration of cytoplasmic material within autophagosomes for bulk degradation by lysosomes. We found that docosahexaenoic acid (DHA), an Omega 3-PUFA, induced not only apoptosis but also autophagy in cancer cells. Autophagy was detected after DHA exposure as indicated by induction of LC3 expression, and formation of autophagic vacuolization. We observed that the DHA-induced autophagy was accompanied by loss of p53. Inhibition of p53 by Pifithrin-α or microRNA-p53 significantly increased the DHA-induced autophagy, suggesting that the DHA-induced autophagy is mediated by downregulation of p53. Further experiments showed that the mechanism of the DHA-induced autophagy associated with p53 attenuation, involved an increase in the active form of AMPK which attenuated the mTOR activity as revealed by p27 sequestration. In addition, compelling evidence for the interplay between autophagy and apoptotic cell death induced by DHA is supported by the findings that autophagy inhibition partially decreased the DHA-induced apoptotic cell death and further autophagy induction by p53 inhibitor enhanced apoptosis in response to treatment with DHA in cancer cells. Our results demonstrate that autophagy may be related to the DHA-induced cytotoxicity in cancer cells. [This work was supported by basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (E00026 and R13-2007-020-01000-0].

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 2867. doi:10.1158/1538-7445.AM2011-2867
Abstract 2873: The pandeacetylase inhibitor panobinostat induces the involvement of autophagy related factors in liver cancer cell death
Authors: Fazio Pietro Di , Montalbano Roberta , Quint Karl , Ocker Matthias
Published: June 2012

Background. Liver cancer still represents an unmet medical need with rising incidence rates also in Western countries. Panobinostat (LBH589, Novartis Oncology), a pandeacetylase inhibitor, represents a new agent with a promising future in liver cancer therapy. We have previously demonstrated that panobinostat is able to induce cell death in two liver cancer cell lines, HepG2 and Hep3B, through activation of ER stress related apoptotic pathways (Cell Oncol 2010;32:285-300). Panobinostat also significantly reduced the growth of HepG2 xenografts in nu/nu mice. We here demonstrate the implication of autophagy mechanisms into cell demise mediated by panobinostat.

Materials and Methods. HepG2 (p53 wt) and Hep3B (p53 null) were treated for 72 hours with panobinostat and cell death was quantified through FACS with propidium ioded staining and CK18 fragmentation staining. Impedance based real-time cell analysis was performed to continuously monitor cell viability after panobinostat treatment. The expression of autophagy related factors was analyzed through RT-qPCR, western blot and immunofluorescence based cytochemistry.

Results. Panobinostat induces cell death in a dose and time dependent manner. Autophagy related factors Atg5, Beclin and its activators Ambra, p62 and UVRAG were up-regulated after 48 hours of treatment. We also observed also significant increase of p73 expression in Hep3B cells after 24 hours of treatment. Immunofluorescence based cytochemistry showed a modification of the ubiquitous cytosolic distribution of Beclin and LC3B in both cell lines to a defined spot formation after treatment with panobinostat, indicating a probable autophagosome formation. Western blot analysis also demonstrated an increase of Beclin and LC3-I and LC3-II.

Conclusion. Panobinostat treatment determines the involvement of autophagy related mechanisms in a ER stress related apoptosis scenario. The wide spectrum of mechanisms of action of panobinostat need to be further investigated.

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 2873. doi:10.1158/1538-7445.AM2011-2873
Abstract 2883: Vimentin downregulation modulates p53 levels and apoptosis in cancer cells
Authors: Fortier Anne-Marie , Cadrin Monique , Asselin Eric
Published: June 2012

Vimentin, a mesenchymal-specific intermediate filament (IF) protein, is often express in epithelial cancers exhibiting a de-differentiated phenotype. Vimentin is associated with increased chemoresistance and metastatic potential. Chemoresistance is a multifactorial phenomenon which is a major cause of treatment failure in human cancer. Some IF proteins are known to promote resistance to stress-induced apoptosis, but the role of vimentin in this process is still unknown. In the present study, we investigate the role of vimentin in response to apoptosis-induced chemotherapeutic drugs in different epithelial carcinoma cell lines. Surprisingly, using siRNA, our results show that vimentin downregulation decreases caspase-3 and PARP cleavage induced by cisplatin, doxorubicin and TNFalpha. This unexpected result could be explained by the decrease of p53 protein levels in vimentin-downregulated cells. Indeed, we observed that vimentin knockdown decreases p53 levels in a proteasome-dependent manner and also modulates p53 ubiquitination. PI3K/Akt pathway is well known to promote chemoresistant phenotype in different cancers and Akt can also regulate p53 function. Our results show that vimentin-induced p53 decrease is associated with an increase of Akt phosphorylation. We propose that vimentin knockdown results in an increase of Akt activity which in turn regulates ubiquitination and proteasomal degradation of p53. We have recently showed that Akt isoforms regulates vimentin and keratins IF expression in epithelial cancer cells, suggesting that IF proteins are directly involved in PI3K/Akt signaling pathway. To our knowledge, these results represent the first indication that vimentin could regulate Akt activity and p53 levels in response to apoptosis-induced chemotherapeutic drugs. These results support the hypothesis that modulation of IF expression plays an active role in cancer progression and that vimentin could be a possible target for cancer therapy.

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 2883. doi:10.1158/1538-7445.AM2011-2883
Abstract 2889: NF-κB inducing kinase modulates melanoma tumorigenesis by regulating pro-survival factors through β-catenin
Authors: Thu Yee Mon , Su Yingjun , Yang Jinming , Splittgerber Ryan , Richmond Ann
Published: June 2012

Nuclear factor-κB (NF-κB) contributes to the progression of many cancers due to its role in regulating the transcription of genes important for inflammation, proliferation, apoptosis, and survival. In melanoma, high NF-κB activity results from constitutive elevation of IKKβ activity, a major kinase of this pathway. Targeted deletion of IKKβ prevents Ras induced melanoma tumor formation in INK4a/ARF-/- mice, underscoring the importance of NF-κB in melanoma. We have demonstrated that the NF-κB inducing kinase (NIK) is elevated in many melanoma cell lines. To determine the role of NIK overexpression in melanoma, we knocked down NIK using shRNA in WM115 and Hs294T human melanoma cell lines. NIK knock-down melanoma cells exhibited increased apoptosis and decreased proliferation. Similarly, NIK depletion substantially decreased tumor burden of WM115 cells in a xenograft model. Analysis of gene expression microarray data suggests that NIK modulates expression of c-Myc, c-Met, CCND2 and CXCR4, genes which are crucial for melanoma progression. Consistent with the apoptotic phenotype of NIK knock-down melanoma cells, expression of survival factors, such as Bcl2, cIAP1 and survivin, were attenuated in NIK knock-down melanoma cells. Intriguingly, Bcl2 and survivin expression levels (mRNA and protein) were also diminished in NIK knock-out mouse embryonic fibroblasts (MEFs), suggesting that these pro-survival genes are under the regulation of NIK. However, NF-κB activity is not reduced in NIK knock down cells, possibly due to the compensatory activation of IKKβ. Thus, we explored the possibility that NIK affects expression of survival factors in a NF-κB independent manner. Our data suggest that NIK regulates subcellular localization of β-catenin and modulates expression of survival genes such as survivin. Our findings reveal a novel mechanism by which NIK modulates pro-survival genes and suggest that NIK may be a promising therapeutic target for melanoma malignancy.

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 2889. doi:10.1158/1538-7445.AM2011-2889
Abstract 290: Use of serum and tissue biomarker analysis embedded in a phase II clinical trial of cytarabine in castration-refractory prostate cancer to investigate prostate cancer biology
Authors: Dhani Neesha , Emmeneger Urban , Sridhar Srikala , Knox Jennifer , Tannock Ian , Day John , Manthe Jonathon , Groskopf Jack , Joshua Anthony M.
Published: June 2012

Background: Other than the androgen receptor, the TMPRSS2-ERG genomic aberrations in prostate cancer provide the first recent opportunity to target therapy in castration refractory prostate cancer (CRPC). We initiated a phase 2 clinical trial of cytarabine in docetaxel refractory CRPC on the basis of microarray, in vitro and case report evidence that cytarabine may be particularly effective in men harbouring abnormalities of the ERG oncogenes. Embedded in this clinical trial was the first use of blood mRNA levels of prostate cancer related genes as biomarkers of response and prognosis. Methods: Patients with docetaxel refractory progressive CRPC received intravenous cytarabine at doses between 1g/m2- 0.25 g/m2 q3 weekly. Responses were defined according to PCWG2C. 10 patients were enrolled between June 2007 and January 2010. TMPRSS2:ERG, PSA and PCA3 mRNA copies in whole blood collected with PAXgene tubes at the beginning of each cycle and at trial termination were quantified using transcription-mediated amplification assays. The prototype TMPRSS2:ERG assay detects the gene fusion isoform TMPRSS2 exon1 to ERG exon4. Results: No patients demonstrated a serum PSA response (PCWG2C). The average number of cycles administered was 2.6. Significant toxicities including grade 3-4 thrombocytopenia (2) and grade 3-4 neutropenia (3). These toxicities necessitated several dose reductions in the protocol, however most patients were removed from trial for serum PSA progression alone. PCA3 and PSA mRNAs were detectable in 8/10 and 9/10 cases, respectively; there was no correlation between serum PSA and PCA3 or PSA mRNA copy levels in blood. Testing for TMPRSS2:ERG in blood was able to predict the presence or absence of the TMPRSS2-ERG rearrangement in 9/10 cases when compared to 3 colour FISH carried out on baseline biopsies/ prostatectomies (2/10 positive for Exon 4:Exon 1 deletion). Conclusions: Cytarabine administation is ineffective in docetaxel refractory CRPC. Blood mRNA levels of prostate cancer genes reveal novel aspects of prostate cancer biology and have implications for the understanding of circulating tumour cells

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 290. doi:10.1158/1538-7445.AM2011-290