48 results on '"Jurado KA"'
Search Results
2. Novel immunomodulatory properties of adenosine analogs promote their antiviral activity against SARS-CoV-2.
- Author
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Monticone G, Huang Z, Hewins P, Cook T, Mirzalieva O, King B, Larter K, Miller-Ensminger T, Sanchez-Pino MD, Foster TP, Nichols OV, Ramsay AJ, Majumder S, Wyczechowska D, Tauzier D, Gravois E, Crabtree JS, Garai J, Li L, Zabaleta J, Barbier MT, Del Valle L, Jurado KA, and Miele L
- Subjects
- Humans, Animals, Immunomodulating Agents pharmacology, Immunomodulating Agents chemistry, Adenosine A2 Receptor Antagonists pharmacology, Adenosine A2 Receptor Antagonists chemistry, Adenosine A2 Receptor Antagonists therapeutic use, Pandemics, COVID-19 Drug Treatment, Chlorocebus aethiops, Virus Replication drug effects, Vero Cells, Betacoronavirus drug effects, Betacoronavirus immunology, Receptor, Adenosine A2A metabolism, Coronavirus Infections drug therapy, Coronavirus Infections immunology, Coronavirus Infections virology, Antiviral Agents pharmacology, Antiviral Agents chemistry, SARS-CoV-2 drug effects, SARS-CoV-2 immunology, Adenosine analogs & derivatives, Adenosine pharmacology, Adenosine chemistry, Adenosine Monophosphate analogs & derivatives, Adenosine Monophosphate pharmacology, Alanine analogs & derivatives, Alanine pharmacology, Alanine chemistry, COVID-19 immunology, COVID-19 virology
- Abstract
The COVID-19 pandemic reminded us of the urgent need for new antivirals to control emerging infectious diseases and potential future pandemics. Immunotherapy has revolutionized oncology and could complement the use of antivirals, but its application to infectious diseases remains largely unexplored. Nucleoside analogs are a class of agents widely used as antiviral and anti-neoplastic drugs. Their antiviral activity is generally based on interference with viral nucleic acid replication or transcription. Based on our previous work and computer modeling, we hypothesize that antiviral adenosine analogs, like remdesivir, have previously unrecognized immunomodulatory properties which contribute to their therapeutic activity. In the case of remdesivir, we here show that these properties are due to its metabolite, GS-441524, acting as an Adenosine A2A Receptor antagonist. Our findings support a new rationale for the design of next-generation antiviral agents with dual - immunomodulatory and intrinsic - antiviral properties. These compounds could represent game-changing therapies to control emerging viral diseases and future pandemics., (© 2024. The Author(s).)
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- 2024
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3. Mitochondrial antioxidants abate SARS-COV-2 pathology in mice.
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Guarnieri JW, Lie T, Albrecht YES, Hewin P, Jurado KA, Widjaja GA, Zhu Y, McManus MJ, Kilbaugh TJ, Keith K, Potluri P, Taylor D, Angelin A, Murdock DG, and Wallace DC
- Subjects
- Animals, Mice, Humans, Angiotensin-Converting Enzyme 2 metabolism, Angiotensin-Converting Enzyme 2 genetics, Lung virology, Lung pathology, Lung metabolism, Reactive Oxygen Species metabolism, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Hypoxia-Inducible Factor 1, alpha Subunit genetics, DNA, Mitochondrial genetics, DNA, Mitochondrial metabolism, Catalase metabolism, Catalase genetics, COVID-19 Drug Treatment, Disease Models, Animal, Immunity, Innate, COVID-19 virology, COVID-19 metabolism, COVID-19 immunology, COVID-19 pathology, Antioxidants metabolism, Antioxidants pharmacology, Mitochondria metabolism, Mitochondria drug effects, SARS-CoV-2 drug effects, Oxidative Phosphorylation drug effects, Mice, Transgenic
- Abstract
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection inhibits mitochondrial oxidative phosphorylation (OXPHOS) and elevates mitochondrial reactive oxygen species (ROS, mROS) which activates hypoxia-inducible factor-1alpha (HIF-1α), shifting metabolism toward glycolysis to drive viral biogenesis but also causing the release of mitochondrial DNA (mtDNA) and activation of innate immunity. To determine whether mitochondrially targeted antioxidants could mitigate these viral effects, we challenged mice expressing human angiotensin-converting enzyme 2 (ACE2) with SARS-CoV-2 and intervened using transgenic and pharmacological mitochondrially targeted catalytic antioxidants. Transgenic expression of mitochondrially targeted catalase (mCAT) or systemic treatment with EUK8 decreased weight loss, clinical severity, and circulating levels of mtDNA; as well as reduced lung levels of HIF-1α, viral proteins, and inflammatory cytokines. RNA-sequencing of infected lungs revealed that mCAT and Eukarion 8 (EUK8) up-regulated OXPHOS gene expression and down-regulated HIF-1α and its target genes as well as innate immune gene expression. These data demonstrate that SARS-CoV-2 pathology can be mitigated by catalytically reducing mROS, potentially providing a unique host-directed pharmacological therapy for COVID-19 which is not subject to viral mutational resistance., Competing Interests: Competing interests statement:D.C.W. is on the scientific advisory boards of Pano Therapeutics, Inc. and Medical Excellent Capital, and published a manuscript with S.E. Schriner in 2022.
- Published
- 2024
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4. Human red blood cells express the RNA sensor TLR7.
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Metthew Lam LK, Oatman E, Eckart KA, Klingensmith NJ, Flowers E, Sayegh L, Yuen J, Clements RL, Meyer NJ, Jurado KA, Vaughan AE, Eisenbarth SC, and Mangalmurti NS
- Subjects
- Humans, Sepsis metabolism, Sepsis blood, Sepsis genetics, Erythrocyte Membrane metabolism, Male, RNA metabolism, RNA genetics, Female, Toll-Like Receptor 7 metabolism, Toll-Like Receptor 7 genetics, Erythrocytes metabolism, COVID-19 virology, COVID-19 metabolism, SARS-CoV-2 metabolism
- Abstract
Red blood cells (RBCs) express the nucleic acid-binding toll-like receptor 9 (TLR9) and bind CpG-containing DNA. However, whether human RBCs express other nucleic acid-binding TLRs is unknown. Here we show that human RBCs express the RNA sensor TLR7. TLR7 is present on the red cell membrane and is associated with the RBC membrane protein Band 3. In patients with SARS-CoV2-associated sepsis, TLR7-Band 3 interactions in the RBC membrane are increased when compared with healthy controls. In vitro, RBCs bind synthetic ssRNA and RNA from ssRNA viruses. Thus, RBCs may serve as a previously unrecognized sink for exogenous RNA, expanding the repertoire of non-gas exchanging functions performed by RBCs., (© 2024. The Author(s).)
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- 2024
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5. Human erythroid progenitors express antigen presentation machinery.
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Clements RL, Kennedy EA, Song D, Campbell A, An HH, Amses KR, Miller-Ensminger T, Addison MM, Eisenlohr LC, Chou ST, and Jurado KA
- Abstract
Early-life immune exposures can profoundly impact lifelong health. However, functional mechanisms underlying fetal immune development remain incomplete. Erythrocytes are not typically considered active immune mediators, primarily because erythroid precursors discard their organelles as they mature, thus losing the ability to alter gene expression in response to stimuli. Erythroid progenitors and precursors circulate in human fetuses and neonates. Although there is limited evidence that erythroid precursors are immunomodulatory, our understanding of the underlying mechanisms remains inadequate. To define the immunobiological role of fetal and perinatal erythroid progenitors and precursors, we analyzed single cell RNA-sequencing data and found that transcriptomics support erythroid progenitors as putative immune mediators. Unexpectedly, we discovered that human erythroid progenitors constitutively express Major Histocompatibility Complex (MHC) class II antigen processing and presentation machinery, which are hallmarks of specialized antigen presenting immune cells. Furthermore, we demonstrate that erythroid progenitors internalize and cleave foreign proteins into peptide antigens. Unlike conventional antigen presenting cells, erythroid progenitors express atypical costimulatory molecules and immunoregulatory cytokines that direct the development of regulatory T cells, which are critical for establishing maternal-fetal tolerance. Expression of MHC II in definitive erythroid progenitors begins during the second trimester, coinciding with the appearance of mature T cells in the fetus, and is absent in primitive progenitors. Lastly, we demonstrate physical and molecular interaction potential of erythroid progenitors and T cells in the fetal liver. Our findings shed light on a unique orchestrator of fetal immunity and provide insight into the mechanisms by which erythroid cells contribute to host defense.
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- 2024
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6. The lipopeptide Pam3CSK4 inhibits Rift Valley fever virus infection and protects from encephalitis.
- Author
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Griesman T, McMillen CM, Negatu SG, Hulahan JJ, Whig K, Dohnalová L, Dittmar M, Thaiss CA, Jurado KA, Schultz DC, Hartman AL, and Cherry S
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- Animals, Mice, Mice, Inbred C57BL, Humans, Immunity, Innate drug effects, Encephalitis, Viral virology, Encephalitis, Viral immunology, Encephalitis, Viral prevention & control, Encephalitis, Viral drug therapy, Antiviral Agents pharmacology, Rift Valley fever virus drug effects, Lipopeptides pharmacology, Rift Valley Fever virology, Rift Valley Fever prevention & control, Neurons metabolism, Neurons virology
- Abstract
Rift Valley fever virus (RVFV) is an encephalitic bunyavirus that can infect neurons in the brain. There are no approved therapeutics that can protect from RVFV encephalitis. Innate immunity, the first line of defense against infection, canonically antagonizes viruses through interferon signaling. We found that interferons did not efficiently protect primary cortical neurons from RVFV, unlike other cell types. To identify alternative neuronal antiviral pathways, we screened innate immune ligands and discovered that the TLR2 ligand Pam3CSK4 inhibited RVFV infection, and other bunyaviruses. Mechanistically, we found that Pam3CSK4 blocks viral fusion, independent of TLR2. In a mouse model of RVFV encephalitis, Pam3CSK4 treatment protected animals from infection and mortality. Overall, Pam3CSK4 is a bunyavirus fusion inhibitor active in primary neurons and the brain, representing a new approach toward the development of treatments for encephalitic bunyavirus infections., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Griesman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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7. Type I interferon alters invasive extravillous trophoblast function.
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Simoni MK, Negatu SG, Park JY, Mani S, Arreguin MC, Amses K, Huh DD, Mainigi M, and Jurado KA
- Abstract
Inappropriate type I interferon (IFN) signaling during embryo implantation and placentation is linked to poor pregnancy outcomes. Here, we evaluated the consequence of elevated type I IFN exposure on implantation using a biomimetic model of human implantation in an organ-on-a-chip device. We found that type I IFN reduced extravillous trophoblast (EVT) invasion capacity. Analyzing single-cell transcriptomes, we uncovered that IFN truncated endovascular EVT emergence in the implantation-on-a-chip device by stunting EVT epithelial-to-mesenchymal transition. Disruptions to the epithelial-to-mesenchymal transition is associated with the pathogenesis of preeclampsia, a life-threatening hypertensive disorder of pregnancy. Strikingly, unwarranted IFN stimulation induced genes associated with increased preeclampsia risk and a preeclamptic gene-like signature in EVTs. These dysregulated EVT phenotypes ultimately reduced EVT-mediated endothelial cell vascular remodeling in the implantation-on-a-chip device. Overall, our work indicates IFN signaling can alter EVT epithelial-to-mesenchymal transition progression which results in diminished EVT-mediated spiral artery remodeling and a preeclampsia gene signature upon sustained stimulation. Our work implicates unwarranted type I IFN as a maternal disturbance that can result in abnormal EVT function that could trigger preeclampsia.
- Published
- 2024
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8. Serotonin reduction in post-acute sequelae of viral infection.
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Wong AC, Devason AS, Umana IC, Cox TO, Dohnalová L, Litichevskiy L, Perla J, Lundgren P, Etwebi Z, Izzo LT, Kim J, Tetlak M, Descamps HC, Park SL, Wisser S, McKnight AD, Pardy RD, Kim J, Blank N, Patel S, Thum K, Mason S, Beltra JC, Michieletto MF, Ngiow SF, Miller BM, Liou MJ, Madhu B, Dmitrieva-Posocco O, Huber AS, Hewins P, Petucci C, Chu CP, Baraniecki-Zwil G, Giron LB, Baxter AE, Greenplate AR, Kearns C, Montone K, Litzky LA, Feldman M, Henao-Mejia J, Striepen B, Ramage H, Jurado KA, Wellen KE, O'Doherty U, Abdel-Mohsen M, Landay AL, Keshavarzian A, Henrich TJ, Deeks SG, Peluso MJ, Meyer NJ, Wherry EJ, Abramoff BA, Cherry S, Thaiss CA, and Levy M
- Subjects
- Humans, COVID-19 complications, Disease Progression, Inflammation, Virus Diseases, Post-Acute COVID-19 Syndrome blood, Post-Acute COVID-19 Syndrome pathology, Serotonin blood
- Abstract
Post-acute sequelae of COVID-19 (PASC, "Long COVID") pose a significant global health challenge. The pathophysiology is unknown, and no effective treatments have been found to date. Several hypotheses have been formulated to explain the etiology of PASC, including viral persistence, chronic inflammation, hypercoagulability, and autonomic dysfunction. Here, we propose a mechanism that links all four hypotheses in a single pathway and provides actionable insights for therapeutic interventions. We find that PASC are associated with serotonin reduction. Viral infection and type I interferon-driven inflammation reduce serotonin through three mechanisms: diminished intestinal absorption of the serotonin precursor tryptophan; platelet hyperactivation and thrombocytopenia, which impacts serotonin storage; and enhanced MAO-mediated serotonin turnover. Peripheral serotonin reduction, in turn, impedes the activity of the vagus nerve and thereby impairs hippocampal responses and memory. These findings provide a possible explanation for neurocognitive symptoms associated with viral persistence in Long COVID, which may extend to other post-viral syndromes., Competing Interests: Declaration of interests E.J.W. is an advisor for Danger Bio, Janssen, New Limit, Marengo, Pluto Immunotherapeutics Related Sciences, Rubius Therapeutics, Santa Ana Bio, Synthekine, and Surface Oncology. E.J.W. is a founder of and holds stock in Surface Oncology, Danger Bio, and Arsenal Biosciences. N.J.M. reports consulting fees from Endpoint Health Inc and AstraZeneca and receives funding from Quantum Leap Healthcare Collaborative outside of the published work., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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9. Endothelial SARS-CoV-2 infection is not the underlying cause of COVID-19-associated vascular pathology in mice.
- Author
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Gao S, Tang AT, Wang M, Buchholz DW, Imbiakha B, Yang J, Chen X, Hewins P, Mericko-Ishizuka P, Leu NA, Sterling S, August A, Jurado KA, Morrisey EE, Aguilar-Carreno H, and Kahn ML
- Abstract
Endothelial damage and vascular pathology have been recognized as major features of COVID-19 since the beginning of the pandemic. Two main theories regarding how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) damages endothelial cells and causes vascular pathology have been proposed: direct viral infection of endothelial cells or indirect damage mediated by circulating inflammatory molecules and immune mechanisms. However, these proposed mechanisms remain largely untested in vivo . In the present study, we utilized a set of new mouse genetic tools developed in our lab to test both the necessity and sufficiency of endothelial human angiotensin-converting enzyme 2 (hACE2) in COVID-19 pathogenesis. Our results demonstrate that endothelial ACE2 and direct infection of vascular endothelial cells do not contribute significantly to the diverse vascular pathology associated with COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Gao, Tang, Wang, Buchholz, Imbiakha, Yang, Chen, Hewins, Mericko-Ishizuka, Leu, Sterling, August, Jurado, Morrisey, Aguilar-Carreno and Kahn.)
- Published
- 2023
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10. Endothelial SARS-CoV-2 infection is not the underlying cause of COVID19-associated vascular pathology in mice.
- Author
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Gao S, Tang AT, Wang M, Buchholz DW, Imbiakha B, Yang J, Chen X, Hewins P, Mericko-Ishizuka P, Leu NA, Sterling S, August A, Jurado KA, Morrisey EE, Aguilar-Carreno H, and Kahn ML
- Abstract
Endothelial damage and vascular pathology have been recognized as major features of COVID-19 since the beginning of the pandemic. Two main theories regarding how Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) damages endothelial cells and causes vascular pathology have been proposed: direct viral infection of endothelial cells or indirect damage mediated by circulating inflammatory molecules and immune mechanisms. However, these proposed mechanisms remain largely untested in vivo. Here, we utilized a set of new mouse genetic tools
1 developed in our lab to test both the necessity and sufficiency of endothelial human angiotensin-converting enzyme 2 (hACE2) in COVID19 pathogenesis. Our results demonstrate that endothelial ACE2 and direct infection of vascular endothelial cells does not contribute significantly to the diverse vascular pathology associated with COVID-19.- Published
- 2023
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11. A nanobody recognizes a unique conserved epitope and potently neutralizes SARS-CoV-2 omicron variants.
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Modhiran N, Lauer SM, Amarilla AA, Hewins P, Lopes van den Broek SI, Low YS, Thakur N, Liang B, Nieto GV, Jung J, Paramitha D, Isaacs A, Sng JDJ, Song D, Jørgensen JT, Cheuquemilla Y, Bürger J, Andersen IV, Himelreichs J, Jara R, MacLoughlin R, Miranda-Chacon Z, Chana-Cuevas P, Kramer V, Spahn C, Mielke T, Khromykh AA, Munro T, Jones ML, Young PR, Chappell K, Bailey D, Kjaer A, Herth MM, Jurado KA, Schwefel D, Rojas-Fernandez A, and Watterson D
- Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) Omicron variant sub-lineages spread rapidly worldwide, mostly due to their immune-evasive properties. This has put a significant part of the population at risk for severe disease and underscores the need for effective anti-SARS-CoV-2 agents against emergent strains in vulnerable patients. Camelid nanobodies are attractive therapeutic candidates due to their high stability, ease of large-scale production, and potential for delivery via inhalation. Here, we characterize the receptor binding domain (RBD)-specific nanobody W25 and show superior neutralization activity toward Omicron sub-lineages in comparison to all other SARS-CoV2 variants. Structure analysis of W25 in complex with the SARS-CoV2 spike glycoprotein shows that W25 engages an RBD epitope not covered by any of the antibodies previously approved for emergency use. In vivo evaluation of W25 prophylactic and therapeutic treatments across multiple SARS-CoV-2 variant infection models, together with W25 biodistribution analysis in mice, demonstrates favorable pre-clinical properties. Together, these data endorse W25 for further clinical development., Competing Interests: D.W., K.C., and P.R.Y are listed as inventors of ‘Molecular Clamp’ patent, US 2020/0040042., (© 2023.)
- Published
- 2023
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12. IL-33 is alarmin the uterus for labor.
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Kennedy EA and Jurado KA
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- Female, Mice, Animals, Uterus, Immunity, Innate, Alarmins, Interleukin-33
- Abstract
Timely labor is critical for both infant and maternal health, yet the mechanisms underlying the initiation of childbirth remain unclear. In this issue of Immunity, Siewiera et al. demonstrate a vital role for innate type 2 immune responses in controlling uterus-intrinsic onset of labor in mice.
1 ., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023. Published by Elsevier Inc.)- Published
- 2023
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13. Positive-strand RNA viruses-a Keystone Symposia report.
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Cable J, Denison MR, Kielian M, Jackson WT, Bartenschlager R, Ahola T, Mukhopadhyay S, Fremont DH, Kuhn RJ, Shannon A, Frazier MN, Yuen KY, Coyne CB, Wolthers KC, Ming GL, Guenther CS, Moshiri J, Best SM, Schoggins JW, Jurado KA, Ebel GD, Schäfer A, Ng LFP, Kikkert M, Sette A, Harris E, Wing PAC, Eggenberger J, Krishnamurthy SR, Mah MG, Meganck RM, Chung D, Maurer-Stroh S, Andino R, Korber B, Perlman S, Shi PY, Bárcena M, Aicher SM, Vu MN, Kenney DJ, Lindenbach BD, Nishida Y, Rénia L, and Williams EP
- Subjects
- Humans, SARS-CoV-2, Positive-Strand RNA Viruses, Antiviral Agents therapeutic use, Pandemics, COVID-19, Zika Virus, Zika Virus Infection epidemiology, Zika Virus Infection prevention & control, Zika Virus Infection drug therapy
- Abstract
Positive-strand RNA viruses have been the cause of several recent outbreaks and epidemics, including the Zika virus epidemic in 2015, the SARS outbreak in 2003, and the ongoing SARS-CoV-2 pandemic. On June 18-22, 2022, researchers focusing on positive-strand RNA viruses met for the Keystone Symposium "Positive-Strand RNA Viruses" to share the latest research in molecular and cell biology, virology, immunology, vaccinology, and antiviral drug development. This report presents concise summaries of the scientific discussions at the symposium., (© 2023 New York Academy of Sciences.)
- Published
- 2023
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14. Cell-autonomous requirement for ACE2 across organs in lethal mouse SARS-CoV-2 infection.
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Tang AT, Buchholz DW, Szigety KM, Imbiakha B, Gao S, Frankfurter M, Wang M, Yang J, Hewins P, Mericko-Ishizuka P, Leu NA, Sterling S, Monreal IA, Sahler J, August A, Zhu X, Jurado KA, Xu M, Morrisey EE, Millar SE, Aguilar HC, and Kahn ML
- Subjects
- Mice, Animals, Angiotensin-Converting Enzyme 2 genetics, SARS-CoV-2 metabolism, Cachexia, Peptidyl-Dipeptidase A genetics, Peptidyl-Dipeptidase A metabolism, Hypoxia, COVID-19
- Abstract
Angiotensin-converting enzyme 2 (ACE2) is the cell-surface receptor for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). While its central role in Coronavirus Disease 2019 (COVID-19) pathogenesis is indisputable, there remains significant debate regarding the role of this transmembrane carboxypeptidase in the disease course. These include the role of soluble versus membrane-bound ACE2, as well as ACE2-independent mechanisms that may contribute to viral spread. Testing these roles requires in vivo models. Here, we report humanized ACE2-floxed mice in which hACE2 is expressed from the mouse Ace2 locus in a manner that confers lethal disease and permits cell-specific, Cre-mediated loss of function, and LSL-hACE2 mice in which hACE2 is expressed from the Rosa26 locus enabling cell-specific, Cre-mediated gain of function. Following exposure to SARS-CoV-2, hACE2-floxed mice experienced lethal cachexia, pulmonary infiltrates, intravascular thrombosis and hypoxemia-hallmarks of severe COVID-19. Cre-mediated loss and gain of hACE2 demonstrate that neuronal infection confers lethal cachexia, hypoxemia, and respiratory failure in the absence of lung epithelial infection. In this series of genetic experiments, we demonstrate that ACE2 is absolutely and cell-autonomously required for SARS-CoV-2 infection in the olfactory epithelium, brain, and lung across diverse cell types. Therapies inhibiting or blocking ACE2 at these different sites are likely to be an effective strategy towards preventing severe COVID-19., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Tang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2023
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15. Being the Alice of academia: lessons from the Red Queen hypothesis.
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Negatu SG, Arreguin MC, Jurado KA, and Vazquez C
- Abstract
Viruses and hosts must navigate environments in which each tries to outcompete the other for survival or to coexist within the same spaces. In Lewis Carrol's Through the Looking Glass, the Red Queen tells Alice, "Now, here, you see, it takes all the running you can do, to keep in the same place. If you want to get somewhere else, you must run at least twice as fast as that!" Borrowing from this idea, the Red Queen hypothesis asserts that organisms, such as viruses, must continuously adapt to environmental pressures to survive. In this commentary, we draw parallels between the Red Queen hypothesis and the experiences scientists of color navigate to thrive in academic spaces. In both phenomena, adapting to environmental pressures is necessary for survival. We identify the various pressures and bottlenecks faced by historically underrepresented groups in academia, as well as the adaptation strategies they must implement to persist in academia., (© The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.)
- Published
- 2022
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16. Neurotropic RNA Virus Modulation of Immune Responses within the Central Nervous System.
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Vazquez C and Jurado KA
- Subjects
- Blood-Brain Barrier, Central Nervous System, Endothelial Cells, Immunity, RNA, RNA Viruses, Viruses
- Abstract
The central nervous system (CNS) necessitates intricately coordinated immune responses to prevent neurological disease. However, the emergence of viruses capable of entering the CNS and infecting neurons threatens this delicate balance. Our CNS is protected from foreign invaders and excess solutes by a semipermeable barrier of endothelial cells called the blood-brain barrier. Thereby, viruses have implemented several strategies to bypass this protective layer and modulate immune responses within the CNS. In this review, we outline these immune regulatory mechanisms and provide perspectives on future questions in this rapidly expanding field.
- Published
- 2022
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17. SARS-CoV-2 infection of olfactory epithelial cells and neurons drives acute lung injury and lethal COVID-19 in mice.
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Tang AT, Buchholz DW, Szigety KM, Imbhiaka B, Gao S, Frankfurter M, Wang M, Yang J, Hewins P, Mericko-Ishizuka P, Adrian Leu N, Sterling S, Monreal IA, Sahler J, August A, Zhu X, Jurado KA, Xu M, Morrisey EE, Millar SE, Aguilar HC, and Kahn ML
- Abstract
Lethal COVID-19 is associated with respiratory failure that is thought to be caused by acute respiratory distress syndrome (ARDS) secondary to pulmonary infection. To date, the cellular pathogenesis has been inferred from studies describing the expression of ACE2, a transmembrane protein required for SARS-CoV-2 infection, and detection of viral RNA or protein in infected humans, model animals, and cultured cells. To functionally test the cellular mechanisms of COVID-19, we generated hACE2
fl animals in which human ACE2 (hACE2) is expressed from the mouse Ace2 locus in a manner that permits cell-specific, Cre-mediated loss of function. hACE2fl animals developed lethal weight loss and hypoxemia within 7 days of exposure to SARS-CoV-2 that was associated with pulmonary infiltrates, intravascular thrombosis and patchy viral infection of lung epithelial cells. Deletion of hACE2 in lung epithelial cells prevented viral infection of the lung, but not weight loss, hypoxemia or death. Inhalation of SARS-CoV-2 by hACE2fl animals resulted in early infection of sustentacular cells with subsequent infection of neurons in the neighboring olfactory bulb and cerebral cortexâ€" events that did not require lung epithelial cell infection. Pharmacologic ablation of the olfactory epithelium or Foxg1Cre mediated deletion of hACE2 in olfactory epithelial cells and neurons prevented lethality and neuronal infection following SARS-CoV-2 infection. Conversely, transgenic expression of hACE2 specifically in olfactory epithelial cells and neurons in Foxg1Cre ; LSL- hACE2 mice was sufficient to confer neuronal infection associated with respiratory failure and death. These studies establish mouse loss and gain of function genetic models with which to genetically dissect viral-host interactions and demonstrate that lethal disease due to respiratory failure may arise from extrapulmonary infection of the olfactory epithelium and brain. Future therapeutic efforts focused on preventing olfactory epithelial infection may be an effective means of protecting against severe COVID-19.- Published
- 2021
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18. Navigating COVID-19: Starting a lab during the pandemic.
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Ordovás-Montañés J, Gerlach C, Moraes-Vieira P, Vento-Tormo R, Zhang C, and Jurado KA
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- Adaptation, Psychological, Biomedical Research trends, COVID-19 psychology, Communication, Humans, Laboratory Personnel psychology, Laboratory Personnel trends, SARS-CoV-2, COVID-19 epidemiology, Laboratories trends
- Abstract
For new principal investigators, the first years are key to getting a laboratory off the ground and running. COVID-19 has changed the world, bringing on unforeseen difficulties and challenges at every level. We asked these investigators to share their experiences in navigating the unique environment since the start of the pandemic-what has changed in their vision for their laboratory, how they have adapted, and what advice they can share with others in a similar situation., (Copyright © 2021. Published by Elsevier Inc.)
- Published
- 2021
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19. SARS-CoV-2 viral proteins NSP1 and NSP13 inhibit interferon activation through distinct mechanisms.
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Vazquez C, Swanson SE, Negatu SG, Dittmar M, Miller J, Ramage HR, Cherry S, and Jurado KA
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- Active Transport, Cell Nucleus genetics, Active Transport, Cell Nucleus immunology, Adaptor Proteins, Signal Transducing genetics, COVID-19 genetics, Cell Nucleus genetics, HeLa Cells, Humans, Interferon Regulatory Factor-3 genetics, NF-kappa B genetics, NF-kappa B immunology, Phosphorylation genetics, Phosphorylation immunology, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases immunology, RNA-Binding Proteins genetics, SARS-CoV-2 genetics, Signal Transduction genetics, Viral Nonstructural Proteins genetics, Adaptor Proteins, Signal Transducing immunology, COVID-19 immunology, Cell Nucleus immunology, Interferon Regulatory Factor-3 immunology, RNA-Binding Proteins immunology, SARS-CoV-2 immunology, Signal Transduction immunology, Viral Nonstructural Proteins immunology
- Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a devastating global pandemic, infecting over 43 million people and claiming over 1 million lives, with these numbers increasing daily. Therefore, there is urgent need to understand the molecular mechanisms governing SARS-CoV-2 pathogenesis, immune evasion, and disease progression. Here, we show that SARS-CoV-2 can block IRF3 and NF-κB activation early during virus infection. We also identify that the SARS-CoV-2 viral proteins NSP1 and NSP13 can block interferon activation via distinct mechanisms. NSP1 antagonizes interferon signaling by suppressing host mRNA translation, while NSP13 downregulates interferon and NF-κB promoter signaling by limiting TBK1 and IRF3 activation, as phospho-TBK1 and phospho-IRF3 protein levels are reduced with increasing levels of NSP13 protein expression. NSP13 can also reduce NF-κB activation by both limiting NF-κB phosphorylation and nuclear translocation. Last, we also show that NSP13 binds to TBK1 and downregulates IFIT1 protein expression. Collectively, these data illustrate that SARS-CoV-2 bypasses multiple innate immune activation pathways through distinct mechanisms., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
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20. ZIKV actively induces calcification in the fetal brain.
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Clements RL and Jurado KA
- Subjects
- Brain diagnostic imaging, Female, Humans, Placenta, Pregnancy, Pregnancy Complications, Infectious, Zika Virus, Zika Virus Infection
- Published
- 2021
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21. mSphere of Influence: Innate Immunity at the Maternal-Fetal Barrier.
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Jurado KA
- Subjects
- Cells, Cultured, Female, Humans, Interferons immunology, Maternal-Fetal Exchange, Pregnancy, Zika Virus pathogenicity, Interferon Lambda, Immunity, Innate, Placenta immunology, Zika Virus Infection immunology
- Abstract
Kellie Ann Jurado works in the field of emerging infectious diseases. In this mSphere of Influence article, she reflects on how the papers "Type III interferons produced by human placental trophoblasts confer protection against Zika virus infection" (https://doi.org/10.1016/j.chom.2016.03.008) and "A three-dimensional culture system recapitulates placental syncytiotrophoblast development and microbial resistance" (https://doi.org/10.1126/sciadv.1501462) by Carolyn Coyne's group have made an impact on her, inspiring her to explore immunity in the placenta by indicating the unique innate immune control elicited at the maternal-fetal barrier as well as by providing physiologically relevant model systems for study., (Copyright © 2020 Jurado.)
- Published
- 2020
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22. A Peptide Derived from Lens Epithelium-Derived Growth Factor Stimulates HIV-1 DNA Integration and Facilitates Intasome Structural Studies.
- Author
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Li M, Chen X, Wang H, Jurado KA, Engelman AN, and Craigie R
- Subjects
- Adaptor Proteins, Signal Transducing chemistry, Adaptor Proteins, Signal Transducing genetics, DNA, Viral genetics, DNA, Viral metabolism, HIV Integrase genetics, Humans, Peptide Fragments genetics, Transcription Factors chemistry, Transcription Factors genetics, Adaptor Proteins, Signal Transducing metabolism, DNA, Viral chemistry, HIV Integrase metabolism, HIV-1 physiology, Peptide Fragments metabolism, Protein Conformation, Transcription Factors metabolism, Virus Integration
- Abstract
The low solubility and aggregation properties of HIV-1 integrase (IN) are major obstacles for biochemical and structural studies. The lens epithelium-derived growth factor (LEDGF) is a cellular factor that binds IN and tethers preintegration complexes to chromatin before integration. The LEDGF also stimulates HIV-1 IN DNA strand transfer activity and improves its solubility in vitro. We show that these properties are conferred by a short peptide spanning residues 178 to 197 of the LEDGF that encompasses its AT-hook DNA-binding elements. The peptide stimulates HIV-1 IN activity both in trans and in cis. Fusion of the peptide to either the N- or C-terminus of IN results in maximal stimulation of concerted integration activity and greatly improves the solubility of the protein and nucleoprotein complexes of IN with viral DNA ends (intasomes). High-resolution structures of HIV-1 intasomes are required to understand the mechanism of IN strand transfer inhibitors (INSTIs), which are front-line drugs for the treatment of HIV-1, and how the virus can develop resistance to INSTIs. We have previously determined the structure of the HIV-1 strand transfer complex intasome. The improved biophysical properties of intasomes assembled with LEDGF peptide fusion IN have enabled us to determine the structure of the cleaved synaptic complex intasome, which is the direct target of INSTIs., (Published by Elsevier Ltd.)
- Published
- 2020
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23. Playing Favorites: Integrin αvβ5 Mediates Preferential Zika Infection of Neural Stem Cells.
- Author
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Vazquez C and Jurado KA
- Subjects
- Humans, Receptors, Vitronectin, SOXB1 Transcription Factors, Glioblastoma, Neural Stem Cells, Zika Virus, Zika Virus Infection
- Abstract
The molecular basis dictating specificity of Zika virus infection in neural stem cells (NSCs) remains elusive. Two recent papers in Cell Stem Cell (Zhu et al., 2020) and Cell Reports (Wang et al., 2020) identify integrin αvβ5 as an internalization factor that increases susceptibility in NSCs and glioblastoma stem cells., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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24. Editorial overview: Viruses in a changing world.
- Author
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Jurado KA and Cherry S
- Subjects
- Communicable Diseases, Emerging epidemiology, Communicable Diseases, Emerging virology, Humans, Virus Diseases epidemiology, Viruses genetics, Virus Diseases virology, Virus Physiological Phenomena
- Published
- 2020
- Full Text
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25. An Antiviral Branch of the IL-1 Signaling Pathway Restricts Immune-Evasive Virus Replication.
- Author
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Orzalli MH, Smith A, Jurado KA, Iwasaki A, Garlick JA, and Kagan JC
- Subjects
- Animals, Cell Line, Chlorocebus aethiops, Female, Fibroblasts immunology, Fibroblasts virology, Gene Expression immunology, HEK293 Cells, Human Umbilical Vein Endothelial Cells, Humans, Male, Mice, Mice, Inbred C57BL, Skin immunology, Skin virology, Vero Cells, Immune Evasion immunology, Interleukin-1 immunology, Signal Transduction immunology, Virus Replication immunology
- Abstract
Virulent pathogens often cause the release of host-derived damage-associated molecular patterns (DAMPs) from infected cells. During encounters with immune-evasive viruses that block inflammatory gene expression, preformed DAMPs provide backup inflammatory signals that ensure protective immunity. Whether DAMPs exhibit additional backup defense activities is unknown. Herein, we report that viral infection of barrier epithelia (keratinocytes) elicits the release of preformed interleukin-1 (IL-1) family cytokines, including the DAMP IL-1α. Mechanistic studies revealed that IL-1 acts on skin fibroblasts to induce an interferon (IFN)-like state that restricts viral replication. We identified a branch in the IL-1 signaling pathway that induces IFN-stimulated gene expression in infected cells and found that IL-1 signaling is necessary to restrict viral replication in human skin explants. These activities are most important to control immune-evasive virus replication in fibroblasts and other barrier cell types. These findings highlight IL-1 as an important backup antiviral system to ensure barrier defense., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
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26. Antiviral CD8 T cells induce Zika-virus-associated paralysis in mice.
- Author
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Jurado KA, Yockey LJ, Wong PW, Lee S, Huttner AJ, and Iwasaki A
- Subjects
- Animals, Blood-Brain Barrier pathology, Blood-Brain Barrier virology, Brain immunology, Brain pathology, Brain virology, Central Nervous System immunology, Central Nervous System pathology, Central Nervous System virology, Disease Models, Animal, Humans, Mice, Mice, Knockout, Neurons virology, Receptor, Interferon alpha-beta genetics, Zika Virus pathogenicity, Zika Virus Infection pathology, Zika Virus Infection virology, CD8-Positive T-Lymphocytes immunology, Zika Virus drug effects, Zika Virus Infection immunology
- Abstract
Zika virus (ZIKV) is an emerging, mosquito-borne RNA virus. The rapid spread of ZIKV within the Americas has unveiled microcephaly
1 and Guillain-Barré syndrome2,3 as ZIKV-associated neurological complications. Recent reports have also indicated other neurological manifestations to be associated with ZIKV, including myelitis4 , meningoencephalitis5 and fatal encephalitis6 . Here, we investigate the neuropathogenesis of ZIKV infection in type I interferon receptor IFNAR knockout (Ifnar1-/- ) mice, an infection model that exhibits high viral burden within the central nervous system. We show that systemic spread of ZIKV from the site of infection to the brain requires Ifnar1 deficiency in the haematopoietic compartment. However, spread of ZIKV within the central nervous system is supported by Ifnar1-deficient non-haematopoietic cells. Within this context, ZIKV infection of astrocytes results in breakdown of the blood-brain barrier and a large influx of CD8+ effector T cells. We also find that antiviral activity of CD8+ T cells within the brain markedly limits ZIKV infection of neurons, but, as a consequence, instigates ZIKV-associated paralysis. Taken together, our study uncovers mechanisms underlying ZIKV neuropathogenesis within a susceptible mouse model and suggests blood-brain barrier breakdown and T-cell-mediated neuropathology as potential underpinnings of ZIKV-associated neurological complications in humans.- Published
- 2018
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27. Type I interferons instigate fetal demise after Zika virus infection.
- Author
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Yockey LJ, Jurado KA, Arora N, Millet A, Rakib T, Milano KM, Hastings AK, Fikrig E, Kong Y, Horvath TL, Weatherbee S, Kliman HJ, Coyne CB, and Iwasaki A
- Subjects
- Animals, Disease Models, Animal, Female, Fetal Growth Retardation immunology, Fetal Growth Retardation virology, Fetus immunology, Fetus virology, Humans, Male, Mice, Mice, Inbred C57BL, Placenta immunology, Placenta virology, Pregnancy, Receptor, Interferon alpha-beta immunology, Uterus immunology, Uterus virology, Zika Virus Infection virology, Fetal Death etiology, Interferon Type I immunology, Pregnancy Complications, Infectious immunology, Pregnancy Complications, Infectious virology, Zika Virus immunology, Zika Virus Infection immunology
- Abstract
Zika virus (ZIKV) infection during pregnancy is associated with adverse fetal outcomes, including microcephaly, growth restriction, and fetal demise. Type I interferons (IFNs) are essential for host resistance against ZIKV, and IFN-α/β receptor (IFNAR)-deficient mice are highly susceptible to ZIKV infection. Severe fetal growth restriction with placental damage and fetal resorption is observed after ZIKV infection of type I IFN receptor knockout ( Ifnar1
-/- ) dams mated with wild-type sires, resulting in fetuses with functional type I IFN signaling. The role of type I IFNs in limiting or mediating ZIKV disease within this congenital infection model remains unknown. In this study, we challenged Ifnar1-/- dams mated with Ifnar1+/- sires with ZIKV. This breeding scheme enabled us to examine pregnant dams that carry a mixture of fetuses that express ( Ifnar1+/- ) or do not express IFNAR ( Ifnar1-/- ) within the same uterus. Virus replicated to a higher titer in the placenta of Ifnar1-/- than within the Ifnar1+/- concepti. Yet, rather unexpectedly, we found that only Ifnar1+/- fetuses were resorbed after ZIKV infection during early pregnancy, whereas their Ifnar1-/- littermates continue to develop. Analyses of the fetus and placenta revealed that, after ZIKV infection, IFNAR signaling in the conceptus inhibits development of the placental labyrinth, resulting in abnormal architecture of the maternal-fetal barrier. Exposure of midgestation human chorionic villous explants to type I IFN, but not type III IFNs, altered placental morphology and induced cytoskeletal rearrangements within the villous core. Our results implicate type I IFNs as a possible mediator of pregnancy complications, including spontaneous abortions and growth restriction, in the context of congenital viral infections., (Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)- Published
- 2018
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28. Zika virus targets blood monocytes.
- Author
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Jurado KA and Iwasaki A
- Subjects
- Female, Humans, Immune Tolerance, Monocytes, Pregnancy, Immunologic Deficiency Syndromes, Zika Virus, Zika Virus Infection
- Published
- 2017
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29. Nlrp9b inflammasome restricts rotavirus infection in intestinal epithelial cells.
- Author
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Zhu S, Ding S, Wang P, Wei Z, Pan W, Palm NW, Yang Y, Yu H, Li HB, Wang G, Lei X, de Zoete MR, Zhao J, Zheng Y, Chen H, Zhao Y, Jurado KA, Feng N, Shan L, Kluger Y, Lu J, Abraham C, Fikrig E, Greenberg HB, and Flavell RA
- Subjects
- Animals, Apoptosis Regulatory Proteins metabolism, CARD Signaling Adaptor Proteins metabolism, Caspase 1 metabolism, DEAD-box RNA Helicases metabolism, Epithelial Cells metabolism, Female, Immunity, Innate, Inflammasomes chemistry, Inflammasomes genetics, Interleukin-18 immunology, Intestinal Mucosa metabolism, Intestines immunology, Intracellular Signaling Peptides and Proteins, Male, Mice, Mice, Inbred C57BL, Phosphate-Binding Proteins, Pyroptosis, RNA, Double-Stranded metabolism, Receptors, G-Protein-Coupled deficiency, Receptors, G-Protein-Coupled immunology, Rotavirus growth & development, Epithelial Cells immunology, Epithelial Cells virology, Inflammasomes metabolism, Intestines cytology, Receptors, G-Protein-Coupled metabolism, Rotavirus immunology, Rotavirus Infections immunology, Rotavirus Infections virology
- Abstract
Rotavirus, a leading cause of severe gastroenteritis and diarrhoea in young children, accounts for around 215,000 deaths annually worldwide. Rotavirus specifically infects the intestinal epithelial cells in the host small intestine and has evolved strategies to antagonize interferon and NF-κB signalling, raising the question as to whether other host factors participate in antiviral responses in intestinal mucosa. The mechanism by which enteric viruses are sensed and restricted in vivo, especially by NOD-like receptor (NLR) inflammasomes, is largely unknown. Here we uncover and mechanistically characterize the NLR Nlrp9b that is specifically expressed in intestinal epithelial cells and restricts rotavirus infection. Our data show that, via RNA helicase Dhx9, Nlrp9b recognizes short double-stranded RNA stretches and forms inflammasome complexes with the adaptor proteins Asc and caspase-1 to promote the maturation of interleukin (Il)-18 and gasdermin D (Gsdmd)-induced pyroptosis. Conditional depletion of Nlrp9b or other inflammasome components in the intestine in vivo resulted in enhanced susceptibility of mice to rotavirus replication. Our study highlights an important innate immune signalling pathway that functions in intestinal epithelial cells and may present useful targets in the modulation of host defences against viral pathogens.
- Published
- 2017
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30. Fetal Growth Restriction Caused by Sexual Transmission of Zika Virus in Mice.
- Author
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Uraki R, Jurado KA, Hwang J, Szigeti-Buck K, Horvath TL, Iwasaki A, and Fikrig E
- Subjects
- Animals, Disease Models, Animal, Female, Male, Mice, Pregnancy, Sexually Transmitted Diseases, Viral virology, Zika Virus Infection virology, Fetal Growth Retardation virology, Pregnancy Complications, Infectious virology, Sexually Transmitted Diseases, Viral transmission, Zika Virus, Zika Virus Infection transmission
- Abstract
Zika virus (ZIKV) can be transmitted by mosquito bite or sexual contact. Using mice that lack the type I interferon receptor, we examined sexual transmission of ZIKV. Electron microscopy analyses showed association of virions with developing sperm within testes as well as with mature sperm within epididymis. When ZIKV-infected male mice were mated with naive female mice, the weight of fetuses at embryonic day 18.5 was significantly reduced compared with the control group. Additionally, we found ocular deformities in a minority of the fetuses. These results suggest that ZIKV causes fetal abnormalities after female mating with an infected male., (© The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)
- Published
- 2017
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31. Inhibition of HIV-1 Maturation via Small-Molecule Targeting of the Amino-Terminal Domain in the Viral Capsid Protein.
- Author
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Wang W, Zhou J, Halambage UD, Jurado KA, Jamin AV, Wang Y, Engelman AN, and Aiken C
- Subjects
- Amino Acid Substitution genetics, Binding Sites genetics, Capsid Proteins genetics, Cell Line, Tumor, Cell Proliferation, Drug Resistance, Viral genetics, HEK293 Cells, HIV-1 genetics, Humans, Protein Structure, Tertiary, Reverse Transcription drug effects, Anti-HIV Agents pharmacology, Benzimidazoles pharmacology, Capsid metabolism, Capsid Proteins metabolism, HIV-1 growth & development, Virus Assembly drug effects
- Abstract
The human immunodeficiency virus type 1 (HIV-1) capsid protein is an attractive therapeutic target, owing to its multifunctionality in virus replication and the high fitness cost of amino acid substitutions in capsids to HIV-1 infectivity. To date, small-molecule inhibitors have been identified that inhibit HIV-1 capsid assembly and/or impair its function in target cells. Here, we describe the mechanism of action of the previously reported capsid-targeting HIV-1 inhibitor, Boehringer-Ingelheim compound 1 (C1). We show that C1 acts during HIV-1 maturation to prevent assembly of a mature viral capsid. However, unlike the maturation inhibitor bevirimat, C1 did not significantly affect the kinetics or fidelity of Gag processing. HIV-1 particles produced in the presence of C1 contained unstable capsids that lacked associated electron density and exhibited impairments in early postentry stages of infection, most notably reverse transcription. C1 inhibited assembly of recombinant HIV-1 CA in vitro and induced aberrant cross-links in mutant HIV-1 particles capable of spontaneous intersubunit disulfide bonds at the interhexamer interface in the capsid lattice. Resistance to C1 was conferred by a single amino acid substitution within the compound-binding site in the N-terminal domain of the CA protein. Our results demonstrate that the binding site for C1 represents a new pharmacological vulnerability in the capsid assembly stage of the HIV-1 life cycle. IMPORTANCE The HIV-1 capsid protein is an attractive but unexploited target for clinical drug development. Prior studies have identified HIV-1 capsid-targeting compounds that display different mechanisms of action, which in part reflects the requirement for capsid function at both the efferent and afferent phases of viral replication. Here, we show that one such compound, compound 1, interferes with assembly of the conical viral capsid during virion maturation and results in perturbations at a specific protein-protein interface in the capsid lattice. We also identify and characterize a mutation in the capsid protein that confers resistance to the inhibitor. This study reveals a novel mechanism by which a capsid-targeting small molecule can inhibit HIV-1 replication., (Copyright © 2017 American Society for Microbiology.)
- Published
- 2017
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32. Zika virus causes testicular atrophy.
- Author
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Uraki R, Hwang J, Jurado KA, Householder S, Yockey LJ, Hastings AK, Homer RJ, Iwasaki A, and Fikrig E
- Subjects
- Animals, Atrophy, Male, Mice, Mice, Knockout, RNA, Viral genetics, Receptor, Interferon alpha-beta genetics, Receptor, Interferon alpha-beta metabolism, RNA, Viral biosynthesis, Testis metabolism, Testis virology, Testosterone blood, Virus Replication physiology, Zika Virus physiology, Zika Virus Infection blood, Zika Virus Infection genetics, Zika Virus Infection pathology
- Abstract
Zika virus (ZIKV) is an emerging mosquito-borne flavivirus that has recently been found to cause fetal infection and neonatal abnormalities, including microcephaly and neurological dysfunction. ZIKV persists in the semen months after the acute viremic phase in humans. To further understand the consequences of ZIKV persistence in males, we infected Ifnar1
-/- mice via subcutaneous injection of a pathogenic but nonlethal ZIKV strain. ZIKV replication persists within the testes even after clearance from the blood, with interstitial, testosterone-producing Leydig cells supporting virus replication. We found high levels of viral RNA and antigen within the epididymal lumen, where sperm is stored, and within surrounding epithelial cells. Unexpectedly, at 21 days post-infection, the testes of the ZIKV-infected mice were significantly smaller compared to those of mock-infected mice, indicating progressive testicular atrophy. ZIKV infection caused a reduction in serum testosterone, suggesting that male fertility can be affected. Our findings have important implications for nonvector-borne vertical transmission, as well as long-term potential reproductive deficiencies, in ZIKV-infected males.- Published
- 2017
- Full Text
- View/download PDF
33. Zika virus infection of Hofbauer cells.
- Author
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Simoni MK, Jurado KA, Abrahams VM, Fikrig E, and Guller S
- Subjects
- Animals, Dengue Virus physiology, Female, Fetal Development, Humans, Infectious Disease Transmission, Vertical, Macrophages virology, Microcirculation, Placenta virology, Pregnancy, Virus Replication, Zika Virus Infection transmission, Congenital Abnormalities immunology, Macrophages immunology, Placenta immunology, Zika Virus physiology, Zika Virus Infection immunology
- Abstract
Recent studies have linked antenatal infection with Zika virus (ZIKV) with major adverse fetal and neonatal outcomes, including microcephaly. There is a growing consensus for the existence of a congenital Zika syndrome (CZS). Previous studies have indicated that non-placental macrophages play a key role in the replication of dengue virus (DENV), a closely related flavivirus. As the placenta provides the conduit for vertical transmission of certain viruses, and placental Hofbauer cells (HBCs) are fetal-placental macrophages located adjacent to fetal capillaries, it is not surprising that several recent studies have examined infection of HBCs by ZIKV. In this review, we describe congenital abnormalities associated with ZIKV infection, the role of HBCs in the placental response to infection, and evidence for the susceptibility of HBCs to ZIKV infection. We conclude that HBCs may contribute to the spread of ZIKV in placenta and promote vertical transmission of ZIKV, ultimately compromising fetal and neonatal development and function. Current evidence strongly suggests that further studies are warranted to dissect the specific molecular mechanism through which ZIKV infects HBCs and its potential impact on the development of CZS., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2017
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- View/download PDF
34. Zika virus productively infects primary human placenta-specific macrophages.
- Author
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Jurado KA, Simoni MK, Tang Z, Uraki R, Hwang J, Householder S, Wu M, Lindenbach BD, Abrahams VM, Guller S, and Fikrig E
- Abstract
The strong association of Zika virus infection with congenital defects has led to questions of how a flavivirus is capable of crossing the placental barrier to reach the fetal brain. Here, we demonstrate permissive Zika virus infection of primary human placental macrophages, commonly referred to as Hofbauer cells, and placental villous fibroblasts. We also demonstrate Zika virus infection of Hofbauer cells within the context of the tissue ex vivo using term placental villous explants. In addition to amplifying infectious virus within a usually inaccessible area, the putative migratory activities of Hofbauer cells may aid in dissemination of Zika virus to the fetal brain. Understanding the susceptibility of placenta-specific cell types will aid future work around and understanding of Zika virus-associated pregnancy complications., Competing Interests: The authors have declared that no conflict of interest exists.
- Published
- 2016
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35. Genetics of War and Truce between Mosquitos and Emerging Viruses.
- Author
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Hwang J, Jurado KA, and Fikrig E
- Subjects
- Alphavirus pathogenicity, Animals, Flavivirus pathogenicity, Humans, Orthobunyavirus pathogenicity, Alphavirus genetics, Culicidae genetics, Culicidae virology, Flavivirus genetics, Insect Vectors genetics, Insect Vectors virology, Orthobunyavirus genetics
- Abstract
Arboviruses have made unexpected reappearances in recent years. Unlike viruses that undergo direct transmission, arboviruses utilize an arthropod vector (e.g., mosquitos, sandflies, and ticks) to spread throughout human populations. Here, we provide a snapshot of mosquito susceptibility to viral infection using flaviviruses, alphaviruses, and bunyaviruses as examples of emerging pathogens of global health relevance., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
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36. VII. GENERAL DISCUSSION.
- Author
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Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Age Factors, Female, Humans, Infant, Longitudinal Studies, Male, Mother-Child Relations, Neurosecretory Systems, Child Development physiology, Cognition drug effects, Hydrocortisone analysis, Hydrocortisone metabolism
- Published
- 2015
- Full Text
- View/download PDF
37. III. JUSTIFICATION FOR CORTISOL RESPONSE CATEGORIES.
- Author
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Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Humans, Hydrocortisone pharmacology, Infant, Infant, Newborn, Models, Statistical, Research Design, Child Development drug effects, Hydrocortisone metabolism
- Published
- 2015
- Full Text
- View/download PDF
38. IV. LONGITUDINAL ANALYSES OF CORTISOL AND MATERNAL SENSITIVITY.
- Author
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Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Age Factors, Child Development drug effects, Female, Humans, Hydrocortisone pharmacology, Infant, Longitudinal Studies, Male, Models, Statistical, Hydrocortisone metabolism, Mother-Child Relations psychology
- Published
- 2015
- Full Text
- View/download PDF
39. II. GENERAL METHODOLOGY FOR ALL PHASES OF THE LONGITUDINAL STUDY.
- Author
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Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Adolescent, Adult, Child Development physiology, Data Collection methods, Hearing, Humans, Hydrocortisone analysis, Infant, Infant, Newborn, Mother-Child Relations psychology, Video Recording, Young Adult, Child Development drug effects, Hydrocortisone metabolism, Hydrocortisone pharmacology, Longitudinal Studies, Research Design
- Published
- 2015
- Full Text
- View/download PDF
40. VI. MATERNAL SENSITIVITY AND DETECTION OF CHANGE IN AUDITORY AND VISUAL STIMULI.
- Author
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Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Age Factors, Child Development drug effects, Female, Humans, Hydrocortisone metabolism, Infant, Auditory Perception, Mother-Child Relations, Visual Perception
- Published
- 2015
- Full Text
- View/download PDF
41. I. INTRODUCTION.
- Author
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Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Child Development physiology, Cognition drug effects, Emotions, Female, Humans, Hydrocortisone pharmacology, Hypothalamo-Hypophyseal System metabolism, Infant, Infant, Newborn, Learning, Male, Mother-Child Relations psychology, Neurosecretory Systems metabolism, Pituitary-Adrenal System metabolism, Self-Control, Sex Factors, Child Development drug effects, Hydrocortisone metabolism
- Published
- 2015
- Full Text
- View/download PDF
42. V. CORTISOL RESPONSE PATTERNS AND DETECTION OF CHANGE IN AUDITORY AND VISUAL STIMULI.
- Author
-
Thompson LA, Morgan G, Jurado KA, and Gunnar MR
- Subjects
- Adult, Child Development drug effects, Cognition drug effects, Female, Humans, Hydrocortisone pharmacology, Infant, Male, Auditory Perception, Hydrocortisone metabolism, Photic Stimulation
- Published
- 2015
- Full Text
- View/download PDF
43. Distribution and Redistribution of HIV-1 Nucleocapsid Protein in Immature, Mature, and Integrase-Inhibited Virions: a Role for Integrase in Maturation.
- Author
-
Fontana J, Jurado KA, Cheng N, Ly NL, Fuchs JR, Gorelick RJ, Engelman AN, and Steven AC
- Subjects
- Cryoelectron Microscopy, HEK293 Cells, HIV-1 metabolism, Humans, Microscopy, Electron, Transmission, Polymerase Chain Reaction, Virion metabolism, HIV Integrase metabolism, HIV-1 physiology, Nucleocapsid Proteins metabolism, Virion physiology, Virus Assembly physiology
- Abstract
Unlabelled: During virion maturation, HIV-1 capsid protein assembles into a conical core containing the viral ribonucleoprotein (vRNP) complex, thought to be composed mainly of the viral RNA and nucleocapsid protein (NC). After infection, the viral RNA is reverse transcribed into double-stranded DNA, which is then incorporated into host chromosomes by integrase (IN) catalysis. Certain IN mutations (class II) and antiviral drugs (allosteric IN inhibitors [ALLINIs]) adversely affect maturation, resulting in virions that contain "eccentric condensates," electron-dense aggregates located outside seemingly empty capsids. Here we demonstrate that in addition to this mislocalization of electron density, a class II IN mutation and ALLINIs each increase the fraction of virions with malformed capsids (from ∼ 12% to ∼ 53%). Eccentric condensates have a high NC content, as demonstrated by "tomo-bubblegram" imaging, a novel labeling technique that exploits the susceptibility of NC to radiation damage. Tomo-bubblegrams also localized NC inside wild-type cores and lining the spherical Gag shell in immature virions. We conclude that eccentric condensates represent nonpackaged vRNPs and that either genetic or pharmacological inhibition of IN can impair vRNP incorporation into mature cores. Supplying IN in trans as part of a Vpr-IN fusion protein partially restored the formation of conical cores with internal electron density and the infectivity of a class II IN deletion mutant virus. Moreover, the ability of ALLINIs to induce eccentric condensate formation required both IN and viral RNA. Based on these observations, we propose a role for IN in initiating core morphogenesis and vRNP incorporation into the mature core during HIV-1 maturation., Importance: Maturation, a process essential for HIV-1 infectivity, involves core assembly, whereby the viral ribonucleoprotein (vRNP, composed of vRNA and nucleocapsid protein [NC]) is packaged into a conical capsid. Allosteric integrase inhibitors (ALLINIs) affect multiple viral processes. We have characterized ALLINIs and integrase mutants that have the same phenotype. First, by comparing the effects of ALLINIs on several steps of the viral cycle, we show that inhibition of maturation accounts for compound potency. Second, by using cryoelectron tomography, we find that ALLINIs impair conical capsid assembly. Third, by developing tomo-bubblegram imaging, which specifically labels NC protein, we find that ALLINIs block vRNP packaging; instead, vRNPs form "eccentric condensates" outside the core. Fourth, malformed cores, typical of integrase-deleted virus, are partially replaced by conical cores when integrase is supplied in trans. Fifth, vRNA is necessary for ALLINI-induced eccentric condensate formation. These observations suggest that integrase is involved in capsid morphogenesis and vRNP packaging., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
- Published
- 2015
- Full Text
- View/download PDF
44. The mechanism of H171T resistance reveals the importance of Nδ-protonated His171 for the binding of allosteric inhibitor BI-D to HIV-1 integrase.
- Author
-
Slaughter A, Jurado KA, Deng N, Feng L, Kessl JJ, Shkriabai N, Larue RC, Fadel HJ, Patel PA, Jena N, Fuchs JR, Poeschla E, Levy RM, Engelman A, and Kvaratskhelia M
- Subjects
- Cell Line, Crystallography, X-Ray, HIV Integrase chemistry, HIV Integrase genetics, Histidine genetics, Histidine metabolism, Humans, Models, Molecular, Mutant Proteins chemistry, Mutant Proteins genetics, Mutant Proteins metabolism, Protein Binding, Protein Conformation, Acetates metabolism, Drug Resistance, Viral, HIV Integrase metabolism, HIV Integrase Inhibitors metabolism, HIV-1 drug effects, HIV-1 enzymology, Mutation, Missense, Quinolines metabolism
- Abstract
Background: Allosteric HIV-1 integrase (IN) inhibitors (ALLINIs) are an important new class of anti-HIV-1 agents. ALLINIs bind at the IN catalytic core domain (CCD) dimer interface occupying the principal binding pocket of its cellular cofactor LEDGF/p75. Consequently, ALLINIs inhibit HIV-1 IN interaction with LEDGF/p75 as well as promote aberrant IN multimerization. Selection of viral strains emerging under the inhibitor pressure has revealed mutations at the IN dimer interface near the inhibitor binding site., Results: We have investigated the effects of one of the most prevalent substitutions, H171T IN, selected under increasing pressure of ALLINI BI-D. Virus containing the H171T IN substitution exhibited an ~68-fold resistance to BI-D treatment in infected cells. These results correlated with ~84-fold reduced affinity for BI-D binding to recombinant H171T IN CCD protein compared to its wild type (WT) counterpart. However, the H171T IN substitution only modestly affected IN-LEDGF/p75 binding and allowed HIV-1 containing this substitution to replicate at near WT levels. The x-ray crystal structures of BI-D binding to WT and H171T IN CCD dimers coupled with binding free energy calculations revealed the importance of the Nδ- protonated imidazole group of His171 for hydrogen bonding to the BI-D tert-butoxy ether oxygen and establishing electrostatic interactions with the inhibitor carboxylic acid, whereas these interactions were compromised upon substitution to Thr171., Conclusions: Our findings reveal a distinct mechanism of resistance for the H171T IN mutation to ALLINI BI-D and indicate a previously undescribed role of the His171 side chain for binding the inhibitor.
- Published
- 2014
- Full Text
- View/download PDF
45. Engineered hyperactive integrase for concerted HIV-1 DNA integration.
- Author
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Li M, Jurado KA, Lin S, Engelman A, and Craigie R
- Subjects
- Archaeal Proteins metabolism, Base Sequence, DNA, Viral genetics, DNA, Viral metabolism, DNA-Binding Proteins metabolism, HIV Infections genetics, HIV Infections metabolism, HIV Infections virology, HIV Integrase metabolism, HIV-1 physiology, Humans, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Sulfolobus solfataricus metabolism, Archaeal Proteins genetics, DNA-Binding Proteins genetics, HIV Integrase genetics, HIV-1 genetics, Protein Engineering, Sulfolobus solfataricus genetics, Virus Integration
- Abstract
The DNA cutting and joining reactions of HIV-1 integration are catalyzed by integrase (IN), a viral protein that functions as a tetramer bridging the two viral DNA ends (intasome). Two major obstacles for biochemical and structural studies of HIV-1 intasomes are 1) the low efficiency of assembly with oligonucleotide DNA substrates, and 2) the non-specific aggregation of both intasomes and free IN in the reaction mixture. By fusing IN with a small non-specific DNA binding protein, Sulfolobus solfataricus chromosomal protein Sso7d (PDB: 1BNZ), we have engineered a highly soluble and hyperactive IN. Unlike wild-type IN, it efficiently catalyzes intasome assembly and concerted integration with oligonucleotide DNA substrates. The fusion IN protein also functions to integrate viral reverse transcripts during HIV-infection. The hyperactive HIV-1 IN may assist in facilitating future biochemical and structural studies of HIV-1 intasomes. Understanding the mechanistic basis of the Sso7d-IN fusion protein could provide insight into the factors that have hindered biophysical studies of wild-type HIV-1 IN and intasomes.
- Published
- 2014
- Full Text
- View/download PDF
46. Multimodal mechanism of action of allosteric HIV-1 integrase inhibitors.
- Author
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Jurado KA and Engelman A
- Subjects
- Allosteric Regulation, Allosteric Site, Amino Acid Sequence, Animals, Catalytic Domain, HIV Integrase metabolism, Humans, Intercellular Signaling Peptides and Proteins chemistry, Intercellular Signaling Peptides and Proteins metabolism, Molecular Sequence Data, Protein Binding, Protein Multimerization drug effects, HIV Integrase chemistry, HIV Integrase Inhibitors pharmacology
- Abstract
Integrase (IN) is required for lentivirus replication and is a proven drug target for the prevention of AIDS in HIV-1-infected patients. While clinical strand transfer inhibitors disarm the IN active site, allosteric inhibition of enzyme activity through the disruption of IN-IN protein interfaces holds great therapeutic potential. A promising class of allosteric IN inhibitors (ALLINIs), 2-(quinolin-3-yl) acetic acid derivatives, engage the IN catalytic core domain dimerisation interface at the binding site for the host integration co-factor LEDGF/p75. ALLINIs promote IN multimerisation and, independent of LEDGF/p75 protein, block the formation of the active IN-DNA complex, as well as inhibit the IN-LEDGF/p75 interaction in vitro. Yet, rather unexpectedly, the full inhibitory effect of these compounds is exerted during the late phase of HIV-1 replication. ALLINIs impair particle core maturation as well as reverse transcription and integration during the subsequent round of virus infection. Recapitulating the pleiotropic phenotypes observed with numerous IN mutant viruses, ALLINIs provide insight into underlying aspects of IN biology that extend beyond its catalytic activity. Therefore, in addition to the potential to expand our repertoire of HIV-1 antiretrovirals, ALLINIs afford important structural probes to dissect the multifaceted nature of the IN protein throughout the course of HIV-1 replication.
- Published
- 2013
- Full Text
- View/download PDF
47. Allosteric integrase inhibitor potency is determined through the inhibition of HIV-1 particle maturation.
- Author
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Jurado KA, Wang H, Slaughter A, Feng L, Kessl JJ, Koh Y, Wang W, Ballandras-Colas A, Patel PA, Fuchs JR, Kvaratskhelia M, and Engelman A
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Allosteric Regulation drug effects, Cell Line, Humans, Transcription Factors genetics, Virion genetics, Virion metabolism, Virus Replication physiology, Adaptor Proteins, Signal Transducing metabolism, HIV Integrase metabolism, HIV Integrase Inhibitors chemistry, HIV Integrase Inhibitors pharmacology, HIV-1 physiology, Transcription Factors metabolism, Virus Replication drug effects
- Abstract
Integration is essential for HIV-1 replication, and the viral integrase (IN) protein is an important therapeutic target. Allosteric IN inhibitors (ALLINIs) that engage the IN dimer interface at the binding site for the host protein lens epithelium-derived growth factor (LEDGF)/transcriptional coactivator p75 are an emerging class of small molecule antagonists. Consistent with the inhibition of a multivalent drug target, ALLINIs display steep antiviral dose-response curves ex vivo. ALLINIs multimerize IN protein and concordantly block its assembly with viral DNA in vitro, indicating that the disruption of two integration-associated functions, IN catalysis and the IN-LEDGF/p75 interaction, determines the multimode mechanism of ALLINI action. We now demonstrate that ALLINI potency is unexpectedly accounted for during the late phase of HIV-1 replication. The compounds promote virion IN multimerization and, reminiscent of class II IN mutations, block the formation of the electron-dense viral core and inhibit reverse transcription and integration in subsequently infected target cells. Mature virions are recalcitrant to ALLINI treatment, and compound potency during virus production is independent of the level of LEDGF/p75 expression. We conclude that cooperative multimerization of IN by ALLINIs together with the inability for LEDGF/p75 to effectively engage the virus during its egress from cells underscores the multimodal mechanism of ALLINI action. Our results highlight the versatile nature of allosteric inhibitors to primarily inhibit viral replication at a step that is distinct from the catalytic requirement for the target enzyme. The vulnerability of IN to small molecules during the late phase of HIV-1 replication unveils a pharmacological Achilles' heel for exploitation in clinical ALLINI development.
- Published
- 2013
- Full Text
- View/download PDF
48. HRP2 determines the efficiency and specificity of HIV-1 integration in LEDGF/p75 knockout cells but does not contribute to the antiviral activity of a potent LEDGF/p75-binding site integrase inhibitor.
- Author
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Wang H, Jurado KA, Wu X, Shun MC, Li X, Ferris AL, Smith SJ, Patel PA, Fuchs JR, Cherepanov P, Kvaratskhelia M, Hughes SH, and Engelman A
- Subjects
- Acetates pharmacology, Animals, Binding Sites, Cell Line, Fibroblasts metabolism, Fibroblasts virology, HIV-1 drug effects, HIV-1 genetics, Humans, Intercellular Signaling Peptides and Proteins genetics, Intercellular Signaling Peptides and Proteins metabolism, Mice, Mice, Knockout, Quinolines pharmacology, HIV Integrase Inhibitors pharmacology, HIV-1 physiology, Intercellular Signaling Peptides and Proteins physiology, Virus Integration
- Abstract
The binding of integrase (IN) to lens epithelium-derived growth factor (LEDGF)/p75 in large part determines the efficiency and specificity of HIV-1 integration. However, a significant residual preference for integration into active genes persists in Psip1 (the gene that encodes for LEDGF/p75) knockout (KO) cells. One other cellular protein, HRP2, harbors both the PWWP and IN-binding domains that are important for LEDGF/p75 co-factor function. To assess the role of HRP2 in HIV-1 integration, cells generated from Hdgfrp2 (the gene that encodes for HRP2) and Psip1/Hdgfrp2 KO mice were infected alongside matched control cells. HRP2 depleted cells supported normal infection, while disruption of Hdgfrp2 in Psip1 KO cells yielded additional defects in the efficiency and specificity of integration. These deficits were largely restored by ectopic expression of either LEDGF/p75 or HRP2. The double-KO cells nevertheless supported residual integration into genes, indicating that IN and/or other host factors contribute to integration specificity in the absence of LEDGF/p75 and HRP2. Psip1 KO significantly increased the potency of an allosteric inhibitor that binds the LEDGF/p75 binding site on IN, a result that was not significantly altered by Hdgfrp2 disruption. These findings help to rule out the host factor-IN interactions as the primary antiviral targets of LEDGF/p75-binding site IN inhibitors.
- Published
- 2012
- Full Text
- View/download PDF
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