| 1. |
- Guimond, S. E., et al.
(författare)
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Synthetic Heparan Sulfate Mimetic Pixatimod (PG545) Potently Inhibits SARS-CoV-2 by Disrupting the Spike-ACE2 Interaction
- 2022
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Ingår i: ACS Central Science. - : American Chemical Society (ACS). - 2374-7943 .- 2374-7951. ; 8:5, s. 527-545
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Tidskriftsartikel (refereegranskat)abstract
- Heparan sulfate (HS) is a cell surface polysaccharide recently identified as a coreceptor with the ACE2 protein for the S1 spike protein on SARS-CoV-2 virus, providing a tractable new therapeutic target. Clinically used heparins demonstrate an inhibitory activity but have an anticoagulant activity and are supply-limited, necessitating alternative solutions. Here, we show that synthetic HS mimetic pixatimod (PG545), a cancer drug candidate, binds and destabilizes the SARS-CoV-2 spike protein receptor binding domain and directly inhibits its binding to ACE2, consistent with molecular modeling identification of multiple molecular contacts and overlapping pixatimod and ACE2 binding sites. Assays with multiple clinical isolates of SARS-CoV-2 virus show that pixatimod potently inhibits the infection of monkey Vero E6 cells and physiologically relevant human bronchial epithelial cells at safe therapeutic concentrations. Pixatimod also retained broad potency against variants of concern (VOC) including B.1.1.7 (Alpha), B.1.351 (Beta), B.1.617.2 (Delta), and B.1.1.529 (Omicron). Furthermore, in a K18-hACE2 mouse model, pixatimod significantly reduced SARS-CoV-2 viral titers in the upper respiratory tract and virus-induced weight loss. This demonstration of potent anti-SARS-CoV-2 activity tolerant to emerging mutations establishes proof-of-concept for targeting the HS-Spike protein-ACE2 axis with synthetic HS mimetics and provides a strong rationale for clinical investigation of pixatimod as a potential multimodal therapeutic for COVID-19.
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| 2. |
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| 3. |
- Bagdonaite, I., et al.
(författare)
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A Strategy for O-Glycoproteomics of Enveloped Viruses-the O-Glycoproteome of Herpes Simplex Virus Type 1
- 2015
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Ingår i: Plos Pathogens. - : Public Library of Science (PLoS). - 1553-7366 .- 1553-7374. ; 11:4
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Tidskriftsartikel (refereegranskat)abstract
- Glycosylation of viral envelope proteins is important for infectivity and interaction with host immunity, however, our current knowledge of the functions of glycosylation is largely limited to N-glycosylation because it is difficult to predict and identify site-specific O-glycosylation. Here, we present a novel proteome-wide discovery strategy for O-glycosylation sites on viral envelope proteins using herpes simplex virus type 1 (HSV-1) as a model. We identified 74 O-linked glycosylation sites on 8 out of the 12 HSV-1 envelope proteins. Two of the identified glycosites found in glycoprotein B were previously implicated in virus attachment to immune cells. We show that HSV-1 infection distorts the secretory pathway and that infected cells accumulate glycoproteins with truncated O-glycans, nonetheless retaining the ability to elongate most of the surface glycans. With the use of precise gene editing, we further demonstrate that elongated O-glycans are essential for HSV-1 in human HaCaT keratinocytes, where HSV-1 produced markedly lower viral titers in HaCaT with abrogated O-glycans compared to the isogenic counterpart with normal O-glycans. The roles of O-linked glycosylation for viral entry, formation, secretion, and immune recognition are poorly understood, and the O-glycoproteomics strategy presented here now opens for unbiased discovery on all enveloped viruses.
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| 4. |
- Bagdonaite, I., et al.
(författare)
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Global Mapping of O-Glycosylation of Varicella Zoster Virus, Human Cytomegalovirus, and Epstein-Barr Virus
- 2016
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Ingår i: Journal of Biological Chemistry. - 0021-9258. ; 291:23, s. 12014-12028
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Tidskriftsartikel (refereegranskat)abstract
- Herpesviruses are among the most complex and widespread viruses, infection and propagation of which depend on envelope proteins. These proteins serve as mediators of cell entry as well as modulators of the immune response and are attractive vaccine targets. Although envelope proteins are known to carry glycans, little is known about the distribution, nature, and functions of these modifications. This is particularly true for O-glycans; thus we have recently developed a "bottom up" mass spectrometry-based technique for mapping O-glycosylation sites on herpes simplex virus type 1. We found wide distribution of O-glycans on herpes simplex virus type 1 glycoproteins and demonstrated that elongated O-glycans were essential for the propagation of the virus. Here, we applied our proteome-wide discovery platform for mapping O-glycosites on representative and clinically significant members of the herpesvirus family: varicella zoster virus, human cytomegalovirus, and Epstein-Barr virus. We identified a large number of O-glycosites distributed on most envelope proteins in all viruses and further demonstrated conserved patterns of O-glycans on distinct homologous proteins. Because glycosylation is highly dependent on the host cell, we tested varicella zoster virus-infected cell lysates and clinically isolated virus and found evidence of consistent O-glycosites. These results present a comprehensive view of herpesvirus O-glycosylation and point to the widespread occurrence of O-glycans in regions of envelope proteins important for virus entry, formation, and recognition by the host immune system. This knowledge enables dissection of specific functional roles of individual glycosites and, moreover, provides a framework for design of glycoprotein vaccines with representative glycosylation.
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| 5. |
- Iversen, M. B., et al.
(författare)
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An innate antiviral pathway acting before interferons at epithelial surfaces
- 2016
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Ingår i: Nature Immunology. - : Springer Science and Business Media LLC. - 1529-2908 .- 1529-2916. ; 17:2, s. 150-158
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Tidskriftsartikel (refereegranskat)abstract
- Mucosal surfaces are exposed to environmental substances and represent a major portal of entry for microorganisms. The innate immune system is responsible for early defense against infections and it is believed that the interferons (IFNs) constitute the first line of defense against viruses. Here we identify an innate antiviral pathway that works at epithelial surfaces before the IFNs. The pathway is activated independently of known innate sensors of viral infections through a mechanism dependent on viral O-linked glycans, which induce CXCR3 chemokines and stimulate antiviral activity in a manner dependent on neutrophils. This study therefore identifies a previously unknown layer of antiviral defense that exerts its action on epithelial surfaces before the classical IFN response is operative.
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| 6. |
- Kanekar, N., et al.
(författare)
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Constraints on changes in the proton-electron mass ratio using methanol lines
- 2015
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Ingår i: Monthly Notices of the Royal Astronomical Society: Letters. - : Oxford University Press (OUP). - 1745-3925 .- 1745-3933. ; 448:1, s. L104-L108
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Tidskriftsartikel (refereegranskat)abstract
- We report Karl G. Jansky Very Large Array (VLA) absorption spectroscopy in four methanol (CH3OH) lines in the z = 0.885 82 gravitational lens towards PKS1830-211. Three of the four lines have very different sensitivity coefficients K-mu to changes in the proton-electron mass ratio mu; a comparison between the line redshifts thus allows us to test for temporal evolution in mu. We obtain a stringent statistical constraint on changes in mu by comparing the redshifted 12.179 and 60.531 GHz lines, [Delta mu/mu] 4 sigma significance). The sensitivity of the above result, and that of all earlier CH3OH studies, is thus likely to be limited by unknown systematic errors, probably arising due to the frequency-dependent structure of PKS1830-211. A robust result is obtained by combining the three lines at similar frequencies, 48.372, 48.377 and 60.531 GHz, whose line profiles are found to be in good agreement. This yields the 2 sigma constraint [Delta mu/mu] less than or similar to 4 x 10(-7), the most stringent current constraint on changes in mu. We thus find no evidence for changes in the proton-electron mass ratio over a lookback time of approximate to 7.5 Gyr.
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| 7. |
- Schulz, A., et al.
(författare)
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The inhomogeneous ISM toward PKS 1830–211 SW: A detailed view of molecular gas at a look-back time of 7.5 Gyr
- 2015
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Ingår i: Astronomy and Astrophysics. - : EDP Sciences. - 0004-6361 .- 1432-0746. ; 574, s. 108-
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Tidskriftsartikel (refereegranskat)abstract
- Based on measurements with the Effelsberg 100-m telescope, a multiline study of molecular species is presented toward the southwestern source of the gravitational lens system PKS 1830–211, which is by far the best known target for studying molecular gas in absorption at intermediate redshift. Determining line parameters and optical depths and performing large velocity gradient radiative transfer calculations, the aims of this study are (1) to evaluate physical parameters of the absorbing foreground gas at z ~ 0.89, in particular its homogeneity; and (2) to monitor the spectroscopic time variability caused by fluctuations in the z ~ 2.5 background continuum source. We find, that the gas is quite inhomogeneous with n(H2) ~ 2 × 103 cm-3 for most molecular species but with higher values for H2CO and lower ones for SO. Measuring the CS J = 1 ← 0 transition during a time interval of more than a decade, from 2001 to 2012, the peak absorption depth of the line remains approximately constant, while the line shape undergoes notable variations. Covering the time between 1996 and 2013, CS, HCO+, and CH3OH data indicate maximum integrated optical depths in ~2001 and 2011/2012. This is compatible with a ~10 yr periodicity, which, however, needs confirmation by substantially longer time monitoring. Comparing molecular abundances with those of different types of Galactic and nearby extragalactic clouds we find that the observed cloud complex does not correspond to one particular type but to a variety of cloud types with more diffuse and denser components as can be expected for an observed region with a transverse linear scale of several parsec and a likely greater depth along the line of sight. A tentative detection of Galactic absorption in the c-C3H2 110−101 line at 18.343 GHz is also reported.
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