bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5547: EF24 attenuates radiation-regulated eNOS→NO-dependent NFκB-mediated matrix metalloproteinases in human neuroblastoma
Authors: Madhusoodhanan Rakhesh , Veeraraghavan Jamunarani , Natarajan Mohan , Herman Terence S. , Awasthi Vibhudutta , Aravindan Natarajan 
Published: February 2011

Abstract
We recently reported that radiation (IR) induced NFκB in neuroblastoma cells and treatment with curcumin impedes the activation of NFκB and enhance IR-induced cell death. However, full potential of curcumin is yet to be realized because of its poor bioavailability. This prompted us to investigate the efficacy of a more potent synthetic analogue, EF24 in inhibiting IR-induced NFκB dependent cell survival and tumor progression and delineated the functional mechanism involved. Human SH-SY5Y and IMR-32 cells, either exposed to 2Gy or treated with 100nM EF24 were exposed to IR and examined after 1h, 3, 6, 24, 48 or 72h. IR profoundly inhibited both eNOS phosphorylation and transcriptional activation. Consistently, IR reduced the bioavailability of NO. EMSA analysis showed a robust and persistent NFκB activation at least up to 72h in IR exposed cells. eNOS inhibition (L-NAME) and NO induction (GSNO) studies demonstrated the influence of IR-regulated eNOS and NO in NFκB activation. Furthermore, IR significantly induced transcriptional levels of MMP-1, 2, 3, 7, 9, 11, 13 and 14 in neuroblastoma cells. NFκB knock-out (using RelA siRNA) or forced expression of NF-kB (p50, p65 over expression) exhibits that IR-induced NFκB mediates MMPs activation. Conversely, EF24 treatment significantly reverted IR-negated eNOS activity and enhanced NO availability in the cells. Moreover, EF24 profoundly inhibited IR-induced NFκB and this induced inhibition was sustained at least up to 72h. eNOS inhibition studies with EF24 clearly portraits that EF24 regulates induced NFκB in an eNOS-NO dependent manner. More importantly, EF24 completely suppressed IR-induced MMPs transcript levels in these cells. NFκB over expression studies validates that EF24 regulates IR-induced MMPs by muting IR-activated NFκB. In addition, MTT analysis revealed that EF24 significantly conferred IR-induced inhibition of cell survival in both SH-SY5Y and IMR-32 cells by targeting IR-induced NFκB. Taken together these results suggests that IR-regulated eNOS and reduction in NO availability activates NFκB and NFκB dependent transcriptional activation of MMPs. More importantly, reverting IR-regulated eNOS→NO with EF24 may significantly inhibit NFκB dependent survival advantage and, may serve as a potential deliverable in mitigating neuroblastoma relapse and progression after radiotherapy.
Funding Support: ACS- IRG-05-066-01; Presbyterian Health Foundation
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5547.
Abstract 5560: Biomarker Discovery and Pathway Mapping using Differential Phosphoprofiling of PI3K-Pathway Inhibitors: PRAS40 Correlates with AKT Activation, but not PTEN Expression in Lung and Breast Cancer
Authors: Paweletz Cloud P. , Andersen Jannik , Sathyanarayanan Sriram , Bacco Alessandra Di , Chi An , Zhang Theresa , Albert Chen , Feldman Igor , Kraus Manfred , Lynch Bethany , Rush John , Blume-Jensen Peter , Hendrickson Ronald C. 
Published: February 2011

Abstract
Protein phosphorylation plays a key role in cell signaling and pathophysiological alterations in protein kinases and phosphatases contribute to human diseases. We used stable isotope labeling by amino acids (SILAC) in cell culture combined with phospho-antibody based enrichment for quantitative mass-spectrometry-based identification of differentially phosphorylated proteins in response to small-molecule PI3K-pathway inhibition. We quantified over 500 non-redundant serine/threonine phosphopeptides (NSTPs) containing either the AKT-, MAPK-substrate, or PDK1-docking motif. Of these NSTPs, 71 phosphoproteins were modulated by inhibitors targeting either AKT, PDK1 or PI3K/mTOR and a common set of 11 were modulated by all three drugs. Bioinformatics analysis of the regulated phosphoproteins identified core components of the canonical PI3K pathway and showed enrichment in adaptor and scaffolding molecules involved in cell polarity (PAK4), cytoskeletal reorganization (Filamin, Stathmin, Dynamin) vesicle transport (LARP1, VPS13D, SLC20A1), protein translation (S6RP and PRAS40), and transcription (EIF4BP-1). These results have guided the rational selection of antibodies for biomarker and patient stratification assays and a newly developed phosphor-specific antibody against PRAS40T246 shows specificity for oncogenic PI3K-pathway activation in PTEN-deficient mouse prostate tumor tissue by immunohistochemistry. In addition, the phospho-PRAS40T246 biomarker was evaluated across a panel of 67 breast and 96 lung cancer cell lines and in triple negative human breast tumor tissue. In these datasets, phopspho-PRAS40T246 positively correlates with phospho-AKTS473, but not PTEN protein expression. As such, we have positioned PRAS40T246 as a clinically relevant biomarker for the identification of PI3K-pathway activated tumors to enable individualized cancer therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5560.
Abstract 5567: Pancreatic disease-specific carriers of the CA 19-9 antigen identified by antibody microarrays and mass spectrometry
Authors: Yue Tingting , Hurley Mary C. , Brand Randel E. , Andrews Philip C. , Haab Brian B. 
Published: February 2011

Abstract
Distinguishing patients with malignant pancreatic tumors from those with benign conditions is critical for doctors to make proper treatment decisions. However, this process is usually costly and invasive due to the lack of a simple detection method such as a blood test. The current standard serological biomarker of pancreatic cancer, the CA 19-9 antigen, is not accurate enough for early diagnosis due to its occasional elevation in benign pancreatic diseases. The CA 19-9 antigen is a carbohydrate epitope found on multiple protein carriers. We sought to determine whether the protein carriers of the CA 19-9 antigen are different between benign and malignant pancreatic disease, and whether these differences could be harnessed to develop improved biomarkers. Two approaches were used to investigate this question. In the first, we used antibody microarrays to capture multiple different mucins from the sera of early-stage pancreatic cancer patients (n = 33), late-stage pancreatic cancer patients (n = 17), pancreatitis patients (n = 38), and healthy control subjects (n = 20), and we probed the level of the CA 19-9 antigen on the captured proteins using a monoclonal antibody. Pancreatic cancer progression was associated with a shift in CA 19-9 carriers from MUC16 to MUC1 and MUC5AC. Certain monoclonal antibodies captured glycoforms of MUC1 that predominantly carried CA 19-9 in the cancer patients. Because of that relationship, the detection of CA 19-9 on specific protein glycoforms improved the discrimination of cancer from control subjects over total CA 19-9 (AUC= 0.93 vs. AUC=0.86. In the second approach, we immunoprecipitated CA19-9 carrier proteins from pooled sera of late-stage cancer, early-stage cancer, pancreatitis, and control subjects, followed by deglycosylation and mass spectrometry (MALDI/TOF/TOF). Several proteins were identified in the sera of the cancer patients but not in the sera of pancreatitis or control subjects, indicating the possible identification of state-specific carriers of CA 19-9. Ongoing studies are aimed at verifying these identifications and testing performance as biomarkers of pancreatic cancer. These findings may lead to more accurate methods of discriminating benign from malignant pancreatic disease, which would improve the ability to render proper care to patients at the earliest possible stages.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5567.
Abstract 5581: Antigen-specific CTLA-4-regulated T-cell responses elicited in patients with prostate cancer treated by DNA immunization
Authors: McNeel Douglas G. , Jankowska-Gan Ewa , Burlingham William J. 
Published: February 2011

Abstract
Background: We and others have been interested in plasmid DNA vaccines as a means of eliciting tumor antigen-specific CD4+ and CD8+ T-cell responses. We have investigated plasmid DNA vaccines encoding prostatic acid phosphatase (PAP), a well-characterized prostate tumor antigen, in rodent models and found these to be effective in eliciting antigen-specific CD8+ T cells with evidence of anti-tumor efficacy. Multiple immunizations at high plasmid doses, however, elicited antigen-specific IL-10 secretion. We have recently completed a phase I clinical trial evaluating a DNA vaccine encoding PAP in patients with minimal residual prostate cancer. In that trial, while in vitro PAP-specific T-cell responses were elicited in multiple patients following six biweekly immunizations, others did not have evidence of an immune response, and some responses were not detectable until months later. The current study was conducted to determine whether immunization might have elicited an antigen-specific regulatory T-cell response in some individuals.
Methods: Peripheral blood mononuclear cells obtained from 21 patients prior to immunization, and two weeks after six biweekly immunizations, were evaluated for PAP-specific immune responses in a SCID mouse trans-vivo delayed-type hypersensitivity (DTH) assay. Specifically, DTH responses to PAP in PBMC were evaluated in the presence or absence of antibodies to regulatory molecules (IL-10, TGFß, CTLA-4, PD-1) to “uncover” antigen-specific regulatory mechanisms.
Results: 8/21 individuals had a PAP-specific T-cell response following immunization that was not detectable pre-immunization and which was “uncovered” by CTLA-4 blockade. This response was found to persist in at least one individual for several months. IL-10- and TGFß-regulated responses were not detected. Baseline regulatory PAP-specific T cells could be detected that suppressed concurrent tetanus-specific DTH responses. 6/8 individuals with a detectable CTLA-4-regulated response experienced a favorable increase in PSA doubling time.
Conclusions: Patients with prostate cancer can have PAP-specific regulatory T cells. Plasmid DNA immunization can elicit effector T cells specific for PAP that in many individuals is regulated by CTLA-4. Further studies are underway to characterize the regulatory T cell population(s), however these findings suggest that the efficacy of DNA vaccines might be further augmented by combination with anti-CTLA-4 blockade. In addition, these studies suggest that these trans-vivo DTH methods, in use for evaluating mechanisms of antigen-specific tolerance, could be more generally applied to the immune monitoring of anti-tumor vaccine clinical trials.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5581.
Abstract 5587: Serum HPV 6, 11, 16 and 18 neutralizing antibody detection by multiplex liquid array system (luminex)
Authors: Bae Heejin , Bae Sumi , Lee Keun Ho , Han Sei Jun , Ahn Woong Shick 
Published: February 2011

Abstract
Recently two prophylactic HPV vaccines including types 6, 11, 16 and 18 are clinically available. Detection of HPV DNA in cervix and neutralizing antibody in serum are important before and after vaccination. A luminex technology facilitates the simultaneous evaluation of multiple immune mediators with advantages of high throughput, small sample volume, lower cost and short time than ELISA. We are already developed HPV DNA in cervix detection system using multiplex liquid array (Cancer science, 2007) and now established detection system using luminex.
For production the antigen, HPV 6, 11, 16 and 18 L1 genes were isolated from their genomic DNA by PCR, cloned into pGEM vector with simian virus 40 (SV 40) large T-antigen and expressed in E. coli. Purified GST fusion recombinant proteins are coupled to microbeads, respectively. The characteristics of the assay were optimized using monoclonal antibody (MAb) specificity, validated with several distinct serum panels and used to determine the serum neutralizing antibody.
Using this system, we tested 40 females who vaccinated quadrivalent prophylactic HPV vaccine 6 month ago (20) and sexually unexposed group in teenager (20). The value of neutralizing antibody was average 1800 (±200) MFI in vaccinated group. This high-throughput multiplex liquid array system is highly sensitive.
We can use population based validation and combination with HPV DNA test using luminex. By using multiplex luminex beads coupled HPV antigen, we can check whether neutralizing antibodies formation or not, and measure the titers of antibodies to each subtype of HPV. We found one possibility to check up the HPV DNA and serum neutralizing antibody before and after HPV prophylactic vaccination.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5587.
Abstract 5592: Lovastatin and low dose celecoxib decrease proliferation and inflammation in a cellular model of lung pre-malignancy
Authors: Grant Jeanette L. , Walser Tonya C. , Yanagawa Jane , Lee Gina , Minna John D. , Prasad Sri , Hazra Saswati , Dubinett Steven M. 
Published: February 2011

Abstract
Statin drugs, such as lovastatin and pravastatin, block the cellular synthesis of cholesterol by inhibiting the function of HMG-CoA reductase. In a prospective study, patients undergoing statin treatment for prevention of atherosclerosis were at a lower risk of developing certain cancers, including lung and breast, suggesting a possible use for lovastatin in the chemoprevention of lung cancer. Here, we show that lovastatin dose-dependently decreases proliferation in both histologically normal and Kras-mutated human bronchial epithelial cells (HBECs) through the inhibition of cholesterol production. However, increased expression of PGE2, which we and others have shown to contribute to lung cancer progression, was noted with lovastatin treatment alone. Lovastatin up-regulates the production of PGE2 by up-regulating COX-2 expression and simultaneously down-regulating metabolism of PGE2 via down-regulation of 15-PGDH expression. Treatment with the cholesterol precursor, mevalonic acid, inhibited up-regulation of PGE2 and blocked dose-dependent anti-proliferative activity of lovastatin, indicating sterol-dependence. Treatment with both lovastatin and low dose (0.1uM) celecoxib, a COX-2 inhibitor, decreased PGE2 protein production in vector control and Kras-mutated HBECs. These findings suggest that the combination of lovastatin and low dose celecoxib overcomes up-regulation of PGE2 by lovastatin alone and that the combination represents a potentially effective lung cancer chemopreventative regimen worthy of further investigation.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5592.
Abstract 5594: Immunotherapy of metastastic renal cell carcinoma reveals a critical role for nitric oxide expression in the control of lung metastases but not primary tumors
Authors: Weiss Jonathan M. , Ridnour Lisa , Hussein Perwez , He Peijun , Back Timothy C. , Scarzello Tony , Maciag Anna E. , Keefer Larry , Murphy William J. , Harris Curtis C. , Wink David A. , Wiltrout Robert H. 
Published: February 2011

Abstract
Using an orthotopic mouse model of renal cell carcinoma, we showed previously that IL-2/αCD40 combination immunotherapy resulted in synergistic anti-tumor responses. In contrast, the use of either IL-2 or αCD40 as single agents mediated only partial, transient anti-tumor effects. We now show that treatment of tumor-bearing mice with IL-2/αCD40, but not either IL-2 or αCD40 alone, induces significant nitric oxide synthase (NOS2) expression within the tumor microenvironment through an interferon gamma-dependent pathway. In saline-control treated mice (low NO levels), NO inhibition reduced lung metastases by itself. However, in the context of immunotherapy (high NO levels), NOS2 inhibition completely abrogated the ability of IL-2/αCD40 treatment to reduce lung metastases but had no effect on the ability of IL-2/αCD40 to reduce primary kidney tumor burden. We further show that IL-2/αCD40 induces the NO-dependent decrease in matrix metalloproteinase (MMP) expression and activity, concomitant with an increase in the expression of tissue-inhibitor of metalloproteinase (TIMP)-1 within the tumor microenvironment. Finally, the treatment of tumor-bearing mice with the NO donor, JS-K significantly reduced lung metastases but had no effect on primary tumor size. These data distinguish the primary anti-tumor effects of IL-2/αCD40 immunotherapy, which are independent of NO expression, from the important ability of IL-2/αCD40 to inhibit lung metastases through the NO-dependent regulation of MMP and TIMP functions. Furthermore, the reduced MMP9 activity may be indicative of M1 polarized macrophages within the tumor microenvironment after IL-2/αCD40 treatment. Our data demonstrate the mechanistic basis for the IL-2/αCD40-mediated control of lung metastases and indicate that the context-dependent application of NO-inducing agents may hold considerable promise for the treatment of metastastic disease.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5594.
Abstract 560: Mesenchymal stem cells promote mammosphere formation in normal and malignant breast cells
Authors: Klopp Ann H. , Lacerda Lara , Gupta Anshul , Debeb Bisrat , Marini Frank , Andreeff Michael , Woodward Wendy A 
Published: February 2011

Abstract
Purpose: Normal and malignant breast tissue contains a rare population of cells with the capacity to self-renew, referred to as stem cells, or tumor initiating cells (TIC) in the case of tumors. These cells can be enriched by growth as “mammospheres” in three-dimensional cultures. We tested the hypothesis that mesenchymal stem cells (MSC), which are known to support tumor growth and metastasis, increase mammosphere formation.
Materials and Methods: Normal human mammary epithelial cells (HMEC), and breast cancer cells (MCF-7, SUM149 and IBC3) were grown as mammospheres, plated at low density in serum-free growth factor supplemented suspension cultures. MSC were added as an increasing percentage of total cells plated (0, 2, 5 and 10%). MSC-conditioned media was generated by growing MSC as spheroids for 5 days.
Results: MSC integrated into mammospheres, resulting in a dose-dependent increase in the number of mammospheres formed. Normal mammary epithelial cells formed 1.6 to 2.4 to 2.7-fold (p&lt; 0.001) more mammospheres in the presence of 2, 5 and 10% MSC. Similarly, the number of MCF-7 derived mammospheres increased 5.9 to 8.5 to14.8-fold (p = ≤ 0.001) in the presence of 2, 5 and 10% MSC. A similar dose-dependent increase in sphere formation was seen for SUM149 cells and inflammatory breast cancer cells in short-term culture derived from pleural effusions. This effect was found to be mediated by secreted factors as MSC-conditioned media also increased sphere formation. SUM149 cells formed 6.4 to 14.2 to 13.7-fold more mammospheres when incubated for 5 days in 5, 25 and 50% MSC conditioned-media (p &lt;0.0001). HMEC derived mammospheres had higher levels of expression of N-cadherin and RhoC and lower levels of expression of E-cadherin and Notch when formed in the presence of MSC conditioned media.
Conclusions: Mesenchymal stem cells increase mammosphere formation of normal breast epithelial cells, established cancer cell lines (MCF-7 and SUM149) and short term primary breast cancer cells (IBC3). This effect is mediated by MSC secreted factors and results in cadherin switching, which may promote breast cancer progression and resistance to therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 560.
Abstract 5602: MSC-mediated intratumoral cytokine delivery augments anti-tumor immunotherapy of glioma
Authors: Gunnarsson Salina , Svensson Andreas , Siesjö Peter , Darabi Anna , Bengzon Johan 
Published: February 2011

Abstract
Glioblastoma multiforme is the most common and aggressive form of primary brain tumor. In spite of surgical resection, combined radiation therapy and chemotherapy, the mean survival is less than 2 years following diagnosis. Using the established N32 animal model of glioma, shown to have many similarities with the human tumor, we are able to study and develop gene therapy strategies based on multipotent mesenchymal stromal cells (MSCs) as an efficient vector system.
MSCs derived from adult rat bone marrow have been shown to selectively target tumors when inoculated intracranially and these cells can be used as cellular vector systems for gene therapy of the tumor. Our group has previously shown that the cells, when transplanted into an established tumor, migrate within the tumor as well as along infiltrating tumor arms and to distant micro-satellites. We have transformed these rat MSCs to produce the cytokine interleukin 7 (IL-7), a growth factor involved in the development of immune cells. When IL-7 secreting MSCs are injected intratumorally following peripheral immunizations with IFNγ-secreting autologous irradiated tumor cells, a synergistic effect on anti tumor immunity is noticed. Tumor size is significantly smaller in treated animals compared to control as recently reported by our group. These studies have now been extended with survival experiments which show that there is an increased survival in the group receiving MSC-IL7 + IFNγ-immunization compared to IFNγ-immunization alone, 45% and 21% (animals alive) respectively at day 120 post tumor inoculation. In addition, MSCs transduced to express IFNγ in combination with peripheral IFNγ-immunization result in a 40% survival in the same study.
The use of MSCs from adult bone marrow has several advantages, it can be retrieved relatively easy and the patient self can donate it. Combining immunization with tumor cells together with MSC-delivered cytokines intratumorally might be further developed into a clinically useful treatment strategy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5602.