18 results on '"Hadas Cohen Dvashi"'
Search Results
2. Longitudinal Isolation of Potent Near-Germline SARS-CoV-2-Neutralizing Antibodies from COVID-19 Patients
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Artem Ashurov, Timm Weber, Alexandra Kupke, Hanna Janicki, Nico Pfeifer, Reinhild Brinker, Hadas Cohen-Dvashi, Jan Mathis Eckert, Simone Lederer, Meryem S. Ercanoglu, Cornelius Rohde, Stephan Becker, Veronica Di Cristanziano, Maria J G T Vehreschild, Clemens M. Wendtner, Manuel Koch, Florian Klein, Sandro Halwe, Christoph Kreer, Philipp Schommers, Matthias Zehner, Kanika Vanshylla, Verena Krähling, Henning Gruell, M. Korenkov, Lutz Gieselmann, Timo Wolf, and Ron Diskin
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Coronavirus disease 2019 (COVID-19) ,Isolation (health care) ,medicine.drug_class ,Somatic cell ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Naive B cell ,Pneumonia, Viral ,Cell ,Immunoglobulin Variable Region ,Monoclonal antibody ,Antibodies, Viral ,Article ,General Biochemistry, Genetics and Molecular Biology ,Germline ,03 medical and health sciences ,Betacoronavirus ,0302 clinical medicine ,Immune system ,medicine ,Humans ,Longitudinal Studies ,Neutralizing antibody ,Pandemics ,030304 developmental biology ,0303 health sciences ,B-Lymphocytes ,biology ,SARS-CoV-2 ,single B cell analysis ,Correction ,COVID-19 ,neutralizing antibody ,Antibodies, Neutralizing ,Virology ,Vaccination ,medicine.anatomical_structure ,monoclonal antibody ,2019-nCoV ,biology.protein ,Somatic Hypermutation, Immunoglobulin ,Antibody ,Coronavirus Infections ,Immunologic Memory ,030217 neurology & neurosurgery - Abstract
Summary The SARS-CoV-2 pandemic has unprecedented implications for public health, social life, and the world economy. Because approved drugs and vaccines are limited or not available, new options for COVID-19 treatment and prevention are in high demand. To identify SARS-CoV-2-neutralizing antibodies, we analyzed the antibody response of 12 COVID-19 patients from 8 to 69 days after diagnosis. By screening 4,313 SARS-CoV-2-reactive B cells, we isolated 255 antibodies from different time points as early as 8 days after diagnosis. Of these, 28 potently neutralized authentic SARS-CoV-2 with IC100 as low as 0.04 μg/mL, showing a broad spectrum of variable (V) genes and low levels of somatic mutations. Interestingly, potential precursor sequences were identified in naive B cell repertoires from 48 healthy individuals who were sampled before the COVID-19 pandemic. Our results demonstrate that SARS-CoV-2-neutralizing antibodies are readily generated from a diverse pool of precursors, fostering hope for rapid induction of a protective immune response upon vaccination., Graphical Abstract, Highlights • Isolation of highly potent SARS-CoV-2-neutralizing antibodies • Longitudinal sampling reveals early class-switched neutralizing response • SARS-CoV-2 S-protein-reactive antibodies show little somatic mutation over time • Potential antibody precursor sequences identified in SARS-CoV-2-naive individuals, In a longitudinal analysis of SARS-CoV-2-infected people, Kreer et al. find highly potent neutralizing antibodies that use a broad spectrum of variable (V) genes and show low levels of somatic mutations. They also identify potential precursor sequences of these SARS-CoV-2-neutralizing antibodies from virus-naive individuals, sampled before the COVID-19 pandemic. This could indicate that neutralizing antibodies can be readily generated from existing germline antibody sequences found in the general population.
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- 2020
3. Rational design of universal immunotherapy for TfR1-tropic arenaviruses
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Thomas W. Geisbert, Vered Padler-Karavani, Sylvain Baize, Hadas Cohen-Dvashi, Ron Amon, Krystle N. Agans, Aliza Borenstein-Katz, Ron Diskin, Robert W. Cross, Mathieu Mateo, Weizmann Institute of Science [Rehovot, Israël], Tel Aviv University (TAU), The University of Texas Medical Branch (UTMB), Biologie des Infections Virales Émergentes - Biology of Emerging Viral Infections (UBIVE), Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] (IP), BSL-4 experiments using MACV were supported by the Pasteur-Weizmann joint research program collaborative research grant S-CR18069-02 to M.M. and R.D. BSL-4 operations of the Galveston National Laboratory and Department of Microbiology and Immunology are supported by NIAID/NIH Grant No. UC7AI094660 to T.W.G. The lab of R.D. is supported by research grants from the Enoch Foundation, from the Abramson Family Center for Young Scientists, from Ms. Rudolfine Steindling, and from the I-CORE Program of the Planning and Budgeting Committee and The Israel Science Foundation (grants no. 1775/12 and 682/16)., Tel Aviv University [Tel Aviv], Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] (IP), and Bodescot, Myriam
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0301 basic medicine ,New World Arenavirus ,Viral protein ,medicine.medical_treatment ,Science ,viruses ,General Physics and Astronomy ,Transferrin receptor ,Biologics ,Biology ,Antibodies, Viral ,medicine.disease_cause ,Article ,General Biochemistry, Genetics and Molecular Biology ,Viral Proteins ,03 medical and health sciences ,0302 clinical medicine ,Virology ,Receptors, Transferrin ,medicine ,Arenaviridae Infections ,Humans ,lcsh:Science ,X-ray crystallography ,[SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Multidisciplinary ,Transmission (medicine) ,Rational design ,virus diseases ,Arenavirus ,General Chemistry ,Immunotherapy ,Antibodies, Neutralizing ,Arenaviruses ,3. Good health ,Hemorrhagic Fevers ,030104 developmental biology ,Drug Design ,[SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology ,Biological warfare ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,[SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology ,Receptors, Virus ,lcsh:Q ,030217 neurology & neurosurgery - Abstract
Certain arenaviruses that circulate in rodent populations can cause life-threatening hemorrhagic fevers when they infect humans. Due to their efficient transmission, arenaviruses pose a severe risk for outbreaks and might be exploited as biological weapons. Effective countermeasures against these viruses are highly desired. Ideally, a single remedy would be effective against many or even all the pathogenic viruses in this family. However, despite the fact that all pathogenic arenaviruses from South America utilize transferrin receptor 1 (TfR1) as a cellular receptor, their viral glycoproteins are highly diversified, impeding efforts to isolate cross-neutralizing antibodies. Here we address this problem using a rational design approach to target TfR1-tropic arenaviruses with high potency and breadth. The pan-reactive molecule is highly effective against all arenaviruses that were tested, offering a universal therapeutic approach. Our design scheme avoids the shortcomings of previous immunoadhesins and can be used to combat other zoonotic pathogens., New World arenaviruses utilize the cellular transferrin receptor 1 (TfR1) to enter host cells. Here, the authors develop a TfR1-mimetic immunoadhesin, Arenacept, that targets viral spike complexes and exerts effective pan-reactive neutralization against pathogenic mammarenaviruses.
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- 2020
4. Variations in Core Packing of GP2 from Old World Mammarenaviruses in their Post-Fusion Conformations Affect Membrane-Fusion Efficiencies
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Michael Katz, Ron Diskin, Hadas Cohen-Dvashi, Harry M. Greenblatt, Anastasiya Shulman, and Yaakov Levy
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Protein Conformation ,Viral protein ,viruses ,Molecular Dynamics Simulation ,Crystallography, X-Ray ,medicine.disease_cause ,Membrane Fusion ,Cell Line ,03 medical and health sciences ,Lassa Fever ,0302 clinical medicine ,Structural Biology ,medicine ,Animals ,Humans ,Lassa virus ,Molecular Biology ,030304 developmental biology ,Cell entry ,chemistry.chemical_classification ,0303 health sciences ,Fusion ,Lipid bilayer fusion ,Virus Internalization ,HEK293 Cells ,Membrane ,chemistry ,Biophysics ,Glycoprotein ,030217 neurology & neurosurgery ,Mammarenavirus - Abstract
Lassa virus (LASV) is a notorious human pathogen in West Africa. Its class I trimeric spike complex displays a distinct architecture, and its cell entry mechanism involves unique attributes not shared by other related viruses. We determined the crystal structure of the GP2 fusion glycoprotein from the spike complex of LASV (GP2LASV) in its post-fusion conformation. GP2LASV adopts a canonical helical bundle configuration similarly to other viruses in its family. The core packing of GP2LASV, however, is more organized compared to GP2 from other viruses reducing the formation of internal hydrophobic cavities. We demonstrate a link between the formation of such unfavorable hydrophobic cavities and the efficiencies of membrane fusion and cell entry. Our study suggests that LASV has evolved a more efficient membrane fusogen compared to other viruses from its family by optimizing the post-fusion configuration of its GP2 module.
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- 2019
5. Cross-reactive antibodies against human coronaviruses and the animal coronavirome suggest diagnostics for future zoonotic spillovers
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Ronen Gabizon, Eilat Shinar, Ron Diskin, Lihee Moss, Roei D Mazor, Sigal Leviatan, Shelley Klompus, Nachum Nathan, Gur Yaari, Hadas Cohen Dvashi, Liat Stoler-Barak, Sharon Kagan Ben Tikva, Anastasia Godneva, Ziv Shulman, Iris N. Kalka, Thomas Vogl, Eran Segal, Nir London, Adina Weinberger, and Ayelet Peres
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Adult ,Male ,0301 basic medicine ,Adolescent ,Coronavirus disease 2019 (COVID-19) ,medicine.drug_class ,viruses ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Immunology ,Cross Reactions ,Biology ,Antibodies, Viral ,Monoclonal antibody ,medicine.disease_cause ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,Peptide Library ,Zoonoses ,medicine ,Animals ,Humans ,Peptide library ,Antigens, Viral ,Aged ,Coronavirus ,Transmission (medicine) ,virus diseases ,General Medicine ,Middle Aged ,Virology ,3. Good health ,030104 developmental biology ,biology.protein ,Female ,Antibody ,Coronavirus Infections ,030217 neurology & neurosurgery - Abstract
The spillover of animal coronaviruses (aCoVs) to humans has caused SARS, MERS, and COVID-19. While antibody responses displaying cross-reactivity between SARS-CoV-2 and seasonal/common cold human coronaviruses (hCoVs) have been reported, potential cross-reactivity with aCoVs and the diagnostic implications are incompletely understood. Here, we probed for antibody binding against all seven hCoVs and 49 aCoVs represented as 12,924 peptides within a phage-displayed antigen library. Antibody repertoires of 269 recovered COVID-19 patients showed distinct changes compared to 260 unexposed pre-pandemic controls, not limited to binding of SARS-CoV-2 antigens but including binding to antigens from hCoVs and aCoVs with shared motifs to SARS-CoV-2. We isolated broadly reactive monoclonal antibodies from recovered COVID-19 patients that bind a shared motif of SARS-CoV-2, hCoV-OC43, hCoV-HKU1, and several aCoVs, demonstrating that interspecies cross-reactivity can be mediated by a single immunoglobulin. Employing antibody binding data against the entire CoV antigen library allowed accurate discrimination of recovered COVID-19 patients from unexposed individuals by machine learning. Leaving out SARS-CoV-2 antigens and relying solely on antibody binding to other hCoVs and aCoVs achieved equally accurate detection of SARS-CoV-2 infection. The ability to detect SARS-CoV-2 infection without knowledge of its unique antigens solely from cross-reactive antibody responses against other hCoVs and aCoVs suggests a potential diagnostic strategy for the early stage of future pandemics. Creating regularly updated antigen libraries representing the animal coronavirome can provide the basis for a serological assay already poised to identify infected individuals following a future zoonotic transmission event.
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- 2021
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6. Structural Basis for a Convergent Immune Response against Ebola Virus
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Stefanie A Ehrhardt, Ron Diskin, Matthias Zehner, Florian Klein, Nadav Elad, Michael Katz, and Hadas Cohen-Dvashi
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Zaire ebolavirus ,Viral protein ,medicine.disease_cause ,Antibodies, Viral ,Microbiology ,Virus ,03 medical and health sciences ,Epitopes ,0302 clinical medicine ,Immune system ,Viral Envelope Proteins ,Antibody Specificity ,Virology ,medicine ,Humans ,Amino Acid Sequence ,Ebola Vaccines ,030304 developmental biology ,Ebolavirus ,0303 health sciences ,Ebola virus ,biology ,Cryoelectron Microscopy ,Antibodies, Monoclonal ,Antibodies, Neutralizing ,Protein Structure, Tertiary ,HEK293 Cells ,Immunization ,biology.protein ,Parasitology ,Antibody ,030217 neurology & neurosurgery ,Protein Binding - Abstract
Ebola virus disease is a severe health problem in Africa. Vaccines that display the Zaire ebolavirus glycoprotein spike complex are a prime component for the effort to combat it. The VH3-15/Vλ1-40-based class of antibodies was recently discovered to be a common response in individuals who received the Ebola virus vaccines. These antibodies display attractive properties, and thus likely contribute to the efficacy of the vaccines. Here, we use cryo-EM to elucidate how three VH3-15/Vλ1-40 antibodies from different individuals target the virus and found a convergent mechanism against a partially conserved site on the spike complex. Our study rationalizes the selection of the VH3-15/Vλ1-40 germline genes for specifically targeting this site and highlights Ebolavirus species-specific sequence divergences that may restrict breadth of VH3-15/Vλ1-40-based humoral response. The results from this study could help develop improved immunization schemes and further enable the design of immunogens that would be efficacious against a broader set of Ebolavirus species.
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- 2019
7. Biomolecular Recognition of the Glycan Neoantigen CA19-9 by Distinct Antibodies
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Ron Amon, Shani Leviatan Ben-Arye, Hai Yu, Sarel J. Fleishman, Nova Tasnima, Maayan Eilon, Shira Warszawski, Aliza Borenstein-Katz, Vered Padler-Karavani, Ron Diskin, Hadas Cohen-Dvashi, and Xi Chen
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Models, Molecular ,Protein Conformation ,design ,Antibody Affinity ,Computational algorithm ,Crystallography, X-Ray ,Mice ,0302 clinical medicine ,Models ,Structural Biology ,Monoclonal ,Cancer ,chemistry.chemical_classification ,0303 health sciences ,Crystallography ,biology ,Antibodies, Monoclonal ,CA19-9 ,Antigen binding ,3. Good health ,Antibody ,Carbohydrate antigen ,Algorithms ,Biochemistry & Molecular Biology ,Glycan ,CA-19-9 Antigen ,Computational biology ,Microbiology ,Antibodies ,Article ,Medicinal and Biomolecular Chemistry ,03 medical and health sciences ,Glycolipid ,Antigen ,Animals ,Humans ,structure ,Molecular Biology ,030304 developmental biology ,Prevention ,Molecular ,Computational Biology ,chemistry ,Mutation ,X-Ray ,biology.protein ,glycans ,Biochemistry and Cell Biology ,Glycoprotein ,030217 neurology & neurosurgery - Abstract
Glycans decorate the cell surface, secreted glycoproteins and glycolipids, and altered glycans are often found in cancers. Despite their high diagnostic and therapeutic potential, however, glycans are polar and flexible molecules that are quite challenging for the development and design of high-affinity binding antibodies. To understand the mechanisms by which glycan neoantigens are specifically recognized by antibodies, we analyze the biomolecular recognition of the tumor-associated carbohydrate antigen CA19-9 by two distinct antibodies using X-ray crystallography. Despite the potential plasticity of glycans and the very different antigen-binding surfaces presented by the antibodies, both structures reveal an essentially identical extended CA19-9 conformer, suggesting that this conformer's stability selects the antibodies. Starting from the bound structure of one of the antibodies, we use the AbLIFT computational algorithm to design a variant with seven core mutations in the variable domain's light-heavy chain interface that exhibits tenfold improved affinity for CA19-9. The results reveal strategies used by antibodies to specifically recognize glycan antigens and show how automated antibody-optimization methods may be used to enhance the clinical potential of existing antibodies.
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- 2021
8. Polyclonal and convergent antibody response to Ebola virus vaccine rVSV-ZEBOV
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Stefanie A, Ehrhardt, Matthias, Zehner, Verena, Krähling, Hadas, Cohen-Dvashi, Christoph, Kreer, Nadav, Elad, Henning, Gruell, Meryem S, Ercanoglu, Philipp, Schommers, Lutz, Gieselmann, Ralf, Eggeling, Christine, Dahlke, Timo, Wolf, Nico, Pfeifer, Marylyn M, Addo, Ron, Diskin, Stephan, Becker, and Florian, Klein
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Adult ,Male ,Volunteers ,B-Lymphocytes ,Vaccination ,Vesiculovirus ,Hemorrhagic Fever, Ebola ,Middle Aged ,Antibodies, Viral ,Ebolavirus ,Antibodies, Neutralizing ,Immunity, Humoral ,Antibody Formation ,Humans ,Female ,Ebola Vaccines - Abstract
Recombinant vesicular stomatitis virus-Zaire Ebola virus (rVSV-ZEBOV) is the most advanced Ebola virus vaccine candidate and is currently being used to combat the outbreak of Ebola virus disease (EVD) in the Democratic Republic of the Congo (DRC). Here we examine the humoral immune response in a subset of human volunteers enrolled in a phase 1 rVSV-ZEBOV vaccination trial by performing comprehensive single B cell and electron microscopy structure analyses. Four studied vaccinees show polyclonal, yet reproducible and convergent B cell responses with shared sequence characteristics. EBOV-targeting antibodies cross-react with other Ebolavirus species, and detailed epitope mapping revealed overlapping target epitopes with antibodies isolated from EVD survivors. Moreover, in all vaccinees, we detected highly potent EBOV-neutralizing antibodies with activities comparable or superior to the monoclonal antibodies currently used in clinical trials. These include antibodies combining the IGHV3-15/IGLV1-40 immunoglobulin gene segments that were identified in all investigated individuals. Our findings will help to evaluate and direct current and future vaccination strategies and offer opportunities for novel EVD therapies.
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- 2019
9. Structural basis for receptor recognition by Lujo virus
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Hadas Cohen-Dvashi, Itay Kilimnik, and Ron Diskin
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0301 basic medicine ,Microbiology (medical) ,Old World ,New World Arenavirus ,Viral protein ,Arenaviridae ,Immunology ,Protein domain ,Virus Attachment ,Human pathogen ,Transferrin receptor ,Context (language use) ,Computational biology ,medicine.disease_cause ,Crystallography, X-Ray ,Applied Microbiology and Biotechnology ,Microbiology ,03 medical and health sciences ,Protein Domains ,Viral Envelope Proteins ,Genetics ,medicine ,Humans ,Dystroglycans ,Pathogen ,Lujo virus ,Binding Sites ,biology ,Mechanism (biology) ,Cell Biology ,biology.organism_classification ,Neuropilin-2 ,030104 developmental biology ,HEK293 Cells ,Mutation ,Protein Binding - Abstract
Lujo virus (LUJV) has emerged as a novel and highly fatal human pathogen. Despite its membership among the Arenaviridae, LUJV does not classify with the known Old and New World groups of that viral family. Likewise, LUJV was recently found to use neuropilin-2 (NRP2) as a cellular receptor instead of the canonical α-dystroglycan (α-DG) or transferrin receptor 1 (TfR1) utilized by Old World (OW) and New World (NW) arenaviruses, respectively. The emergence of a deadly new pathogen into human populations using an unprecedented entry route raises many questions regarding the mechanism of cell recognition and the risk that Arenaviruses are further diversifying their infection strategies. To provide the basis for combating LUJV in particular, and to increase our general understanding of the molecular changes that accompany an evolutionary switch to a new receptor for Arenaviruses, we used X-ray crystallography to reveal how the GP1 receptor-binding domain of LUJV (LUJVGP1) recognizes NRP2. Our structural data imply that LUJV is evolutionary closer to OW than to NW arenaviruses. Structural analysis supported by experimental validation further suggests that NRP2 recognition is metal ion dependent and that the complete NRP2 binding is formed in the context of the trimeric spike. Taken together, our data provide the mechanism for the cell attachment step of LUJV, the evolutionary relationship between the GP1 domain of this novel pathogen and other arenaviruses, and indispensable information for combating LUJV.
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- 2018
10. Molecular Mechanism for LAMP1 Recognition by Lassa Virus
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Ron Diskin, Hadas Cohen-Dvashi, Hadar Israeli, and Nadav Cohen
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Models, Molecular ,Viral protein ,viruses ,Molecular Sequence Data ,Immunology ,Plasma protein binding ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,Cell Line ,Lassa Fever ,Viral Envelope Proteins ,Virology ,medicine ,Humans ,Amino Acid Sequence ,Lassa virus ,Lassa fever ,Mutation ,Structure and Assembly ,Lysosome-Associated Membrane Glycoproteins ,virus diseases ,medicine.disease ,Hemorrhagic Fevers ,Insect Science ,biology.protein ,Receptors, Virus ,Antibody ,Sequence Alignment ,Protein Binding - Abstract
Lassa virus is a notorious human pathogen that infects many thousands of people each year in West Africa, causing severe viral hemorrhagic fevers and significant mortality. The surface glycoprotein of Lassa virus mediates receptor recognition through its GP1 subunit. Here we report the crystal structure of GP1 from Lassa virus, which is the first representative GP1 structure for Old World arenaviruses. We identify a unique triad of histidines that forms a binding site for LAMP1, a known lysosomal protein recently discovered to be a critical receptor for internalized Lassa virus at acidic pH. We demonstrate that mutation of this histidine triad, which is highly conserved among Old World arenaviruses, impairs LAMP1 recognition. Our biochemical and structural data further suggest that GP1 from Lassa virus may undergo irreversible conformational changes that could serve as an immunological decoy mechanism. Together with a variable region that we identify on the surface of GP1, those could be two distinct mechanisms that Lassa virus utilizes to avoid antibody-based immune response. IMPORTANCE Structural data at atomic resolution for viral proteins is key for understanding their function at the molecular level and can facilitate novel avenues for combating viral infections. Here we used X-ray protein crystallography to decipher the crystal structure of the receptor-binding domain (GP1) from Lassa virus. This is a pathogenic virus that causes significant illness and mortality in West Africa. This structure reveals the overall architecture of GP1 domains from the group of viruses known as the Old World arenaviruses. Using this structural information, we elucidated the mechanisms for pH switch and binding of Lassa virus to LAMP1, a recently identified host receptor that is critical for successful infection. Lastly, our structural analysis suggests two novel immune evasion mechanisms that Lassa virus may utilize to escape antibody-based immune response.
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- 2015
11. Mapping of the Lassa virus LAMP1 binding site reveals unique determinants not shared by other old world arenaviruses
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Hadas Cohen-Dvashi, Ron Diskin, Anastasiya Shulman, Amir Shimon, and Hadar Israeli
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0301 basic medicine ,RNA viruses ,Models, Molecular ,viruses ,medicine.disease_cause ,Pathology and Laboratory Medicine ,Biochemistry ,Membrane Fusion ,Binding Analysis ,Morogoro virus ,Medicine and Health Sciences ,Amino Acids ,Lassa fever ,lcsh:QH301-705.5 ,Crystallography ,Organic Compounds ,Physics ,virus diseases ,Condensed Matter Physics ,Chemistry ,Medical Microbiology ,Filoviruses ,Viral Pathogens ,Physical Sciences ,Viruses ,Crystal Structure ,Lassa Virus ,Basic Amino Acids ,Pathogens ,Ebola Virus ,Research Article ,Protein Binding ,lcsh:Immunologic diseases. Allergy ,Cell Binding ,Cell Physiology ,Viral Entry ,Viral protein ,Immunology ,Receptors, Cell Surface ,Biology ,Research and Analysis Methods ,Microbiology ,03 medical and health sciences ,Lassa Fever ,Viral envelope ,Species Specificity ,Viral entry ,Virology ,Genetics ,medicine ,Solid State Physics ,Animals ,Arenaviridae Infections ,Humans ,Histidine ,Amino Acid Sequence ,Molecular Biology ,Microbial Pathogens ,Chemical Characterization ,Ebola virus ,Binding Sites ,Hemorrhagic Fever Viruses ,Organic Chemistry ,Chemical Compounds ,Organisms ,Lipid bilayer fusion ,Biology and Life Sciences ,Proteins ,Lysosome-Associated Membrane Glycoproteins ,Cell Biology ,medicine.disease ,Arenaviruses ,Models, Structural ,030104 developmental biology ,Lassa virus ,lcsh:Biology (General) ,Parasitology ,lcsh:RC581-607 ,Arenaviruses, Old World ,Sequence Alignment ,Viral Transmission and Infection - Abstract
Cell entry of many enveloped viruses occurs by engagement with cellular receptors, followed by internalization into endocytic compartments and pH-induced membrane fusion. A previously unnoticed step of receptor switching was found to be critical during cell entry of two devastating human pathogens: Ebola and Lassa viruses. Our recent studies revealed the functional role of receptor switching to LAMP1 for triggering membrane fusion by Lassa virus and showed the involvement of conserved histidines in this switching, suggesting that other viruses from this family may also switch to LAMP1. However, when we investigated viruses that are genetically close to Lassa virus, we discovered that they cannot bind LAMP1. A crystal structure of the receptor-binding module from Morogoro virus revealed structural differences that allowed mapping of the LAMP1 binding site to a unique set of Lassa residues not shared by other viruses in its family, illustrating a key difference in the cell-entry mechanism of Lassa virus that may contribute to its pathogenicity., Author summary To infect, enveloped viruses need to fuse their membrane with the host membrane. Fusion is mediated by special glycoprotein machineries that must be triggered only at the right time and at the right place. A major cue that viruses utilize for triggering is acidic pH. Until recently, such pH-induced triggering was assumed to be the only mechanism used by the Arenaviridae family. However, Lassa virus, a notorious pathogenic member of this family, was shown to use the binding to an intracellular receptor named LAMP1 to potentiate its pH-induced triggering. This two-step mechanism was a surprising finding that raised critical questions regarding the cell-entry mechanisms of other viruses from this family. Here we used a structure-guided approach to investigate whether other Arenaviridae utilize LAMP1 for cell entry. We mapped the LAMP1 binding site on the Lassa-derived glycoprotein and confirmed its identity using grafting experiments. This mapping revealed the unique sequence signature needed for LAMP1 binding. Sequence analysis suggests that no other members of the Arenaviridae bind LAMP1.
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- 2017
12. EGF induces microRNAs that target suppressors of cell migration: miR-15b targets MTSS1 in breast cancer
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Cindy Körner, Noa Bossel Ben-Moshe, Maicol Mancini, Fernando Schmitt, Sara Lavi, Silvia Carvalho, Giovanni Blandino, Yosef Yarden, Nir Ben-Chetrit, Francesca Biagioni, Mattia Lauriola, Merav Kedmi, Hadas Cohen-Dvashi, Stefan Wiemann, Kedmi, Merav, Ben-Chetrit, Nir, Körner, Cindy, Mancini, Maicol, Ben-Moshe, Noa Bossel, Lauriola, Mattia, Lavi, Sara, Biagioni, Francesca, Carvalho, Silvia, Cohen-Dvashi, Hada, Schmitt, Fernando, Wiemann, Stefan, Blandino, Giovanni, and Yarden, Yosef
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Breast Neoplasms ,Mice, SCID ,Biology ,Biochemistry ,Metastasis ,Mice ,Breast cancer ,Cell Movement ,Epidermal growth factor ,Cell Line, Tumor ,microRNA ,medicine ,Animals ,Humans ,EGFR pathway ,MICRORNA (MIRNA) ,Molecular Biology ,GENE EXPRESSION PROFILE ,Epidermal Growth Factor ,Microfilament Proteins ,Cancer ,Cell migration ,Cell Biology ,medicine.disease ,Molecular biology ,Neoplasm Proteins ,MicroRNAs ,Invadopodia ,Cancer research ,Heterografts ,Female ,Neoplasm Transplantation ,Metastasis Suppressor Protein - Abstract
Growth factors promote tumor growth and metastasis. We found that epidermal growth factor (EGF) induced a set of 22 microRNAs (miRNAs) before promoting the migration of mammary cells. These miRNAs were more abundant in human breast tumors relative to the surrounding tissue, and their abundance varied among breast cancer subtypes. One of these miRNAs, miR-15b, targeted the 3' untranslated region of MTSS1 (metastasis suppressor protein 1). Although xenografts in which MTSS1 was knocked down grew more slowly in mice initially, longer-term growth was unaffected. Knocking down MTSS1 increased migration and Matrigel invasion of nontransformed mammary epithelial cells. Overexpressing MTSS1 in an invasive cell line decreased cell migration and invasiveness, decreased the formation of invadopodia and actin stress fibers, and increased the formation of cellular junctions. In tissues from breast cancer patients with the aggressive basal subtype, an inverse correlation occurred with the high expression of miRNA-15b and the low expression of MTSS1. Furthermore, low abundance of MTSS1 correlated with poor patient prognosis. Thus, growth factor-inducible miRNAs mediate mechanisms underlying the progression of cancer.
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- 2015
13. Navigator-3, a modulator of cell migration, may act as a suppressor of breast cancer progression
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Lee Roth, Silvia Carvalho, Assif Yitzhaky, Mattia Lauriola, Yosef Yarden, Sophia Nisani, Hadas Cohen-Dvashi, Raya Eilam, Ziv Porat, Amit Zeisel, Nir Ben-Chetrit, Fernando Schmitt, Wolfgang J. Köstler, Gabi Tarcic, Rainer Will, Jacques Zylberg, Roslin Russell, Sara Ricardo, Nishanth Belugali Nataraj, Sara Lavi, Carlos Caldas, Stefan Wiemann, Merav Kedmi, Maicol Mancini, Yoav Wigelman, DIPARTIMENTO DI MEDICINA SPECIALISTICA, DIAGNOSTICA E SPERIMENTALE, AREA MIN. 05 - Scienze biologiche, and Hadas Cohen-Dvashi1, Nir Ben-Chetrit1, Roslin Russell2, Silvia Carvalho1, Mattia Lauriola1,‡, Sophia Nisani1, Maicol Mancini1, Nishanth Nataraj1, Merav Kedmi1, Lee Roth1, Wolfgang Köstler1,†, Amit Zeisel3, Assif Yitzhaky3, Jacques Zylberg4, Gabi Tarcic1, Raya Eilam1, Yoav Wigelman1, Rainer Will5, Sara Lavi1, Ziv Porat6, Stefan Wiemann5, Sara Ricardo7, Fernando Schmitt7, Carlos Caldas2 & Yosef Yarden1
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cell migration ,medicine.medical_treatment ,Regulator ,Breast Neoplasms ,Nerve Tissue Proteins ,Biology ,Bioinformatics ,Metastasis ,microtubules ,Mice ,Cell Movement ,medicine ,Gene silencing ,Animals ,Humans ,cancer ,Neoplasm Metastasis ,Research Articles ,cytoskeleton ,growth factor ,Growth factor ,Cancer ,Membrane Proteins ,Cell migration ,medicine.disease ,Primary tumor ,Disease Models, Animal ,Cancer research ,Molecular Medicine ,Ectopic expression - Abstract
none 27 si Dissemination of primary tumor cells depends on migratory and invasive attributes. Here, we identify Navigator-3 (NAV3), a gene frequently mutated or deleted in human tumors, as a regulator of epithelial migration and invasion. Following induction by growth factors, NAV3 localizes to the plus ends of microtubules and enhances their polarized growth. Accordingly, NAV3 depletion trimmed microtubule growth, prolonged growth factor signaling, prevented apoptosis and enhanced random cell migration. Mathematical modeling suggested that NAV3-depleted cells acquire an advantage in terms of the way they explore their environment. In animal models, silencing NAV3 increased metastasis, whereas ectopic expression of the wild-type form, unlike expression of two, relatively unstable oncogenic mutants from human tumors, inhibited metastasis. Congruently, analyses of > 2,500 breast and lung cancer patients associated low NAV3 with shorter survival. We propose that NAV3 inhibits breast cancer progression by regulating microtubule dynamics, biasing directionally persistent rather than random migration, and inhibiting locomotion of initiated cells. Hadas Cohen-Dvashi1, Nir Ben-Chetrit1, Roslin Russell2, Silvia Carvalho1, Mattia Lauriola1,‡, Sophia Nisani1, Maicol Mancini1, Nishanth Nataraj1, Merav Kedmi1, Lee Roth1, Wolfgang Köstler1,†, Amit Zeisel3, Assif Yitzhaky3, Jacques Zylberg4, Gabi Tarcic1, Raya Eilam1, Yoav Wigelman1, Rainer Will5, Sara Lavi1, Ziv Porat6, Stefan Wiemann5, Sara Ricardo7, Fernando Schmitt7, Carlos Caldas2 & Yosef Yarden1,* Hadas Cohen-Dvashi1, Nir Ben-Chetrit1, Roslin Russell2, Silvia Carvalho1, Mattia Lauriola1,‡, Sophia Nisani1, Maicol Mancini1, Nishanth Nataraj1, Merav Kedmi1, Lee Roth1, Wolfgang Köstler1,†, Amit Zeisel3, Assif Yitzhaky3, Jacques Zylberg4, Gabi Tarcic1, Raya Eilam1, Yoav Wigelman1, Rainer Will5, Sara Lavi1, Ziv Porat6, Stefan Wiemann5, Sara Ricardo7, Fernando Schmitt7, Carlos Caldas2 & Yosef Yarden1,*
- Published
- 2015
14. Synaptojanin 2 is a druggable mediator of metastasis and the gene is overexpressed and amplified in breast cancer
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Ronen Alon, Hava Gil-Henn, Rotem Ben-Hamo, Marcelo Ehrlich, Nir Ben-Chetrit, Yosef Yarden, Tomer Itkin, Wolfgang J. Koestler, Dalia Seger, Silvia Carvalho, Carlos Caldas, Maicol Mancini, Ali Abdul-Hai, Daniela Aleida Ferraro, David Chetrit, Fresia Pareja, Mattia Lauriola, Marc Symons, Kirti Shukla, Ziv Shulman, Tsvee Lapidot, Hadas Cohen-Dvashi, Cindy Körner, Fernanda Milanezi, Moshit Lindzen, Merav Kedmi, Fernando Schmitt, Stefan Wiemann, Haim Barr, Sol Efroni, Roslin Russell, Ben-Chetrit, Nir, Chetrit, David, Russell, Roslin, K('o)rner, Cindy, Mancini, Maicol, Abdul-Hai, Ali, Itkin, Tomer, Carvalho, Silvia, Cohen-Dvashi, Hada, Koestler, Wolfgang J., Shukla, Kirti, Lindzen, Moshit, Kedmi, Merav, Lauriola, Mattia, Shulman, Ziv, Barr, Haim, Seger, Dalia, Ferraro, Daniela A., Pareja, Fresia, Gil-Henn, Hava, Lapidot, Tsvee, Alon, Ronen, Milanezi, Fernanda, Symons, Marc, Ben-Hamo, Rotem, Efroni, Sol, Schmitt, Fernando, Wiemann, Stefan, Caldas, Carlo, Ehrlich, Marcelo, and Yarden, Yosef
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Immunoblotting ,Gene Dosage ,Endocytic recycling ,Fluorescent Antibody Technique ,Breast Neoplasms ,Synaptojanin ,Mice, SCID ,Biochemistry ,Statistics, Nonparametric ,Metastasis ,Mice ,Breast cancer ,Growth factor receptor ,BREAST CANCER ,Cell Movement ,Cell Line, Tumor ,Drug Discovery ,medicine ,Image Processing, Computer-Assisted ,Animals ,Humans ,Epidermal growth factor receptor ,EGFR pathway ,Pseudopodia ,Neoplasm Metastasis ,RNA, Small Interfering ,Molecular Biology ,biology ,Cell migration ,Cell Biology ,medicine.disease ,Immunohistochemistry ,Phosphoric Monoester Hydrolases ,ErbB Receptors ,Gene Expression Regulation, Neoplastic ,Immunology ,Invadopodia ,METASTASIS ,Podosomes ,Cancer research ,biology.protein ,Microscopy, Electron, Scanning ,Female - Abstract
Amplified HER2, which encodes a member of the epidermal growth factor receptor (EGFR) family, is a target of effective therapies against breast cancer. In search for similarly targetable genomic aberrations, we identified copy number gains in SYNJ2, which encodes the 5'-inositol lipid phosphatase synaptojanin 2, as well as overexpression in a small fraction of human breast tumors. Copy gain and overexpression correlated with shorter patient survival and a low abundance of the tumor suppressor microRNA miR-31. SYNJ2 promoted cell migration and invasion in culture and lung metastasis of breast tumor xenografts in mice. Knocking down SYNJ2 impaired the endocytic recycling of EGFR and the formation of cellular lamellipodia and invadopodia. Screening compound libraries identified SYNJ2-specific inhibitors that prevented cell migration but did not affect the related neural protein SYNJ1, suggesting that SYNJ2 is a potentially druggable target to block cancer cell migration.
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- 2015
15. Diurnal suppression of EGFR signalling by glucocorticoids and implications for tumour progression and treatment
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Morris E. Feldman, Lee Roth, Hadas Cohen-Dvashi, Moshit Lindzen, Fernando Schmitt, Rossella Solmi, Michal Sharon-Sevilla, Amit Zeisel, Stefan Wiemann, Yehoshua Enuka, Gabriele D'Uva, Mattia Lauriola, Nir Ben-Chetrit, Yosef Yarden, Nava Nevo, Eytan Domany, Silvia Carvalho, Alon Chen, Kirti Sharma, Merav Kedmi, Lauriola M, Enuka Y, Zeisel A, D'Uva G, Roth L, Sharon-Sevilla M, Lindzen M, Sharma K, Nevo N, Feldman M, Carvalho S, Cohen-Dvashi H, Kedmi M, Ben-Chetrit N, Chen A, Solmi R, Wiemann S, Schmitt F, Domany E, and Yarden Y.
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medicine.medical_specialty ,MAP Kinase Signaling System ,EGFR ,Circadian clock ,General Physics and Astronomy ,Biology ,Ligands ,Article ,General Biochemistry, Genetics and Molecular Biology ,Receptor tyrosine kinase ,Mice ,Receptors, Glucocorticoid ,Cell Movement ,Cell Line, Tumor ,Neoplasms ,Oscillometry ,Internal medicine ,Negative feedback ,medicine ,Animals ,Humans ,Receptor ,Glucocorticoids ,Mice, Knockout ,Multidisciplinary ,Kinase ,General Chemistry ,Circadian Rhythm ,ErbB Receptors ,Mice, Inbred C57BL ,Treatment Outcome ,Endocrinology ,Nuclear receptor ,Disease Progression ,Cancer research ,biology.protein ,Female ,Signal transduction ,Signal Transduction ,Hormone - Abstract
Signal transduction by receptor tyrosine kinases (RTKs) and nuclear receptors for steroid hormones is essential for body homeostasis, but the cross-talk between these receptor families is poorly understood. We observed that glucocorticoids inhibit signalling downstream of EGFR, an RTK. The underlying mechanism entails suppression of EGFR’s positive feedback loops and simultaneous triggering of negative feedback loops that normally restrain EGFR. Our studies in mice reveal that the regulation of EGFR’s feedback loops by glucocorticoids translates to circadian control of EGFR signalling: EGFR signals are suppressed by high glucocorticoids during the active phase (night-time in rodents), while EGFR signals are enhanced during the resting phase. Consistent with this pattern, treatment of animals bearing EGFR-driven tumours with a specific kinase inhibitor was more effective if administered during the resting phase of the day, when glucocorticoids are low. These findings support a circadian clock-based paradigm in cancer therapy., Glucocorticoids are released in a diurnal pattern. Here, the authors show that EGF receptor (EGFR) signalling is negatively regulated by glucocorticoids, and that EGFR inhibitor has stronger antitumour effects when administered during the resting phase, when glucocorticoids are low, offering potential optimization of cancer therapy regimens.
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- 2014
16. Epidermal growth-factor - induced transcript isoform variation drives mammary cell migration
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Ulrich Tschulena, Gideon Rechavi, Stefan Wiemann, Jasmine Jacob-Hirsch, Hadas Cohen-Dvashi, Jonathan M. Tsai, Kirti Sharma, Yosef Yarden, Wolfgang J. Köstler, Amit Zeisel, Nir Ben-Chetrit, Eric Lader, Cindy Körner, Assif Yitzhaky, and Eytan Domany
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Gene isoform ,Small interfering RNA ,Science ,Biology ,Polyadenylation ,Cell Line ,Transcriptome ,03 medical and health sciences ,Exon ,Cell Movement ,Epidermal growth factor ,Humans ,RNA, Messenger ,RNA, Small Interfering ,Mammary Glands, Human ,Oligonucleotide Array Sequence Analysis ,030304 developmental biology ,0303 health sciences ,Multidisciplinary ,Epidermal Growth Factor ,030302 biochemistry & molecular biology ,Alternative splicing ,Genetic Variation ,Epithelial Cells ,Cell migration ,Exons ,Molecular biology ,Alternative Splicing ,Medicine ,Female ,Signal transduction ,Research Article ,Signal Transduction - Abstract
Signal-induced transcript isoform variation (TIV) includes alternative promoter usage as well as alternative splicing and alternative polyadenylation of mRNA. To assess the phenotypic relevance of signal-induced TIV, we employed exon arrays and breast epithelial cells, which migrate in response to the epidermal growth factor (EGF). We show that EGF rapidly – within one hour – induces widespread TIV in a significant fraction of the transcriptome. Importantly, TIV characterizes many genes that display no differential expression upon stimulus. In addition, similar EGF-dependent changes are shared by a panel of mammary cell lines. A functional screen, which utilized isoform-specific siRNA oligonucleotides, indicated that several isoforms play essential, non-redundant roles in EGF-induced mammary cell migration. Taken together, our findings highlight the importance of TIV in the rapid evolvement of a phenotypic response to extracellular signals.
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- 2013
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17. Inhibition of triple-negative breast cancer models by combinations of antibodies to EGFR
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Daniela Aleida Ferraro, Ruth Maron, Hadas Cohen-Dvashi, Nadège Gaborit, Sara Lavi, Nir Ben-Chetrit, Ziv Porat, Fresia Pareja, Moshit Lindzen, Michael Sela, and Yosef Yarden
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medicine.drug_class ,Receptor, ErbB-2 ,Immunoblotting ,Cetuximab ,Mice, Nude ,Breast Neoplasms ,Biology ,Monoclonal antibody ,Antibodies, Monoclonal, Humanized ,Cell Line ,Mice ,Cell Movement ,Cell Line, Tumor ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Panitumumab ,Animals ,Humans ,Kinase activity ,Receptor ,Triple-negative breast cancer ,EGFR inhibitors ,Cell Proliferation ,Multidisciplinary ,Antibodies, Monoclonal ,Biological Sciences ,Molecular biology ,Xenograft Model Antitumor Assays ,Tumor Burden ,ErbB Receptors ,Receptors, Estrogen ,Monoclonal ,Proteolysis ,Cancer research ,Female ,Receptors, Progesterone ,medicine.drug ,HeLa Cells - Abstract
Breast tumors lacking expression of human epidermal growth factor receptor 2 (HER2) and the estrogen and the progesterone receptors (triple negative; TNBC) are more aggressive than other disease subtypes, and no molecular targeted agents are currently available for their treatment. Because TNBC commonly displays EGF receptor (EGFR) expression, and combinations of monoclonal antibodies to EGFR effectively inhibit other tumor models, we addressed the relevance of this strategy to treatment of TNBC. Unlike a combination of the clinically approved monoclonal antibodies, cetuximab and panitumumab, which displaced each other and displayed no cooperative effects, several other combinations resulted in enhanced inhibition of TNBC’s cell growth both in vitro and in animals. The ability of certain antibody mixtures to remove EGFR from the cell surface and to promote its intracellular degradation correlated with the inhibitory potential. However, unlike EGF-induced sorting of EGFR to lysosomal degradation, the antibody-induced pathway displayed independence from the intrinsic kinase activity and dimer formation ability of EGFR, and it largely avoided the recycling route. In conclusion, although TNBC clinical trials testing EGFR inhibitors reported lack of benefit, our results offer an alternative strategy that combines noncompetitive antibodies to achieve robust degradation of EGFR and tumor inhibition.
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- 2013
18. Deubiquitination of EGFR by Cezanne-1 contributes to cancer progression
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Sara Lavi, Fan Zhang, Zohar Yakhini, Peter Sinn, Nicola Crosetto, Hadas Cohen-Dvashi, Nir Ben-Chetrit, Gur Pines, Gordon B. Mills, Daniela Aleida Ferraro, Fernando Schmitt, Sebastian Aulmann, Ivan Dikic, Chanan Rubin, Silvia Carvalho, Yosef Yarden, Wolfgang J. Köstler, Fresia Pareja, and Roy Navon
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Cancer Research ,gene amplification ,medicine.medical_treatment ,Molecular oncology ,Article ,Catalysis ,Deubiquitinating enzyme ,03 medical and health sciences ,0302 clinical medicine ,Ubiquitin ,Growth factor receptor ,RNA interference ,Neoplasms ,Endopeptidases ,Genetics ,medicine ,endocytosis ,Humans ,Epidermal growth factor receptor ,Phosphorylation ,RNA, Small Interfering ,Molecular Biology ,deubiquitination ,030304 developmental biology ,0303 health sciences ,biology ,Growth factor ,Ubiquitination ,growth factor ,ErbB Receptors ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Disease Progression ,Deubiquitination - Abstract
Once stimulated, the epidermal growth factor receptor (EGFR) undergoes self-phosphorylation, which, on the one hand, instigates signaling cascades, and on the other hand, recruits CBL ubiquitin ligases, which mark EGFRs for degradation. Using RNA interference screens, we identified a deubiquitinating enzyme, Cezanne-1, that opposes receptor degradation and enhances EGFR signaling. These functions require the catalytic- and ubiquitin-binding domains of Cezanne-1, and they involve physical interactions and transphosphorylation of Cezanne-1 by EGFR. In line with the ability of Cezanne-1 to augment EGF-induced growth and migration signals, the enzyme is overexpressed in breast cancer. Congruently, the corresponding gene is amplified in approximately one third of mammary tumors, and high transcript levels predict an aggressive disease course. In conclusion, deubiquitination by Cezanne-1 curtails degradation of growth factor receptors, thereby promotes oncogenic growth signals.
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- 2012
- Full Text
- View/download PDF
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