bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 50: An informatic approach to rational selection of cell lines modeling complex clinicopathologic phenotypes based on gene expression: The COXEN algorithm
Authors: Smith Steven C. , Theodorescu Dan
Published: June 2012

A perennial issue in biomedical research regards validation of the relevance of experimentally tractable model systems to disease characteristics of interest. In the study of many types of cancer, this often operationally means selection of a cell line or a number of cell line models for perturbation within in vitro or in vivo models of disease progression or treatment. However, many classic cell lines are of uncertain provenance, may have been isolated from tumors staged and graded decades ago in a fashion of limited interpretability with regards to current care, or have exhibited drifting phenotypes in intervening years. One example is that of the classic bladder cancer cell line, T24, isolated from a sample of “verified grade III papillomatosis of the bladder” in 1973, which has given rise to isogenically verified metastatic variants like T24T (1). We recently have reported an algorithm, called COXEN (COeXpression ExtrapolatioN), which uses concordant gene expression signatures of complex clinicopathologic phenotypes to infer interpretable and predictable relationships between disparate cell line and tumor populations, despite significantly different global transcriptional differences. While prior reports have used COXEN to predict chemotherapeutic response in clinical trials a priori based on signatures from cell lines (2,3), herein we report an application of the reverse analysis, using signatures of clinicopathologic characteristics in tumors including invasion, nodal metastasis, and poor survival to select from a panel of forty bladder cancer cell lines models reflecting with highest fidelity the transcriptional patterns underlying the parameter of interest. We find that this approach validates some cell line model systems of bladder cancer and proposes new uses for others. This approach, widely adaptable to other tumor histologies, may provide important guidance for selection of appropriate models to test therapies targeted at clinically important phenotypes.

(1) Harding MA et al. Cancer Res 2002.

(2) Lee JK et al. PNAS 2007.

(3) Williams et al. Cancer Res 2009.

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 50. doi:10.1158/1538-7445.AM2011-50
Abstract 5007: WT1 promotes tumor growth and vascularization by regulating matrix metalloproteinase 9 in Ewing's sarcoma xenografts
Authors: Katuri Varalakshmi , Hammers Hans- Joerg , Gerber Stephanie , Qiu Xiaofei , Montgomery Elizabeth , Loeb David M.
Published: June 2012

WT1, a zinc-finger transcription factor, is expressed in tumors of varied origins including sarcomas. High level WT1 expression often correlates with poor patient survival. The role of WT1 in sarcoma biology and the mechanism by which high WT1 expression confers a poor prognosis are both unclear. The aim of this study is to investigate the effect of WT1 expression on sarcoma growth and angiogenesis. We transfected the WT1-null Ewing's sarcoma cell line SK-ES-1 with one of two major WT1 isoforms, and used shRNA to silence WT1 expression in the WT1-positive Ewing's sarcoma cell line MHH-ES. Cells were injected into the flanks of immune deficient mice and tumor growth was monitored. Tumors arising fromWT1-expressing SK-ES-1 cells grew faster than control, while tumors arising from WT1-silenced MHH-ES cells grew more slowly than control. Because our laboratory and others have shown that WT1 regulates VEGF expression, we evaluated tumor vascularity using the endothelial cell marker CD31 and the pericyte marker αSMA. In xenografted SK-ES-1 cells, quantification of CD31 positive area in WT1 expressing tumors showed 60 -70 % more compared to the controls. In contrast, CD31 expression is abolished in tumors arising from WT1-silenced MHH-ES cells. To investigate the mechanism by which WT1 affects tumor growth and vascularity, we performed a gene expression profiling in SK-ES-1 cells expressing different isoforms of WT1. We identified a number of genes up- or downregulated by WT1, including matrix metalloproteinase-9 (MMP9). Luciferase reporter assays showed that WT1 can upregulate MMP9 promoter activity in transiently transfected NIH3T3 cells. Also, immunohistochemical analysis showed increased MMP9 expression in WT1-expressing tumors. Finally, imunohistochemical analysis of primary Ewing's sarcoma samples showed correlated expression of WT1, VEGF, and MMP9. Taken together, our data suggest that WT1 promotes tumor growth and angiogenesis, perhaps via upregulation of MMP9. Increased tumor angiogenesis may lead to more aggressive tumors and a worse outcome for patients, explaining the effect of WT1 on prognosis.

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 5007. doi:10.1158/1538-7445.AM2011-5007
Abstract 5024: Effects of aerobic training to improve cardiovascular function and prevent cardiac remodeling after cytotoxic therapy in early breast cancer
Authors: Jones Lee , Dolinsky Vernon , Haykowsky Mark , Patterson Ian , Jones Lee , Allen Jason , Scott Jessica , Rogan Kyle , Khouri Michael , Hornsby Whitney , Young Martin , Peppercorn Jeffrey , Kimmick Gretchen , Dyck Jason
Published: June 2012

Background: It is well-established that doxorubicin (DOX)-containing chemotherapy causes progressive cardiomyopathy clinically manifest as decreased left ventricular ejection fraction (LVEF). The effects of DOX on the other organ components of oxygen (O2) transport and utilization that govern global aerobic reserve capacity are poorly characterized.

Methods and Results: To test our research questions, we established a mouse model of DOX in which female mice were subjected to DOX (8mg.kg.wk-1) or placebo (CON) (0.9% saline) for 4 weeks. Transthoracic echocardiography was performed to assess left ventricular wall thickness, volumes and LVEF, determined from M-mode tracings at baseline and 8 weeks. As expected, LV systolic volumes were higher and fractional shortening and LVEF were lower in the DOX group compared to CON after 8 weeks (p's <0.05). There was no change in effective arterial elastance (i.e. peripheral resistance, total lumped vascular compliance and characteristic impedance) following DOX however these mice had significantly decreased expression of proteins responsible for oxidative phosphorylation / mitochondrial biogenesis in the solelus muscle relative to CON mice (p<0.05). Injury to O2 transport organs led to significant reductions in global aerobic capacity (as measured by time to exhaustion on a motorized treadmill) in DOX compared with CON animals (p<0.05). Next, we tested whether aerobic training could prevent DOX-induced injury. Mice were subjected treadmill running, 5 d.wk-1, 18m.min-1, 45 mins/session for 8 wks during concurrent DOX (8mg.kg.wk-1) therapy (DOX+AT), DOX (8mg.kg.wk-1) only, or CON (0.9% saline). DOX-induced injury was completely abrogated by chronic aerobic training, which prevented injury in all organ components of O2 transport / utilization cumulating in preserved aerobic capacity. In patients with newly diagnosed operable breast cancer (n=20), DOX-containing chemotherapy caused a significant decline in aerobic capacity (p<0.05) despite normal LVEF and endothelial function. Supervised aerobic training completely averted this effect.

Conclusions: Our findings have important implications for the identification and protection against DOX-induced acute and late-occurring injury as well as optimizing therapeutic dosing.

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 5024. doi:10.1158/1538-7445.AM2011-5024
Abstract 5027: Steadily improving survival in lung cancer
Authors: Dillman Robert O. , McClure Stephanie E. , Zusman Douglas R. , VanderMolen Louis A. , Mahdavi Khosrow , Hafer Russell A. , Cox Craig A. , Joyo Colin I. , Barth Neil M.
Published: June 2012

Background and Purpose: There have been many improvements in the diagnosis and management of lung cancer during the past 25 years, including better imaging and the potential for earlier diagnosis, better surgical staging and introduction of minimally invasive surgery, improved radiation technology, and introduction of several systemic cancer drugs, including targeted therapies, and acceptance of multidisciplinary approaches rather than focusing on single modalities of therapy. We examined how treatment has changed in successive eras and how this relates to survival. Patients and Methods: The Hoag Cancer Institute data base was used to identify patients with lung cancer diagnosed during 1986-2009, who were diagnosed and/or received some or all of their lung cancer treatment at Hoag Hospital within four months of diagnosis. For the four successive 5-year periods, data was summarized for patient demographics and treatment, and observed and relative 5-year survival rates were calculated. Results: Data is summarized in the table below.

Conclusion: Over time there have been steady increases in median age, and in the proportion that have adenocarcinoma, are female, have local stage disease, undergo surgery alone as therapy, and receive systemic therapy with surgery as initial treatment. These changes have been associated with a 13.3 percentage point absolute increase in observed survival and an 81% relative increase. Relative 5-year survival increased from 18% to 34%, which contrasts with national data showing an increase from only 14% to 16% in the U.S.

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 5027. doi:10.1158/1538-7445.AM2011-5027
Abstract 504: Epithelial-mesenchymal transition confers stem-like properties and migratory ability to lung tumor cells
Authors: Bertolini Giulia , Roz Luca , Tortoreto Monica , Boeri Mattia , Pastorino Ugo , Sozzi Gabriella
Published: June 2012

We previously demonstrated that lung cancer CD133+ cells are highly tumorigenic, display stem-like features and are spared by cisplatin treatment. Recent studies proposed that the process of EMT could generate cancer cells endowed with migratory and invasive abilities and with characteristics of cancer stem cells (CSCs)

To test this hypothesis in lung cancer we induced EMT by transforming growth factor β (TGFβ1) in a lung cancer cell line established from primary tumor (LT73). EMT program activation in LT73 cell line resulted in cellular morphological changes, repression of E-cadherin expression and up-regulation of EMT-related genes, as evaluated by Real Time PCR. We also demonstrated that EMT-induced cells expressed high levels of stemness genes, similarly to mesenchymal stem cells. We furthermore found that LT73 cells undergone EMT exhibited an enhanced migrating potential and an increased drug resistance to cisplatin treatment than parental cell line, indicating that EMT activation provided cells with mesenchymal traits as well as stem-like properties.

FASC analysis revealed an enrichment of CD133+ lung cancer initiating cells in EMT-induced LT73 cells. Interestingly in EMT-generated CD133+ cells we identified a fraction of CD133+ /ESA− cells, not detectable in parental cell line, that express high level of pro-metastatic chemokine receptor CXCR4.

To validate the results obtained in the in vitro model, we analyzed primary lung cancer and corresponding lymph nodes metastasis, as well as spontaneous lung metastasis generated from subcutaneous xenografts tumor of H460 cell line. Preliminary results confirmed the existence of CD133+/ESA− /CXCR4High cells also in some primary lung cancer. Interestingly we found that lymph node metastasis contained a higher amount of this subpopulation of cells compared to corresponding primary tumors. Moreover the phenotypic analysis of spontaneous lung metastasis, derived from subcutaneous H460 xenograft tumor, demonstrated an enrichment for CD133/CXCR4 double positive population compared to parental tumors.

In conclusion these data indicate that EMT process could generate a subpopulation of CD133+/ESA− /CXCR4High cells, endowed with the potential to initiate tumor and disseminate, that possibly may represent the migrating-CSC fraction, involved in metastatic process. The existence of EMT-generated cancer cells could provide new insights into the tumor metastatic progression and lead to novel approaches for selective targeting of CSC.

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 504. doi:10.1158/1538-7445.AM2011-504
Abstract 5045: AUY922, a heat shock protein 90 inhibitor, has antitumor effects in human hepatocellular carcinoma cells
Authors: Gao Ming , Yeh Pei Yen , Lu Yen-Shen , Yu Huei-Chieh , Lin Hsiu-Mei , Chen Jun-Wei , Lin Shiou-Der , Cheng Ann-Lii
Published: June 2012

Hepatocellular carcinoma (HCC) is the fifth most common cancer and third leading cause of cancer death worldwide. Recently, molecular targeted therapy (MTT) agent, sorafenib, was shown efficacious in this dreadful cancer. However, HCC appears to be a cancer without single driver mutation, targeting multiple proteins in cancer cells might be an alternative strategy for better efficacy. Heat shock protein 90 (HSP90) is a ubiquitously expressed molecular chaperone regulating various important proteins associated with pivotal signal transduction and cancer progression of HCC. This study sought to explore whether HSP90 can be a potential target of HCC treatment. AUY922, a highly potent non-geldanamycin HSP90 inhibitor, suppressed cell growth of Huh7 and HepG2 cells with IC50 below 10nM. Flow cytometry analysis showed that G2/M arrest and sub G1 increase were induced after AUY922 treatment. AUY922 downregulated the expressions of Akt and cell cycle regulatory proteins, including Chk1, cdc2, Aurora B, and cyclin B1. Morphologic observation of AUY922-treated Huh7 cells demonstrated the formation of giant, micronucleated cells and multipolar mitosis, a picture compatible with mitotic cell death. Finally, treatment of BALB/c nude mice with 50mg/kg of AUY922 three times a week significantly suppressed the growth of Huh7 tumor xenografts. Our data provide the evidence that AUY922 inhibit HCC cells growth at least partially through mitotic cell death, and inhibiting HSP90 by AUY922 is a potentially useful therapeutic strategy for HCC.

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 5045. doi:10.1158/1538-7445.AM2011-5045
Abstract 5058: Effects of honokiol on UACC-62 malignant melanoma cell line
Authors: Guillermo Ruth F. , Stevens Jonathan , Kaushik Radhey S. , Young Alan , Dwivedi Chandradhar
Published: June 2012

Studies from our laboratory have indicated the chemopreventive effects of honokiol on UVB-induced squamous cell carcinoma in mouse skin. The purpose of this study is to investigate the effects of honokiol on UACC-62 malignant melanoma cell and elucidate the mechanisms of action. UACC-62 cells were grown in RPMI media in cell culture conditions. The effects of honokiol on cell viability and cell proliferation were studied by the MTT and BrdU assays. To investigate the effects of honokiol on cell cycle, UACC-62 cells were treated with honokiol, stained with propidium iodide and analyzed by flow cytometry. Cell lysates were prepared in similar conditions as the previous experiments to assess how honokiol affected the expressions of proteins involved in different pathways. The MTT assay showed a significant decrease in cell viability starting at 75 μM honokiol at 24 hours. The BrdU assay results indicated decreased cell proliferation as early as 12 hours and 50 μM honokiol. The cell cycle analysis indicated that honokiol caused G0/G1 phase arrest in melanoma cells (p < 0.05) at 24h and 50 μM. Western blots of the cell lysates prepared showed that honokiol decreased the expressions of cyclin E, CDK4, cyclin D2, NF-kβ, and p-MEK starting at 50 μM honokiol at 24 hours. The expressions of apoptotic proteins cleaved caspase-3, cleaved caspase-9, caspase 8, and cleaved PARP were increased by honokiol treatments. This data indicated that honokiol decreases cell proliferation and induces apoptosis of UACC-62 cells thus could be a candidate for pre-clinical studies as an agent for treating or preventing melanoma skin cancer.

Supported by Translational Cancer Research Center funded as a 2010 Research Initiative Center by the State of South Dakota.

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 5058. doi:10.1158/1538-7445.AM2011-5058
Abstract 506: Mechanisms of microvesicle-mediated communication in cancer
Authors: Svensson Katrin J. , Christianson Helena C. , Wittrup Anders , Johansson Maria C. , Mörgelin Matthias , Belting Mattias
Published: June 2012

The many functional roles of microvesicles (MVs) in tumor development and progression have lately become a subject of intense investigation. Cancer cell derived genetic material and signaling proteins residing in MVs can be transferred to recipient cells by as yet ill-defined mechanisms. We have found that heparan sulfate proteoglycans (HSPGs), which previously have been implicated as receptors of various macromolecular ligands, also can mediate the uptake of MVs. Our data suggest that MV uptake involves membrane raft, non-caveolin dependent endocytosis. These findings are consistent with recent data from our group on HSPG-mediated uptake of antibody-conjugated nanoparticles. Furthermore, MVs were found to be sorted to CD63-containing, vesicular compartments through intracellular transport that appeared highly dependent on an intact microtubule network. Intriguingly, preliminary findings support the concept of intracellular exchange of MV cargo, thus giving rise, in a single transporting vesicle, to a signal that integrates information from multiple cells. The presented results reveal a novel receptor that mediates MV uptake, and advance our general understanding of MV-dependent transfer of molecular information. Finally, the finding that MVs can potentially harbor molecules from multiple cells residing in the tumor environment opens up new, exciting roles of MVs as information carriers in 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 506. doi:10.1158/1538-7445.AM2011-506
Abstract 5075: An integrated in silico and proteomic approach towards the identification of novel cancer biomarkers based on tissue specificity
Authors: Prassas Ioannis , Makawita Shalini , Diamandis Eleftherios
Published: June 2012

Serological biomarkers represent a non-invasive and cost-effective means to aid in clinical management of cancer patients, particularly in areas of disease detection, prognosis, monitoring and therapeutic stratification. Several notable biomarkers currently in clinical use are proteins that demonstrate high tissue specificity to a single tissue, such as prostate specific antigen (PSA) for prostate cancer and human chorionic gonadotropin (hCG) for testicular cancer, the former being highly expressed only in the prostate and the latter in the placenta. Recent advances in high-throughput technologies (e.g. high-content microarray chips, serial analysis of gene expression platforms, thorough expressed sequence tag annotation) has enabled creation of publicly available gene and/or protein databases which describe the expression of thousands of genes (and/or proteins) in multiple tissues. Furthermore, over the past five years, our laboratory has characterized the proteomes of over 35 cell lines from numerous cancer sites as well as relevant biological fluids using a 2D-LC-MS/MS approach, identifying between 1000-4000 proteins per cancer site.

In this respect, the present study represents an integrative analysis of existing databases in combination with our in-house generated proteomic datasets, in pursuit of highly tissue specific proteins, which could be further explored as cancer biomarkers and/or therapeutic targets. Through stringent bioinformatic mining of the Human Protein Atlas, BioGPS, Unigene, TiGER, TiSGeD and other related databases, a list of proteins highly specific to normal lung, colon, prostate, testis and ovaries was generated. Interestingly, only a few proteins from each tissue were commonly reported as highly specific across all of these databases and these were selected for further analysis. Subsequent integration and comparison to our in-house generated proteomes: prostate cancer (PC3, LNCaP, 22Rv1 and seminal plasma from prostate cancer patients; 1958 proteins), lung cancer (H1688, H23, H460, and H520; 1856 proteins), ovarian cancer (HTB75, TOV-112D, TOV-21G and RMUG-S; 1123 proteins) and colon cancer (HCT116, LoVo, LS174T, LS180, SW1116, SW480 and SW620; 2124 proteins), helped to corroborate our candidate list.

Three, 4, 5, 10 and 7 highly specific proteins based on the applied criteria were prioritized for colon, testis, ovaries, prostate and lung, respectively. The identification of known biomarkers within these lists (e.g. PSA, KLK2, MSMB) warrants the analysis of the remaining proteins in serum, as potential cancer biomarkers and/or therapeutic targets.

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 5075. doi:10.1158/1538-7445.AM2011-5075
Abstract 5078: The NCI-NIST-FDA effort to develop fitness-for-purpose biospecimen assessment standards: Development of quality assessment metrics and standards for optimal biospecimen utilization
Authors: Aragon Richard , Dickherber Anthony , Litwack David , Vaught Jimmie , Compton Carolyn
Published: June 2012

The US Congress, through recent health care reform legislation, has directed the National Institutes of Health (NIH) to formulate a more aggressive strategy by which to translate scientific discoveries directly into the development of novel therapeutic and diagnostic capabilities. The National Cancer Institute (NCI) seeks to facilitate these efforts by targeting specific bottlenecks caused by the lack of standardization, quality metrics, and comparative benchmarks for research and clinical biospecimens used throughout the relevant discovery and validation pipelines. Many of the bottlenecks that currently impede routes to more effective biomarker discovery as well as drug, biologic and device development are impeded by a current lack of standards surrounding the research biospecimens used to advance this type of research.

The National Institutes of Standards and Technology (NIST), the Food and Drug Administration (FDA) and the NCI have engaged in an interagency effort to develop fitness-for-purpose quality assessment tools and metrics for the research biospecimens used by the clinical research, diagnostics, and technology development communities. An interagency workshop was convened consisting of biomedical and biospecimen scientists, standards/metrology scientists, commercial assay developers, experimental design and research methods experts, statisticians, and regulatory scientists that are among the leaders in their field. Subsequent to that workshop, a number of specimen quality assessment metric development projects were initiated to target metrics and standards that would have a significant impact on the development pipelines for diagnostics and therapeutics development, including the incorporation of certain regulatory efficiencies that could potentially accelerate development.

The NCI would like to present these ongoing projects and share ideas for future biospecimen quality assessment standards development projects for the cancer research, diagnostics, and technology development communities. Progress in designing such projects and preliminary results from experiments discussed in the October workshop will thus be described.

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 5078. doi:10.1158/1538-7445.AM2011-5078
Abstract 5081: Inhibiting DNA double strand break repair for alpha radioimmunotherapy
Authors: Song Hong , Hobbs Robert F. , Shao Chunbo , Sgouros George
Published: June 2012

Metastasis to distant organs is the major cause of mortality in breast cancer patients. Median overall survival remains poor (5 year survival 26.1%) despite recent advances in new targeted biological therapy. Radiolabeled antibody offers an effective approach to deliver cytotoxic radiation doses to eliminate micrometastatic tumors. Alpha particle emitters are promising against small micrometastases because of their high potency and specificity (high linear energy transfer, LET>80 keV/µm, particle range ∼ 80 μm). Alpha particles typically cause DNA double strand breaks (DSBs) when they traverse the cell nucleus. Our experience with alpha particle emitter labeled antibody showed that it significantly prolonged survival in a mouse model of metastatic breast cancer with eventual recurrence at estimated micromet dose of 4.4 Gy. Understanding cellular repair of alpha radiation induced DNA DSBs could help us rationally design more effective strategy by inhibiting components of the DNA DSB repair pathways to sensitize cancer cells to alpha radiation. Under this hypothesis, we evaluated the role of DNA dependent protein kinase catalytic subunit (DNA-PKcs) inhibition on targeted alpha-emitter therapy. Inhibition, with small molecule inhibitor NU7441, of this key enzyme in the non-homologous end joining (NHEJ) pathway sensitizes triple negative breast cancer cells to alpha particle emitter 213Bi labeled anti-EGFR antibody. Immuno-fluorescent staining of γH2AX foci confirms a reduction in the DNA DSB repair rate in these cells. Using a cell-line deficient in the homologous recombination (HR) pathway (BRCA-1 deficient), we also have preliminary evidence suggesting that HR deficiency reduces the incremental effect of NHEJ inhibition but the combination increases overall cell kill. This data suggest inhibiting components of DNA double strand repair pathways is an effective approach to enhance the efficacy of alpha particle emitter labeled monoclonal antibody.

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 5081. doi:10.1158/1538-7445.AM2011-5081
Abstract 5082: c-Abl function in TPA- and radiation-induced apoptosis in human prostate cancer cells
Authors: Mino Akihisa , Penate-Medina Tuula , Truman Jean-Philip , Mesicek Judith , Lee Hyunmi , Kolesnick Richard , Fuks Zvi , Haimovitz-Friedman Adriana
Published: June 2012

We previously reported that 12-O-tetradecanoylphorbol 13-acetate (TPA) decreased transcription and protein levels of Ataxia-Telangiectasia Mutated (ATM) in human prostate cancer cell lines LNCaP and CWR22-Rv1, de-repressing the enzyme ceramide synthase (CerS) to promote apoptosis. Here we analyzed the expression and subcellular distribution of the six mammalian CerS isoforms and their specific ceramide species generated within mitochondrial-associated membranes (MAM) and within the mitochondria in these cells before and after TPA±radiation treatments. This analysis revealed that TPA±radiation induces de novo synthesis of ceramide by specifically activating CerS isoforms 5 and/or 6 generating pro-apoptotic C16-ceramide species in mitochondrial membranes. In addition we demonstrate that LNCaP cells transfected with a c-Abl kinase dead (KD) mutant showed a significant decrease in both TPA and TPA + radiation-induced apoptosis compared with c-Abl (WT) transfected LNCaP cells. Thus, this result indicates that the kinase activity of c-Abl is necessary for TPA±radiation-induced apoptosis in LNCaP cells. In LNCaP cells transfected with WT-c-Abl, ceramide elevation was observed in response to TPA coupled to a synergistic increase of ceramide after TPA+radiation treatment. However, ceramide elevation was abolished in LNCaP transfected with the KD-c-Abl, for both TPA and TPA+radiation treatments. These findings identify kinase active c-Abl as a pro-apoptotic factor in these cells, which utilizes the activation of the CerS pathway to transduce this apoptotic response. Whether c-Abl directly or indirectly activates CerS5 and/or 6 activities, responsible for the generation of the pro-apoptotic ceramide species in response to TPA±radiation, remains to be elucidated.

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 5082. doi:10.1158/1538-7445.AM2011-5082
Abstract 5083: Nanoaerosol delivery of beclin1 combined with radiation suppressed tumor growth in the lungs of K-rasLA1 mice
Authors: Shin Ji Young , Cho Myung Haing
Published: June 2012

As incidence and mortality due to lung cancer increase, great effort has been put into developing feasible therapeutics in medical field. Radiation, together with surgery and chemotherapy, is the must in treating lung cancer, however, side effects have to be taken into consideration.

To minimize the side effect of radiotherapy, repeated exposures (5 times, 24 hours interval) with low dose (2Gy) of radiation to restricted area (thorax, 2cm) were attempted. After sensitizing lungs of K-rasLA1 mice with radiation, beclin1, known to be a tumor suppressor, involved in autophagy, complexed with nano-sized biodegradable poly(ester amine), was prepared and delivered into K-rasLA1 mice via aerosol 3 times for 4 weeks. Seventeen weeks old K-rasLA1 lung cancer model mice (n=20, 5 mice/group) were sacrificed and collected samples were analysed.

K-rasLA1 mice treated with beclin1 and radiation showed significant tumor regression with low progression to adenocarcinoma and increment in number of autophagic vacuoles. Precedent event of beclin1-bcl2 dissociation stimulated synergistic anti-tumor effect via inhibition of protein translation and angiogenesis.

Taken together, non-invasive treatment, radiation with PEA-beclin1 aerosol delivery, can be a feasible candidate for developing novel therapeutics in lung cancer treatment.

This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government (MEST) (NRF-2010-0000784).

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 5083. doi:10.1158/1538-7445.AM2011-5083
Abstract 5085: Radiosensitization of castration-resistant prostate cancer via modulating autophagic cell death
Authors: Qu Yang , Lian Jiqin , Wu Xiaoqing , Tang Wenhua , He Fengtian , Ji Min , Xu Liang
Published: June 2012

Radioresistance markedly impair the efficacy of cancer therapy. Anti-apoptotic Bcl-2 family proteins such as Bcl-xL, Bcl-2 and Mcl-1 are overexpressed in castration-resistant prostate cancer (CRPC) and contribute to prostate tumor initiation, progression and resistance to radiotherapy. A natural BH3-mimetic, small molecule inhibitor of Bcl-2, (−)-gossypol, shows promise in ongoing Phase II clinical trials for human prostate cancer. We have recently shown that (−)-gossypol preferentially induces autophagy in CRPC cells that have high levels of Bcl-2 and are resistant to apoptosis, both in vitro and in vivo, but not in androgen-dependent cells with low Bcl-2 and sensitive to apoptosis. Our results demonstrate for the first time that (−)-gossypol can interrupt the interaction between Beclin1 and Bcl-2/Bcl-xL at the endoplasmic reticulum, thus releasing the BH3-only pro-autophagic protein Beclin1, which in turn triggers the autophagic cascade. (−)-Gossypol-induced autophagy is Beclin1- and Atg5-dependent, together with Bcl-2 downregulation and Beclin1 upregulation. (−)-Gossypol increased autophagic cell death induced by X-ray radiation in the CRPC cells. Orally administered (−)-gossypol achieved a much greater efficacy with long-term tumor regression when used in combination with ionizing radiation. We have also explored a series of gossypol derivatives and identified two novel BH3-mimetic lead compounds that are more potent than the parental compound in inducing autophagic cell death in CRPC. Taken together, (−)-gossypol significantly enhances the anti-tumor activity of radiotherapy in vitro and in vivo, and represents a promising new regime for treatment of castration-resistant human prostate cancer with overexpression of Bcl-2. Our data provide new insights into the mode of cell death induced by Bcl-2 inhibitors, which would facilitate the rational design of clinical trials by selecting patients who are most likely to benefit from the Bcl-2-targeted molecular 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 5085. doi:10.1158/1538-7445.AM2011-5085
Abstract 5092: Variants at 6q21 implicate PRDM1 in the etiology of therapy-induced second malignancies after Hodgkin's lymphoma
Authors: Onel Kenan , Best Timothy , Skol Andrew D. , Li Dalin , Kirchhoff Tomas , Yasui Yutaka , Bhatia Smita , Strong Louise C. , Domchek Susan M. , Nathanson Katherine L. , Olopade Olufunmilayo , Mack Thomas M. , Conti David V. , Offit Kenneth , Cozen Wendy , Robison Leslie L.
Published: June 2012

Survivors of Hodgkin's lymphoma (HL) are at increased risk for second malignant neoplasms (SMNs) induced by exposure to radiation therapy (RT). Although SMNs are a leading cause of death in survivors of HL, it is not possible to identify those patients at greatest risk. We performed a genome-wide association study (GWAS) of 99 HL survivors who developed SMNs and 86 who did not, all of whom were treated for HL with RT. We then tested three SNPs in a replication set of 120 RT-treated HL patients with SMNs and 112 without SMNs. The functional effects of risk-associated variants on gene expression were assessed in lymphoblastoid cell lines before and after exposure to radiation. Two variants at 6q21 between PRDM1 and ATG5 were associated with SMNs in younger but not older HL patients treated with RT [rs4946728: P = 1.26×10-9, and rs1040411: P = 4.68×10-8]; individuals homozygous for the rs4946728 risk allele comprise 50% of the general population and were found to be at an eleven-fold increased risk for SMNs as compared to those homozygous for the protective allele. The risk variants comprise a haplotype that was associated with decreased PRDM1 expression (p=0.03) and protein levels. Furthermore, the risk variants were associated with a marked attenuation in the induction of PRDM1 following radiation exposure (p=0.002). Thus, we identified a new risk locus for SMNs following radiation treatment for HL at 6q21. This locus is associated with both basal and radiation-induced expression of PRDM1. These findings demonstrate a novel gene-exposure interaction associated with radiation-induced carcinogenesis and suggest that patients homozygous for the risk variants may benefit from early cancer screening.

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 5092. doi:10.1158/1538-7445.AM2011-5092
Abstract 5096: Identification of biomarkers and therapeutic targets in oral squamous cell carcinomas through comparative proteomic analyses
Authors: Tan Kee-Koon , Tsay Yeou-Guang , Lo Jeng-Fan
Published: June 2012

Background: Oral squamous carcinoma cell (OSCC) is the sixth most common worldwide cancer and responsible for about 640,000 new cases each year worldwide. Recently, a small subpopulation of oral cancer tumor cells with self-renewal capacity, or oral cancer-tumor initiating cells (OC-TICs), have been characterized in our lab. Accumulating evidence suggests that these OC-TICs contribute to tumor initiation, maintenance and metastasis while they are resistant to radiotherapy and chemotherapy. Thus, identification of the biomarkers and therapeutic targets for these cells should help the development of new diagnostics and therapeutics for oral cancers.

Methods: Sphere formation assay was used to enrich the OC-TIC population, whose proteins were resolved in a SDS polyacrylamide gel along with those from parental cells. The proteins within the gels were subjected to tryptic digestion and subsequent 18O peptide labeling. Liquid chromatography-mass spectrometry was programed to collect the tandem mass spectra (MS/MS) of peptides from proteins with >2-fold increase in OC-TICs. These MS/MS were matched against the NCBI protein database with TurboSequest search engine and the results were interpreted by bioinformatics tools.

Results: With such a new platform, we found that 61 proteins were up-regulated in OC-TICs proteomes. In order to define collective functional roles of the identified proteins, these proteins were categorized based on the information in Gene Ontology database. Molecular function analyses showed that, besides binding, catalytic activity and transporter activity, a majority of proteins appear to participate in protein metabolism and modification, protein/vesicle trafficking and carbohydrate metabolism. Meanwhile, cellular compartment studies revealed that most proteins appear to be from plasma membrane and mitochondria.

Conclusion: Our results indicate that OC-TICs behave differently in protein metabolism/trafficking and carbohydrate metabolism. This data set will be used to identify novel biomarkers or therapeutic targets for oral cancer-tumor initiating cells (OC-TICs).

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 5096. doi:10.1158/1538-7445.AM2011-5096
Abstract 5099: Identification of lung cancer biomarkers using lung cancer in female never smokers
Authors: Park Yun Gyu
Published: June 2012

On a worldwide level, lung cancer in never smokers (LCINS) occupies 25% of total lung cancer incidence and this disease shows the 7th leading cause of cancer death. It is known that LCINS occurs frequently in asian women. It has been reported that 75% of female lung cancer occurred in never smokers in Republic of Korea. The clinico-pathological and molecular differences as well as geographic and gender variations between lung cancers in never smokers and smokers, indicate that two types of lung cancers should be regarded as a different disease entity. However, data for molecular understanding of LCINS are limited. We designed this study to identify the protein differentially expressed in lung cancer in female never smokers through proteomic analysis including 2-DE and MALDI-TOF-MS. To increase the probability of finding LCINS-associated proteins, the clinical specimens including sera and cancer tissues were classified as the following: normal female smokers, normal female never-smokers, lung cancer in female smokers, and lung cancer in female never smokers. Out of the proteins differentially expressed in LCINS tissues, levels of 10 proteins were increased and those of 8 proteins were decreased. Levels of 6 proteins were increased and those of 5 proteins were decreased in serum of patients with LCINS. The identified proteins were verified by performing Western blot analysis not only in the same sample used in proteomic analysis but also in samples obtained from lung cancer in female smokers. The serum protein, which was difficult to be identified in Western blotting, was verified by ELISA. Seven proteins identified in tissues and serum of patients with LCINS were verified by Western blot analysis and ELISA. These identified proteins might be applicable to biomarker development for diagnosis of LCINS and provide a more comprehensive understanding of tumor maintenance or 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 5099. doi:10.1158/1538-7445.AM2011-5099
Abstract 5104: Serum metabolomic profiles acquired by gas chromatography-mass spectrometry (GC-MS) distinguish patients with pancreatic adenocarcinoma from those with benign pancreatic disease
Authors: McConnell Yarrow J. , Weljie Aalim M. , Kopciuk Karen A. , Dixon Elijah , Sutherland Francis , Dunse Nicole , Bathe Oliver F.
Published: June 2012

Introduction: Pancreatic adenocarcinoma is often difficult to accurately distinguish from benign pancreatic lesions such as pancreatitis. Accurate preoperative identification of patients with benign disease may reduce the number of highly invasive and costly pancreatic resections conducted in this group. This study aims to identify differences in the metabolomic profile of serum between patients with pancreatic adenocarcinoma versus benign pancreatic disease.

Methods: Fasting serum samples were collected, as part of an institutional biorepository program (IRB#E20846), from patients with pancreatic adenocarcinoma or benign pancreatic disease. Accompanying clinical data were collected prospectively. Gas chromatography-mass spectrometry (GC-MS) spectra were acquired for aqueous metabolites and analyzed using multivariate methods (orthogonal partial least squares-discriminant analysis, OPLS-DA) using SIMCA-P+ (V12.0.1) software. Metabolite identification was conducted using the TargetSearch approach.

Results: Of the 136 included patients, 101 had pancreatic adenocarcinoma and 35 had benign pancreatic disease (26 pancreatitis/pseudocyst, 7 serous cystic neoplasm, 2 other). Median patient age was 66 years, 51.4% were male, 22.8% were jaundiced, and 79.4% presented with a common bile duct stricture or pancreatic mass. The metabolomic profile of serum from patients with pancreatic adenocarcinoma was significantly different from that of patients with benign pancreatic disease based on OPLS-DA modelling (p=0.0004). Overall, the profile contained 168 targeted metabolites, of which 14 were significantly different between malignant and benign pancreatic disease on multivariate modeling.

Conclusion: The serum metabolomic profiles of pancreatic adenocarcinoma and benign pancreatic disease differ significantly. Further analysis of these differences may yield a novel serum test to distinguish these lesions in clinical practice.

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 5104. doi:10.1158/1538-7445.AM2011-5104
Abstract 5109: Proteomics of murine BRCA1 deficient breast tumors identifies DNA repair proteins with prognostic value in human breast cancer
Authors: Warmoes Marc O. , Jaspers Janneke , Massink Maarten P.G. , Pham Thang , Piersma Sander R. , Rottenberg Sven , Boven Epie , Meijers-Heijboer Hanne , Waisfisz Quinten , Jonkers Jos , Jimenez Connie R.
Published: June 2012

Background: Breast cancer is the most common malignancy in women in the western world. The outcome of breast cancer would be strongly improved if patients could be diagnosed and treated early. This especially holds for patients with hereditary breast cancer.

Aim: Our aim is to identify proteins that are associated with BRCA1 deficient hereditary breast cancer. These proteins could have potential use as screening, prognostic or predictive biomarkers.

Approach: In-depth proteome profiling of tumor tissues of mouse breast cancer models was employed to identify BRCA1-associated proteins. To this end, tumor tissue lysates of five BRCA1 deficient and five proficient mouse models (Liu et al., 2007) were fractionated using SDS-PAGE followed by tryptic in-gel digestion, nanoLC-MS/MS and database searching. Normalized spectral counting was used for protein quantification and beta binomial statistics to discover significantly regulated proteins. Ingenuity Pathway Analysis and COFECO protein complex analysis was used to support data interpretation. The prognostic power was assessed by the use of publicly available gene expression human breast cancer data sets.

Results: We identified a total of 3614 proteins, of which 804 were differential between the genomic instable BRCA1-deficient and the genomic stable BRCA1-proficient breast tumors. Pathway analysis and protein complex identified DNA-repair and associated function like chromatin modeling and RNA processing as the major functions associated with the upregulated proteins of the BRCA1 deficient tumors. We created a core DNA-repair protein signature using pathway and protein Complex analysis. When the signature proteins were mapped to human breast cancer gene-expression data sets, they were shown to have prognostic power.

Conclusion: Proteomics of genetic mouse models for familial breast cancer is a powerful strategy to discover novel candidate BRCA1 deficiency proteins with human relevance. The major upregulation of DNA-repair complexes may indicate a compensatory mechanism for loss of BRCA1 DNA-repair. Further validation studies are required to investigate prognostic candidates in tissue microarrays. Integration with data on tumor and cancer cell-secreted proteins will pinpoint which of the BRCA1 deficiency associated proteins may have potential for screening of BRCA1 tumors and whether they have predictive value for sensitivity to PARP1 inhibition and Cisplatin treatment in BRCA1 as well as non-BRCA1-deficient tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5109. doi:10.1158/1538-7445.AM2011-5109