bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3022: PAR-2 activation is required for cancer stem cells maintenance through upregulation of POSTN
Authors: Ma Yiming , Wang Hongying 
Published: May 2015

Abstract
Backgrounds: PARs are seven transmembrane-spanning domain G-protein coupled receptors (GPCRs) which act as targets of certain serine proteinases, such as thrombin and trypsin. PAR2 is the only member in the family which cannot be activated by thrombin. The studies in knockout mice revealed that PAR2 plays more important roles in tumor formation compared to other PARs.
Aims: To investigate the roles of PAR2 activation in cancer stem cell maintenance and possible mechanisms.
Methods and Results: In human colorectal cancer samples, PAR2 was significantly correlated with CD133 at mRNA level measured with real time PCR. With six colorectal cancer cell lines, it was found that the cell line with higher level of PAR2 expression presents higher rate of CD133+ cells and vice versa. To test whether PAR2 is required for cancer stem cell, we developed cell lines with knock down of PAR2 stablely. It was found that inhibition of PAR2 expression in cancer cells not only reduced the level of CD133 but also significantly blocked the ability of sphere formation in vitro. In addition, we found that PAR2 activation induced the expression of POSTN, which is a well established secreted protein related to cancer stem cell maintenance. Deficiency of PAR2 in cancer cell also reduced the level of POSTN. More importantly, recombinant POSTN protein was able to restore the ability of sphere formation decreased by PAR2 deficiency in cancer cells. Furthermore, Activation of PAR2 caused TGFβ activation without affecting levels of total TGFβ. Given that TGFβ(5ng/ml) only could stimulate POSTN expression, it is very possible that PAR2 stimulates POSTN expression through TGFβ-related pathway. Finally, to test the role of PAR2-induced POSTN in cancer stem cell maintenance in vivo, we measured the correlation of PAR2 and POSTN in paired primary and metastatic ovarian cancer samples. Not surprisingly, significant correlation of PAR2 and POSTN expression was only observed in metastatic ovarian cancer group, which express significant higher level of cancer stem cell-related genes compared with primary group.
Conclusion: Our study strongly indicated that PAR2 activation was involved in cancer stem cell maintenance through TGFβ- mediated POSTN expression.
Citation Format: Yiming Ma, Hongying Wang. PAR-2 activation is required for cancer stem cells maintenance through upregulation of POSTN. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3022. doi:10.1158/1538-7445.AM2014-3022
Abstract 3150: Exploring the role of Rap1Gap in the progression from DCIS to invasive breast carcinoma
Authors: Shah Seema , Osuala Kingsley , Mao Shihong , Li Quanwen , Sloane Bonnie , Krawetz Stephen , Mattingly Raymond R. 
Published: May 2015

Abstract
Understanding the underlying mechanisms that drive malignant progression of breast cancer is important. The purpose of this study is to define the role of Rap1Gap in the progression of ductal carcinoma in-situ (DCIS) to invasive cancer. We employed an in-vitro three-dimensional (3D) overlay model that provides a physiologically relevant microenvironment to study mechanisms of mammary gland development and malignant progression. We previously employed next generation sequencing (NGS) and identified 63 consistently upregulated genes in three DCIS lines (SUM102, SUM225, MCF10.DCIS) compared to non-transformed mammary epithelial cells. To gain insight into common transcriptional elements of co-regulated genes that may be critical to the DCIS phenotype, we have mined our NGS data using Frameworker (Genomatix). The 63 upregulated genes are associated with 244 candidate promoters out of a total of 82,703 promoters in the entire human genome. Our analysis shows that the common framework RXRF-ZF02-ZF02-PLAG-HDBP is only present in the promoters of three genes: RAP1GAP, SPRY4 and PDGFB. All of these genes are upregulated in our DCIS signature, which means that this framework is hugely enriched (336-fold) over what would be expected by chance. Rap1Gap has tumor suppressor properties; its functions, via regulation of Rap1 activity, include regulation of cell adhesion, suppression of cell proliferation and metastasis. In other epithelial cancers, loss of Rap1Gap has been linked to epithelial to mesenchymal transition (EMT) and invasion. To begin study of Rap1Gap in breast cancer progression, we performed immunohistochemistry. We show strong Rap1Gap staining in DCIS with little to no expression in myoepithelial and stromal components. Immunoblotting results show that Rap1Gap levels in MCF10.CA1d cells (a model of invasive carcinoma) are reduced compared to those in MCF10.DCIS cells. Rap1Gap expression is low in additional basal breast cancer cell lines, but high in luminal ER+ /PR+ cell lines. We propose that over-expression of Rap1Gap may serve as a tumor suppressive mechanism in DCIS; subsequent reduction of Rap1Gap may be a switch for EMT and progression to an invasive phenotype.
Citation Format: Seema Shah, Kingsley Osuala, Shihong Mao, Quanwen Li, Bonnie Sloane, Stephen Krawetz, Raymond R. Mattingly. Exploring the role of Rap1Gap in the progression from DCIS to invasive breast carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3150. doi:10.1158/1538-7445.AM2014-3150
Abstract 4475: IL-4 receptor-targeted delivery of liposomal doxorubicin and siRNA to tumor
Authors: Murugan Poongkavithai Vadevoo Sri , Rangaswamy Gunassekaran Gowri , Chi Lianhua , Padmanaban Guruprasath , Baek Moon-Chang , Kim In-San , Park Rang-Woon , Lee Byung-Heon 
Published: May 2015

Abstract
Targeted delivery of imaging agents and therapeutics to tumors would provide early detection and increased therapeutic efficacy against cancer. Using phage displayed-random peptide libraries, we have identified IL4RPep-1 (IL-4 receptor-binding peptide-1), CRKRLDRNC, that binds to IL-4 receptor (IL4R). IL4R is over-expressed on many types of cancer cells including lung cancer and breast cancer. Peptides have smaller size and in turn may exert better tissue penetration than bulky antibodies. IL4RPep-1 bound to H226 lung tumor cells that over-express IL4R, while little binding was observed in H460 lung tumor cells that express IL4R at low levels. When injected intravenously into nude mice bearing a subcutaneous H226 tumor, Cy7.5 near-infrared fluorescence (NIRF) dye-conjugated IL4RPep-1 selectively homed to tumor. IL4RPep-1-labeled liposomes containing doxorubicin showed more efficient anti-tumor growth activity than untargeted liposomes. In addition, IL4RPep-1-labeled, branched-type polyethyleneimine nanoparticles containing Bcl-xL siRNA more efficiently knocked down Bcl-xL expression and showed higher cytotoxic effect on cancer cells over-expressing IL4R than cancer cells expressing IL4R at low levels. These results suggest that IL4RPep-1 could be a useful probe for affinity-based imaging and drug delivery to tumor that over-expressing IL4R.
Citation Format: Poongkavithai Vadevoo Sri Murugan, Gunassekaran Gowri Rangaswamy, Lianhua Chi, Guruprasath Padmanaban, Moon-Chang Baek, In-San Kim, Rang-Woon Park, Byung-Heon Lee. IL-4 receptor-targeted delivery of liposomal doxorubicin and siRNA to tumor. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4475. doi:10.1158/1538-7445.AM2014-4475
Abstract 4520: Inhibition of melanoma growth by small molecules that attenuate PI3K/PDK1 signaling and promote ATF2 mitochondrial localization
Authors: Varsano Tal , Feng Yongmei , Claps Giuseppina , Scortegagna Marzia , Lau Eric , Leonard Marilyn , Pinkerton Anthony , Pellecchia Maurizio , Davies Michael , Bosenberg Marcus , Ronai Ze'ev 
Published: May 2015

Abstract
Phosphoinositide-dependent kinase-1 (PDK-1) phosphorylates members of the AGC kinase superfamily, including AKT and PKC. We recently demonstrated the importance of PDK1 in melanoma development and progression (Oncogene 2013). Genetic ablation of PDK1 in melanocytes effectively delayed melanoma development and inhibited metastasis in a Braf/Cdkn2a model with WT or mutant Pten. In an effort to identify small molecule inhibitors that could affect the PDK1 signaling we characterized SBI-0089410, a small molecule identified in a screen for nuclear exclusion of the transcription factor ATF2, which attenuates its oncogenic activity while enabling its function at the mitochondrial membrane to promote apoptosis. SBI-0089410 reduced melanoma cell viability, inhibited colony formation and spheroid growth, decreased mitochondrial membrane permeability, decreased ATF2 phosphorylation by PKCϵ and altered ATF2 transcriptional activity. Notably, the effect of SBI-0089401 was attenuated by the overexpression of constitutively active PKCϵ or ATF2T52E, which carries a phosphomimetic mutation at the PKCϵ phosphorylation site. Reverse phase protein array (RPPA), validated by western blot analysis, revealed that SBI-408910 attenuates PI3K/AKT/PDK1 signaling pathways. In vivo, SBI-089410 attenuated the growth of Braf/Cdkn2a/Pten tumors in a syngeneic B6 mouse model, as well as prevented their resistance to BRAFi. Our studies substantiate the importance of targeting PI3K/PDK1 signaling in melanoma as a means for effective therapy and prevention of chemoresistance.
Citation Format: Tal Varsano, Yongmei Feng, Giuseppina Claps, Marzia Scortegagna, Eric Lau, Marilyn Leonard, Anthony Pinkerton, Maurizio Pellecchia, Michael Davies, Marcus Bosenberg, Ze'ev Ronai. Inhibition of melanoma growth by small molecules that attenuate PI3K/PDK1 signaling and promote ATF2 mitochondrial localization. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4520. doi:10.1158/1538-7445.AM2014-4520
Abstract 458: Regulation of HIF1a by Set9 lysine methyltransferase
Authors: Liu Qiong , Geng Hao , Xue Changhui , Beer Tomasz M. , Qian David Z. 
Published: May 2015

Abstract
HIF-1 is consisted of an oxygen-sensitive HIF-1α and a constitutive HIF-1β subunit. The activity of HIF-1 is implicated in human cancer development. In normoxia, the HIF-1α subunit is constantly synthesized, but rapidly degraded by the oxygen-dependent mechanism. In hypoxia due to tumor growth and anti-angiogenic therapy, HIF-1α is stabilized and dimerizes with HIF-1β to form HIF-1, which transactivates genes promoting cancer hallmarks, including angiogenesis, metabolic reprogramming, migration and invasion. Notably, HIF-1 increases multiple genes on the glycolytic pathway, which mediates the metabolic adaptation of hypoxic stress. Therefore, HIF-1α is an intriguing anticancer target. However, how HIF-1α is regulated in hypoxia is poorly understood. Here, we report that the histone methyltransferase Set9 promotes HIF-1α protein stability in hypoxia and plays an important role in cancer cell glycolytic adaptation. Specifically, Set9 interacts with HIF-1α to maintain its stability in hypoxia. Silencing of Set9 by siRNA in hypoxia significantly reduces HIF-1α protein level, reduces HIF-1 transcriptional activity, and attenuates expressions of HIF-1-responsive glycolytic genes as well as hypoxia-induced glycolysis. Further, we have found that Set9 binds to the chromatins of hypoxia response elements within HIF-1-responsive glycolytic genes. The results suggest that Set9 epigenetically contributes to the glycolytic phenotype of cancer through HIF-1.
Citation Format: Qiong Liu, Hao Geng, Changhui Xue, Tomasz M. Beer, David Z. Qian. Regulation of HIF1a by Set9 lysine methyltransferase. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 458. doi:10.1158/1538-7445.AM2014-458
Abstract 4588: Polylysine linker for successful manufacturing of hydrophobic peptides
Authors: Shakoori Firuz , Gangakhedkar Archana 
Published: May 2015

Abstract
Hydrophobic peptides are common to be used in cancer vaccinations. Hydrophobic peptides, continue to present challenges for large scale production in peptide industry. Experimental Procedures: Full sequence prepared by Solid phase Synthesis:The peptide was synthesized with Fmoc-Lys(Boc)-Wang resin by using standard Fmoc/tBu chemistry. Protecting groups used for amino acids are: tBu for Thr, Boc for Trp and Lys and Boc-Ser- OH was used as a hydrolysis-labile ester provider. Fmoc-Leu-OH was coupled on the side chain of Ser by DIC/DMAP (Amino acid/DIC/DMAP, 3/1.5/0.3 eq). The other amino acids were assembled by repetitive removal of the Fmoc protecting group by treating with 2% DBU/2% piperidine in DMF 30 minutes and coupling of protected amino acid by using DIC/HOBt/NMM (4:4:4 eq) in 180 minutes to overnight. Ninhydrin test was performed after each coupling to check the coupling efficiency. After the last coupling,resin was washed and dried, then treated with reagent K for cleavage and removal of the side chain protecting groups. Crude peptide was precipitated from cold ether and collected by filtration. Table.1
Summary of 4 synthesize results
Syn. NoLinker StructurePurity of CrudeDKP deletion: Product Hydrolysis YieldOverall Yield1No Linker61.4%12.3%2(O)-Ser56.5%1:252.8%12.5%3(O)-Ac-Ser40.8%1:370.6%28.2%4HMBA78.1%&lt;1:2451.0%61.2%
Conclusion: American Peptide demonstrated the utility of a hydrophilic peptide ‘tail’ in purification of a hydrophobic peptide on both small (20-30 mmol) research scale and large (800 mmol) cGMP scales to provide the peptide in hundreds-of-grams quantities. The approach described allowed for both time-and cost-competitive manufacturing of this difficult peptide for clinical trials.
Citation Format: Firuz Shakoori, Archana Gangakhedkar. Polylysine linker for successful manufacturing of hydrophobic peptides. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4588. doi:10.1158/1538-7445.AM2014-4588
Abstract 481: Dualistic regulation of NF-kB signaling by CD99 and EWS-FLI1 in Ewing sarcoma: impact on cell differentiation
Authors: Ventura Selena , Manzalini Chiara , Aryee Dave , Picci Piero , Kovar Heinrich , Scotlandi Katia 
Published: May 2015

Abstract
Introduction: Ewing sarcoma (ES) is a high-grade rare malignant bone tumor in children and adolescents. This disease is primarily characterized by: i) Chromosomal translocations that fuse EWS gene and a gene of the ETS family of transcription factors (around 85% EWS-FLI1 and 10% EWS-ERG, respectively) which are the oncogenic drivers of this tumor; ii) High expression of CD99, a cell surface glycoprotein virtually present in all of ES cells. EWS-FLI1 and CD99 were reported to have divergent roles in modulating ES malignancy and neural differentiation In this study we investigated the possible role of nuclear factor-kappa B (NF-kB) signaling pathway on the neural differentiation of ES cells and its relationship with EWS-FLI1 and CD99.
Materials and Methods: Transcriptional activity of several transcription factors (AP1, SRE, CREB, E2F and NF-kB) were evaluated by gene reporter assay. Using a knockdown approach in stable and transient conditions we silenced CD99, EWS-FLI1 and NF-kB itself, alone or in combination, in three ES cell lines (TC-71, IOR/CAR and ASP14). NF-kBp65 was also transiently overexpressed in TCshCD99 and CARshCD99 clones with pCMVp65 construct. Neural differentiation was detected evaluating β-III tubulin by immunofluorescence microscopy.
Results: NF-kB was found heterogeneously expressed in a panel of ES cell lines. Stable CD99 silencing was found to decrease NF-kB activity (pValue &lt;0.05) without any differences at RNA and protein levels. On the contrary, EWS-FLI1 knockdown increased NF-kB transactivation (pValue &lt;0.01). CD99 affects NF-kBp65 activity and its effect are dominant with respect to EWS-FLI1. Accordingly, EWS-FLI1 silencing was not able to revert the reduction of NF-kB activity observed after CD99 deprivation. In addition cells silenced for CD99 but not for EWS-FLI1 (TCshCD99 and CARshCD99 clones) showed a neural differentiated phenotype, which was lost when cells were induced to re-express NF-kB.
Conclusion: Our findings support the concept that inhibition of NF-kB is able to induce terminal neural differentiation of ES cells. This identifying NF-kB as a potential therapeutic target against Ewing sarcoma progression.
(Supported by AIRC_2013_IG 14049 and Liddy Shriver Foundation)
Citation Format: Selena Ventura, Chiara Manzalini, Dave Aryee, Piero Picci, Heinrich Kovar, Katia Scotlandi. Dualistic regulation of NF-kB signaling by CD99 and EWS-FLI1 in Ewing sarcoma: impact on cell differentiation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 481. doi:10.1158/1538-7445.AM2014-481
Abstract 4811: Soluble biomarkers identify mCRC patients subgroups showing increased benefit from the VEGF signaling inhibitor cediranib in combination with chemotherapy
Authors: Pommier Aurelien J.C. , Spencer Stuart , Morgan Shethah , Womack Chris , Jurgensmeier Juliane M. , Critchlow Susan , Barry Simon T. 
Published: May 2015

Abstract
Identifying potential predictive biomarkers differentiating sensitivity/resistance to VEGF signaling inhibitors (VEGFi) will enhance use of these therapies. We used &gt;580 pre and on-treatment serum samples and 259 tumours diagnostic biopsies from mCRC patients enrolled in a Phase III trial (HORIZON II) treated with chemotherapy ± cediranib(a VEGFi), to screen the potential predictive value of multiple serum proteins and tumour genes.
At baseline leptin was the most significant marker associated with OS of patients treated with chemo-ced (Pinteraction&lt;0.001). Leptin concentration correlated with the body mass index (BMI) and obese patients (BMI&gt;30) showed improved OS when treated with chemo-ced compared to non-obese patients (HR=0.55, 0.36-0.85, p=0.008). In the same patients, we identified two gene expression signatures associated with benefit on chemo-ced. Patients characterized by a high expression of GAHGL (Glucose metabolism, Angiogenesis, Hypoxia, Glutamine metabolism, Leptin signaling) (Pinteraction=0.028) and low expression of LOPS (Lipid metabolism, Oxidative activity, Proliferation and Serine synthesis) gene signature (Pinteraction=0.002) had improved OS in chemo-ced treated patients. Serum leptin concentration was found positively correlated with expression of genes involved in GAHGL in patients.
In addition a baseline signature (BS) of 47 biomarkers including VEGFA, VEGFD, VEGFR2, VEGFR3 and TIE-2, defined two distinct subgroups of patients. Patients treated with chemotherapy plus cediranib who had ‘high’ BS had shorter PFS (HR=1.82, p=0.003) than patients with ‘low’ BS. This BS had no effect on PFS of the patients treated with chemotherapy plus placebo.
Finally, we identified a profile of 16 pharmacodynamic (PD) proteins on treatment associated with PFS (HR=0.58, p&lt;0.001) and OS (HR=0.52, p&lt;0.001) in patients treated with chemotherapy plus cediranib. This PD profile had no effect on PFS and OS in patients treated with chemotherapy plus placebo.
This work provides original candidate biomarkers to select patients most likely to benefit from VEGFi therapies.
Citation Format: Aurelien J.C. Pommier, Stuart Spencer, Shethah Morgan, Chris Womack, Juliane M. Jurgensmeier, Susan Critchlow, Simon T. Barry. Soluble biomarkers identify mCRC patients subgroups showing increased benefit from the VEGF signaling inhibitor cediranib in combination with chemotherapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4811. doi:10.1158/1538-7445.AM2014-4811
Abstract 4843: Neutrophil elastase enhances adaptive immunity via increase of peptide presentation and downregulation of T cell inhibitory molecules
Authors: Chawla Akhil , Philips Anne V. , Qiao Na , Sukhumalchandra Pariya , Kerros Celine , Alatrash Gheath , Molldrem Jeffrey J. , Mittendorf Elizabeth A. 
Published: May 2015

Abstract
Introduction: Neutrophil elastase (NE) is an inflammatory mediator that is taken up by breast cancer cells. We have previously shown that NE uptake increases susceptibility to lysis by cytotoxic T lymphocytes (CTL) targeting the tumor antigens PR1 and cyclin E. We investigated whether NE uptake alters adaptive immunity by affecting MHC class I antigen presentation and T cell inhibitory molecules.
Methods: MDA-MB-231 breast cancer cells were maintained in standard media ± NE. Healthy donor HLA-A2+ peripheral blood mononuclear cells were used to generate antigen specific CTL by pulsing dendritic cells with E75, a HER2-derived epitope we are currently investigating as a vaccine in clinical trials. Calcein-AM cytotoxicity assays evaluated E75-specific lysis. Flow cytometry was used to show NE uptake, HLA-A2, pan-HLA class I (HLA-ABC) and PD-L1 surface expression. Western blot analysis was used to show total MHC class I heavy chain, B2M, LMP2, TAP1, calnexin and tapasin expression. Relative PD-L1 mRNA levels were determined using qPCR.
Results: NE uptake enhanced tumor cell lysis by E75-specific CTL at all effector to target (E:T) ratios (lysis for 20 to 1 E:T ratio was 50% ± 7 for NE treated vs 28% ± 7 for untreated, p &lt; 0.05), and led to an increase in HLA-A2 and HLA-ABC surface expression (p &lt; 0.05; Table). However, NE uptake did not change total expression of MHC class I or components of the MHC class I pathway. Additionally, NE uptake led to a decrease in PD-L1 surface expression and transcript levels (p &lt; 0.05; Table).
Conclusion: NE increases tumor antigen presentation as well as MHC class I on the cell surface. However, NE uptake does not change total MHC class I expression or expression of MHC class I antigen processing components, suggesting that NE increases peptide availability as a mechanism of increased MHC class I antigen presentation. NE also leads to the downregulation of PD-L1. Together, our data indicate that NE uptake enhances adaptive immune responses.
HLA-A2 Surface Expression (MFI)HLA-ABC Surface Expression (MFI)PD-L1 Surface Expression (MFI)PD-L1 mRNA Expression (RQ)NE Treated (Mean ± SD)7341 ± 8671317 ± 1201723 ± 550.274 ± 0.02Untreated (Mean ± SD)5144 ± 272802 ± 1336966 ± 4511
Citation Format: Akhil Chawla, Anne V. Philips, Na Qiao, Pariya Sukhumalchandra, Celine Kerros, Gheath Alatrash, Jeffrey J. Molldrem, Elizabeth A. Mittendorf. Neutrophil elastase enhances adaptive immunity via increase of peptide presentation and downregulation of T cell inhibitory molecules. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4843. doi:10.1158/1538-7445.AM2014-4843