The Experts below are selected from a list of 15369 Experts worldwide ranked by ideXlab platform
Sohail Z Husain - One of the best experts on this subject based on the ideXlab platform.
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Pancreatic Cell tracing, lineage tagging and targeted genetic manipulations in multiple Cell types using Pancreatic ductal infusion of adeno-associated viral vectors and/or Cell-tagging dyes
Nature Protocols, 2014Co-Authors: Xiangwei Xiao, Krishna Prasadan, Chiyo Shiota, Lauren Peirish, Shane Fischbach, Zewen Song, Iljana Gaffar, John Wiersch, Yousef El-gohary, Sohail Z HusainAbstract:Pancreatic ductal infusion enables the delivery of dyes or viruses directly to specific Pancreatic Cell types, permitting their manipulation. This manipulation is helpful for the study of diabetes, pancreatitis and Pancreatic cancer. Genetic manipulations, with or without lineage tracing for specific Pancreatic Cell types, are very powerful tools for studying diabetes, pancreatitis and Pancreatic cancer. Nevertheless, the use of Cre/ loxP systems to conditionally activate or inactivate the expression of genes in a Cell type– and/or temporal-specific manner is not applicable to Cell tracing and/or gene manipulations in more than one lineage at a time. Here we report a technique that allows efficient delivery of dyes for Cell tagging into the mouse pancreas through the duct system, and that also delivers viruses carrying transgenes or siRNA under a specific promoter. When this technique is applied in genetically modified mice, it enables the investigator to perform either double lineage tracing or Cell lineage tracing combined with gene manipulation in a second lineage. The technique requires
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Pancreatic Cell tracing, lineage tagging and targeted genetic manipulations in multiple Cell types using Pancreatic ductal infusion of adeno-associated viral vectors and/or Cell-tagging dyes
Nature Protocols, 2014Co-Authors: Xiangwei Xiao, Krishna Prasadan, Chiyo Shiota, Lauren Peirish, Shane Fischbach, Zewen Song, Iljana Gaffar, John Wiersch, Yousef El-gohary, Sohail Z HusainAbstract:Genetic manipulations, with or without lineage tracing for specific Pancreatic Cell types, are very powerful tools for studying diabetes, pancreatitis and Pancreatic cancer. Nevertheless, the use of Cre/ loxP systems to conditionally activate or inactivate the expression of genes in a Cell type– and/or temporal-specific manner is not applicable to Cell tracing and/or gene manipulations in more than one lineage at a time. Here we report a technique that allows efficient delivery of dyes for Cell tagging into the mouse pancreas through the duct system, and that also delivers viruses carrying transgenes or siRNA under a specific promoter. When this technique is applied in genetically modified mice, it enables the investigator to perform either double lineage tracing or Cell lineage tracing combined with gene manipulation in a second lineage. The technique requires
Kimberly A Kelly - One of the best experts on this subject based on the ideXlab platform.
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abstract 1558 therapeutic targeting of Cell surface plectin induces anti cancer immune response and Pancreatic cancer regression
Cancer Research, 2019Co-Authors: Julien Dimastromatteo, Amanda Poisonnier, Samantha Perez, Lisa M Coussens, Kimberly A KellyAbstract:Background: Pancreatic ductal adenocarcinoma (PDA) is the 3rd deadliest cancer, diagnosed typically in advanced stages, with only an 8% 5-year survival rate, thus demonstrating the need for novel therapeutic approaches that significantly enhance chemo- and/or immune-therapy. Our team previously identified a promising functional target for cancer therapy in PDA, Cell surface plectin 1 (CSP1) that is aberrantly expressed on PDA Cells and thus a Cell surface-associated biomarker of cancer. CSP1 expression first becomes apparent in high grade dysplasias, remaining high in early and advanced cancers and in metastases. Our first-in-human imaging trial in PDA patients using a CSP1-targeted imaging agent revealed that CSP is an available target and accessible for binding, a potentially a target for cancer therapy. We hypothesized that a monoclonal antibody (mAb) against CSP1 could lead to novel Pancreatic cancer treatment options, thus, we developed a therapeutic mAb, e.g., ZB131, representing a first-in-class antibody selectively targeting CSP1. Methods: ZB131 is a humanized mAb targeted against human plectin 1 (rhSec8) that also binds murine CSP1. ZB131 affinity and its effect on cancer Cells including proliferation, cytotoxicity, and migration were tested in vitro using saturation binding, SRB-based survival assays, flow cytometry, and migration assays on various Pancreatic Cell types and homeostatic “normal” controls. In vivo validation was performed using two nu/nu mouse models bearing subcutaneous MiaPACA2 or Yapc PDA Cells, and a syngeneic KPC-derived tumor model to also evaluate immune responses to tumors treated with ZB131 or IgG control. Results: ZB131 exhibits high specificity and high affinity (0.4±0.1nM) to CSP1, and functionally induces G0 growth arrest followed by necrotic Cell death of PDA Cells in culture, and is synergistic with gemcitabine resulting in a 50-fold decrease in IC50. In vivo, in subcutaneous xenograft models, ZB131 monotherapy decreased PDA tumor volume 5-fold as compared to controls, and in combination with cisplatin resulted in sustained tumor reduction with greater than 85% tumor necrosis. In subcutaneous syngeneic PDA models, ZB131 induced complete tumor regression within 35 days mediated by an anti-tumor immune response as upon tumor rechallenge, full tumor regression was again achieved without additional ZB131 therapy. Leukocyte complexity analysis of regressing PDA tumors versus controls revealed an ~3-fold increase in effector and central memory T Cells. Conclusion: CSP1 is a first in class anti-cancer target expressed on the Cell surface of PDA, as well as other cancers including ovarian, esophageal and head neck. ZB131, an anti-CSP1 mAb, induces tumor Cell intrinsic Cell death, as well as a robust anti-tumor T Cell response leading to complete tumor regression indicating the potential therapeutic efficacy of ZB131 in late-stage cancers. Citation Format: Julien Dimastromatteo, Amanda Poisonnier, Samantha Perez, Lisa Coussens, Kimberly Kelly. Therapeutic targeting of Cell surface plectin induces anti-cancer immune response and Pancreatic cancer regression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1558.
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plectin 1 targeted aav vector for the molecular imaging of Pancreatic cancer
Frontiers in Oncology, 2013Co-Authors: Prasad R Konkalmatt, Defeng Deng, Stephanie Thomas, Michael T Wu, Craig D Logsdon, Brent A French, Kimberly A KellyAbstract:Pancreatic ductal adenocarcinoma (PDAC) is highly malignant disease that is the 4th leading cause of cancer-related death in the US. Gene therapy using AAV vectors to selectively deliver genes to PDAC Cells is an attractive treatment option for Pancreatic cancer. However, most AAV serotypes display a broad spectrum of tissue tropism and none of the existing serotypes specifically target PDAC Cells. This study tests the hypothesis that AAV2 can be genetically re-engineered to specifically target PDAC Cells by modifying the capsid surface to display a peptide that has previously been shown to bind plectin-1. Towards this end, a Plectin-1 Targeting Peptide (PTP) was inserted into the loop IV region of the AAV2 capsid, and the resulting capsid (AAV-PTP) was used in a series of in vitro and in vivo experiments. In vitro, AAV-PTP was found to target all five human PDAC Cell lines tested (PANC-1, MIA PaCa-2, HPAC, MPanc-96 and BxPC-3) preferentially over two non-neoplastic human Pancreatic Cell lines (HPDE and hPSC). In vivo, mice bearing subcutaneous tumor xenografts were generated using the PANC-1 Cell line. Once tumors reached a size of ~1-2 mm in diameter, the mice were injected intravenously with luciferase reporter vectors packaged in the either AAV-PTP or wild type AAV2 capsids. Luciferase expression was then monitored by bioluminescence imaging on days 3, 7 and 14 after vector injection. The results indicate that the AAV-PTP capsid displays a 37-fold preference for PANC-1 tumor xenographs over liver and other tissues; whereas the wild type AAV2 capsid displays a complementary preference for liver over tumors and other tissues. Together, these results establish proof-of-principle for the ability of PTP-modified AAV capsids to selectively target gene delivery to PDAC Cells in vivo, which opens promising new avenues for the early detection, diagnosis and treatment of Pancreatic cancer.
Raul Urrutia - One of the best experts on this subject based on the ideXlab platform.
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zymophagy a novel selective autophagy pathway mediated by vmp1 usp9x p62 prevents Pancreatic Cell death
Journal of Biological Chemistry, 2011Co-Authors: Daniel Grasso, Raul Urrutia, Alejandro Ropolo, Andrea Lo Re, Veronica Boggio, Maria Ines Molejon, Juan L Iovanna, Claudio Gonzalez, Maria I VaccaroAbstract:Autophagy has recently elicited significant attention as a mechanism that either protects or promotes Cell death, although different autophagy pathways, and the Cellular context in which they occur, remain to be elucidated. We report a thorough Cellular and biochemical characterization of a novel selective autophagy that works as a protective Cell response. This new selective autophagy is activated in Pancreatic acinar Cells during pancreatitis-induced vesicular transport alteration to sequester and degrade potentially deleterious activated zymogen granules. We have coined the term "zymophagy" to refer to this process. The autophagy-related protein VMP1, the ubiquitin-protease USP9x, and the ubiquitin-binding protein p62 mediate zymophagy. Moreover, VMP1 interacts with USP9x, indicating that there is a close cooperation between the autophagy pathway and the ubiquitin recognition machinery required for selective autophagosome formation. Zymophagy is activated by experimental pancreatitis in genetically engineered mice and cultured Pancreatic acinar Cells and by acute pancreatitis in humans. Furthermore, zymophagy has pathophysiological relevance by controlling pancreatitis-induced intraCellular zymogen activation and helping to prevent Cell death. Together, these data reveal a novel selective form of autophagy mediated by the VMP1-USP9x-p62 pathway, as a Cellular protective response.
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TIEG proteins join the Smads as TGF-β-regulated transcription factors that control Pancreatic Cell growth
American Journal of Physiology-gastrointestinal and Liver Physiology, 2000Co-Authors: Tiffany Cook, Raul UrrutiaAbstract:The control of epithelial Cell proliferation, differentiation, and apoptosis requires a balance between signaling and transcriptional regulation. Recent developments in Pancreatic Cell research have revealed that transforming growth factor-β (TGF-β) signaling is important for the regulation of each of these phenomena. More importantly, perturbations in this pathway are associated with Pancreatic cancer. A chief example of these alterations is the mutation in the TGF-β-regulated transcription factor Smad4/DPC4 that is found in a large percentage of Pancreatic tumors. Surprisingly, studies on transcription factors have remained an underrepresented area of Pancreatic research. However, the discovery of Smad4/DPC4 as a transcription factor fueled further studies aimed at characterizing transcription factors involved in normal and neoplastic Pancreatic Cell growth. Our laboratory recently described the existence of a novel family of zinc finger transcription factors, TGF-β-inducible early-response gene (TIEG)1 and TIEG2, from the exocrine pancreas that, similarly to Smads, participate in the TGF-β response and inhibit epithelial Cell proliferation. This review therefore focuses on describing the structure and function of these two families of transcription factor proteins that are becoming key players in the regulation of Pancreatic Cell growth.
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Exploring the role of homeobox and zinc finger proteins in Pancreatic Cell proliferation, differentiation, and apoptosis
International Journal of Gastrointestinal Cancer, 1997Co-Authors: Raul UrrutiaAbstract:Transcription factors are DNA binding proteins that regulate gene expression in response to a large variety of extraCellular stimuli, and thereby act as key molecular switches for controlling Cell differentiation, proliferation, and apoptosis. During the last decade, a myriad of these proteins have been identified and classified into different structural families, including homeobox, zinc finger, leucine zipper, and helix-loop-helix transcription factors. Members of the homeobox and zinc finger superfamilies are among the best-characterized transcription factors known to act as potent regulators of normal development in organisms ranging from insects to humans. In addition, mutations or aberrant expression in genes encoding these proteins can result in neoplastic transformation in several different Cell types, further supporting their role as “guardians” of normal Cell growth and differentiation. Therefore, the purpose of this article is to review this field of research with a particular emphasis on the role of homeobox- and zinc finger-containing transcription factors in Pancreatic Cell growth, Cell differentiation, and apoptosis. The potential participation of these proteins in neoplastic transformation is also discussed.
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Identification and characterization of zinc finger encoding genes from the tumoral exocrine Pancreatic Cell line AR42J
Cancer Letters, 1996Co-Authors: Kristin Mesa, Tiffany Cook, Brian Gebelein, Raul UrrutiaAbstract:Abstract Zinc finger transcription factors are DNA-binding proteins that are known to determine the identity of Cells by regulating Cell-specific gene expression. In addition, mutations in some members of this family have been found to be associated with several neoplasias, including Wilms' tumor and leukemias. Because the mechanisms that regulate normal, as well as neoplastic, Pancreatic Cell differentiation are poorly understood, we are searching for pancreas-enriched transcription factors that may determine the identity of Pancreatic Cells. Towards this end, we have used the polymerase chain reaction and degenerate primers against the highly conserved (Cys 2 -His 2 ) zinc finger domain to amplify novel transcription factor encoding cDNAs from the well-characterized Pancreatic acinar Cell line AR42J. Using this approach, we have identified 17 different zinc finger encoding cDNAs (AZF-1 to -17). Sequence analysis shows that all of these clones encode for different zinc finger peptides which share the consensus DNA binding motif with the Drosophila transcription factor kruppel . As a first step in the characterization of these genes, the purified PCR products were used to determine their spatial pattern of expression by northern blot analysis. Using these techniques, we find that numerous zinc finger encoding genes are expressed in AR42J acinar Cells as well as in normal adult rat pancreas and suggest that they may play a role as transcription factors in exocrine Pancreatic Cells.
Xiangwei Xiao - One of the best experts on this subject based on the ideXlab platform.
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Pancreatic Cell tracing, lineage tagging and targeted genetic manipulations in multiple Cell types using Pancreatic ductal infusion of adeno-associated viral vectors and/or Cell-tagging dyes
Nature Protocols, 2014Co-Authors: Xiangwei Xiao, Krishna Prasadan, Chiyo Shiota, Lauren Peirish, Shane Fischbach, Zewen Song, Iljana Gaffar, John Wiersch, Yousef El-gohary, Sohail Z HusainAbstract:Pancreatic ductal infusion enables the delivery of dyes or viruses directly to specific Pancreatic Cell types, permitting their manipulation. This manipulation is helpful for the study of diabetes, pancreatitis and Pancreatic cancer. Genetic manipulations, with or without lineage tracing for specific Pancreatic Cell types, are very powerful tools for studying diabetes, pancreatitis and Pancreatic cancer. Nevertheless, the use of Cre/ loxP systems to conditionally activate or inactivate the expression of genes in a Cell type– and/or temporal-specific manner is not applicable to Cell tracing and/or gene manipulations in more than one lineage at a time. Here we report a technique that allows efficient delivery of dyes for Cell tagging into the mouse pancreas through the duct system, and that also delivers viruses carrying transgenes or siRNA under a specific promoter. When this technique is applied in genetically modified mice, it enables the investigator to perform either double lineage tracing or Cell lineage tracing combined with gene manipulation in a second lineage. The technique requires
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Pancreatic Cell tracing, lineage tagging and targeted genetic manipulations in multiple Cell types using Pancreatic ductal infusion of adeno-associated viral vectors and/or Cell-tagging dyes
Nature Protocols, 2014Co-Authors: Xiangwei Xiao, Krishna Prasadan, Chiyo Shiota, Lauren Peirish, Shane Fischbach, Zewen Song, Iljana Gaffar, John Wiersch, Yousef El-gohary, Sohail Z HusainAbstract:Genetic manipulations, with or without lineage tracing for specific Pancreatic Cell types, are very powerful tools for studying diabetes, pancreatitis and Pancreatic cancer. Nevertheless, the use of Cre/ loxP systems to conditionally activate or inactivate the expression of genes in a Cell type– and/or temporal-specific manner is not applicable to Cell tracing and/or gene manipulations in more than one lineage at a time. Here we report a technique that allows efficient delivery of dyes for Cell tagging into the mouse pancreas through the duct system, and that also delivers viruses carrying transgenes or siRNA under a specific promoter. When this technique is applied in genetically modified mice, it enables the investigator to perform either double lineage tracing or Cell lineage tracing combined with gene manipulation in a second lineage. The technique requires
Bernard Peers - One of the best experts on this subject based on the ideXlab platform.
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Transcriptome analysis of Pancreatic Cells across distant species highlights novel important regulator genes
BMC Biology, 2017Co-Authors: Estefanía Tarifeño-saldivia, Arnaud Lavergne, Alice Bernard, Keerthana Padamata, David Bergemann, Isabelle Manfroid, Bernard PeersAbstract:Background Defining the transcriptome and the genetic pathways of Pancreatic Cells is of great interest for elucidating the molecular attributes of pancreas disorders such as diabetes and cancer. As the function of the different Pancreatic Cell types has been maintained during vertebrate evolution, the comparison of their transcriptomes across distant vertebrate species is a means to pinpoint genes under strong evolutionary constraints due to their crucial function, which have therefore preserved their selective expression in these Pancreatic Cell types.
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Transcriptome analysis of Pancreatic Cells across distant species highlights novel important regulator genes
BMC Biology, 2017Co-Authors: Estefanía Tarifeño-saldivia, Arnaud Lavergne, Alice Bernard, Keerthana Padamata, David Bergemann, Isabelle Manfroid, Bernard PeersAbstract:Background Defining the transcriptome and the genetic pathways of Pancreatic Cells is of great interest for elucidating the molecular attributes of pancreas disorders such as diabetes and cancer. As the function of the different Pancreatic Cell types has been maintained during vertebrate evolution, the comparison of their transcriptomes across distant vertebrate species is a means to pinpoint genes under strong evolutionary constraints due to their crucial function, which have therefore preserved their selective expression in these Pancreatic Cell types. Results In this study, RNA-sequencing was performed on Pancreatic alpha, beta, and delta endocrine Cells as well as the acinar and ductal exocrine Cells isolated from adult zebrafish transgenic lines. Comparison of these transcriptomes identified many novel markers, including transcription factors and signaling pathway components, specific for each Cell type. By performing interspecies comparisons, we identified hundreds of genes with conserved enriched expression in endocrine and exocrine Cells among human, mouse, and zebrafish. This list includes many genes known as crucial for Pancreatic Cell formation or function, but also pinpoints many factors whose Pancreatic function is still unknown. A large set of endocrine-enriched genes can already be detected at early developmental stages as revealed by the transcriptomic profiling of embryonic endocrine Cells, indicating a potential role in Cell differentiation. The actual involvement of conserved endocrine genes in Pancreatic Cell differentiation was demonstrated in zebrafish for myt1b , whose invalidation leads to a reduction of alpha Cells, and for cdx4 , selectively expressed in endocrine delta Cells and crucial for their specification. Intriguingly, comparison of the endocrine alpha and beta Cell subtypes from human, mouse, and zebrafish reveals a much lower conservation of the transcriptomic signatures for these two endocrine Cell subtypes compared to the signatures of pan-endocrine and exocrine Cells. These data suggest that the identity of the alpha and beta Cells relies on a few key factors, corroborating numerous examples of inter-conversion between these two endocrine Cell subtypes. Conclusion This study highlights both evolutionary conserved and species-specific features that will help to unveil universal and fundamental regulatory pathways as well as pathways specific to human and laboratory animal models such as mouse and zebrafish.
