bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2441: Engineering and mapping difficult proteins using comprehensive mutagenesis
Authors: Paes Cheryl , Goodman Jason , Wescott Melanie , Thaker Yana , Thomas Anu , Couto Joseph , Rucker Joseph , Doranz Benjamin J. 
Published: August 2015

Abstract
Alanine (Ala) scanning is a widely used mutagenesis approach for systematic evaluation of protein function, which for large targets can be laborious and time-consuming. We have developed a platform technology, Shotgun Mutagenesis, for rapid and comprehensive alanine scanning mutagenesis for protein mapping and engineering applications. Using this technology, every residue in a target protein is individually mutated, expressed in human cells, and assayed for desired functions. Critical mutations are rapidly identified using high-throughput 384-well assays including flow cytometry. Here we used Shotgun Mutagenesis to create comprehensive mutation libraries for different classes of targets including the GPCR CXCR4, bitter taste receptor TAS2R16 and the therapeutic antibody Palivizumab (Synagis). For the cancer target CXCR4, we identified critical residues for antibody binding (epitope mapping) and ligand-dependent activation (functional domain mapping). We also discovered TAS2R16 residues that can be altered for increased expression (protein optimization) and Palivizumab residues which tolerate germline substitutions permitting engineering of a maximally humanized antibody variant (antibody humanization). This platform allows comprehensive protein mapping and engineering of challenging targets previously considered prohibitively time-consuming or expensive, enabling discovery pathways for many untapped therapeutic targets.
Citation Format: Cheryl Paes, Jason Goodman, Melanie Wescott, Yana Thaker, Anu Thomas, Joseph Couto, Joseph Rucker, Benjamin J. Doranz. Engineering and mapping difficult proteins using comprehensive mutagenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2441. doi:10.1158/1538-7445.AM2015-2441
Abstract 2453: The role of p21 in the toxicity of mitomycin C and decarbamoylmitomycin C
Authors: Champeil Elise , Cheng Shu-Yuan , Huang Bik Tzu , Seo Jiwon 
Published: August 2015

Abstract
The mitomycins are a family of antitumor antibiotics made by Streptomyces. One member of this family, mitomycin C (MC), is currently used to treat certain cancers. Its mode of action has been extensively examined. In comparison, 10-decarbamoyl mitomycin C (DMC) has not been the object of such intense investigation. It has recently been found to form similar or identical adducts with DNA as mitomycin C (MC). In particular, DMC generates a unique stereoisomeric interstrand crosslink (beta isomer, β-ICL). Although the DNA-adducts of both drugs share common structural features, the biochemical responses to the two drugs are different. In particular, contrary to MC, the DNA-adducts generated by DMC treatment (β-ICL) rapidly activate a p53-independent signal transduction pathway. Thus, the study MC-DMC provides an ideal model for identifying structural features determining the cell signaling outcome in the presence or the absence of a functioning p53 pathway. In particular, we are interested in correlating MC and DMC-adducts structures with the role of p21 in the toxicity of the α-ICL and β-ICL. In this report, we present the following preliminary results 1) Synthesis of the beta DNA monoadduct of DMC via post-oligomerization methods 2) Role of p21 in the upstream p53-independent signaling pathway in response to MC/DMC and the α-ICL.
Citation Format: Elise Champeil, Shu-Yuan Cheng, Bik Tzu Huang, Jiwon Seo. The role of p21 in the toxicity of mitomycin C and decarbamoylmitomycin C. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2453. doi:10.1158/1538-7445.AM2015-2453
Abstract 2454: Targeting the translational enhancer complex MCT-1:DenR to disrupt NHL survival
Authors: Stowe Sean D. , Cavalier Michael C. , Godoy-Ruiz Raquel , Varney Kristen J. , Wilder Paul T. , Gartenhaus Ronald B. , Weber David J. 
Published: August 2015

Abstract
Despite the vast number of therapeutic advances that have been made, Non-Hodgkin Lymphoma (NHL) remains a leading cause of cancer-related deaths in the United States. In order to increase the NHL cure rate, new targeted options are needed. One such option that has been the focus of much recent interest is the post-transcriptional regulation in anti-apoptotic phenotypes of NHL, particularly the translational enhancer complex of MCT-1 and DENR. Involved in translation initiation and ribosomal recycling for transcripts regulated by 5′-uORF’s, the MCT-1:DENR complex has been shown to increase cell proliferation and the translation of survival-related transcripts through the combination of two critical RNA-binding domains. The goal of this project has been to biophysically characterize the formation and activity of this complex and to develop a method for evaluating the effects of disrupting its formation in vivo through the combination of NMR, X-ray crystallography, cell-free and cell-based assays. Ultimately, the information gained from this study will be used to evaluate a library of perturbagens that interfere with dimer formation and the development of MCT-1 driven anti-apoptotic phenotypes.
Citation Format: Sean D. Stowe, Michael C. Cavalier, Raquel Godoy-Ruiz, Kristen J. Varney, Paul T. Wilder, Ronald B. Gartenhaus, David J. Weber. Targeting the translational enhancer complex MCT-1:DenR to disrupt NHL survival. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2454. doi:10.1158/1538-7445.AM2015-2454
Abstract 2468: Antibody targeting soluble NKG2D ligand sMIC induces regression of primary tumors and eliminates metastasis in multiple pre-clinical cancer models
Authors: Wu Jennifer D. , Basher Fahmin , Rubinstein Mark 
Published: August 2015

Abstract
Expression of the MHC I-chain related molecules A and B (MICA/B) on epithelial cell surface in response to transformation or DNA damage can signal the immune system of the abnormality and thus initiate active immune surveillance by Natural Killer (NK) cells and cytotoxicity T cells. We and others have shown that malignant tumor cells can shed MICA/B to down regulate NKG2D expression and negatively impact NK and CD8 T cell function in cancer patients. Increased tumor-specific shedding of cell surface NKG2D ligand, MHC I chain related molecule (MIC), is associated with advanced stage and metastasis in many types of epithelial cancer. High serum levels of soluble MIC (sMIC) insults the immune system not only by down-regulating NKG2D expression on natural killer (NK) cells and effector T cells but also perturbing NK cell peripheral maintenance. Whether sMIC is an effective cancer therapeutic target has not been addressed due to the lack of specific sMIC-blocking reagent and clinically relevant pre-clinical animal models. Using a “humanized” clinically relevant spontaneous prostate carcinoma TRAMP/MIC bi-transgenic mouse model and multiple engineered syngeneic transplantable tumor models, we unprecedentedly show that therapy with a sMIC-specific monoclonal antibody induced rapid regression of primary tumors and metastasis without systemic toxicity. The therapy revamped a myriad of anti-tumor immune responses including NK cell homeostatic renewal and function, CD4 T cells to Th1 and Th17 responses. Using the well-defined melanoma model, we further show that anti-sMIC therapy augments Pmel-1 antigen-specific CD8 T cell responses. Notably, depletion of NK cells mitigated the therapeutic effect of anti-sMIC antibody and the effector function of both CD4 and CD8 T cells. We conclude that sMIC is an effective therapeutic target to potentiate innate and adoptive immune responses against MIC+ malignancies.
Citation Format: Jennifer D. Wu, Fahmin Basher, Mark Rubinstein. Antibody targeting soluble NKG2D ligand sMIC induces regression of primary tumors and eliminates metastasis in multiple pre-clinical cancer models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2468. doi:10.1158/1538-7445.AM2015-2468
Abstract 2487: The antitumor efficacy of anti-BST2 antibody is significantly increased in combination with CpG oligodeoxynucleotide in endometrial cancer
Authors: Takahashi Yusuke , Serada Satoshi , Hiramatsu Kosuke , Kobiyama Kouji , Fujimoto Minoru , Ishii Ken , Naka Tetsuji 
Published: August 2015

Abstract
Background:
CpG oligodeoxynucleotides (ODN) has been shown to exhibit a potent immunostimulatory activity. Therefore, CpG ODN has been considered to be a promising material for enhancing the therapeutic effect of anticancer monoclonal antibody with antibody-dependent cell-mediated cytotoxicity (ADCC) activity. We have previously identified bone marrow stromal antigen 2 (BST2) as a therapeutic target of endometrial cancer cells and also demonstrated that an anti-BST2 antibody showed a potent anti-tumor effect by ADCC and complement-dependent cytotoxicity (CDC) activities against BST2 positive cells. The aim of this study was to evaluate the synergistic anti-tumor effect by combination of anti-BST2 antibody and CpG ODN, and to clarify the mechanisms of its anti-tumor effect, using xenograft model of BST2 positive endometrial cancer cells.
Material and Method:
We implanted 7-8 week-old female SCID mice with 5×106 HEC-88 nu endometrial cancer cells subcutaneously. Mice with tumor were treated by intra-peritoneal injection of an anti-BST2 mAb alone (12.5 and 200 μg/mouse, twice a week for four weeks), intra-tumoral injection of CpG ODN alone (10 μg/mouse, twice a week for four weeks) or combination therapy (12.5 μg/mouse of anti-BST2 mAb and 10 μg/mouse of CpG ODN). Anti-tumor effect was evaluated by monitoring the changes of tumor volume. Finally, population of NK cells and macrophages infiltrated into the tumor were analyzed by the FACS analysis, how CpG ODN induces immunopotentiative action.
Results:
Intraperitoneal treatment by anti-BST2 antibody showed a significant anti-tumor effect, dose dependently. The combination therapy of anti-BST2 antibody and CpG ODN was more effective than the single-agent therapy (p = 0.0002). In addition, the combination therapy also resulted high population of NK cells and macrophages in tumor significantly (p = 0.00018).
Conclusion:
We showed that CpG ODN increases the efficacy of anti-tumor effect of anti-BST2 mAb induced ADCC activity via potentiating NK cells and macrophages mediated cancer cell injury. Combination of GpG ODN would be a promising approach for enhancing the activity of ADCC based monoclonal antibodies, which targeting anti-tumor antigen.
Citation Format: Yusuke Takahashi, Satoshi Serada, Kosuke Hiramatsu, Kouji Kobiyama, Minoru Fujimoto, Ken Ishii, Tetsuji Naka. The antitumor efficacy of anti-BST2 antibody is significantly increased in combination with CpG oligodeoxynucleotide in endometrial cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2487. doi:10.1158/1538-7445.AM2015-2487
Abstract 2505: Therapeutic efficacy of the ZVex™ and GLAAS™ platforms in a B16-F10/hCAIX melanoma mouse model
Authors: Campbell David J. , Reeves Rebecca S. , Flynn Patrick A. , Robbins Scott H. , Berglund Peter , ter Meulen Jan H. 
Published: August 2015

Abstract
ZVex™ and GLAAS™ are two dendritic-cell (DC) targeting platform technologies designed to enhance immune responses through the in vivo induction of antigen specific CD8 and CD4 T-cells, respectively. ZVex™ is a lentiviral vector pseudotyped with a modified Sindbis virus envelope engineered to deliver tumor antigen-encoding nucleic acids to dendritic cells in vivo. GLAAS™(Glucopyranosyl Lipid A Adjuvant System) activates DC by binding to the TLR-4 receptor and inducing strong Th1 type CD4 responses against co-delivered recombinant proteins. Currently both platform technologies are being investigated in phase I clinical trials in cancer patients.
Human carbonic anhydrase 9 (hCAIX) is a tumor-associated transmembrane antigen that is over-expressed on various cancer cell types. We mapped hCAIX specific, multi-functional CD8 and CD4 T-cell epitopes within the extracellular and transmembrane regions of the protein for the mouse haplotype H-2b by intracellular cytokine staining. Mice lethally challenged s.c. on the flank with a B16-F10 tumor cell line expressing the hCAIX protein (designated BC.12) fully controlled large tumors (&gt;100 mm2) when therapeutically immunized (subcutaneously at the base of tail) with ZVex™ encoding hCAIX or the recombinant hCAIX protein with GLAAS™ (either s.c. or i.m.). In both models, tumor control was dose-dependent. Additionally, the presence of a strong transmembrane H-2b restricted CD8 T-cell epitope was required for tumor control and regression. hCAIX-specific CD8 T-cell responses were detectable as far out as day 67 post challenge in mice displaying full regression of tumor. These results demonstrate proof of concept for ZVex™ and GLAAS™ platform technologies in an aggressive murine melanoma model.
Citation Format: David J. Campbell, Rebecca S. Reeves, Patrick A. Flynn, Scott H. Robbins, Peter Berglund, Jan H. ter Meulen. Therapeutic efficacy of the ZVex™ and GLAAS™ platforms in a B16-F10/hCAIX melanoma mouse model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2505. doi:10.1158/1538-7445.AM2015-2505
Abstract 2506: Preclinical characterization of LV305, a lentiviral vector targeting tumors expressing NY-ESO-1
Authors: Albershardt Tina C. , Campbell David J. , Parsons Andrea J. , ter Meulen Jan H. , Berglund Peter 
Published: August 2015

Abstract
The cancer-testis antigen, NY-ESO-1, is an attractive tumor antigen target for cancer immunotherapy, because it is expressed in numerous cancer types (e.g., melanoma, breast cancer, sarcoma, lung cancer) but not in normal adult tissues except the testis or placenta. Although recent studies using various NY-ESO-1 formulations show promising results, an optimal immunotherapeutic strategy to generate T cell responses for controlling tumor growth is still lacking. Lentiviral vectors (LVs) are proven tools for delivering nucleic acid payloads into cells due to their ability to transduce not only dividing cells but also quiescent cells, such as dendritic cells (DCs). Because DCs are essential for the initiation of T cell responses, we have engineered LV305 (a product candidate from ZVex™, our LV-based discovery platform) to selectively deliver the antigen-encoding NY-ESO-1 gene to human DCs in vivo. To develop tools to evaluate LV305 in a preclinical setting, we 1) generated a cell line, CIN.23, which is a CT26 murine colon cancer cell line that stably expresses human NY-ESO-1 protein; and 2) mapped and confirmed NY-ESO-1 CD8 and CD4 T cell epitopes recognized by H2d. Mice immunized with LV305 developed strong, dose-dependent, poly-functional, and cytotoxic NY-ESO-1-specific CD8 T cells within 14 days post-immunization and could be boosted with LV305 at least three times to recall peak-level CD8 T cell responses. Immunization with LV305 protected mice against CIN.23 tumor challenge, which was abrogated upon depletion of CD8 T cells. Adoptive transfer of CD8 T cells, together with CD4 T cells or NK cells, from LV305-immunized donor mice to tumor-bearing mice conferred significant protection against metastatic tumor growth. This protection was not observed when CD8 T cells alone were adoptively transferred. Taken together, these findings suggest that CD8 T cells are required but not sufficient for mediating LV305-induced anti-tumor efficacy. We have demonstrated the anti-tumor therapeutic potential of LV305-mediated in vivo DC immunization in murine models and are currently investigating LV305 in a phase I clinical trial in cancer patients with tumors expressing NY-ESO-1.
Citation Format: Tina C. Albershardt, David J. Campbell, Andrea J. Parsons, Jan H. ter Meulen, Peter Berglund. Preclinical characterization of LV305, a lentiviral vector targeting tumors expressing NY-ESO-1. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2506. doi:10.1158/1538-7445.AM2015-2506
Abstract 252: Antibody-mediated phosphatidylserine blockade significantly enhances the efficacy of immune checkpoint blockades in K1735 and B16 mouse melanoma models
Authors: Freimark Bruce , Gong Jian , Ye Dan , Brekken Rolf , Yin Shen , Hutchins Jeff , Nguyen Van , Hughes Chris , Huang Xianming 
Published: August 2015

Abstract
Phosphatidylserine (PS) is a phospholipid normally residing in the inner leaflet of the plasma membrane and becomes exposed abundantly on tumor endothelial cells, tumor-secreted microvesicles, and tumor cells in response to chemo- and radiotherapy. Extensive studies have shown that PS is a global immune checkpoint and major contributor to tumor immunosuppression, which promotes the expansion of immunosuppressive cells, such MDSC and M2 macrophages, inhibits DCs maturation, while stimulates them to secrete immunosuppressive mediators. We have shown that PS targeting can override PS-mediated immunosuppression, reactivate innate tumor immunity, and evoke adaptive antitumor immunity. In the present study, we assessed the antitumor effect of the combination of PS blockade and anti-CTLA-4 or anti-PD-1 antibodies in B16 and K1735 melanoma models. Both combinations showed significantly superior tumor growth inhibition over single treatment, with many mice achieving complete tumor regression. Flow cytometry analysis showed that the combination treatment had significantly greater total and functional tumor-infiltrating CD8+ T, more IL-2- and IFNγ-producing splenic T cells, and lower number of splenic MDSCs than did single treatment. In addition, the ratio of M2 to M1 in the tumor was significantly lower in the combination treatment than that in single treatment. Finally, no toxicity was observed in any of the treatment groups following multiple treatment doses. These data suggest that combination of PS blockade with immune checkpoint blockade promotes strong, localized, enhanced therapeutic efficacy without the side-effects of systemic immune activation and represents a promising combinatorial strategy for cancer immunotherapy. A chimeric PS-targeting antibody, bavituximab, is being used in combination with chemotherapy to treat patients with solid tumors in multiple late-stage clinical trials. These data support our Phase I IST evaluation of our PS-targeting antibody bavituximab in combination with ipilimumab in advanced melanoma patients.
Citation Format: Bruce Freimark, Jian Gong, Dan Ye, Rolf Brekken, Shen Yin, Jeff Hutchins, Van Nguyen, Chris Hughes, Xianming Huang. Antibody-mediated phosphatidylserine blockade significantly enhances the efficacy of immune checkpoint blockades in K1735 and B16 mouse melanoma models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 252. doi:10.1158/1538-7445.AM2015-252
Abstract 2520: Synergistic combination of lurbinectedin and PARP inhibitors in breast cancer tumor cell lines
Authors: Santamaría Gema , Avila Sonia , Moneo Victoria , Cuevas Carmen , García-Fernández Luis F. , Galmarini Carlos M. 
Published: August 2015

Abstract
Lurbinectedin (PM1183) is a new tetrahydroisoquinoline derivative currently evaluated in phase II trials in BRCA1-mutated breast cancer and NSCLC patients. Two pivotal trials (in platinum-resistant ovarian cancer and in SCLC) will be launched in 2015. PM1183 shows antitumor activity against a wide variety of tumor cells with mean IC50 values in the low nanomolar range. In living cells, PM1183 inhibits trans-activated transcription and blocks NER-dependent DNA repair inducing the formation of dsDNA breaks and the collapse of replication forks that need to be repaired by homologous recombination (HR). As expected, PM1183 is more active against homologous recombination (HR)-deficient cell lines. In this work, we evaluated the in vitro antitumor effect of PM1183 when combined with the PARP inhibitors ABT-88 (Veliparib, PARP-1 and -2), AZD-2281 (Olaparib, PARP-1), BSI-201 (Iniparib, PARP-1) and BMN-673 (PARP-1 and -2) (Selleck Chemicals, Houston, TX, USA) in a panel of human breast carcinoma cells with different BRCA1 status, including MCF-7 (BRCA1 +/+), MDA-MB-231 (BRCA1 +/-), HCC-1937 (BRCA1 -/-) and MDA-MB-436 (BRCA1 -/-). The combination index (CI) of each different drug-drug combination was calculated by applying the Chou and Talalay method. Different degrees of synergism (CI &lt; 1) were recorded when PM1183 was combined with AZD-2281 (Olaparib) and BMN-673, depending on the tumor cell line and the conditions tested. The highest degree of synergism with Olaparib was observed in MDA-MB-436 cells followed by MDA-MB-231, MCF7 and HCC-1937 cells. The highest degree of synergism with BMN-673 was observed in MCF-7 cells followed by MDA-MB-231 and HCC-1937 cells. In MDA-MB-231 cells, both combinations led to a synergistic induction of γ-H2AX expression and PARP-1 cleavage, as analyzed by western blot. Collectively, our results indicate that the combination of PM1183 and PARP inhibitors (Olaparib and BMN673) induces an artificial synthetic lethality that can be advantageously used to kill several breast cancer cells, independently from their BRCA1 status.
Citation Format: Gema Santamaría, Sonia Avila, Victoria Moneo, Carmen Cuevas, Luis F. García-Fernández, Carlos M. Galmarini. Synergistic combination of lurbinectedin and PARP inhibitors in breast cancer tumor cell lines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2520. doi:10.1158/1538-7445.AM2015-2520