bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 87: SULF2 methylation is a prognostic biomarker for lung cancer survival and increases sensitivity to camptothecin analogues via expression of ISG15
Authors: Tessema Mathewos Z. , Yingling Christin M. , Thomas Cynthia L. , Klinge Donna M. , Bernauer Amanda M. , Stidley Chris , Joste Nancy , Dahlberg Suzanne , Schiller Joan , Belinsky Steven A.
Published: June 2012

Heparan sulfate proteoglycans (HSPGs) serve as co-receptors for numerous signaling molecules including growth factors and cytokines. The sulfation pattern on the heparan sulfate (HS) motif is central to the interaction between HSPGs and the myriads of molecules including interferon (INF). SULF2 encodes for HS 6-O-endosulfatase enzyme that removes the 6-O-sulfate moieties from HS and regulates the specificity of this interaction. Increased expression of SULF2 promotes angiogenic and oncogenic signaling. In hepatocellular carcinoma higher expression of SULF2 is associated with worse prognosis after resection. Conversely, we recently reported that methylation-mediated SULF2 silencing is associated better survival of resected lung cancer patients. Here we show that SULF2 methylation is also a favorable survival marker in unresectable lung cancer. Furthermore, epigenetic silencing of SULF2 suppresses cell migration and induces the expression of multiple INF-inducible genes. Knocking down 80% of SULF2 expression in Calu-3 cells using siRNA resulted in 7-fold increase in the expression of INF-inducible regulator of ubiquitination, ISG15. Similarly, lung cancer cell lines with methylated SULF2 (SULF2M) on average express 60-fold higher ISG15 than cell lines with unmethylated SULF2 (SULF2U). ISG15 is a key marker for increased sensitivity of cancer cells to topoisomerase-1 inhibitors such as camptothecin and topotecan. In vitro treatment of lung cancer cell lines with camptothecin revealed a 65-fold higher sensitivity in cell lines with SULF2M and ISG15 high (ISG15H) phenotype compared to SULF2U and ISG15 low (ISG15L) cell lines. In nude mice, 10mg/kg/week topotecan treatment dramatically arrested the growth of SULF2M and ISG15H lung cancer xenografts but had no effect on SULF2U and ISG15L xenografts. Heterogeneity for ISG15 expression was also seen in primary tumors. One third of primary lung tumors have ISG15H phenotype with an average of 5-fold higher expression than the remaining 2/3rd tumors in which the ISG15 level is comparable to normal lung. Taken together these results indicate SULF2 methylation is not only a favorable prognostic marker for lung cancer, but through its association with increased expression of ISG15 could indicate a higher sensitivity to topoisomerase-1 inhibitors.

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 87. doi:10.1158/1538-7445.AM2011-87
Abstract 881: Polymorphisms of PUMA, a BH3-only pro-apoptotic gene, modify risk of squamous cell carcinoma of head and neck associated with HPV16 infection
Authors: Zhou ZiYuan , Li Guojun , Liu Zhensheng , Wang Li-E , Sturgis Erich M. , Wei Qingyi
Published: June 2012

PUMA (p53 upregulated modulator of apoptosis) and Noxa (Latin for damage), as well as their common target MCL-1 (myeloid cell leukemia 1), respond to human papillomavirus 16 (HPV16) oncoprotein E6-mediated p53 degradation or E7-mediated pRb inactivation, and therefore may alter cancer susceptibility through the PUMA (Noxa)-MCL1-apoptosis mechanism.

In this case-control study of 380 cases and 335 cancer-free matched controls, we evaluated HPV16 seropositivity and two single nucleotide polymorphisms (SNPs) in the promoter region for each gene (rs3810294 and rs2032809 for PUMA, rs4558496 and rs9957673 for Noxa, rs9803935 and rs3738485 for MCL1) to examine their possible associations with risk of squamous cell carcinoma of head and neck (SCCHN).

Of the 6 SNPs, both two SNPs of PUMA were significantly associated with 1.5-fold increased SCCHN risk in a dominant model. When we include the HPV16 seropositivity in the model, the risk of SCCHN was significantly increased in an allele-dose effect manner (Ptrend< 0.0001 for each SNP, respectively). Notably, in the site-stratified analysis, oropharyngeal cancer risk was dramatically increased, particularly among minor alleles carriers having the HPV16 L1 seropositivity with OR (95%CI) of 13.8 (5.4-35.5) for rs3810294 and 8.1 (4.3-15.0) for rs2032809 (both Ptrend < 0.0001). These data suggest some synergistic effects between PUMA variants and HPV16 seropositivity on susceptibility to SCCHN, especially in those cases with HPV-related oropharyngeal cancer. Additionally, the modifying effect of PUMA variants on oropharyngeal cancer associated with the HPV16 L1 seropositivity was more predominant in never-smokers, never-drinkers and young subjects. Our results suggest that PUMA promoter polymorphisms, synergistically interacting with HPV16 L1 seropositivity, modify risk of HPV-related oropharyngeal cancer. Further molecular and epidemiologic studies with larger samples are needed to validate our findings.

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 881. doi:10.1158/1538-7445.AM2011-881
Abstract 902: Serum antibodies to the HPV16 proteome as biomarkers for head and neck cancer
Authors: Anderson Karen S. , Wong Jessica , D'Souza Gypsyamber , Riemer Angelika B. , Lorch Jochen , Haddad Robert , Pai Sarah , Longtine Janina , Bryan Janine , McClean Michael , Labaer Joshua , Kelsey Karl , Posner Marshall R.
Published: June 2012

Purpose: Human papillomavirus (HPV) type 16 is associated with oropharyngeal carcinomas (OPC). Antibodies (Abs) to HPV16 E6 and E7 oncoproteins have been detected in patient sera, however Abs to other early HPV-derived proteins have not been well explored.

Experimental Design: Abs to the HPV16 proteome were quantified using a novel multiplexed bead assay, using C-terminal GST fusion proteins captured onto Luminex beads. Sera were obtained from two cohorts of untreated OPC patients with known Abs to HPV16 Virus-Like Particles (VLP) (N=10), HPV18 VLP (N=10), healthy controls (N=20), and from a validation cohort of untreated patients with OPC (N=30), partners of patients with HPV16+ OPC (N=11), and healthy controls (N=30).

Results: OPC patients in the known VLP+ cohort had elevated serum Abs to HPV16 E1, E2, E4, E6, and E7, and L1 antibody levels but not E5. The ratios of specific median fluorescent intensity (MFI) to p21-GST compared to controls were: E1 50.7 vs. 2.1; E4 14.6 vs. 1.3; E6 11.3 vs. 2.4; E7 43.1 vs. 2.6; and L1 10.3 vs. 2.6 (each p<0.01). No cross-reactivity was detected between HPV16 E7 and HPV18 E7 Abs. In the validation cohort, HPV16 E1, E2, and E7 antibody levels were most significantly elevated compared to healthy control samples (p<0.0001) and partners of OPC patients (p<0.01).

Conclusion: A novel multiplexed bead assay for the detection of HPV16-specific Abs demonstrates that patients with HPV16+ OPC have detectable Abs to multiple early genes, in particular E1, E2 and E7. These Abs are potential biomarkers for HPV-associated OPC.

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 902. doi:10.1158/1538-7445.AM2011-902
Abstract 912: The tumor promoting activity of the prostaglandin E2 EP4 receptor in mouse skin
Authors: Simper Melissa S. , Bowen Rebecca A. , Surh Inok , Fischer Susan M.
Published: June 2012

Chronic inflammation has recently been suggested to be involved in cancer development, and cyclooxygenase-2 (COX-2), a key mediator of the inflammatory response, is known to be upregulated in several cancers. COX-2 is responsible for the production of prostaglandins, and prostaglandin E2 (PGE2) is the major prostaglandin involved in inflammation. PGE2 binds to four receptors (EP1-EP4), and the EP4 receptor has recently been shown to be important in carcinogenesis, depending on the tissue in which it is expressed. To determine the role of EP4 in skin carcinogenesis, transgenic mice were generated that overexpress EP4 in the basal layer of the epidermis under the control of the bovine keratin 5 promoter (BK5.EP4 Tg). Using a two-stage chemical carcinogenesis protocol, the BK5.EP4 Tg mice were shown to develop both papillomas and squamous cell carcinomas earlier than wild type (WT) mice. However, a tumor observed before 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment on one BK5.EP4 Tg mouse suggested that TPA was not necessary for the promotion of the tumors. Therefore a 7,12-dimethylbenz[a]anthracene (DMBA)-only carcinogenesis protocol was performed, and by 30 weeks, the BK5.EP4 Tg mice developed more carcinomas compared to none on the WT mice. Immunohistochemistry performed on DMBA-treated mice demonstrated that the BK5.EP4 Tg epidermis began to detach 48 hours after treatment with total detachment occurring 5 days after treatment, while WT mice had only increased epidermal thickening. Caspase-3, MMP-9, and MMP-7 expression also were upregulated in the BK5.EP4 Tg mice at the same time detachment of the epidermis was occurring. Ki67 analysis showed an initial increase in proliferation in WT mice that slowly declined back to normal levels, while the BK5.EP4 Tg mice instead had a sustained increase in proliferation, which occurred during reepithelialization of the epidermis. These data suggest that overexpression of EP4 in the mouse skin can increase tumorigenesis, possibly through the wounding of the skin and the subsequent epidermal regeneration acting as a tumor promoter. Supported by CA100140.

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 912. doi:10.1158/1538-7445.AM2011-912
Abstract 930: Reprogramming the transcriptional response to hormone through functionally-distinct classes of gene enhancers in prostate cancer cells
Authors: Garcia-Bassets Ivan , Wang Dong , Benner Chris , Wenbo Li , Su Xue , Glass Christopher K. , Rosenfeld Michael G. , Fu Xiang-Dong
Published: June 2012

Mammalian genomes are populated with thousands of transcriptional enhancers that orchestrate cell type-specific gene expression programs; however, the potential that there are pre-established enhancers that permit alternative signal-dependent transcriptional responses has remained unexplored. Here we present evidence that cell lineage-specific factors, such as FoxA1, which contribute to the activation of cell-type specific enhancers by selectively favoring the recruitment of key regulated transcription factors such as the androgen receptor (AR) in prostate cancer cells, can simultaneously restrict these same factors from activating additional (alternative) pre-established but yet inactive enhancers, thus licensing specific signal-activated responses and restricting others. Importantly, these alternative hormonal responses are linked to distinct structural and functional classes of enhancers. Consequently, FoxA1 down-regulation, an unfavorable prognostic sign in some sets of advanced prostate tumors, causes a massive switch in AR binding from one functional enhancer class to another, which massively reprograms the hormonal response. The molecular basis for this switch lies in the release of FoxA1-mediated “restriction” of AR binding to the new enhancer class that requires no apparent nucleosome remodeling for their activation.

Multiple global analyses of enhancer formation, structure, looping, and activation were combined to support these conclusions, including: H3K4me1/me3, H3K27ac, and H4K4ac mapping (ChIP-seq); nucleosomal H3K4me2 mapping (nucleosomal ChIP-seq); FoxA1, AR, p300, and MED12 mapping (ChIP-seq), also supported by the 3C assay; enhancer ncRNA or eRNAs generation analysis (GRO-seq); and gene expression analysis (microarray and GRO-seq).

Together, these findings reveal a large repository of pre-determined enhancers in the human genome that can be dynamically tuned to induce alternative gene expression programs, which -we speculate- may underlie many sequential gene expression events in development or during cancer progression.

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 930. doi:10.1158/1538-7445.AM2011-930
Abstract 942: Protein kinase CK2 contributes to 1, 25-dihydroxyvitamin D3-mediated CYP24A1 gene expression in prostate cancer cells
Authors: Luo Wei , Karpf Adam R. , Chernov Mikhail Chernov , Johnson Candace , Trump Donald
Published: June 2012

The active vitamin D metabolite 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) plays a major role in regulating calcium homeostasis and bone mineralization. 1,25(OH)2D3 also modulates cellular proliferation and differentiation in a variety of cell types through the activation of vitamin D receptor (VDR), by both genomic and nongenomic pathways. 1,25-(OH)2D3 has anti-tumor effects in several tumor models. 24-hydroxylase, encoded by the CYP24A1 gene, plays a key role in degrading 1,25(OH)2D3. Suppression of CYP24A1 function enhances the antitumor activity of vitamin D. In order to isolate regulators of CYP24A1 expression in prostate cancer cells, we established the prostate cancer cell line PC3 stably transfected with CYP24A1 promoter driving luciferase expression and used this cell line to screen for compounds that inhibit CYP24A1 promoter activity, using a drug library. From this screening, we identified, 4,5,6,7-tetrabromobenzimidazole (TBBz), a protein kinase CK2 selective inhibitor as a disruptor of CYP24A1 promoter activity. We show that TBBz specifically inhibits CYP24A1 promoter activity induced by 1,25(OH)2D3 in prostate cancer DU145 and PC3 cells. In addition, TBBz downregulates endogenous CYP24A1 mRNA level in TBBz treated PC3 cells. Furthermore, siRNA-mediated CK2 knockdown reduces 1,25(OH)2D3 induced CYP24A1 mRNA expression in PC3 cells. This inhibitory effect on CYP24A1 expression by CK2 siRNA is enhanced by the addition of TBBz. These results suggest that CK2 signaling contributes to 1,25(OH)2D3 mediated induction of CYP24A expression. This study provides among the first line of evidence that CK2 is linked to vitamin D metabolism. Future studies will elucidate the mechanism(s) underlying the mechanism by which CK2 regulates CYP24A1 expression. In summary, our findings reveal that protein kinase CK2 is involved in the regulation of CYP24A1 expression by 1,25(OH)2D3, and highlight a novel strategy to facilitate the optimization of antitumor therapy of vitamin D.

This study was supported by NIH/NCI grants 5R01CA067267 and 5R01CA095045

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 942. doi:10.1158/1538-7445.AM2011-942
Abstract 98: Differential methylation of promoters in ductal adenocarcinoma and cancers of the ampulla vateri of the pancreas
Authors: Pilarsky Christian , Biewusch Katja , Heyne Marie , Grützmann Robert , Saeger Hans Detlev
Published: June 2012

Background: Prognosis for patients with pancreatic carcinoma (PDAC) remains poor. Despite of increasing knowledge about the molecular basis of PDAC neither a specific marker for early diagnosis nor a target protein for a new therapeutic approach have been identified so far. Determination of DNA methylation events is a promising technology for the early detection and classification of cancer. We were therefore interested to elucidate the significance of DNA methylation to distinct between ductal adenocarcinoma and cancers of the ampulla vateri (AVT) of the pancreas.

Methods: DNA isolation using the Qiamp mini kit and modification using the Zymo EZ DNA Methylation-Gold system was performed. Primers for methylation specific PCR (MSP) for RELN, ADAMTS8, SFRP1, BNIP3, ID4 and TFPI2 were selected from the literature and tested for their performance on bisulfite modified fully methylated human DNA. After successfully establishing the assays, DNA samples isolated from formalin fixed and paraffin embedded tissue from patients with diagnosed ductal adenocarcinoma (n = 14) and tumors of the ampulla vateri (n=7) were investigated. MSP was performed and the amount of produced fragment was scored semiquantitatively using agarose gelelectrophoresis.

Results: MSP analysis of the bisulfite modified DNA showed methylation differences in RELN (PDAC: 63%, AVT: 37%), ADAMTS8 (PDAC: 37%, AVT: 14%), BNIP3 (PDAC: 85%, AVT: 50%), SFRP1 (PDAC: 84%, AVT: 100%) and ID4 (PDAC: 93%, AVT: 62,5). Interestingly, for TFPI2 both tumors types showed nearly the same incidence of methylation events (PDAC: 86%; AVT: 87%).

Conclusion: Detection of DNA methylation is able to indentify differences between tumors of the pancreas. This might resemble crucial changes during tumor development, which are also reflected in gene expression and histopathologic appearance. For the establishment of a methylation-based discrimination of tumors further samples and promoters have to be tested.

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 98. doi:10.1158/1538-7445.AM2011-98
Abstract LB-106: Expression of stem cell biomarkers in benign breast tissue from patients with triple negative breast cancer
Authors: Atkinson Rachel L. , Yang Wei T. , Landis Melissa D. , Wong Helen , Creighton Chad , Hilsenbeck Susan G. , Lewis Michael T. , Woodward Wendy A. , Brewster Abenaa , Sahin Aysegul A. , Chang Jenny C.
Published: June 2012

Numerous studies suggest that breast cancers contain a subpopulation of cancer stem cells that are thought to maintain the heterogeneity and self-renewing potential of the tumor. In our previously published data, we had described a breast cancer stem cell signature comprised of 493 statistically significant differentially expressed genes, which were present in a tumorigenic subpopulation of cells. Cells from patient samples with the cell surface markers CD44+CD49f+CD133/2+ have been shown to have self-renewal and tumorigenic properties – characteristics of breast cancer stem cells. In this study, we investigated the benign tissue from 42 patients with breast cancer treated with mastectomy to examine the incidence CD44+CD49f+CD133/2+ positive cells. Benign tissue was obtained at a distant site, at least 5 cm from the primary tumor, and examined with hematoxylin-eosin by a breast pathologist (AS), only samples containing normal breast tissue with at least 60% epithelial content were selected. A significant increase in expression of CD44+CD49f+CD133/2+ by immunofluorescence was identified in the benign tissue associated with patients with triple negative (ER negative/PR negative/Her2 negative) disease (8 out of 8), compared with ER positive and Her2 positive tumors combined (5 out of 36), P<0.001, Wilcoxon rank sum test. The definition of a positive score was more then 1% positive staining or negative for less then 1% triple positive cells, scoring approximately 1000 cells per sample. We further examined whether global RNA expression from these benign tissues correlated to our previously published tumorigenic microarray signature. We found that array data from benign tissue from patients with triple negative breast cancer was significantly correlated with our previously published tumorigenic signature (P=.01 two sided t test). Thus, these pathologically defined benign cells bearing cancer stem cell markers may be cells of origin for tumor development in triple negative breast cancer, and their presence in benign tissue (including tissue in some cases from the uninvolved contralateral breast) implies a systemic etiology to triple negative breast cancer. In conclusion, putative cancer stem cell markers are enriched in the benign breast tissue of women who have developed triple negative breast cancer, and their presence may represent a biomarker for increased breast cancer risk.

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 LB-106. doi:10.1158/1538-7445.AM2011-LB-106
Abstract LB-157: Overcoming T-regulatory effects towards developing a more effective immunotherapy targeting multiple myeloma
Authors: Chang Lung-Ji , Han Shuhong , Okada Starlyn , Patel Ekta , Yang Li-jun , Moreb Jan S. , Pircher Hanspeter , Perez Lia E. , Alsina Melissa
Published: June 2012

Recent progress in dendritic cell (DC) and T cell therapies has offered a new avenue in treating multiple myeloma (MM). We have reported that MM patients' DCs cannot effectively present MM antigens and, instead, preferrentially activate T regulatory cells (Tregs). Furthermore, the MM cells actively suppress patients' DC and T cell functions (Han et al. 2008 Mol Ther. 16, 269–279). Therefore, an effective immunotherapy must overcome immune tolerance to the cancer cells and effectively expand functional MM-specific effector cells. To obtain critical data about MM patient's immune status, a longitudinal study of Treg cells before and after treatments of twenty MM patients was conducted. Patients' bone marrow (BM) and peripheral blood (PB) specimens were collected in the first visit and the follow up visits during treatment. Treg cell populations were examined using antibodies against CD4, CD25, CD127 and Foxp3. At the same time, naïve versus memory phenotype was assessed using antibodies against CD45RA, CCR7 and CD27. Besides the above markers, the surface molecule KLRG1 reflecting T cell senescence and longivity was also detected by flow cytometry. Analysis of Tregs at different time points showed a clear trend of gradual increase in Treg population both in the BM and PB of the MM patients; compared with healthy donors, MM patients' Tregs expand more than two to three folds. Surface marker analysis suggests that the expanded Tregs are mostly memory effector cells, which coincides with a decrease in the ratio of naïve Tregs. Analysis of MM-specific T cell responses by examining effector cytokines and cytolytic activities against autologous tumor cells revealed that BM contained more cytotoxic T cells than did PB. Increased expansion of MM-specific immune effector cells in culture was observed when Treg activities were suppressed using various immune modulators. Further understanding of Treg-mediated cancer tolerance will be essential to develop clinically more effective immunotherapy towards improved survival of MM patients.

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 LB-157. doi:10.1158/1538-7445.AM2011-LB-157
Abstract LB-158: MEDI4736: Delivering effective blockade of immunosupression to enhance tumour rejection: Monoclonal antibody discovery and preclinical development
Authors: Stewart Ross A. , Morrow Michelle , Chodorge Matthieu , Marcus Danielle , Boyle Melanie , Mulgrew Kathy , Hammond Scott , Wang Suping , Alimzhanov Marat , Robbins Paul , Lanning Karen , Bedian Vahe , McCourt Matthew , Lo Mathew
Published: June 2012

Cancerous cells emerge within the body following accumulation of deleterious genetic mutations. These mutations alter the phenotype of a cancer cell marking it as distinct from the surrounding host; an immunological state termed “altered self”. These cells, like other non-self entities such as viruses and bacteria, are recognised by the immune system and marked for destruction, a process known as “immune surveillance”. B7-H1 expression by tumour cells is believed to aid tumours in evading detection and elimination by the immune system. B7-H1 functions in this respect via several alternative mechanisms including driving exhaustion and anergy of tumour infiltrating T lymphocytes, stimulating secretion of immune repressive cytokines into the tumour micro-environment, stimulating repressive regulatory T cell function and protecting B7-H1 expressing tumour cells from lysis by tumour cell specific cytotoxic T cells.

Using hybridoma technology and high throughput screening MedImmune has identified a series of fully human antibodies specific for human B7-H1. Further characterisation of these antibodies led to the identification of a single high affinity antibody, MEDI 4736, with the ability to relieve B7-H1 mediated suppression of T cell activation in vitro and to enhance sub-optimal T cell activation in a mixed lymphocyte reaction. In vitro testing shows that MEDI 4736 does not trigger non-specific cytokine release in whole blood, and is only able to activate T cells in the context of an active T cell receptor signal.

A surrogate anti-mouse B7-H1 antibody shows significant anti-tumour activity in a syngeneic model when dosed in combination with chemotherapy. Similarly MEDI 4736 is able to inhibit tumour growth in a novel in vivo xenograft model, via a mechanism that is dependent on the presence of tumour specific human T 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 LB-158. doi:10.1158/1538-7445.AM2011-LB-158
Abstract LB-224: Development of a novel breast cancer segregation panel assay for multiple genetic subtyping based on gene expression modules developed from Oncomine™
Authors: Oades Kahuku , Chattopadhyay Sukla , Kim Hyunsoo , Vo Lien , Telford David , Wang Yipeng , Lee Byung-in , Monforte Joseph , Vansant Gordon , Freshley John , Wyngaard Peter , Rhodes Daniel , Tomlins Scott
Published: June 2012

Breast cancer is a highly heterogeneous disease as evidenced by comprehensive genetic studies which have revealed multiple subtypes using gene expression profiling and cell lineage classifier analyses. Previous studies have characterized different subtypes including normal breast-like, luminal epithelial A, luminal epithelial B, Her 2 over-expression and basal type carcinoma. However, the genetic variation within breast cancer is far more diverse than these core subtypes, and it is necessary to fully characterize this diversity in order to move beyond simple prognosis and to specifically predict drug sensitivity.

In a review of global gene expression and SNP-based cytogenetic data of more than 5,000 breast cancer patients in the Oncomine™ database, we have been able to characterize approximately 30 different genetic variations that are shared by 1% or more of the breast cancer population. These core, independent variables reflect diverse elements of the disease at a molecular level including cell lineage, dysregulated core biological functions, factors of cell growth, and importantly, the tumor microenvironment. Further genetic subtypes are characterized within the various large and focal genomic amplifications, such as Her2 and Myc, as well as focal expression events present subpopulations of patients. In aggregate these genetic variables represent all of the major genetic factors that present within breast cancer.

Currently biomarker/diagnostic approaches have tended to be over-tailored to specific clinical questions and therefore have lacked broad applicability, with every diagnostic test requiring a custom gene set and tailored signature and in some cases, requiring separate validated assays using multiple technologies and consequent splitting of clinical samples. To overcome these limitations, we have developed a single, 96-gene qRT-PCR test for rapid breast cancer companion diagnostics development using FFPE tumor tissue. All 30 of the core variables or “modules” are represented by this test which reports on both gene expression and chromosomal amplification events. We demonstrate in this study that this single test, with its multiple modules, can report on standard histopathological parameters, such as ER, PR and Her2, and reproduce existing prognostic and predictive genomic signatures. Data will be presented on prediction of overall survival, neoadjuvant chemotherapy response, and in-vitro sensitivity to MEK and PI3K inhibitors.

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 LB-224. doi:10.1158/1538-7445.AM2011-LB-224
Abstract LB-236: Identifying novel kinase fusions via targeted DNA capture and next-generation sequencing
Authors: Chmielecki Juliann , Peifer Martin , Viale Agnes , Socci Nicolas , Jia Peilin , Zhao Zhongming , Thomas Roman , Pao William
Published: June 2012

Tyrosine kinase (TK) fusion proteins result from genomic rearrangements that juxtapose TK signaling domains with various upstream partners. Since tumor cells can become dependent upon the resultant constitutive kinase activity for tumorigenesis and survival, TK fusions serve as attractive targets for anti-cancer therapy. In the past, identification of TK fusions in cancers has been hindered by experimental techniques that lack sufficient sensitivity and throughput. To overcome these challenges, we developed a screen to systematically identify TK fusions in tumor DNA based upon observed conservation of genomic fusion breakpoints. The approach involves selective capture of regions upstream of GXGXXG kinase motifs where TK fusion breakpoints are likely to occur followed by next generation sequencing and custom computational analysis. The feasibility of this approach (v1.0) was previously validated using cell line DNA (Chmielecki et al, 2010). We have since developed an improved version of our platform (v2.0) and used it to identify a novel c6orf204-PDGFRbeta fusion using DNA isolated from whole blood of a patient with T-cell acute lymphoblastic leukemia (T-ALL). The patient had an elevated white blood cell count with hypereosinophilia (HES) that was responsive to imatinib therapy. Multiple tumor samples (e.g. EGFR/KRAS/ALK wildtype lung adenocarcinomas, lung squamous cell carcinomas, ‘triple-negative’ breast carcinomas, and head and neck squamous cells carcinomas, etc.) are currently undergoing screening for other novel TK fusions, and candidate fusions are undergoing validation.

This work was supported by an SU2C-AACR Innovative Research Grant to WP.

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 LB-236. doi:10.1158/1538-7445.AM2011-LB-236
Abstract LB-238: Regulation of signaling pathways by endogenous small molecules
Authors: Nachtergaele Sigrid , Mydock Laurel , Krishnan Kathiresan , Covey Douglas , Rohatgi Rajat
Published: June 2012

A major goal in cancer biology is the comprehensive understanding of signals that drive the growth and spread of cancer cells. My long term goal is to develop methods to isolate new small molecules that play a role in cancer signaling and then to identify the proteins that interact with these small molecules. Such small molecule-protein pairs are likely to be particularly good drug targets in oncology. To develop tools for this endeavor, we are focusing on the identification of small molecules that regulate the “Hedgehog” signaling circuit. Damage to this circuit has been shown to drive the development of a large number of adult and childhood cancers. We are taking advantage of the previous observation that molecules related to cholesterol, called oxysterols, can activate the Hedgehog pathway. To understand how these enigmatic molecules work, we are attempting to find the protein targets through which oxysterols function and also generating analogs of these molecules as novel modulators of this important cancer signaling pathway.

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 LB-238. doi:10.1158/1538-7445.AM2011-LB-238
Abstract LB-70: Breaking the Silence
Authors: Hall Shaundra
Published: June 2012

The National Cervical Cancer Coalition (NCCC) is a grassroots organization that exists solely to educate and support patients, caregivers, family members, and communities about cervical cancer and other HPV-related diseases. NCCC is responsible for the development of the nation's first hotline devoted to helping women and family members resolve issues they have related to cervical cancer and HPV. The NCCC also developed the Cervical Cancer/HPV Quilts Project that travels the country to help put a personal face on the disease. The NCCC began the nationally-recognized FREE Pap Test Day held on the second Friday of January to enable disadvantaged women who have not received a Pap test in the past three years. The NCCC runs an award winning cervical cancer website and is currently developing a national network of local state affiliates. The NCCC cosponsored two fundraising conferences on cervical cancer issues in Malawi, Africa, and has held many prevention education fundraisers over the years as well. The coalition is developing a national grassroots effort in all 50 states. Through the efforts of our coalition members, we are addressing issues of cervical cancer/HPV prevention with education about vaccination and regular Pap and HPV testing. The organization always works to reduce the stigma associated with HPV infection and cervical cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-70. doi:10.1158/1538-7445.AM2011-LB-70
Abstract SY14-03: Inferring causality for oncology drug discovery and development
Authors: Khalil Iya
Published: June 2012

Decisions in drug development are made based upon determinations of cause and effect from experimental observations that span all development phases. Despite major advances in our powers of observation due to the advent of multi-omic technologies, the ability to determine disease mechanisms, biomarkers, and effective combination therapies from large-scale and diverse data sets in the context of ill-defined and complex biological systems continues to be a major bottleneck in the drug development process. Current methods of statistical analysis do not allow for causal inferences, leaving this impossible task to the human mind. This major bottleneck can only be overcome by utilizing automated computational learning methods that identify from compound data the circuits and connections between drug-affected molecular constituents and physiological observables. We will present several case studies in oncology drug development where we employ massively parallel machines to reverse engineer populations of models that predict the ‘gearing’ between drug, genetic, and genomic molecular components with respect to oncology phenotypic outcomes. Since a single topology or model cannot capture the reality of a biological system under investigation, Monte Carlo simulation strategies over the population of models reveal how the drug, genetic, gene expression and clinical variables work together to impact phenotypic outcomes. We will show how the marriage of multi-omic technologies within the network inference approach is helping to fill in the missing insights needed to improve drug development success rates.

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 SY14-03. doi:10.1158/1538-7445.AM2011-SY14-03
Abstract SY16-04: Contrasting effects of TGF-β signaling in the proangiogenic tumor stroma
Authors: Pietras Kristian
Published: June 2012

Cancer results from the concerted performance of tumor cells interacting with their microenvironment. The formation of a reactive stroma evidently contributes to tumor initiation, growth and progression by providing growth-regulatory factors, supporting tumor angiogenesis, and facilitating invasion. As a rich source of cancer-promoting factors, the tumor stroma is an underestimated target for the development of novel therapeutics.

Members of the transforming growth factor (TGF)-β family have been linked to vascular formation through mouse genetic studies. However, contradictory reports on the role of TGF-β signaling in physiological and pathological angiogenesis make the need for mechanistic studies apparent. Here, we describe functional studies on the role of the TGF-β receptor activin receptor-like kinase (ALK) 1 and its co-receptor endoglin in tumor angiogenesis. We demonstrate by genetic and pharmacological means that diminution of ALK1 gene dosage, or systemic treatment with an ALK1-Fc fusion protein, retarded tumor growth and progression by inhibition of angiogenesis. Moreover, we uncovered an unexpected synergy through which the combined action of different ALK1 ligands primed endothelial cells to respond to prototypical angiogenic stimuli both in vitro and in vivo. In parallel studies, genetic targeting of endoglin was revealed to give rise to an initial block in the angiogenic switch of tumors, which was overcome through acquired resistance characterized by an increased metastatic spread. The rise in metastatic dissemination was consequential to a loss of junctional patency in the endoglin-deficient vasculature due to ongoing endothelial-to-mesenchymal transition.

Taken together, our studies delineate a decisive role for signaling by TGF-β family members in tumor angiogenesis and offer mechanistic insight for the forthcoming clinical development of drugs blocking TGF-β signaling in oncology.

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 SY16-04. doi:10.1158/1538-7445.AM2011-SY16-04
Abstract SY22-01: Interrogating gene expression programs from preclinical analyses of genetically engineered mouse models
Authors: Abate-Shen Cory , Meneses Alvaro Aytes , Kinkade Carolyn W. , Mitrofanova Antonina , Lefebvre Celine , Chua Chee Wai , Castillo-Martin Mireia , Gelmann Edward , Shen Michael M. , Califano Andrea
Published: June 2012

Our laboratories have been investigating novel mechanisms of cancer progression, as well as new therapeutic approaches for early intervention and treatment of advanced disease by integrating analyses of human clinical data with molecular and functional studies of genetically engineered mutant (GEM) mice. In our analyses of prostate cancer, we have generated GEM mice that recapitulate the entire spectrum of disease progression from preinvasive lesions, termed prostate intraepithelial neoplasia (PIN), to castration-resistant and metastatic disease, which are the lethal forms of the disease. These GEM models are based on perturbation of the Akt/mTOR and MAP kinase signaling pathways, which are often co-activated in human prostate cancer and which function synergistically to promote castration-resistant metastatic prostate cancer. We have found that combinatorial inhibition of these signaling pathways in GEM mice that display castration-resistant metastatic prostate cancer results in abrogation of primary tumors, improved survival and reduced metastasis. Using computational approaches for comparative analyses of large-scale gene expression profiling data for mouse and human prostate cancer, we have been assembling molecular networks, called interactomes, to elucidate conserved cancer pathways. We have been investigating the consequences of drugs perturbations on the transcriptional network with the ultimate goal of identifying new druggable targets. In summary, our comparative investigations of prostate cancer in humans and GEM mice have revealed promising new molecular targets and new therapuetic approaches for the treatment of human cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr SY22-01. doi:10.1158/1538-7445.AM2011-SY22-01
Abstract SY31-01: Combinatorial development of biomaterials for tissue engineering and drug delivery
Authors: Anderson Daniel G.
Published: June 2012

High throughput, combinatorial approaches have revolutionized small molecule drug discovery. Here we describe our work on high throughput methods for developing and characterizing biomaterials for drug delivery and tissue engineering. We have developed automated methods allowing for rapid nanoliter scale synthesis of 1000's of biomaterials, as well as the testing of their chemical, material, and biological properties. These methods have also been applied towards the development of new methods to control stem cell behavior, as well as vehicles for drug delivery. In particular, these combinatorial libraries of different biomaterials have enabled new methods for microparticulate drug delivery, non-viral gene therapy, siRNA delivery, and vaccines.

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 SY31-01. doi:10.1158/1538-7445.AM2011-SY31-01
Abstract A43: Inhibition studies of Siah2, a ubiquitin ligase, involved in hypoxia induced cancer
Authors: Gopalsamy Anupriya
Published: June 2012

Ubiquitin ligase siah2 has been shown to regulate the hypoxia response by regulating prolyl hydroxylase3 (PHD3) involved in a hypoxia inducible factor 1α (Hif-1α) dependent pathway which up regulates the genes like vascular endothelial growth factor (VEGF), resulting in tumor cell survival. Thus it has been suggested that siah2 may act as a target for developing inhibitors. In this study we have proved the interaction of siah2 and PHD3 using co-immunoprecipitation and GST pull down assay. Interaction studies indicate that C-terminal substrate binding domain (SBD) of Siah2 is involved in the binding to PHD3 irrespective of the N-terminal ring domain. The structure-based high throughput virtual screening (HTVS) method was used to identify potential small molecule inhibitors and we have identified five drug-like lead molecules specific for the Siah2 using 14400 drug-like subsets obtained from the Maybridge database, which might inhibit the interaction between Siah2 and PHD3. The lead molecules from screening were then tested in silico for their pharmacokinetic properties and percent human oral absorption using QikProp. Further, pharmacokinetic analysis based on Lipinski's rule of five which is based on ADMET (absorption, distribution, metabolism, excretion and toxicity) properties suggested potential utility of these compounds as drug leads. Docking studies were carried out to analyze the binding conformations of the identified lead molecules towards the Siah2 which reveals that Ser39 residue of SBD may be the key amino acid residue interacting with these ligands necessary for the induction of Siah2 activity. These results set the stage for the development of a workflow for selection of domain specific drug development. Five potential drug-like molecules specific to the SBD domain were eventually selected on the basis of binding characteristics, docking energy and ADMET properties. Further in-vivo and in-vitro experimental studies to investigate the inhibitory potential of these lead drug-like small molecules on this ligase and its binding are under progress.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr A43.
Abstract A54: Downregulation of hepatocyte growth factor activator inhibitor-2 is involved in the progression of prostate cancer cells
Authors: Lan Shao-Wei , Cheng Tai-Shan , Wu Shang-Ru , Shyu Hsin-Yi , Ko Chun-Jung , Hsiao Pei-Wen , Lee Ming-Shyue
Published: June 2012

Dysregulation of pericellular proteolysis has been strongly involved in tumorigenesis. In this study, we delineated the role of hepatocyte growth factor activator inhibitor-2 (HAI-2) in prostate cancer cell invasion and tumorigenicity within a human prostate cancer progression model (103E, N1, and N2 cells). This model was newly established by a serial of intraprostatic propagation of a human prostate cancer 103E cell line and isolation of the metastatic cells from nearby lymph nodes. The invasion capability of these cells was gradually increased after the serial isolations (103E<N1 <N2). In this series of cells, the expression of HAI-2 was significantly decreased during the progression. Following the course of orthotopic tumor growth in mice, the expression level and activity of a HAI-2 target, matriptase, increased while the HAI-2 protein level decreased. Restoration of HAI-2 expression in N2 cells down-regulated the matriptase activity and cell proliferation, thus decreased the tumorigenicity. These results show that during the progression of human prostate cancer, matriptase was primarily controlled by HAI-2 expression. The imbalance in HAI-2 and matriptase expression led to matriptase activation, therefore increased cell invasion and tumorigenicity.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr A54.