bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4540: Development of a biochemical High Throughput Screening (HTS) assay for chemical inhibitors of MALT1, a target for lymphoma therapeutics.
Authors: Zhai Dayong , Shu Chih-Wen , Diaz Paul , Reed John C. 
Published: August 2014

Abstract
MALT1 plays critical role in antigen receptor-mediated lymphocyte activation and lymphomagenesis. MALT1 acts both as a scaffold protein to recruit upstream signal transducers and as an intracellular cysteine protease with proteolytic activity that cleaves substrate proteins involved in NF-kB induction. Dysregulation of MALT1 activity occurs in lymphoma as a result of chromosomal translocations that fuse the catalytic domain of MALT1 with portions of BirC3 (c-IAP2). Previous studies have shown that MALT1’s proteolytic activity is required for its participation in NF-kB induction and for promoting cell viability of certain types of lymphomas, particularly the activated B-cell (ABC) type of diffuse large B-cell lymphoma.
To lay a foundation for discovery of drug-like chemical inhibitors of MALT1, we developed a fluorescence-based, biochemical assay for measuring protease activity of MALT1 in vitro in 384 and 1,536 well formats. The assay components included recombinant purified full-length MALT1 protein, the MALT1-binding protein, Bcl-10, and the fluorigenic substrate Ac-LRSR-AMC (Ac-Leu-Arg-Ser-Arg-7-amino-4-methylcoumarin). Experiments showed that addition of Bcl-10 increases MALT1‘s protease activity up to 20-40 fold. The high throughput screening (HTS) assay was validated with excellent Z’ factors (&gt;0.7) in both 384 and 1,536 well formats, with performance remaining robust for more than 6 hours, and thus suitable for screening of large chemical libraries. The HTS assay was validated by performing pilot screens with two small collections of compounds enriched in bioactive molecules (n = 1280 for LopacTM and 2000 for SpectrumTM library), yielding confirmed hit rates of 0.2 % and 0.7%, respectively. Counter-screening assays were also generated using other types of intracellular cysteine proteases, specifically Caspase-3 and Autophagin-1 (ATG4B), for eliminating non-specific inhibitors. Various supporting cell-based assays have also been generated to support hit characterization. In conclusion, we have generated a robust biochemical HTS assays for detection of chemical inhibitors of the MALT1 protease. We plan to utilize the HTS assay for screening large compound libraries as a starting point for discovery of novel therapeutics for treatment of lymphoma, autoimmunity, and allograft rejection.
Citation Format: Dayong Zhai, Chih-Wen Shu, Paul Diaz, John C. Reed. Development of a biochemical High Throughput Screening (HTS) assay for chemical inhibitors of MALT1, a target for lymphoma therapeutics. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4540. doi:10.1158/1538-7445.AM2013-4540
Abstract 4548: Identify novel molecular structures targeting X-Linked Inhibitor of Apoptosis Protein by ligand-based virtual screening.
Authors: Wang Jin , Duke Charles B. , Li Wei 
Published: August 2014

Abstract
Resistance to apoptosis is believed as one of the mechanism underlying a number of human malignancies notorious resistances to current chemotherapeutics. The X-linked inhibitor of apoptosis protein (XIAP) inhibits apoptosis by directly binding and deactivating caspase-9, caspase-3/7. Elevated XIAP expression in tumor tissue is closely associated with metastatic progression. Mimicking the XIAP-binding motif of second mitochondria-derived activator of caspase (SMAC) is the most widely used strategy to discover or design therapeutic agents targeting XIAP. Here we screened a large database of 362K compounds (the University of Cincinnati's Drug Discovery Center Library) in silico to find molecules with novel structures that can strongly interact with the third BIR domain of XIAP. We chose AVPI tetrapeptide from the SMAC as the lead structure and run the shape similarity search using program Canvas, version 1.4 (Schrodinger, LLC, New York, USA). Then top 10,000 structures of the similarity rank were docked into SMAC binding sites in two different SMAC-BIR3 domain complexes crystal structure (PDB ID: 1TW1 and 1G73) separately. The highest docking score of the screened structures is up to -9.0 in both of complexes while the SMAC AVPI tetrapeptide showed docking score at -9.4. Considering the docking scores in both complexes and the flexibility of chemical scaffolds, we manually selected 72 compounds with docking scores varied from -8.4 to -5.5 for in vitro anti-proliferation and caspase functional testing. Four compounds which haven't been reported with any anti-cancer activity, inhibited the cell growth by over 70% in two human melanoma cell lines (A375 and MDA-MB-435) and two human prostate cell lines (PC-3 and LNCap) at the concentration of 10uM after 48hr incubation. IC50 values of those compounds were at least 4 folds smaller than that of Embelin, a well-recognized XIAP inhibitor, in all four cell lines tested. The best of them, compound 402112, have IC50 values of 650nM to 2.5uM against the four tested cancer cell lines. Caspase functional assay showed that compound 402112 activates capase-9 activity at 4 μM, significantly better than Embelin did at a 10-fold higher concentration (40 μM) after 48hr treatment in PC-3 and A375 cells. Taken together, our study identified novel molecule structures which are amicable to further extensive modification for the development of high potent XIAP inhibitors.
Citation Format: Jin Wang, Charles B. Duke, Wei Li. Identify novel molecular structures targeting X-Linked Inhibitor of Apoptosis Protein by ligand-based virtual screening. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4548. doi:10.1158/1538-7445.AM2013-4548
Abstract 4554: A docking domain directed inhibitor targeting ERK regulated F-site substrates.
Authors: Zhang Jun , Oashi Taiji , Samadani Ramin , Nevels Kerrick , Burkhard Kimberly , Priyakumar Deva , Raman Prabhu , Fletcher Steven , Pozharsky Edvin , Shapiro Paul , MacKerell Alexander 
Published: August 2014

Abstract
Extracellular signal-regulated kinases 1 and 2 (ERK1/2) are ubiquitously expressed in almost all tissues and cell types. Accordingly, ERK1/2 regulate a diversity of cellular processes by phosphorylating more than 150 substrate proteins. With its constitutive activation being frequently implicated in human malignancies, ERK has been an appealing target in anti-cancer drug development. However, currently available ERK1/2 inhibitors are ATP-competitive. Since these ERK inhibitors block all ERK functions, the potential for side effects in normal tissues and cells may limit their potential of being developed into therapeutic agents. To overcome this limitation, we used a combination of computational and experimental methods to develop small molecular weight inhibitors that target ERK-substrate docking domains. Here we report the identification, characterization and optimization of a compound with a thienyl benzenesulfonate scaffold that specifically inhibits substrates containing an F-site or DEF (docking site for ERK, FXF) motif. Our X-ray crystallography data showed that this compound bound to a pocket in the vicinity of the F-site docking domain on ERK. Biological analysis further demonstrated that this compound and its analogs could preferentially inhibit phosphorylation and function of F-site-containing ERK substrates, and that these compounds preferentially inhibited growth of melanoma cells harboring a V600E B-Raf mutant. Additionally, this compound was shown to inhibit the phosphorylation of specific PKC isoforms via an ERK dependent mechanism. These findings represent the first identification and validation of chemicals that selectively inhibit ERK substrates and signaling events through ATP-independent mechanisms. These compounds have potential utility for elucidating the complex biological roles of ERK1/2 and development into novel anti-cancer agents.
Citation Format: Jun Zhang, Taiji Oashi, Ramin Samadani, Kerrick Nevels, Kimberly Burkhard, Deva Priyakumar, Prabhu Raman, Steven Fletcher, Edvin Pozharsky, Paul Shapiro, Alexander MacKerell. A docking domain directed inhibitor targeting ERK regulated F-site substrates. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4554. doi:10.1158/1538-7445.AM2013-4554
Abstract 4565: Whole exome sequencing links MAPK1 mutation to exquisite sensitivity to brief erlotinib monotherapy in head and neck cancer.
Authors: Lui Vivian WY , Van Allen Eliezer , Li Hua , Zhang Fan , Zeng Yan , Johnson Jonas T. , Garraway Levi A. , Mills Gordon B. , Grandis Jennifer Rubin 
Published: August 2014

Abstract
Activating mutations of EGFR, primarily found in a subgroup of lung cancers, serve as predictive biomarkers for EGFR tyrosine kinase inhibitor (TKI) sensitivity. Clinical responses to EGFR TKIs in other cancers are only moderate and predictive biomarkers are lacking. Here, we report a head and neck cancer patient with no EGFR genetic alterations, who experienced a complete histologic clinical response after 14 days of erlotinib monotherapy on a window-of-opportunity presurgical trial. To determine the somatic genetic events underlying the extraordinary and unexpected response to erlotinib alone, whole exome sequencing was performed. A bi-allelic missense mutation of MAPK1(E322K) was detected in the tumor in the absence of other mutations reported to be correlated with EGFR TKI sensitivity in preclinical models or cancer patients. Combined bioinformatics analyses of genomics-drug databases revealed, for the first time, that loss-of-function of MAPK1 or MAPK1 mutation correlates with erlotinib sensitivity. Functional studies confirmed heightened sensitivity to erlotinib by loss-of-function of MAPK1 in HNSCC cells. Our preliminary findings suggest that this MAPK1 mutation, which is also found at low frequencies in HNSCC and cervical cancers, may identify a subgroup of cancer patients who may be cured with EGFR TKI monotherapy.
Citation Format: Vivian WY Lui, Eliezer Van Allen, Hua Li, Fan Zhang, Yan Zeng, Jonas T. Johnson, Levi A. Garraway, Gordon B. Mills, Jennifer Rubin Grandis. Whole exome sequencing links MAPK1 mutation to exquisite sensitivity to brief erlotinib monotherapy in head and neck cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4565. doi:10.1158/1538-7445.AM2013-4565
Abstract 457: Matrix metalloproteinase-2 modulates antitumor immunity.
Authors: Godefroy Emmanuelle , Gallois Anne , Manches Olivier , Ensanyat Shaheen , Jotereau Francine , Bhardwaj Nina 
Published: August 2014

Abstract
Background: Cancer cells, including melanoma, can be highly resistant to traditional treatments such as chemotherapy and radiotherapy. As a result, a lot of effort has been put into developing immune therapies to treat cancer patients. The main barrier to design effective immunotherapies is the immunosuppression induced by the tumor. Matrix metalloproteinase-2 (MMP-2) is over-expressed in most cancers including melanoma, and its expression is associated with increased dissemination and poorer survival/prognosis. Here, we not only uncovered an immunosuppressive role of MMP-2 in inducing ineffective/detrimental TH2 immune responses, but also the underlying mechanisms.
Methods: Human dendritic cells (DCs) were cultured in the presence of MMP-2 and/or various TLR agonists. DCs responses were monitored using immunostaining, ELISA or cytometric bead array methods. Autologous CD4+ T cells were stimulated using these conditioned tumor-associated antigen (TAA)-pulsed DCs. TAA-specific CD4+ T cells were cloned and characterized using mainly the same techniques as for DCs.
Results: Here we showed that MMP-2-conditioned human DCs primed naïve CD4+ T cells to differentiate into an inflammatory TH2 phenotype, i.e. mainly secreting TNFα, IL-4 and IL-13 and expressing GATA3. Accordingly, we detected MMP-2-specific CD4+ T cells displaying the same inflammatory TH2 profile in tumor-infiltrating lymphocytes from melanoma patients. We revealed the underlying mechanisms: on the one hand, MMP-2 was found to degrade the type-I IFN receptor IFNAR1, thereby preventing STAT1 phosphorylation, which is necessary for IL-12 production. On the other hand, MMP-2 triggers the Toll-like receptor (TLR)-2 on DCs, which leads to NF-κB activation and OX40L expression. Both lack of IL-12 and over-expression of OX40L were found responsible for skewing T cell responses towards a detrimental TH2 phenotype.
Conclusions: MMP-2, therefore, acts as an endogenous TH2 "conditioner" and may underlie the prevalence of detrimental TH2 responses in cancer. Our findings also described the responsible MMP-2-dependent mechanisms, which open the way to novel therapeutic strategies and/or targets to skew anti-tumor CD4+ T cell responses towards a more efficient anti-tumor TH1 phenotype to treat cancer patients.
Citation Format: Emmanuelle Godefroy, Anne Gallois, Olivier Manches, Shaheen Ensanyat, Francine Jotereau, Nina Bhardwaj. Matrix metalloproteinase-2 modulates antitumor immunity. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 457. doi:10.1158/1538-7445.AM2013-457
Abstract 4573: STINGVAX - A novel tumor vaccine with cyclic dinucleotides - can induce potent anti-tumor responses in vivo.
Authors: Dubensky Thomas W. , Leong Meredith L. , Kanne David B. , Lemmens Edward E. , Metchette Ken , Liu Weiqun , Fasso Marcella , Fu Juan , Woodward Joshua J. , Pardoll Drew , Portnoy Daniel A. , Kim Young J. 
Published: August 2014

Abstract
Cyclic dinucleotides (CDN) are ubiquitous bacterial intracellular messengers, and they have recently been shown to also function as a pathogen associated molecular pattern (PAMP) molecules that are sensed by eukaryotic host Stimulator of Interferon Genes (STING) that can activate the TBK-1/IRF-3 signaling pathway to induce type I interferon and other co-regulated genes. CDN was found to induce STING dependent augmentation of T-cell priming in multiple vaccination models, potentially by activating dendritic cells. We formulated CDN with a GM-CSF secreting tumor cell vaccine (STINGVAX) to mobilize and as well as activate dendritic cells both in vitro and in vivo. STINGVAX was tested in a stringent B16 melanoma treatment model, and we demonstrated significant reduction of tumor growth rate in vivo. STINGVAX's anti-tumor response was correlated with increased tumor infiltrating CD8 T-cells as well as increased number of p15E tumor-specific cytotoxic T-cells. STINGVAX's in vivo anti-tumor response was T-cell dependent as well as STING dependent. When we combined STINGVAX with multiple TLR 4/7/8 agonists signaling through the MyD88/TRIF pathway that is distinct from CDN/STING/TBK-1 signaling treatment of palpable B16 tumor resulted in regression of 30-60% of these non-immunogenic tumors. When the STINGVAX treated tumor microenvironment was examined, both IFNγ+CD8+ and PD-L1 was upregulated, potentially demonstrating an adaptive immune resistance mechanism which would render STINGVAX an excellent candidate to be combined with anti-PD-1 blockade. Cumulatively, STINGVAX is a novel tumor vaccine with a high potential for translation in clinical oncology.
Citation Format: Thomas W. Dubensky, Meredith L. Leong, David B. Kanne, Edward E. Lemmens, Ken Metchette, Weiqun Liu, Marcella Fasso, Juan Fu, Joshua J. Woodward, Drew Pardoll, Daniel A. Portnoy, Young J. Kim. STINGVAX - A novel tumor vaccine with cyclic dinucleotides - can induce potent anti-tumor responses in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4573. doi:10.1158/1538-7445.AM2013-4573
Abstract 4576: Immunization with CPG in combination with MHC class I and class II peptides stimulates rapid and strong tumor antigen-specific CTL responses in melanoma patients.
Authors: Fourcade Julien , Sun Zhaojun , Pagliano Ornella , Chauvin Joe-Marc , Sander Cindy , Moschos Stergios , Tarhini Ahmad , Tawbi Hussein , Kirkwood John M. , Zarour Hassane M. 
Published: August 2014

Abstract
There is now ample evidence that the toll-like receptor 9 ligand CPG/PF-3512676 represents a potent adjuvant for peptide/protein-based cancer vaccines capable of stimulating ex vivo detectable tumor antigen (TA)-specific CD8+ T cell responses in patients with advanced melanoma. Although CD4+ T cells appeared to promote the expansion and functions of CD8+ T cells in experimental models, melanoma-associated helper peptides have been shown to paradoxically decrease CD8+ T cell responses to a multipeptide melanoma vaccine emulsified in Montanide, possibly by inducing antigen-specific Tregs. To further investigate this question, we have performed one Phase I study of immunization where patients with advanced melanoma were immunized with CPG, Montanide and the HLA-A2-restricted analog peptide NY-ESO-1 157-165V either alone (Arm 1, n=5) or in combination with the pan-DR CD4 helper peptide NY-ESO-1 119-143 (Arm 2, n=7). Patients in both arms exhibited a rapid increase of NY-ESO-1-specific CD8+ T cells that were detectable ex vivo by flow cytometry. Strikingly, the mean frequency of vaccine-induced NY-ESO-1-specific CD8+ T cells after 6 biweekly immunizations was five times higher in patients who also received the helper peptide. Melanoma patients immunized with the helper peptide (Arm 2) developed a rapid increase of Foxp3− NY-ESO-1-specific CD4+ T cells that produced Th-1 type cytokines and IL-21 ex vivo. As compared to Arm 1, melanoma patients included in Arm 2 developed vaccine-induced NY-ESO-1-specific CD8+ T cells that were more differentiated, produced more IFN-γ and exhibited higher cytolytic functions in ex vivo assays. Collectively, our data support the inclusion of well-defined MHC class II peptides in combination with CPG/PF-3512676 and MHC class I peptides in multipeptide melanoma vaccines to better augment the expansion, differentiation, IFN-γ production and lytic function of TA-specific CTLs in patients with advanced melanoma.
This work was supported by the National Institutes of Health/National Cancer Institute grants R01CA90360 and R01CA157467 (to H.M. Zarour).
Citation Format: Julien Fourcade, Zhaojun Sun, Ornella Pagliano, Joe-Marc Chauvin, Cindy Sander, Stergios Moschos, Ahmad Tarhini, Hussein Tawbi, John M. Kirkwood, Hassane M. Zarour. Immunization with CPG in combination with MHC class I and class II peptides stimulates rapid and strong tumor antigen-specific CTL responses in melanoma patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4576. doi:10.1158/1538-7445.AM2013-4576
Abstract 4584: Bone marrow-derived Gr1+ cells can generate a metastasis-resistant microenvironment via induced secretion of thrombospondin-1.
Authors: Catena Raul , Bhattacharya Nandita , El Rayes Tina , Choi Hyejin , Wang Suming , Gao Dingcheng , Joshi Natasha , Ryu Seongho , Bielenberg Diane , Lee Sharrell , Haukaas Svein , Gravdal Karsten , Halvorsen Ole , Akslen Lars , Watnick Randolph , Mittal Vivek 
Published: August 2014

Abstract
Previous studies have shown that metastatic tumors establish premetastatic niches comprised of recruited bone marrow (BM)-derived cells in potential distant target metastatic organs. These premetatatic niches serve as permissive hubs for supporting future metastases. However, little is known about whether tumors that lack metastatic potential establish such niches, or generate niches that inhibit metastasis. Here, we show that metastasis-incompetent tumors are also capable of systemically generating BM-derived niches in the metastatic organs. However, by secreting the glycoprotein prosaposin these tumors systemically stimulate expression of antitumorigenic factor thrombospondin-1 (Tsp-1) in the Gr1+ myeloid cells converting them it into a metastasis-refractory niche. Genetic deletion of Tsp-1 abolished the inhibition of metastasis, which was restored by BM transplant from Tsp-1+ donors.
We developed a 5-amino acid peptide from prosaposin as a pharmacological inducer of Tsp-1 in Gr1+ BM cells. Strikingly, the peptide suppresses metastasis. These results provide mechanistic insights into why certain tumors are deficient in metastatic potential and implicate recruited Gr1+ myeloid cells as the main source of Tsp-1, contrary to previous notion. These results also underscore the plasticity of Gr1+ cells, which, depending on the context and the nature of paracrine factor/s can either promote or inhibit metastasis. Clinically, examination of a tumor tissue microarray (TMA) compiled from 103 prostate cancer patients with long and complete follow up, revealed that prosaposin expression positively correlated with increased overall survival, suggesting that the prosaposin peptide may be a potential therapeutic agent against metastatic cancer.
Citation Format: Raul Catena, Nandita Bhattacharya, Tina El Rayes, Hyejin Choi, Suming Wang, Dingcheng Gao, Natasha Joshi, Seongho Ryu, Diane Bielenberg, Sharrell Lee, Svein Haukaas, Karsten Gravdal, Ole Halvorsen, Lars Akslen, Randolph Watnick, Vivek Mittal. Bone marrow-derived Gr1+ cells can generate a metastasis-resistant microenvironment via induced secretion of thrombospondin-1. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4584. doi:10.1158/1538-7445.AM2013-4584