bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4046: Kinetic quantification of caspase-3/7 mediated apoptosis using live-cell imaging
Authors: Appledorn Daniel M. , Nelson Thomas , Dale Tim , Alcantara Susana , Artymovich Katherine , Endsley Eric W. , Trezise Derek J. , Groppi Vince E. 
Published: August 2012

Abstract
Apoptosis, the biological process by which cells undergo programmed cell death, is required for normal tissue maintenance and development. However, aberrations in apoptotic signaling networks are implicated in numerous human diseases including neurodegeneration and cancer. Stimulation of either the extrinsic or intrinsic apoptotic pathways triggers a signaling cascade that typically results in the activation of executioner caspase-3. Numerous in vitro assays have been designed to measure the activation of caspase-3. The majority of these assays utilize reagent substrates that incorporate the DEVD motif which is recognized by both activated caspases 3 and 7. Such assays result in single, user-defined time point measurements of caspase-3/7 activity and typically require multiple wash steps or cell lifting prior to data collection; potentially resulting in the loss of cells or critical data in experiments where cells undergo apoptosis at different rates according to treatment conditions. In this study, we optimized an assay system incorporating a bi-functional reagent containing the DEVD motif linked to a DNA dye for use on the IncuCyte FLR imaging system. When added to cell culture growth medium, this relatively inert, non-fluorescent molecule crosses the cell membrane where it is available to be a substrate for activated caspase-3/7. The reaction product is a green fluorescent DNA dye that labels the nuclei of cells undergoing apoptosis. Our data show that this strategy can quantitatively measure the kinetic activation of caspase-3/7 through extrinsic stimulation of human tumor derived MDA-MB-231 breast adenocarcinoma cells, HeLa cervical adenocarcinoma cells, A549 lung epithelial carcinoma cells, and HT 1080 fibrosarcoma cells following treatment with TNFα and cycloheximide. Moreover, we exemplify the use of this strategy in 96-well plate screening protocols using both HT 1080 and MDA-MB-231 cells to measure the induction of caspase-3/7 activity in the presence of staurosporine, a well-known general protein kinase inhibitor and inducer of caspase-3/7 mediated apoptosis with Z’ values α0.6. We provide evidence that this stable, kinetic, mix-and-read assay can provide both quantitative and qualitative assessments of caspase-3/7 activity using fluorescent images and high-definition phase contrast images, respectively.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4046. doi:1538-7445.AM2012-4046
Abstract 4049: Early prediction of response to targeted drugs treatment in malignant lymphoma and sarcoma using in vivo proliferation marker 18F-FLT
Authors: Li Zhoulei , Graf Nicolas , Herrmann Ken , Pirsig Sabine , Schwaiger Markus , Scheidhauer Klemens , Buck Andreas , Keller Ulrich , Dechow Tobias 
Published: August 2012

Abstract
Purpose: Last year we have shown that 18F-FLT is superior surrogate marker than 18F-FDG for very early response assessment to therapy in malignant Lymphoma. In this study, we use FLT-PET to predict the response of ewing sarcoma to different cancer therapy and investigate its relation to cancer cell death, especially apoptosis. Methods: FLT and/ or FDG PET were performed prior and early in each therapeutic course on SCID mice bearing with various tumor cells. Ewing sarcoma A673 was applied for investigation of the response to sorafenib treatment or Radiotherapy. GFP labelled Lymphoma cells (Myc449, Myc665) with or without transportation of Bcl2 gene was employed for the observation of cell death especially apoptosis after the treatment with aura kinase inhibitor or chemotherapy. Tumor-to-background ratios (TBR) of FLT-PET were compared to that of FDG-PET. PET findings were correlated with optical Imaging, histopathology and in vitro data comprising cellular tracer uptake, cell cycle related protein expression, cell cycle distribution and viability assessment. Results: In vitro study shows that sarcoma A673 cells were sensitive to all therapy. For in vivo studies, tumor volume of sorafenib treated animals showed modest increase of 0.1cm3 (range 0.02cm3 to 0.2cm3, SD=0.09) within the first week as opposed to a 15-fold increase in control mice. Median TBR of FLT-PET decreased significantly to 30.1% compared to baseline as early as 5d after therapy (range 11.3-53.5%, n=12, p=0.0004). On the other side, TBR in FDG-PET also showed the similar tendency (median TBR 74.6%, range 28.1%-131.8%, n=12, p=0.02). But the reduction of TBR in FLT was significantly stronger than that in FDG (p&lt;0.001). In vitro study of lymphoma cells with Bcl2 transduction showed a strong anti- cell death ability, which will be served for the link between FLT PET and cell death. Conclusion: Compared with FDG-PET, FLT-PET is able to predict the response in the early therapeutic course and has a high specificity for the detection of ewing sarcoma. Continuing studies are intensively carried out to investigate FLT and FDG PET in relation to therapy-induced cell death.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4049. doi:1538-7445.AM2012-4049
Abstract 4051: A mouse model of head and neck cancer with infrared-based molecular imaging
Authors: Mouchli Anas , Han Ziying , Zheng Jun , Lajud Shayanne A. , O'Malley Bert W. , Li Daqing 
Published: August 2012

Abstract
Introduction Head and neck cancer (HNC) is notoriously difficult to treat and the five-year survival rate has improved very little over the past thirty years. Head and neck squamous cell carcinomas (HNSCC) mouse models are invaluable tools for furthering mechanistic HNC research. Molecular imaging (MI) is a promising new modality that may improve current HNC mouse models. Visible florescent proteins (VFPs) have been widely used with animal models for tumor detection using MI. However, the low emission wavelength of VFP results in its overlapping with the auto-fluorescence of many tissues and causes insufficient penetration into deeper tissues, preventing the application of VFP to track local and metastatic tumor progressions. The recent development of infrared-fluorescent protein (IFP) with Biliverdin as a chromophore has greatly advanced in vivo molecular imaging of deep tissues in animals. Hypothesis The ultimate goal of this study is to develop a novel orthotopic mouse model of HNC with regional spreading and distant metastasis tumors that closely mimics human HNC and can be accurately detected by a noninvasive IFP-based molecular imaging (MI) system. Methods A stable human HNC cell line expressing IFP was established. This stable cell line was used for generating a mouse model with HNC. The tumor formation and tumor progression with this model were accurately monitored by a MI system. Results The IFP signal was clearly detected in our mouse model of HNC. Scanned imaging demonstrated a progression pattern of tumor growth in this model. The tumor can be detected earlier with molecular imaging than by conventional external caliper measurement. Unlike surgical measurement, the tumor can be quantified without disturbing the tumor environment. More important, our data suggest that local-regional metastasis tumors can be detected with this model. Conclusion This novel animal model represents an orthotopic human HNC model. The IFP signals penetrate tissues well and can be detected in the deep tissues with this model. Thus, our model has the potential to be used for monitoring tumor spreading and metastasis of advanced stage HNC.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4051. doi:1538-7445.AM2012-4051
Abstract 4052: Highlighting intratumoral heterogeneity and peritumoral areas in human melanoma biopsies by infrared spectral microimaging
Authors: Sebiskveradze David , Gobinet Cyril , CARDOT-LECCIA Nathalie , ORTONNE Jean-Paul , MANFAIT Michel , JEANNESSON Pierre , PIOT Olivier 
Published: August 2012

Abstract
Infrared (IR) spectral microimaging is an efficient label-free optical method to analyze biological samples and to provide information about their intrinsic biochemical composition. Recent studies have shown its potential to detect and characterize cancerous tissues in their early stages, independently of visual morphology. IR microimaging appears more informative than conventional histology and could thus be developed as a sensitive, non-destructive and objective diagnostic tool in clinical oncology. The discrimination between tumoral and surrounding tissues relies on highlighting subtle IR spectral differences. An innovative fuzzy clustering based algorithm was thus developed in order to introduce the notion of nuance into the clustering of IR image pixels and to automatically determine the optimal clustering parameters, particularly the number of clusters. IR spectral imaging has been directly acquired on formalin-fixed paraffin-embedded tissue sections of human cutaneous melanomas with different degrees of invasiveness (n=10 patients). After the application of the fuzzy algorithm, the reconstructed color-coded clustering images allow recovering different histological structures automatically, particularly to precisely localize tumoral areas and their normal counterparts. However, more than reproducing classical histology, our algorithm can give access to additional information about the tissular structures. The notion of nuance introduced by fuzzy clustering is valuable for the pixels located at the interface between tumoral tissue and its microenvironment. Heterogeneous transitional areas between tumor and marginal normal tissue are identified, what is not possible on hematoxylin-eosin stained section or by conventional clustering. In addition, this automatic approach of IR data processing permits to highlight the interconnectivity between heterogeneous intratumoral areas and between tumoral and peritumoral structures. These preliminary results show significant potential for probing tumor progression and for early determination of tumor aggressiveness in cutaneous melanomas.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4052. doi:1538-7445.AM2012-4052
Abstract 4056: Gaussia luciferase complementation demonstrates that secreted CXCL12 (SDF-1) forms dimer under physiologic conditions
Authors: Ray Paramita , Lewin Sarah A. , Mihalko Laura Anne , Lesher-Perez Sasha Cai , Takayama Shuichi , Luker Kathryn E. , Luker Gary D. 
Published: August 2012

Abstract
Chemokine CXCL12 signaling through receptors CXCR4 and/or CXCR7 promotes tumor growth and metastasis in more than 20 different human malignancies, establishing CXCL12 as a promising therapeutic target in cancer. Under physiologic conditions, many chemokines form homodimers, which may activate signaling pathways distinct from those regulated by monomers of the same chemokine. Prior studies with recombinant, mutant forms of CXCL12 engineered to form either monomers or dimers show that monomeric and dimeric CXCL12 preferentially activate different downstream signaling molecules. However, the extent to which CXCL12 forms homodimers under physiologic conditions and resultant effects on CXCR4 and CXCR7 signaling remain to be determined. To analyze dimerization of CXCL12 secreted by mammalian cells, we used bioluminescence imaging of CXCL12 fused to either full length or complementation fragments of Gaussia luciferase (GL). Using column chromatography and GL protein fragment complementation, we demonstrated that CXCL12 is secreted from mammalian cells as both monomers and homodimers, and the homodimer remains stable in the extracellular environment. In 3D spheroid cultures, we also established that CXCL12 secreted from two independent cell populations can form homodimers in the extracellular space. Relative to dimeric CXCL12, monomeric CXCL12 preferentially signaled via CXCR4 to Gαi proteins to activate AKT and suppress intracellular cAMP. By comparison, dimeric CXCL12 was more potent at recruiting α-arrestin 2 to CXCR4 and regulating chemotaxis CXCR4+ breast cancer cells. In cell-based assays and an orthotopic tumor xenograft model of human breast cancer, we also found that CXCR7 preferentially sequestered monomeric CXCL12 and had minimal accumulation of the dimer. From these studies, we conclude that CXCL12 secreted from mammalian cells forms homodimers under physiologic conditions, and monomeric versus homodimeric CXCL12 regulate distinct signaling pathways. These results inform ongoing efforts to target CXCL12 and its receptors for cancer therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4056. doi:1538-7445.AM2012-4056
Abstract 4058: Multispectral imaging of mismatch repair proteins in colorectal cancers and tumor cells isolated from blood
Authors: Zhou Lanlan , Yang Zhaohai , Baker Maria , Dicker David T. , El-Deiry Wafik S. 
Published: August 2012

Abstract
DNA mismatch repair (MMR) is a system for recognizing and repairing erroneous insertion, deletion and misincorporation of bases that can arise during DNA replication and recombination, or some forms of DNA damage. MMR is required for maintaining genomic stability and is highly conserved from prokaryotes to eukaryotes. MMR defects are found in about 15% of sporadic colorectal cancer and in more than 90% of hereditary non-polyposis colorectal cancer (HNPCC)-associated tumors. Detection of defective MMR through analysis of loss of MMR protein immunostaining in colorectal carcinomas stratifies the risk of having an inherited cancer predisposition syndrome and identifies patients who might benefit from subsequent genetic testing and counseling. Multispectral imaging of colorectal cancer (CRC) HCT116, HT29, SW480 and SW620 cells cultured in chambered slides with multiplexed different color quantum dots (Qdots) conjugated MMR proteins MLH1, MSH2, MSH6 and PMS2 shows that HCT116 cells harbor defective MLH1 and PMS2, which is consistent with the finding from conventional single protein immunofluorescence staining. Multispectral imaging of CRC xenografted tumor sections with Qdot-conjugated antibodies to MMR proteins demonstrates that the human colorectal HCT116 tumor xenograft has MLH1 and PMS2 defects, consistent with individual protein immunohistochemical staining. To explore the application of multispectral imaging of MMRs in circulating tumor cells (CTCs), we spiked CRC cells into human blood and recovered CRC cells with the FDA-approved Veridex CellSearch system. Multispectral imaging of recovered CRC cells displayed similar MMR expression spectrum to conventional single protein immunofluorescence staining. The methodology may provide an adjunctive screening tool for detection of MMR defects in CTCs from patients with advanced disease. The utility of this novel assay for detection of MMR gene expression abnormalities will be further examined in the colorectal cancer clinic.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4058. doi:1538-7445.AM2012-4058
Abstract 4063: Image-guided fluorescent endomicroscopy of glioblastoma with SVV-GFP viral labeling
Authors: Wong Kelvin , Alvarado Miguel Valdivia y , Huang Yulun , Zhao Xiumei , Wong Stephen , Li Xiao-nan 
Published: August 2012

Abstract
We propose a novel method to improve detection of infiltrative microscopic GBM tumors during surgery, by labeling the infiltrative tumor cells in vivo by a single intravenous injection of a highly specific tumor seeking fluorescence virus, Seneca Valley Virus (SVV-GFP). The feasibility of this approach was demonstrated in a permissive pediatric GBM (IC-2305GBM) and an adult GBM (IC-A46GBM) orthotopic xenograft mouse model. Both models were derived from primary tissue of GBM from patients to preservethe histopathological and immunophenotypical characteristics of the original tumors. After establishing the xenograft tumors, SVV-GFP was injected intravenously at a dose of 5x1014 viral particles/kg. The animals were imaged at day 3 after SVV-GFP virus injection. During the imaging procedure, the animals were euthanized through anesthesia overdose and a small window was immediately drilled in the skull to introduce a 1mm outer diameter fiberoptic fluorescence probe through the tumor. Fluorescent labeled cells were recorded as a real-time video sequence along the track of the probe. Image quantification starts with low frequency temporal drift correction using Fast Fourier Transform based lowpass filtering. Air image frames (before probe insertion) were used as a zero reference and the intensity range of the video is shifted accordingly. A normal tissue fluorescence image was then estimated using a median statistics of all image frames acquired in normal brain tissue. This image was used to correct for lower detection sensitivity near the probe edge or any field inhomogeneity. The video sequence was then normalized to the normal tissue image to create a TBR map. By definition, air is used as reference with TBR=0 and normal tissue is at TBR=1. A cutoff of TBR&gt;1.1 is used and tumor cells exceeding that threshold were classified as GFP-positive tumor cells. Fractional area occupied by the virus-labeled microscopic tumors were computed with the TBR exceeding 1.1 within the field of view. A low fractional area (&lt;0.01%) was found in normal brain tissue using this method, demonstrating that our method is robust. To summarize, we demonstrated an innovative approach for microscopic tumor detection in glioblastoma that is two to three orders of magnitude smaller than those currently detected with in vivo imaging modalities.This method can potentially be applied to detect microscopic tumors in metastatic retinoblastoma, medulloblastoma, endocrine tumors and small cell lung cancer which are shown to be permissive to infection of this virus.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4063. doi:1538-7445.AM2012-4063
Abstract 4068: A novel microfluidic platform for isolation of circulating tumor cells intended to facilitate downstream biomarker analyses
Authors: Conant Carolyn G. , Nevill Tanner , Ionescu-Zanetti Cristian , Schwartz Micahel 
Published: August 2012

Abstract
Circulating tumor cells (CTCs) inherently hold important biomarker information potentially pertinent to cancer patient therapy and prognosis. Many existent technologies capture CTCs inside of a ‘chip’. Because of the difficulties in efficiently removing very small numbers of CTCs from ‘chips’, these technologies have limited potential for downstream (off-chip) analysis opportunities. We have developed a novel microfluidic system for high purity CTC isolation and analysis specifically for post-capture CTC use. Using our device, analytical studies were performed with cell lines (HT29, SKBR3, PC3) to demonstrate recovery and purity capabilities. We isolated CTCs with high purity (from 1.5- 50%) and high capture efficiency (up to 96%) in volumes as low as 5 µl. Clinical samples from stage IV carcinoma patients of several different pathologies were processed and benchmarked against the predicate system. We found, in all cases, CTCs could be identified even when not identified on the predicate system. Further clinical samples indicated in some cases CTCs could be detected as early as stage II. A technique designed to identify somatic mutations in the KRAS oncogene was employed successfully to characterize samples isolated on the device from colon carcinoma patients. Fluorescence in situ hybridization (FISH) was used to characterize Her-2 and Top2A gene status in CTCs isolated from breast carcinoma patients. The use of this device is likely extensible to a wide variety of molecular biology and biochemistry-based biomarker studies which previously have been precluded due to insufficient CTC sample quality.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4068. doi:1538-7445.AM2012-4068
Abstract 407: Loss of Smad3 alters host-microbial interactions, predisposing the colonic epithelium to inflammation
Authors: Ball Corbie , Ramireddy Bommireddy , Keenan Michael , Stern Stephen , Azhar Mohamad , Gard Constance , Besselsen David G. , Doetschman Thomas 
Published: August 2012

Abstract
Introduction: The TGFβ pathway is mutated in up to 30% of human colon cancers. Genetically Engineered Mouse Models (GEMs) with deficient TGFβ signaling model several characteristics of IBD associated human colon cancers. Introduction of Helicobacter sp. into the Smad3−/− mouse model is necessary for the development of inflammatory lesions which progress to adenoma and carcinoma. The exact role of TGFβ1 signaling and bacterial-associated inflammation has yet to be elucidated and offers a potential target for the prevention of colon cancer. Methods: To determine the function of TGFβ1 signaling on colonic bacterial composition we used the Smad3−/− GEM. We designed primers specific to the 16s rRNA subunit of different Bacteroides species, and using the Roche LightCycler preformed quantitative Real Time-PCR (qRT-PCR) on cecal DNA extracted from Smad3−/-− and Smad3+/+ mice. To further examine the host-microbial interaction we extracted RNA and protein from the cecum of Smad3−/− and Smad3+/+ mice and ran Real Time-PCR or Western Blot for different Toll-like Receptor Pathway components. Results: qRT-PCR showed that Smad3−/-− mice have a significantly lower quantity of Bacteroides sp; the introduction of Helicobacter hepaticus results in a significant increase in B. distasonis and a significant decrease in B. thetaiotaomicron in Smad3−/− mice. These data suggest that loss of SMAD3 and the addition of Helicobacter hepaticus alters the colonic microflora. Previous studies illustrated the importance of epithelial signaling in microbial induced inflammation. To examine if Smad3 is altering the Toll-like receptor pathway we examined RT-PCR of the cecum and found a significant increase in Tlr4, Irak4, Cd14, Myd88, Nfkb, Cox2, and Nos2 mRNA in the Smad3−/−− mice compared to Smad3+/+ mice. Western blots confirmed these data by showing that cecal tissue from Smad3−/− mice has increased IRAK4 and pNF-κB. Conclusion: Smad3−/− mice show heightened Toll-like receptor pathway activity which leads to a hyper-inflammatory response of the mucosa to endogenous microbes. Further, loss of SMAD3 results in dysbiosis of the gut microbiota. These findings suggest that SMAD3 plays a role in microbial-host homeostasis.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 407. doi:1538-7445.AM2012-407