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Covadonga Alonso M.D. Ph.D. is a Professor of research at the Dpt. Biotechnology of INIA-CSIC, Madrid, Spain. Her group is interested in finding new targets for antivirals against highly pathogenic viruses from a One Health perspective. We currently focus on finding common targets for SARS-CoV-2, Ebola, and African swine fever virus, based on host-pathogen interactions and innate immunity studies. Our working model has been the African swine fever virus (ASFV) for several years. We have identified cellular molecules that are crucial for a viral infection at several levels, including viral entry/ uncoating and transport, replication, autophagy, and lipid regulation. This agent causes an acute deadly infection in domestic pigs with a high socio-economic impact in Europe and Asia nowadays. Our goal is to increase knowledge on the mechanisms required for infection in order to find targets for new vaccination and antiviral strategies. I have been Chair of Asfarviridae study group of the ICTV, CEO of the Global African swine fever Research Alliance (GARA), and Vicepresident of the Board of the Spanish Society for Virology, among others.
Niemann-Pick type C1 (NPC1) receptor is an endosomal membrane protein that regulates intracellular cholesterol traffic. This protein has been shown to play an important role for several viruses. It has been reported that SARS-CoV-2 enters the cell through plasma membrane fusion and/or endosomal entry upon availability of proteases. However, the whole process is not fully understood yet and additional viral/host factors might be required for viral fusion and subsequent viral replication. Here, we report a novel interaction between the SARS-CoV-2 nucleoprotein (N) and the cholesterol transporter NPC1. Furthermore, we have found that some compounds reported to interact with NPC1, carbazole SC816 and sulfides SC198 and SC073, were able to reduce SARS-CoV-2 viral infection with a good selectivity index in human cell infection models. These findings suggest the importance of NPC1 for SARS-CoV-2 viral infection and a new possible potential therapeutic target to fight against COVID-19.
Isabel García-Dorival; Miguel Ángel Cuesta-Geijo; Lucía Barrado-Gil; Inmaculada Galindo; Urtzi Garaigorta; Jesús Urquiza; Ana del Puerto; Nuria E. Campillo; Ana Martínez; Pablo Gastaminza; Carmen Gil; Covadonga Alonso. Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target. Antiviral Research 2021, 194, 105167 .
AMA StyleIsabel García-Dorival, Miguel Ángel Cuesta-Geijo, Lucía Barrado-Gil, Inmaculada Galindo, Urtzi Garaigorta, Jesús Urquiza, Ana del Puerto, Nuria E. Campillo, Ana Martínez, Pablo Gastaminza, Carmen Gil, Covadonga Alonso. Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target. Antiviral Research. 2021; 194 ():105167.
Chicago/Turabian StyleIsabel García-Dorival; Miguel Ángel Cuesta-Geijo; Lucía Barrado-Gil; Inmaculada Galindo; Urtzi Garaigorta; Jesús Urquiza; Ana del Puerto; Nuria E. Campillo; Ana Martínez; Pablo Gastaminza; Carmen Gil; Covadonga Alonso. 2021. "Identification of Niemann-Pick C1 protein as a potential novel SARS-CoV-2 intracellular target." Antiviral Research 194, no. : 105167.
African swine fever virus (ASFV) infectious cycle starts with the viral adsorption and entry into the host cell. The virus is internalized via clathrin/dynamin mediated endocytosis and macropinocytosis. As several other viruses, ASF virion is then internalized and incorporated into the endocytic pathway. Endosomal maturation entails luminal acidification and the lowering of pH acting on the multi-layered virion structure dissolves the outer capsid. Upon decapsidation, the inner viral membrane is exposed to interact with the limiting membrane of the late endosome for fusion. Egress from endosome is related to cholesterol efflux, but it remains an intriguing process albeit essential for infection, specifically for the viral nucleic acid exit to the cytoplasm for replication. ASFV proteins E248R and E199L, with structural homology to the VACV proteins of the fusion complex, seem to have similar functions in ASFV. A direct interaction between these ASFV proteins with the cholesterol transporter protein NPC1 (Niemann-Pick C type 1) was observed, which was also shared by the E248R homologous protein L1R of VACV. Binding occurs between the transmembrane domain of E248R with the loop C of NPC1 at the same domain than EBOV binding site. These interactions suggest that these ASFV proteins are crucial for membrane fusion. CRISPR NPC1 KO Vero cells lacking NPC1 protein that were resistant to EBOV, reduced ASFV infection levels significantly. Reductions on ASFV infectivity and replication in NPC1 KO cells were accompanied by lesser viral factories of smaller size and lacking the typical cohesive morphology between endosomes and viral proteins. We observed a compensatory effect in NPC1 KO cells, elevating NPC2 levels while silencing NPC2 in Vero cells with shRNA, also reduced ASFV infection. Our findings pave the way to understand the role of these proteins at the membrane viral fusion step for several viruses. Author Summary African swine fever virus (ASFV) causes a deadly disease of pigs and wild boars that was endemic in Africa but have extended over the last years to Europe, Asia and Oceania with high socioeconomic impact. ASFV enters the cell by endocytosis and has adapted to the endosomal conditions to acquire infectivity. Viral infectivity is dependent on cholesterol traffic at the endosomes, especially at the fusion step. Fusion of the internal viral membrane with the endosomal membrane is required for the exit of the viral DNA to the cytoplasm to start replication. ASF virion internal membrane proteins E248R and E199L were found to bind the Niemann Pick C1 (NPC1) receptor at the endosome. These proteins are highly conserved among ASFV isolates and resemble proteins of the VACV entry/fusion complex. The function of NPC1 is to regulate the efflux of dietary cholesterol efflux from the endosome to the endoplasmic reticulum, which appears to be necessary for viral fusion. NPC1 knockout cells by CRISPR reduced infection affecting infectivity and early replication. Also, removing the associated endosomal protein NPC2, further declined infectivity. These results show the relevance of NPC1 receptor in the viral infection actually shared by unrelated important viral families.
Miguel Ángel Cuesta-Geijo; Jesús Urquiza; Ana del Puerto; Isabel Garcia-Dorival; Fátima Lasala; Lucía Barrado-Gil; Inmaculada Galindo; Rafael Delgado; Covadonga Alonso. Endosomal proteins NPC1 and NPC2 at African swine fever virus entry/fusion. 2021, 1 .
AMA StyleMiguel Ángel Cuesta-Geijo, Jesús Urquiza, Ana del Puerto, Isabel Garcia-Dorival, Fátima Lasala, Lucía Barrado-Gil, Inmaculada Galindo, Rafael Delgado, Covadonga Alonso. Endosomal proteins NPC1 and NPC2 at African swine fever virus entry/fusion. . 2021; ():1.
Chicago/Turabian StyleMiguel Ángel Cuesta-Geijo; Jesús Urquiza; Ana del Puerto; Isabel Garcia-Dorival; Fátima Lasala; Lucía Barrado-Gil; Inmaculada Galindo; Rafael Delgado; Covadonga Alonso. 2021. "Endosomal proteins NPC1 and NPC2 at African swine fever virus entry/fusion." , no. : 1.
Niemann-Pick C1 (NPC1) receptor is an intracellular protein located in late endosomes and lysosomes whose main function is to regulate intracellular cholesterol trafficking. Besides being postulated as necessary for the infection of highly pathogenic viruses in which the integrity of cholesterol transport is required, this protein also allows the entry of the Ebola virus (EBOV) into the host cells acting as an intracellular receptor. EBOV glycoprotein (EBOV-GP) interaction with NPC1 at the endosomal membrane triggers the release of the viral material into the host cell, starting the infective cycle. Disruption of the NPC1/EBOV-GP interaction could represent an attractive strategy for the development of drugs aimed at inhibiting viral entry and thus infection. Some of the today available EBOV inhibitors were proposed to interrupt this interaction, but molecular and structural details about their mode of action are still preliminary thus more efforts are needed to properly address these points. Here, we provide a critical discussion of the potential of NPC1 and its interaction with EBOV-GP as a therapeutic target for viral infections.
Marcos Morales-Tenorio; Tiziana Ginex; Miguel Ángel Cuesta-Geijo; Nuria E. Campillo; César Muñoz-Fontela; Covadonga Alonso; Rafael Delgado; Carmen Gil. Potential pharmacological strategies targeting the Niemann-Pick C1 receptor and Ebola virus glycoprotein interaction. European Journal of Medicinal Chemistry 2021, 223, 113654 .
AMA StyleMarcos Morales-Tenorio, Tiziana Ginex, Miguel Ángel Cuesta-Geijo, Nuria E. Campillo, César Muñoz-Fontela, Covadonga Alonso, Rafael Delgado, Carmen Gil. Potential pharmacological strategies targeting the Niemann-Pick C1 receptor and Ebola virus glycoprotein interaction. European Journal of Medicinal Chemistry. 2021; 223 ():113654.
Chicago/Turabian StyleMarcos Morales-Tenorio; Tiziana Ginex; Miguel Ángel Cuesta-Geijo; Nuria E. Campillo; César Muñoz-Fontela; Covadonga Alonso; Rafael Delgado; Carmen Gil. 2021. "Potential pharmacological strategies targeting the Niemann-Pick C1 receptor and Ebola virus glycoprotein interaction." European Journal of Medicinal Chemistry 223, no. : 113654.
African swine fever virus (ASFV) is an acute and persistent swine virus with a high economic burden that encodes multiple genes to evade host immune response. In this work, we have revealed that early viral protein UBCv1, the only known conjugating enzyme encoded by a virus, modulates innate immune and inflammatory signaling. Transient overexpression of UBCv1 impaired activation of NF-κB and AP-1 transcription factors induced by several agonists of these pathways. In contrast, activation of IRF3 and ISRE signaling upon stimulation with TRIFΔRIP, cGAS/STING or RIG-I-CARD remained unaltered. Experiments aimed at mapping UBCv1 inhibitory activity indicated that this viral protein acts upstream or at the level step of IKKβ. In agreement with this, UBCv1 was able to block p65 nuclear translocation upon cytokine stimulation, a key event in NF-ĸB signaling. Additionally, A549 stably transduced for UBCv1 showed a significant decrease in the levels of NF-ĸB dependent genes. Interestingly, despite the well-defined capacity of UBCv1 to conjugate ubiquitin chains, a mutant disabled for ubiquitylation activity retained similar immunomodulatory activity as the wild-type enzyme, suggesting that the two functions are segregated. Altogether these data suggest that ASFV UBCv1 manipulates the innate immune response targeting the NF-κB and AP-1 pathways and opens new questions about the multifunctionality of this enzyme.
Lucía Barrado-Gil; Ana del Puerto; Inmaculada Galindo; Miguel Cuesta-Geijo; Isabel García-Dorival; Carlos de Motes; Covadonga Alonso. African Swine Fever Virus Ubiquitin-Conjugating Enzyme Is an Immunomodulator Targeting NF-κB Activation. Viruses 2021, 13, 1160 .
AMA StyleLucía Barrado-Gil, Ana del Puerto, Inmaculada Galindo, Miguel Cuesta-Geijo, Isabel García-Dorival, Carlos de Motes, Covadonga Alonso. African Swine Fever Virus Ubiquitin-Conjugating Enzyme Is an Immunomodulator Targeting NF-κB Activation. Viruses. 2021; 13 (6):1160.
Chicago/Turabian StyleLucía Barrado-Gil; Ana del Puerto; Inmaculada Galindo; Miguel Cuesta-Geijo; Isabel García-Dorival; Carlos de Motes; Covadonga Alonso. 2021. "African Swine Fever Virus Ubiquitin-Conjugating Enzyme Is an Immunomodulator Targeting NF-κB Activation." Viruses 13, no. 6: 1160.
Despite the efforts to develop new treatments against Ebola virus (EBOV) there is currently no antiviral drug licensed to treat patients with Ebola virus disease (EVD). Therefore, there is still an urgent need to find new drugs to fight against EBOV. In order to do this, a virtual screening was done on the druggable interaction between the EBOV glycoprotein (GP) and the host receptor NPC1 with a subsequent selection of compounds for further validation. This screening led to the identification of new small organic molecules with potent inhibitory action against EBOV infection using lentiviral EBOV-GP-pseudotype viruses. Moreover, some of these compounds have shown their ability to interfere with the intracellular cholesterol transport receptor NPC1 using an ELISA-based assay. These preliminary results pave the way to hit to lead optimization programs that lead to successful candidates.
Fátima Lasala; Alfonso García-Rubia; Carlos Requena; Inmaculada Galindo; Miguel Angel Cuesta-Geijo; Isabel García-Dorival; Paula Bueno; Nuria Labiod; Joanna Luczkowiak; Ana Martinez; Nuria E. Campillo; Covadonga Alonso; Rafael Delgado; Carmen Gil. Identification of potential inhibitors of protein-protein interaction useful to fight against Ebola and other highly pathogenic viruses. Antiviral Research 2021, 186, 105011 -105011.
AMA StyleFátima Lasala, Alfonso García-Rubia, Carlos Requena, Inmaculada Galindo, Miguel Angel Cuesta-Geijo, Isabel García-Dorival, Paula Bueno, Nuria Labiod, Joanna Luczkowiak, Ana Martinez, Nuria E. Campillo, Covadonga Alonso, Rafael Delgado, Carmen Gil. Identification of potential inhibitors of protein-protein interaction useful to fight against Ebola and other highly pathogenic viruses. Antiviral Research. 2021; 186 ():105011-105011.
Chicago/Turabian StyleFátima Lasala; Alfonso García-Rubia; Carlos Requena; Inmaculada Galindo; Miguel Angel Cuesta-Geijo; Isabel García-Dorival; Paula Bueno; Nuria Labiod; Joanna Luczkowiak; Ana Martinez; Nuria E. Campillo; Covadonga Alonso; Rafael Delgado; Carmen Gil. 2021. "Identification of potential inhibitors of protein-protein interaction useful to fight against Ebola and other highly pathogenic viruses." Antiviral Research 186, no. : 105011-105011.
Niemann-Pick type C1 (NPC1) receptor is an endosomal membrane protein that regulates intracellular cholesterol trafficking, which is crucial in the Ebola virus (EBOV) cycle. The severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2) enters the cell by binding of the viral spike (S) protein to the ACE2 receptor. This requires S-protein processing either by the surface transmembrane serine protease TMPRSS2 for plasma membrane fusion or cathepsin L for endosomal entry. Additional host factors are required for viral fusion at endosomes. Here, we report a novel interaction of the SARS-CoV-2 nucleoprotein (N) with the cholesterol transporter NPC1. Moreover, small molecules interfering with NPC1 that inhibit EBOV entry, also inhibited human coronavirus. Our findings suggest an important role for NPC1 in SARS-CoV-2 infection, a common strategy shared with EBOV, and a potential therapeutic target to fight against COVID-19.
Isabel Garcia-Dorival; Miguel Angel Cuesta-Geijo; Lucia Barrado-Gil; Inmaculada Galindo; Jesus Urquiza; Ana Del Puerto; Carmen Gil; Nuria Campillo; Ana Martinez; Covadonga Alonso. Identification of NPC1 as a novel SARS-CoV-2 intracellular target. 2020, 1 .
AMA StyleIsabel Garcia-Dorival, Miguel Angel Cuesta-Geijo, Lucia Barrado-Gil, Inmaculada Galindo, Jesus Urquiza, Ana Del Puerto, Carmen Gil, Nuria Campillo, Ana Martinez, Covadonga Alonso. Identification of NPC1 as a novel SARS-CoV-2 intracellular target. . 2020; ():1.
Chicago/Turabian StyleIsabel Garcia-Dorival; Miguel Angel Cuesta-Geijo; Lucia Barrado-Gil; Inmaculada Galindo; Jesus Urquiza; Ana Del Puerto; Carmen Gil; Nuria Campillo; Ana Martinez; Covadonga Alonso. 2020. "Identification of NPC1 as a novel SARS-CoV-2 intracellular target." , no. : 1.
African Swine Fever virus (ASFV) causes one of the most relevant emerging diseases affecting swine, now extended through three continents. The virus has a large coding capacity to deploy an arsenal of molecules antagonizing the host functions. In the present work, we have studied the only known E2 viral-conjugating enzyme, UBCv1 that is encoded by the I215L gene of ASFV. UBCv1 was expressed as an early expression protein that accumulates throughout the course of infection. This versatile protein, bound several types of polyubiquitin chains and its catalytic domain was required for enzymatic activity. High throughput mass spectrometry analysis in combination with a screening of an alveolar macrophage library was used to identify and characterize novel UBCv1-host interactors. The analysis revealed interaction with the 40S ribosomal protein RPS23, the cap-dependent translation machinery initiation factor eIF4E, and the E3 ubiquitin ligase Cullin 4B. Our data show that during ASFV infection, UBCv1 was able to bind to eIF4E, independent from the cap-dependent complex. Our results provide novel insights into the function of the viral UBCv1 in hijacking cellular components that impact the mTORC signaling pathway, the regulation of the host translation machinery, and the cellular protein expression during the ASFV lifecycle.
Lucía Barrado-Gil; Ana Del Puerto; Raquel Muñoz-Moreno; Inmaculada Galindo; Miguel Ángel Cuesta-Geijo; Jesús Urquiza; Estanislao Nistal-Villán; Carlos Maluquer De Motes; Covadonga Alonso. African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis. Frontiers in Microbiology 2020, 11, 1 .
AMA StyleLucía Barrado-Gil, Ana Del Puerto, Raquel Muñoz-Moreno, Inmaculada Galindo, Miguel Ángel Cuesta-Geijo, Jesús Urquiza, Estanislao Nistal-Villán, Carlos Maluquer De Motes, Covadonga Alonso. African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis. Frontiers in Microbiology. 2020; 11 ():1.
Chicago/Turabian StyleLucía Barrado-Gil; Ana Del Puerto; Raquel Muñoz-Moreno; Inmaculada Galindo; Miguel Ángel Cuesta-Geijo; Jesús Urquiza; Estanislao Nistal-Villán; Carlos Maluquer De Motes; Covadonga Alonso. 2020. "African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis." Frontiers in Microbiology 11, no. : 1.
African Swine Fever virus (ASFV) causes one of the most relevant emerging diseases affecting swine, now extended through three continents. The virus has a large coding capacity to deploy an arsenal of molecules antagonizing the host functions. In the present work, we have studied the only known E2 viral-conjugating enzyme, UBCv1 that is encoded by the I215L gene of ASFV. UBCv1 was expressed as an early expression protein that accumulates throughout the course of infection. This versatile protein, bound several types of polyubiquitin chains and its catalytic domain was required for enzymatic activity. High throughput mass spectrometry analysis in combination with a screening of an alveolar macrophage library was used to identify and characterize novel UBCv1-host interactors. The analysis revealed interaction with the 40S ribosomal protein RPS23, the cap-dependent translation machinery initiation factor eIF4E, and the E3 ubiquitin ligase Cullin 4B. Our data show that during ASFV infection, UBCv1 was able to bind to eIF4E, independent from the cap-dependent complex. Our results provide novel insights into the function of the viral UBCv1 in hijacking cellular components that impact the mTORC signaling pathway, the regulation of the host translation machinery, and the cellular protein expression during the ASFV lifecycle.
Lucía Barrado-Gil; Ana Del Puerto; Raquel Muñoz-Moreno; Inmaculada Galindo; Miguel Ángel Cuesta-Geijo; Jesús Urquiza; Estanislao Nistal-Villán; Carlos Maluquer De Motes; Covadonga Alonso. African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis. 2020, 11, 1 .
AMA StyleLucía Barrado-Gil, Ana Del Puerto, Raquel Muñoz-Moreno, Inmaculada Galindo, Miguel Ángel Cuesta-Geijo, Jesús Urquiza, Estanislao Nistal-Villán, Carlos Maluquer De Motes, Covadonga Alonso. African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis. . 2020; 11 ():1.
Chicago/Turabian StyleLucía Barrado-Gil; Ana Del Puerto; Raquel Muñoz-Moreno; Inmaculada Galindo; Miguel Ángel Cuesta-Geijo; Jesús Urquiza; Estanislao Nistal-Villán; Carlos Maluquer De Motes; Covadonga Alonso. 2020. "African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis." 11, no. : 1.
The endocytic pathway is a common strategy that several highly pathogenic viruses use to enter into the cell. To demonstrate the usefulness of this pathway as a common target for the development of broad-spectrum antivirals, the inhibitory effect of drug compounds targeting endosomal membrane proteins were investigated. This study entailed direct comparison of drug effectiveness against animal and human pathogenic viruses, namely Ebola (EBOV), African swine fever virus (ASFV), and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A panel of experimental and FDA-approved compounds targeting calcium channels and PIKfyve at the endosomal membrane caused potent reductions of entry up to 90% in SARS-CoV-2 S-protein pseudotyped retrovirus. Similar inhibition was observed against transduced EBOV glycoprotein pseudovirus and ASFV. SARS-CoV-2 infection was potently inhibited by selective estrogen receptor modulators in cells transduced with pseudovirus, among them Raloxifen inhibited ASFV with very low 50% inhibitory concentration. Finally, the mechanism of the inhibition caused by the latter in ASFV infection was analyzed. Overall, this work shows that cellular proteins related to the endocytic pathway can constitute suitable cellular targets for broad range antiviral compounds.
I. Galindo; U. Garaigorta; F. Lasala; M.A. Cuesta-Geijo; P. Bueno; C. Gil; R. Delgado; P. Gastaminza; C. Alonso. Antiviral drugs targeting endosomal membrane proteins inhibit distant animal and human pathogenic viruses. Antiviral Research 2020, 186, 104990 -104990.
AMA StyleI. Galindo, U. Garaigorta, F. Lasala, M.A. Cuesta-Geijo, P. Bueno, C. Gil, R. Delgado, P. Gastaminza, C. Alonso. Antiviral drugs targeting endosomal membrane proteins inhibit distant animal and human pathogenic viruses. Antiviral Research. 2020; 186 ():104990-104990.
Chicago/Turabian StyleI. Galindo; U. Garaigorta; F. Lasala; M.A. Cuesta-Geijo; P. Bueno; C. Gil; R. Delgado; P. Gastaminza; C. Alonso. 2020. "Antiviral drugs targeting endosomal membrane proteins inhibit distant animal and human pathogenic viruses." Antiviral Research 186, no. : 104990-104990.
Currently, humans are immersed in a pandemic caused by the emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which threatens public health worldwide. To date, no drug or vaccine has been approved to treat the severe disease caused by this coronavirus, COVID-19. In this paper, we will focus on the main virus-based and host-based targets that can guide efforts in medicinal chemistry to discover new drugs for this devastating disease. In principle, all CoV enzymes and proteins involved in viral replication and the control of host cellular machineries are potentially druggable targets in the search for therapeutic options for SARS-CoV-2. This Perspective provides an overview of the main targets from a structural point of view, together with reported therapeutic compounds with activity against SARS-CoV-2 and/or other CoVs. Also, the role of innate immune response to coronavirus infection and the related therapeutic options will be presented.
Carmen Gil; Tiziana Ginex; Inés Maestro; Vanesa Nozal; Lucía Barrado-Gil; Miguel Ángel Cuesta-Geijo; Jesús Urquiza; David Ramírez; Covadonga Alonso; Nuria E. Campillo; Ana Martinez. COVID-19: Drug Targets and Potential Treatments. Journal of Medicinal Chemistry 2020, 63, 12359 -12386.
AMA StyleCarmen Gil, Tiziana Ginex, Inés Maestro, Vanesa Nozal, Lucía Barrado-Gil, Miguel Ángel Cuesta-Geijo, Jesús Urquiza, David Ramírez, Covadonga Alonso, Nuria E. Campillo, Ana Martinez. COVID-19: Drug Targets and Potential Treatments. Journal of Medicinal Chemistry. 2020; 63 (21):12359-12386.
Chicago/Turabian StyleCarmen Gil; Tiziana Ginex; Inés Maestro; Vanesa Nozal; Lucía Barrado-Gil; Miguel Ángel Cuesta-Geijo; Jesús Urquiza; David Ramírez; Covadonga Alonso; Nuria E. Campillo; Ana Martinez. 2020. "COVID-19: Drug Targets and Potential Treatments." Journal of Medicinal Chemistry 63, no. 21: 12359-12386.
Animal diseases constitute a continuing threat to animal health, food safety, national economy, and the environment. Among those, African swine fever (ASF) is one of the most devastating viruses affecting pigs and wild suids due to the lack of vaccine or effective treatment. ASF is endemic in countries in sub-Saharan Africa, but since its introduction to the Caucasus region in 2007, a highly virulent strain of ASF virus (ASFV) has continued to circulate and spread into Eastern Europe and Russia, and most recently into Western Europe, China, and various countries of Southeast Asia. Given the importance of this disease, this review will highlight recent discoveries in basic virology with special focus on proteomic analysis, replication cycle, and some recent data on genes involved in cycle progression and viral–host interactions, such as I215L (E2 ubiquitin-conjugating enzyme), EP402R (CD2v), A104R (histone-like protein), QP509L, and Q706L (RNA helicases) or P1192R (Topoisomerase II). Taking into consideration the large DNA genome of ASFV and its complex interactions with the host, more studies and new approaches are to be taken to understand the basic virus–host interaction for ASFV. Proteomic studies are just paving the way for future research.
Axel Karger; Daniel Pérez-Núñez; Jesús Urquiza; Patricia Hinojar; Covadonga Alonso; Ferdinando B. Freitas; Yolanda Revilla; Marie-Frédérique Le Potier; Maria Montoya. An Update on African Swine Fever Virology. Viruses 2019, 11, 864 .
AMA StyleAxel Karger, Daniel Pérez-Núñez, Jesús Urquiza, Patricia Hinojar, Covadonga Alonso, Ferdinando B. Freitas, Yolanda Revilla, Marie-Frédérique Le Potier, Maria Montoya. An Update on African Swine Fever Virology. Viruses. 2019; 11 (9):864.
Chicago/Turabian StyleAxel Karger; Daniel Pérez-Núñez; Jesús Urquiza; Patricia Hinojar; Covadonga Alonso; Ferdinando B. Freitas; Yolanda Revilla; Marie-Frédérique Le Potier; Maria Montoya. 2019. "An Update on African Swine Fever Virology." Viruses 11, no. 9: 864.
African swine fever (ASF) is a hemorrhagic fever of wild and domestic pigs with a high rate of mortality. Originally endemic in Africa, this disease is currently disseminating in Europe and China, causing a large socioeconomic impact. ASF is caused by a DNA virus, African swine fever virus (ASFV). There is no vaccine available against ASFV, limiting the options for disease control. ASFV reorganizes intracellular membranes to generate viral factories (VFs) in order to amplify its genome. However, little is known about the process involved in the formation of these viral replication organelles. Membrane contact sites (MCSs) allow nonvesicular lipids and ion exchange between organelles. Lipid exchange to form VFs apparently requires a number of proteins at MCSs, such as the oxysterol-binding protein (OSBP), the acyl-coenzyme A binding domain containing 3 (ACBD3) and the phosphatidylinositol-phosphate-4-kinase III beta (PI4Kβ). Itraconazole (ITZ) is an antifungal agent that targets sterol-transport molecules such as OSBP and OSBP-related protein 4 (ORP4). 25-Hydroxycholesterol (25-HC) inhibits lipid transport by high affinity binding OSBP. In this work, we analyzed the antiviral function of ITZ and 25-HC against ASFV in Vero cell cultures using the cell-adapted Ba71V isolate. ITZ and 25-HC decreased significantly ASFV replication. Our study revealed OSBP distribution in cytoplasmic membranes in uninfected Vero cells and to the periphery of VFs in infected cells. In addition, we showed that OSBP and OSBP-related proteins, PI4Kβ and ACBD3 were recruited to VFs in the context ASFV infection.
Inmaculada Galindo; Miguel Ángel Cuesta-Geijo; Ana Del Puerto; Eva Soriano; Covadonga Alonso. Lipid Exchange Factors at Membrane Contact Sites in African Swine Fever Virus Infection. Viruses 2019, 11, 199 .
AMA StyleInmaculada Galindo, Miguel Ángel Cuesta-Geijo, Ana Del Puerto, Eva Soriano, Covadonga Alonso. Lipid Exchange Factors at Membrane Contact Sites in African Swine Fever Virus Infection. Viruses. 2019; 11 (3):199.
Chicago/Turabian StyleInmaculada Galindo; Miguel Ángel Cuesta-Geijo; Ana Del Puerto; Eva Soriano; Covadonga Alonso. 2019. "Lipid Exchange Factors at Membrane Contact Sites in African Swine Fever Virus Infection." Viruses 11, no. 3: 199.
The family Asfarviridae includes the single species African swine fever virus, isolates of which have linear dsDNA genomes of 170–194 kbp. Virions have an internal core, an internal lipid membrane, an icosahedral capsid and an outer lipid envelope. Infection of domestic pigs and wild boar results in an acute haemorrhagic fever with transmission by contact or ingestion, or by ticks of the genus Ornithodoros. Indigenous pigs act as reservoirs in Africa, where infection is endemic, and from where introductions occur periodically to Europe. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Asfarviridae, which is available at www.ictv.global/report/asfarviridae.
Covadonga Alonso; Manuel Borca; Linda Dixon; Yolanda Revilla; Fernando Rodriguez; Jose M. Escribano; ICTV Report Consortium. ICTV Virus Taxonomy Profile: Asfarviridae. Journal of General Virology 2018, 99, 613 -614.
AMA StyleCovadonga Alonso, Manuel Borca, Linda Dixon, Yolanda Revilla, Fernando Rodriguez, Jose M. Escribano, ICTV Report Consortium. ICTV Virus Taxonomy Profile: Asfarviridae. Journal of General Virology. 2018; 99 (5):613-614.
Chicago/Turabian StyleCovadonga Alonso; Manuel Borca; Linda Dixon; Yolanda Revilla; Fernando Rodriguez; Jose M. Escribano; ICTV Report Consortium. 2018. "ICTV Virus Taxonomy Profile: Asfarviridae." Journal of General Virology 99, no. 5: 613-614.
Dynein is a cytoskeletal molecular motor protein that transports cellular cargoes along microtubules. Biomimetic synthetic peptides designed to bind dynein have been shown to acquire dynamic properties such as cell accumulation and active intra- and inter-cellular motion through cell-to-cell contacts and projections to distant cells. On the basis of these properties dynein-binding peptides could be used to functionalize nanoparticles for drug delivery applications. Here, we show that gold nanoparticles modified with dynein-binding delivery sequences become mobile, powered by molecular motor proteins. Modified nanoparticles showed dynamic properties, such as travelling the cytosol, crossing intracellular barriers and shuttling the nuclear membrane. Furthermore, nanoparticles were transported from one cell to another through cell-to-cell contacts and quickly spread to distant cells through cell projections. The capacity of these motor-bound nanoparticles to spread to many cells and increasing cellular retention, thus avoiding losses and allowing lower dosage, could make them candidate carriers for drug delivery.
Inmaculada Dalmau-Mena; Pablo del Pino; Beatriz Pelaz; Miguel Ángel Cuesta-Geijo; Inmaculada Galindo; María Moros; Jesús M. de la Fuente; Covadonga Alonso. Nanoparticles engineered to bind cellular motors for efficient delivery. Journal of Nanobiotechnology 2018, 16, 33 .
AMA StyleInmaculada Dalmau-Mena, Pablo del Pino, Beatriz Pelaz, Miguel Ángel Cuesta-Geijo, Inmaculada Galindo, María Moros, Jesús M. de la Fuente, Covadonga Alonso. Nanoparticles engineered to bind cellular motors for efficient delivery. Journal of Nanobiotechnology. 2018; 16 (1):33.
Chicago/Turabian StyleInmaculada Dalmau-Mena; Pablo del Pino; Beatriz Pelaz; Miguel Ángel Cuesta-Geijo; Inmaculada Galindo; María Moros; Jesús M. de la Fuente; Covadonga Alonso. 2018. "Nanoparticles engineered to bind cellular motors for efficient delivery." Journal of Nanobiotechnology 16, no. 1: 33.
Rigid amphipathic fusion inhibitors (RAFIs) are a family of nucleoside derivatives that inhibit the infectivity of several enveloped viruses by interacting with virion envelope lipids and inhibiting fusion between viral and cellular membranes. Here we tested the antiviral activity of two RAFIs, 5-(Perylen-3-ylethynyl)-arabino-uridine (aUY11) and 5-(Perylen-3-ylethynyl)uracil-1-acetic acid (cm1UY11) against African swine fever virus (ASFV), for which no effective vaccine is available. Both compounds displayed a potent, dose-dependent inhibitory effect on ASFV infection in Vero cells. The major antiviral effect was observed when aUY11 and cm1UY11 were added at early stages of infection and maintained during the complete viral cycle. Furthermore, virucidal assay revealed a significant extracellular anti-ASFV activity for both compounds. We also found decrease in the synthesis of early and late viral proteins in Vero cells treated with cm1UY11. Finally, the inhibitory effect of aUY11 and cm1UY11 on ASFV infection in porcine alveolar macrophages was confirmed. Overall, our study has identified novel anti-ASFV compounds with potential for future therapeutic developments.
Astghik Hakobyan; Inmaculada Galindo; Almudena Nañez; Erik Arabyan; Zaven Karalyan; Alexey Chistov; Philipp P. Streshnev; Vladimir A. Korshun; Covadonga Alonso; Hovakim Zakaryan. Rigid amphipathic fusion inhibitors demonstrate antiviral activity against African swine fever virus. Journal of General Virology 2018, 99, 148 -156.
AMA StyleAstghik Hakobyan, Inmaculada Galindo, Almudena Nañez, Erik Arabyan, Zaven Karalyan, Alexey Chistov, Philipp P. Streshnev, Vladimir A. Korshun, Covadonga Alonso, Hovakim Zakaryan. Rigid amphipathic fusion inhibitors demonstrate antiviral activity against African swine fever virus. Journal of General Virology. 2018; 99 (1):148-156.
Chicago/Turabian StyleAstghik Hakobyan; Inmaculada Galindo; Almudena Nañez; Erik Arabyan; Zaven Karalyan; Alexey Chistov; Philipp P. Streshnev; Vladimir A. Korshun; Covadonga Alonso; Hovakim Zakaryan. 2018. "Rigid amphipathic fusion inhibitors demonstrate antiviral activity against African swine fever virus." Journal of General Virology 99, no. 1: 148-156.
Several viruses manipulate the ubiquitin-proteasome system (UPS) to initiate a productive infection. Determined viral proteins are able to change the host’s ubiquitin machinery and some viruses even encode their own ubiquitinating or deubiquitinating enzymes. African swine fever virus (ASFV) encodes a gene homologous to the E2 ubiquitin conjugating (UBC) enzyme. The viral ubiquitin-conjugating enzyme (UBCv1) is expressed throughout ASFV infection and accumulates at late times post infection. UBCv is also present in the viral particle suggesting that the ubiquitin-proteasome pathway could play an important role at early ASFV infection. We determined that inhibition of the final stage of the ubiquitin-proteasome pathway blocked a post-internalization step in ASFV replication in Vero cells. Under proteasome inhibition, ASF viral genome replication, late gene expression and viral production were severely reduced. Also, ASFV enhanced proteasome activity at late times and the accumulation of polyubiquitinated proteins surrounding viral factories. Core-associated and/or viral proteins involved in DNA replication may be targets for the ubiquitin-proteasome pathway that could possibly assist virus uncoating at final core breakdown and viral DNA release. At later steps, polyubiquitinated proteins at viral factories could exert regulatory roles in cell signaling.
Lucía Barrado-Gil; Inmaculada Galindo; Diego Martínez-Alonso; Sergio Viedma; Covadonga Alonso. The ubiquitin-proteasome system is required for African swine fever replication. PLOS ONE 2017, 12, e0189741 -e0189741.
AMA StyleLucía Barrado-Gil, Inmaculada Galindo, Diego Martínez-Alonso, Sergio Viedma, Covadonga Alonso. The ubiquitin-proteasome system is required for African swine fever replication. PLOS ONE. 2017; 12 (12):e0189741-e0189741.
Chicago/Turabian StyleLucía Barrado-Gil; Inmaculada Galindo; Diego Martínez-Alonso; Sergio Viedma; Covadonga Alonso. 2017. "The ubiquitin-proteasome system is required for African swine fever replication." PLOS ONE 12, no. 12: e0189741-e0189741.
African swine fever virus (ASFV) is a large DNA virus that replicates predominantly in the cell cytoplasm and is the only member of the Asfarviridae family. The virus causes an acute haemorrhagic fever, African swine fever (ASF), in domestic pigs and wild boar resulting in the death of most infected animals. Apoptosis is induced at an early stage during virus entry or uncoating. However, ASFV encodes anti-apoptotic proteins which facilitate production of progeny virions. These anti-apoptotic proteins include A179L, a Bcl-2 family member; A224L, an inhibitor of apoptosis proteins (IAP) family member; EP153R a C-type lectin; and DP71L. The latter acts by inhibiting activation of the stress activated pro-apoptotic pathways pro-apoptotic pathways. The mechanisms by which these proteins act is summarised. ASF disease is characterised by massive apoptosis of uninfected lymphocytes which reduces the effectiveness of the immune response, contributing to virus pathogenesis. Mechanisms by which this apoptosis is induced are discussed.
Linda K. Dixon; Pedro Sánchez Cordón; Inmaculada Galindo; Covadonga Alonso. Investigations of Pro- and Anti-Apoptotic Factors Affecting African Swine Fever Virus Replication and Pathogenesis. Viruses 2017, 9, 241 .
AMA StyleLinda K. Dixon, Pedro Sánchez Cordón, Inmaculada Galindo, Covadonga Alonso. Investigations of Pro- and Anti-Apoptotic Factors Affecting African Swine Fever Virus Replication and Pathogenesis. Viruses. 2017; 9 (9):241.
Chicago/Turabian StyleLinda K. Dixon; Pedro Sánchez Cordón; Inmaculada Galindo; Covadonga Alonso. 2017. "Investigations of Pro- and Anti-Apoptotic Factors Affecting African Swine Fever Virus Replication and Pathogenesis." Viruses 9, no. 9: 241.
African swine fever virus (ASFV) infection causes endosomal reorganization. Here, we show that the virus causes endosomal congregation close to the nucleus as the infection progresses, which is necessary to build a compact viral replication organelle. ASFV enters the cell by the endosomal pathway and reaches multivesicular late endosomes. Upon uncoating and fusion, the virus should exit to the cytosol to start replication. ASFV remodels endosomal traffic and redistributes endosomal membranes to the viral replication site. Virus replication also depends on endosomal membrane phosphoinositides (PtdIns) synthesized by PIKfyve. Endosomes could act as platforms providing membranes and PtdIns, necessary for ASFV replication. Our study has revealed that ASFV reorganizes endosome dynamics, in order to ensure a productive infection.
Miguel Ángel Cuesta-Geijo; Lucía Barrado-Gil; Inmaculada Galindo; Raquel Muñoz-Moreno; Covadonga Alonso. Redistribution of Endosomal Membranes to the African Swine Fever Virus Replication Site. Viruses 2017, 9, 133 .
AMA StyleMiguel Ángel Cuesta-Geijo, Lucía Barrado-Gil, Inmaculada Galindo, Raquel Muñoz-Moreno, Covadonga Alonso. Redistribution of Endosomal Membranes to the African Swine Fever Virus Replication Site. Viruses. 2017; 9 (6):133.
Chicago/Turabian StyleMiguel Ángel Cuesta-Geijo; Lucía Barrado-Gil; Inmaculada Galindo; Raquel Muñoz-Moreno; Covadonga Alonso. 2017. "Redistribution of Endosomal Membranes to the African Swine Fever Virus Replication Site." Viruses 9, no. 6: 133.
African swine fever (ASF) is a highly contagious viral disease of swine which causes high mortality, approaching 100%, in domestic pigs. ASF is caused by a large, double stranded DNA virus, ASF virus (ASFV), which replicates predominantly in the cytoplasm of macrophages and is the only member of the Asfarviridae family, genus Asfivirus. The natural hosts of this virus include wild suids and arthropod vectors of the Ornithodoros genus. The infection of ASFV in its reservoir hosts is usually asymptomatic and develops a persistent infection. In contrast, infection of domestic pigs leads to a lethal hemorrhagic fever for which there is no effective vaccine. Identification of ASFV genes involved in virulence and the characterization of mechanisms used by the virus to evade the immune response of the host are recognized as critical steps in the development of a vaccine. Moreover, the interplay of the viral products with host pathways, which are relevant for virus replication, provides the basic information needed for the identification of potential targets for the development of intervention strategies against this disease.
Inmaculada Galindo; Covadonga Alonso. African Swine Fever Virus: A Review. Viruses 2017, 9, 103 .
AMA StyleInmaculada Galindo, Covadonga Alonso. African Swine Fever Virus: A Review. Viruses. 2017; 9 (5):103.
Chicago/Turabian StyleInmaculada Galindo; Covadonga Alonso. 2017. "African Swine Fever Virus: A Review." Viruses 9, no. 5: 103.
Current challenges in global immunization indicate the demand for new delivery strategies, which could be applied to the development of new vaccines against emerging diseases, as well as to improve safety and efficacy of currently existing vaccine formulations. Here, we report a novel antigen nanocarrier consisting of an oily core and a protamine shell, further stabilized with pegylated surfactants. These nanocarriers, named protamine nanocapsules, were rationally designed to promote the intracellular delivery of antigens to immunocompetent cells and to trigger an efficient and long-lasting immune response. Protamine nanocapsules have nanometric size, positive zeta potential and high association capacity for H1N1 influenza hemagglutinin, a protein that was used here as a model antigen. The new formulation shows an attractive stability profile both, as an aqueous suspension or a freeze-dried powder formulation. In vitro studies showed that protamine nanocapsules were efficiently internalized by macrophages without eliciting significant toxicity. In vivo studies indicate that antigen-loaded nanocapsules trigger immune responses comparable to those achieved with alum, even when using significantly lower antigen doses, thus indicating their adjuvant properties. These promising in vivo data, alongside with their versatility for the loading of different antigens and oily immunomodulators and their excellent stability profile, make these nanocapsules a promising platform for the delivery of antigens. Chemical compounds Protamine sulphate (PubChem SID: 7849283), Sodium Cholate (PubChem CID: 23668194), Miglyol (PubChem CID: 53471835), α tocopherol (PubChem CID: 14985), Tween® 20(PubChem CID: 443314), Tween® 80(PubChem CID: 5281955), TPGS (PubChem CID: 71406).
José Vicente González-Aramundiz; Elena Presas; Inmaculada Dalmau-Mena; Susana Martínez-Pulgarín; Covadonga Alonso; José M Escribano; María J. Alonso; Noemi Stefánia Csaba. Rational design of protamine nanocapsules as antigen delivery carriers. Journal of Controlled Release 2017, 245, 62 -69.
AMA StyleJosé Vicente González-Aramundiz, Elena Presas, Inmaculada Dalmau-Mena, Susana Martínez-Pulgarín, Covadonga Alonso, José M Escribano, María J. Alonso, Noemi Stefánia Csaba. Rational design of protamine nanocapsules as antigen delivery carriers. Journal of Controlled Release. 2017; 245 ():62-69.
Chicago/Turabian StyleJosé Vicente González-Aramundiz; Elena Presas; Inmaculada Dalmau-Mena; Susana Martínez-Pulgarín; Covadonga Alonso; José M Escribano; María J. Alonso; Noemi Stefánia Csaba. 2017. "Rational design of protamine nanocapsules as antigen delivery carriers." Journal of Controlled Release 245, no. : 62-69.