bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5393: The advantages of acoustic liquid handling for drug sensitivity screening
Authors: Edwards Bonnie 
Published: May 2015

Abstract
Recently identified associations between variants of cancer genes and drug resistance have increased the value for comprehensive drug sensitivity screening in combination with molecular profiling of cancer cells. This in turn, has led to a demand for improvements to screening throughput and efficiency. Echo® Liquid Handlers use acoustic energy to provide high throughput, non-contact, liquid handling for a range of applications. Non-contact transfer avoids the risk of cross-contamination, eliminates tip costs, and facilitates the progression to high density assay formats. Echo Liquid handlers precisely and accurately transfer 2.5 nL droplets of sample and reagent, which enables the miniaturization of biochemical screens. This poster discusses the implementation of miniaturized drug sensitivity screening, at the Institute of Molecular Medicine in Finland (FIMM), with assay-ready plates produced by the Echo liquid handler.
Citation Format: Bonnie Edwards. The advantages of acoustic liquid handling for drug sensitivity screening. [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 5393. doi:10.1158/1538-7445.AM2014-5393
Abstract 5407: Milk derived exosomes: Scalable source of biologically active drug delivery nanoparticles
Authors: Aqil Farrukh , Munagala Radha , Jeyabalan Jeyaprakash , Gupta Ramesh C. 
Published: May 2015

Abstract
Exosomes are biological nanoparticles and found in almost all the body fluids. They shuttle in and out of cells, are comprised of lipid bilayers, and carry a cargo of macromolecules. Because of the nano size (30-100 nm) these natural vesicles are being explored as nanodevices for the development of new therapeutic applications. Here we show bovine milk as a scalable source of exosomes and as a drug delivery vehicle for small molecules - both lipophilic and hydrophilic. The exosomes were isolated by differential centrifugation with the size generally ranging from 30-100 nm as analyzed by NanoSight and confirmed by electron microscopy and atomic force microscopy. The identity of exosomes was confirmed by surface protein markers (CD63, CD81), buoyant floatation density (1.12 g/ml) determined by sucrose density gradient, and high content of immune-related miRNAs and mRNA as determined by PCR. Exosomes, suspended in PBS, and stored frozen (-80°C) were found to be stable in terms of their size and biological activities. Milk exosomes loaded with fluorescent dye were readily taken up by macrophages and human cancer cells in vitro and distributed in the various tissues of nude mice following treatment with gavage and i.v. routes. Imaging of various tissues collected after euthanasia showed a broad tissue distribution of the exosomes delivered via the oral route, whereas exosome concentration predominated in the liver when administered by the i.v. route. Both lipophilic and hydrophilic chemopreventive (curcumin, withaferin A, anthocyanidins, and punicalagins, etc.) and chemotherapeutic (paclitaxel and docetaxel) agents could be loaded onto exosomes with 8%-60% drug load with respect to exosomal protein content. Drug-exosomal formulations showed significantly higher antiproliferative activity against lung, breast and ovarian cancer cells versus the free drugs. The drug formulation also showed significantly higher tissue drug levels, and enhanced anti-inflammatory and antitumor activity versus the free drug. Intriguingly, in the absence of any drug, the milk exosomes showed anti-proliferative and anti-inflammatory activities in cancer cells, suggesting the presence of protective factors in the milk exosomes (e.g., miRNAs); this notion corroborates the epidemiological and experimental findings of the presence of built-in immune factors in milk. (Supported from USPHS grants CA-118114 and CA-125152, Kentucky Lung Cancer Research Program, Agnes Brown Duggan Endowment, and Helmsley Funds).
Citation Format: Farrukh Aqil, Radha Munagala, Jeyaprakash Jeyabalan, Ramesh C. Gupta. Milk derived exosomes: Scalable source of biologically active drug delivery nanoparticles. [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 5407. doi:10.1158/1538-7445.AM2014-5407
Abstract 5416: Development of a nanoparticle platform for the targeted delivery of siRNA to HER2-positive breast cancers
Authors: Goodyear Shaun M. 
Published: May 2015

Abstract
Successful siRNA based therapy has the potential to revolutionize cancer therapy by mediating the silencing of any gene deemed important for disease progression. However, unprotected siRNA has a short half-life in blood and lacks the ability to selectively identify target cells. Our group is developing an effective siRNA based nanoparticle platform that overcomes these shortcomings by utilizing a mesoporous silica nanoparticle electrostatically loaded with siRNA and conjugated with an antibody for target homing. The human epidermal growth receptor type 2 (HER2) is a highly validated therapeutic target in breast cancer due to its prognostic role in cancer aggressiveness and drug resistance when overexpressed in tumors. Suppression of HER2 with siRNA based therapy (siHER2) offers a new treatment modality and provides a testable system for the development of our nanoparticle platform. Incorporation of Herceptin, a monoclonal antibody which binds to HER2, serves to selectively target HER2-positive cancer cells. In vitro evaluation of the nanoconstruct loaded with a highly optimized siHER2 sequence against a panel of HER2-positive (HCC1954, JIMT-1, BT474) and HER2-negative (MCF-7, MCF-10A, HDFa) cell lines demonstrated efficient knockdown of HER2 gene expression (&gt; 75% reduction of HER2 mRNA and protein). Concomitant cell viability assays showed increased cell death in HER2-positive and no HER2-negative cell lines further highlighting treatment specificity. Importantly, treatment of cells with the siHER2-nanoconstruct effectively inhibited growth in HER2-positive cell lines resistant to Herceptin and Lapatinib. Subsequent in vivo tumor modeling studies using orthotopic transplantation of HCC1954 in the mammary fat pad of SCID mice revealed that i.v. injection of the siHER2-nanocontruct reduced overall tumor burden. HER2 knockdown in the tumors was also significant. These findings provide the foundation for a novel translational platform for facilitating therapeutic siRNA.
Citation Format: Shaun M. Goodyear. Development of a nanoparticle platform for the targeted delivery of siRNA to HER2-positive breast cancers. [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 5416. doi:10.1158/1538-7445.AM2014-5416
Abstract 5419: Computationally-inspired engineering of supramolecular taxane nanoparticles
Authors: Kulkarni Ashish A. , Rao Poornima , Goldman Aaron , Sengupta Shiladitya 
Published: May 2015

Abstract
Clinical efficacy of long circulating lipid nanoparticles depend on high drug to lipid ratio, stability during circulation and sustained drug release after tumor accumulation. Achieving this is the holy grail of paclitaxel (PTX) lipid nanoparticles due to its incompatible physicochemical properties such as bulkiness, structural asymmetry, hydrophobicity that leads to low drug loading efficiency and tendency to crystallize leading to significant instability. Effective strategy to overcome these limitations could be devised by understanding molecular interactions of PTX with lipid bilayer at spatiotemporal level but these remain poorly understood. Here, using computer simulations, we have investigated the interactions of PTX with the model lipid bilayer at low and high drug contents. We found that PTX at low concentrations occupied the space in both hydrophobic and hydrophilic regions of the bilayer and did not exhibit significant destabilizing effect whereas at high concentrations it exclusively accumulated at the hydrophilic tail region causing significant rifts in the bilayer. In silico dynamic simulations showed that conjugation of PTX to cholesterol using a flexible spacer (Suprataxel, STX) stabilized the lipid bilayer at high concentrations. To confirm the theoretical observations, we engineered supramolecular nanoparticles of STX (STX-NP) and characterized the physicochemical and mechanical stability. In vitro efficacy studies showed that STX-NPs is comparable to PTX in multiple cancer cell lines. We demonstrated that STX-NPs were more efficacious than PTX in inhibiting tumor growth in 4T1 breast cancer and K-RasLSL/+/Ptenfl/fl ovarian cancer models. This study shows that the effective Supramolecular nanoparticles could be engineered by understanding the interactions between the drug and the lipid layer.
Note: This abstract was not presented at the meeting.
Citation Format: Ashish A. Kulkarni, Poornima Rao, Aaron Goldman, Shiladitya Sengupta. Computationally-inspired engineering of supramolecular taxane nanoparticles. [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 5419. doi:10.1158/1538-7445.AM2014-5419
Abstract 5425: L-amino acid oxidase from Crotalas adamanteus venom induces caspase-independent apoptosis in human NUGC-3 gastric cancer cells
Authors: Ding Jian , Bay Boon-Huat , Gopalakrishnakone P 
Published: May 2015

Abstract
L-amino acid oxidases (LAAOs), which are largely present in snake venoms, are a group of flavoenzymes, known to catalyze oxidative deamination of L-amino acid substrates, resulting in the formation of corresponding α-keto acids, hydrogen peroxide and ammonia. LAAOs have attracted interest in biomedical and pharmacological research because of their antimicrobial, anti-HIV, anticoagulant, inhibition of platelet aggregation and apoptotic-inducing activities. The aim of this study is to evaluate the anticancer effects of LAAO from Crotalas adamanteus snake venom in human NUGC-3 gastric cancer cells in vitro. Cell viability and cytotoxicity were determined by Alamar Blue and Lactate Dehydrogenase assays. Cell cycle phases were analyzed by flow cytometry. Apoptosis was identified by flow cytometric analysis with Annexin-V and PI staining. The morphological changes were observed by fluorescence microscopy, transmission electron microscopy and scanning electron microscopy. The ROS level was quantified by DCFDA staining as well as malondialdehyde formation. The results showed that LAAO reduced cell viability and induced cytotoxicity in a dose-dependent manner. There was associated cell cycle arrest at the G2/M phase. Morphological changes induced by LAAO treatment included membrane blebbing, cell shrinkage and chromatin condensation. Apoptosis induced by LAAO in NUGC-3 gastric cancer cells as evidenced by Annexin-V and PI positive staining, was found to be mediated via a caspase-independent pathway without activating caspase-3 and Poly ADP-ribose polymerase. Moreover, a concomitant increase of intracellular oxidative stress and the translocation of the apoptosis inducing factor (AIF) from mitochondria to the nucleus were observed. It would appear that LAAO derived from the Crotalas adamanteus venom, induced a caspase-independent apoptotic pathway which was incited by ROS and AIF translocation. This study provides new biological insights into the application of LAAO from a snake venom in cancer therapeutics.
Citation Format: Jian Ding, Boon-Huat Bay, P Gopalakrishnakone. L-amino acid oxidase from Crotalas adamanteus venom induces caspase-independent apoptosis in human NUGC-3 gastric cancer cells. [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 5425. doi:10.1158/1538-7445.AM2014-5425
Abstract 5430: Frizzled-1 receptor, a potential therapeutic target for atypical teratoid rhabdoid tumor
Authors: Chakravadhanula Madhavi , Ozols Victor , Hampton Chris , Bhardwaj Ratan 
Published: May 2015

Abstract
Atypical Teratoid Rhabdoid Tumors (ATRTs) are aggressive tumor types in the field of pediatric neuro-oncology that need new therapeutic strategies since they are associated with significantly worse overall prognosis than other pediatric tumors. A characteristic feature of ATRTs is usually an aberration of SMARCB1/INI1/hSNF5 gene. One of the important developmental pathways for cellular development is the Wnt pathway. Wnt proteins are a family of 19 secreted glycoproteins and act as ligands that bind transmembrane receptors such as the Frizzled receptors. There are 10 known Frizzled receptors, and the binding of a specific Wnt ligand to a specific Frizzled receptor plays a crucial role in the regulation of diverse cellular processes. Perturbation of the activities of Wnt ligands can result in defects in embryonic development and diseased states such as cancer. In our preliminary transcriptome analyses results, Wnt5b and Frizzled-1 receptor (Fz-1) were significantly upregulated in 20 ATRT samples and 3 ATRT cell lines. The binding of Wnt5b with Fz-1 was further confirmed by immunoprecipitation and Western blotting analyses. Treatment of ATRT cells with Fz-1 specific siRNA did not indicate a significant decrease in cell viability. Similarly treatment of ATRT cells with a Fz-1 specific monoclonal antibody (Fz-1Mab) also did not indicate significant decrease in cell viability. However, in this study treatment of cells with a combination of Fz-1 specific siRNA and Fz-1Mab indicated a significant decrease in cell viability. A dose response analysis was performed over time to test various doses of Fz-1Mab with constant Fz-1 siRNA concentrations; a cell titre glo assay was then performed to assess cell viability. The physiologically relevant Fz-1Mab dose was then used to assess the effects of using this combination treatment on Fz-1 and Wnt5b protein levels. A decrease in Fz-1 and Wnt5b protein levels when Fz-1 receptor is blocked by siRNA and Mab indicates that Wnt5b bound to Fz-1 receptor plays an important role in ATRT disease etiology and could be a therapeutic target.
Citation Format: Madhavi Chakravadhanula, Victor Ozols, Chris Hampton, Ratan Bhardwaj. Frizzled-1 receptor, a potential therapeutic target for atypical teratoid rhabdoid 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 5430. doi:10.1158/1538-7445.AM2014-5430
Abstract 5437: Dual targeting of ErbB-2 and ErbB-3: A new potential strategy for the treatment of pancreatic cancer
Authors: Sala Gianluca , Ghasemi Reza , Capone Emily , Rapposelli Ilario Giovanni , Traini Sara , Rossi Cosmo , Natali Pier Giorgio , Iacobelli Stefano 
Published: May 2015

Abstract
Background: Pancreatic cancer (PC) is one of the most lethal cancers and there is an urgent need to find out new therapeutic approaches. Deregulated signaling through ErbB-1 and ErbB-2, members of the epidermal growth factor receptor family, frequently occurs in PC. However, attempts aiming at inhibiting ErbB-1 and ErbB-2 have revealed only a modest impact on disease outcome. A growing body of evidence suggests that ErbB-3 and its ligand NRG-1β are key players in driving oncogenic signaling and resistance to targeted therapy in PC. We have recently developed a novel humanized ErbB-3 antibody, named EV20, which displayed a potent antitumor activity by promoting receptor downregulation in vitro and in vivo. Here we hypothesized that targeting of ErbB-3 with EV20 would enhance the therapeutic effect of ErbB-2 inhibitors in PC.
Materials and Methods: Three different PC cell lines (BxPC-3, HPAF II and SW1990) were chosen based on their metastatic origin, genetic background (KRAS status) and responsiveness to erlotinib. The expression of ErbB-1, ErbB-2 and ErbB-3 was analyzed by FACS and WB. The effects of trastuzumab, cetuximab, EV20 and their combinations on NRG-1β-induced activation of the ErbB3/PI3K/Akt axis, cell proliferation, and in vivo tumor growth were evaluated by standard procedures.
Results: ErbB-1 was overexpressed by all the cell lines, whereas the expression levels of ErbB-2 and ErbB-3 were moderate in HPAFII cells and low in BxPC-3 and SW1990 cells. NRG-1β was more effective than EGF (an ErbB-1 ligand) in activating ErbB-3 downstream signaling, which was not inhibited by cetuximab, despite a high ErbB-1 expression in the cells. Conversely, inhibition of ErbB-2 and/or ErbB-3 resulted in a marked suppression of NRG-1β-driven Akt activation. Finally, EV20 in combination with trastuzumab exerted a superior antitumor activity compared to single agent alone in terms of cell proliferation and growth of PC xenografts in nude mice.
Conclusions: Dual targeting of ErbB-2 and ErbB-3 could represent a newer therapeutic approach in PC.
Note: This abstract was not presented at the meeting.
Citation Format: Gianluca Sala, Reza Ghasemi, Emily Capone, Ilario Giovanni Rapposelli, Sara Traini, Cosmo Rossi, Pier Giorgio Natali, Stefano Iacobelli. Dual targeting of ErbB-2 and ErbB-3: A new potential strategy for the treatment of pancreatic cancer. [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 5437. doi:10.1158/1538-7445.AM2014-5437
Abstract 5440: Activity of mesothelin-targeted immunotoxin RG7787 against triple-negative breast cancer
Authors: Alewine Christine , Maon-Osann Emily , Pastan Ira 
Published: May 2015

Abstract
Mesothelin (MSLN) is a cell surface glycoprotein normally expressed by mesothelial cells of the pleura, pericardium and peritoneum, but also highly expressed in mesothelioma, pancreatic and ovarian cancers. It was recently reported that many triple negative breast cancers (TNBCs) also express MSLN. RG7787 is a next generation, deimmunized recombinant immunotoxin (RIT) consisting of a high-affinity anti-MSLN Fab fragment for targeting, fused to a 24 kD fragment of Pseudomonas exotoxin A for cell killing. Here, we demonstrate using flow cytometry that the TNBC cell lines HCC70 and SUM149 express significant MSLN on the cell surface. HCC70 and SUM149 are sensitive to RG7787 and related RITs with IC50 values in the subnanomolar range. RG7787 has less off-target toxicity in mice than earlier mesothelin-targeted immunotoxins and could be safely administered to mice at six-fold higher doses. RG7787 shrinks HCC70 tumor xenografts grown in athymic nude mice by &gt;40% and produces a 10 day delay in tumor progression compared to vehicle (p &lt; 0.02). Tumor volume decreased by &gt;85% when RG7787 was given following paclitaxel, a statistically significant improvement over paclitaxel alone. Together these data demonstrate that RG7787 has significant activity against TNBC and warrants further study as a treatment for this malignancy.
Citation Format: Christine Alewine, Emily Maon-Osann, Ira Pastan. Activity of mesothelin-targeted immunotoxin RG7787 against triple-negative breast cancer. [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 5440. doi:10.1158/1538-7445.AM2014-5440
Abstract 5446: FPA144: A therapeutic antibody for treating patients with gastric cancers bearing FGFR2 gene amplification
Authors: Gemo Abigael T. , Deshpande Amit M. , Palencia Servando , Bellovin David I. , Brennan Thomas J. , Patil Namrata S. , Huang Carol , Los Gerrit , Pierce Kristen L. 
Published: May 2015

Abstract
A subset of patients with gastric cancer have an amplification of the receptor tyrosine kinase fibroblast growth factor receptor 2 (FGFR2) gene. The amplification is most common in the diffuse type of gastric cancer and its presence correlates with poor patient prognosis. Although it has been reported that there is high expression of FGFR2 protein in patients with the amplification, it has been unknown which of the two major FGFR2 isoforms, FGFR2b or 2c, is expressed. In this study, we demonstrate, by both qPCR and immunohistochemistry using FGFR2b-selective antibodies, that it is the FGFR2b isoform, and not FGFR2c, that is overexpressed in gastric cancer tumors that contain the FGFR2 amplification.
Five Prime Therapeutics has developed an FGFR2b-specific antibody, FPA144, to treat patients with gastric cancers bearing amplification of the FGFR2 gene. FPA144 is glycoengineered for enhanced antibody-dependent cell cytotoxicity (ADCC). FPA144 causes tumor growth inhibition by 72% to 100% in gastric cancer xenograft models with FGFR2 gene amplification that overexpress the FGFR2b protein isoform. In the SNU-16 model, FPA144 reduces the levels of FGFR2b protein expressed in the tumors by approximately 50%, and decreases both FGFR2b phosphorylation and phosphorylation of the downstream effector, FRS2. The anti-tumor effect of FPA144 is additive with the standard of care chemotherapy regimens of 5-fluoruracil/cisplatin and paclitaxel in the OCUM-2 gastric cancer xenograft model. Since FPA144, unlike small molecule FGFR tyrosine kinase inhibitors, blocks signaling by only the FGFR2b and not the other FGFRs, we expect a favorable toxicology profile, either alone or in combination with chemotherapy. In addition, by engaging immune effector cells for direct tumor cell killing, FPA144 has a second mechanism of action that small molecule FGFR tyrosine kinase inhibitors do not have. We anticipate starting clinical trials of FPA144 in the last quarter of 2014.
Citation Format: Abigael T. Gemo, Amit M. Deshpande, Servando Palencia, David I. Bellovin, Thomas J. Brennan, Namrata S. Patil, Carol Huang, Gerrit Los, Kristen L. Pierce. FPA144: A therapeutic antibody for treating patients with gastric cancers bearing FGFR2 gene amplification. [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 5446. doi:10.1158/1538-7445.AM2014-5446