Active Components of Commonly Prescribed Medicines Affect Influenza A Virus-Host Cell Interaction : A Pilot Study

Background: Every year, millions of people are hospitalized and thousands die from influenza A virus (FLUAV) infection. Most cases of hospitalizations and death occur among the elderly. Many of these elderly patients are reliant on medical treatment of underlying chronic diseases, such as arthritis, diabetes, and hypertension. We hypothesized that the commonly prescribed medicines for treatment of underlying chronic diseases can affect host responses to FLUAV infection and thus contribute to the morbidity and mortality associated with influenza. Therefore, the aim of this study was to examine whether commonly prescribed medicines could affect host responses to virus infection in vitro. Methods: We first identified 45 active compounds from a list of commonly prescribed medicines. Then, we constructed a drug-target interaction network and identified the potential implication of these interactions for FLUAV-host cell interplay. Finally, we tested the effect of 45 drugs on the viability, transcription, and metabolism of mock- and FLUAV-infected human retinal pigment epithelial (RPE) cells. Results: In silico drug-target interaction analysis revealed that drugs such as atorvastatin, candesartan, and hydroxocobalamin could target and modulate FLUAV-host cell interaction. In vitro experiments showed that at non-cytotoxic concentrations, these compounds affected the transcription and metabolism of FLUAV- and mock-infected cells. Conclusion: Many commonly prescribed drugs were found to modulate FLUAV-host cell interactions in silico and in vitro and could therefore affect their interplay in vivo, thus contributing to the morbidity and mortality of patients with influenza virus infections.Peer reviewe

Leaky severe combined immunodeficiency in mice lacking non-homologous end joining factors XLF and MRI

Non-homologous end-joining (NHEJ) is a DNA repair pathway required to detect, process, and ligate DNA double-stranded breaks (DSBs) throughout the cell cycle. The NHEJ pathway is necessary for V(D)J recombination in developing B and T lymphocytes. During NHEJ, Ku70 and Ku80 form a heterodimer that recognizes DSBs and promotes recruitment and function of downstream factors PAXX, MRI, DNA-PKcs, Artemis, XLF, XRCC4, and LIG4. Mutations in several known NHEJ genes result in severe combined immunodeficiency (SCID). Inactivation of Mri, Paxx or Xlf in mice results in normal or mild phenotype, while combined inactivation of Xlf/Mri, Xlf/Paxx, or Xlf/Dna-pkcs leads to late embryonic lethality. Here, we describe three new mouse models. We demonstrate that deletion of Trp53 rescues embryonic lethality in mice with combined deficiencies of Xlf and Mri. Furthermore, Xlf-/-Mri-/-Trp53+/- and Xlf-/-Paxx-/-Trp53+/- mice possess reduced body weight, severely reduced mature lymphocyte counts, and accumulation of progenitor B cells. We also report that combined inactivation of Mri/Paxx results in live-born mice with modest phenotype, and combined inactivation of Mri/Dna-pkcs results in embryonic lethality. Therefore, we conclude that XLF is functionally redundant with MRI and PAXX during lymphocyte development in vivo. Moreover, Mri genetically interacts with Dna-pkcs and Paxx

Mono- and combinational drug therapies for global viral pandemic preparedness

Broadly effective antiviral therapies must be developed to be ready for clinical trials, which should begin soon after the emergence of new life-threatening viruses. Here, we pave the way towards this goal by reviewing conserved druggable virus-host interactions, mechanisms of action, immunomodulatory properties of available broad-spectrum antivirals (BSAs), routes of BSA delivery, and interactions of BSAs with other antivirals. Based on the review, we concluded that the range of indications of BSAs can be expanded, and new pan- and cross-viral mono- and combinational therapies can be developed. We have also developed a new scoring algorithm that can help identify the most promising few of the thousands of potential BSAs and BSA-containing drug cocktails (BCCs) to prioritize their development during the critical period between the identification of a new virus and the development of virus-specific vaccines, drugs, and therapeutic antibodies.Peer reviewe

Evidence for Warped Disks of Young Stars in the Galactic Center

The central parsec around the super-massive black hole in the Galactic Center hosts more than 100 young and massive stars. Outside the central cusp (R~1") the majority of these O and Wolf-Rayet (WR) stars reside in a main clockwise system, plus a second, less prominent disk or streamer system at large angles with respect to the main system. Here we present the results from new observations of the Galactic Center with the AO-assisted near-infrared imager NACO and the integral field spectrograph SINFONI on the ESO/VLT. These include the detection of 27 new reliably measured WR/O stars in the central 12" and improved measurements of 63 previously detected stars, with proper motion uncertainties reduced by a factor of four compared to our earlier work. We develop a detailed statistical analysis of their orbital properties and orientations. Half of the WR/O stars are compatible with being members of a clockwise rotating system. The rotation axis of this system shows a strong transition as a function of the projected distance from SgrA*. The main clockwise system either is either a strongly warped single disk with a thickness of about 10 degrees, or consists of a series of streamers with significant radial variation in their orbital planes. 11 out of 61 clockwise moving stars have an angular separation of more than 30 degrees from the clockwise system. The mean eccentricity of the clockwise system is 0.36+/-0.06. The distribution of the counter-clockwise WR/O star is not isotropic at the 98% confidence level. It is compatible with a coherent structure such as stellar filaments, streams, small clusters or possibly a disk in a dissolving state. The observed disk warp and the steep surface density distribution favor in situ star formation in gaseous accretion disks as the origin of the young stars.Comment: ApJ in pres

Mmp17-deficient mice exhibit heightened goblet cell effector expression in the colon and increased resistance to chronic Trichuris muris infection

Intestinal epithelial homeostasis is maintained by intrinsic and extrinsic signals. The extrinsic signals include those provided by mesenchymal cell populations that surround intestinal crypts and is further facilitated by the extracellular matrix (ECM), which is modulated by proteases such as matrix metalloproteinases (MMPs). Extrinsic signals ensure an appropriate balance between intestinal epithelial proliferation and differentiation. This study explores the role of MMP17, which is preferentially expressed by smooth muscle cells in the intestine, in intestinal homeostasis and during immunity to infection. Mice lacking MMP17 expressed high levels of goblet-cell associated genes and proteins, such as CLCA1 and RELM-β, which are normally associated with immune responses to infection. Nevertheless, Mmp17 KO mice did not have altered resistance during a bacterial Citrobacter rodentium infection. However, when challenged with a low dose of the helminth Trichuris muris, Mmp17 KO mice had increased resistance, without a clear role for an altered immune response during infection. Mechanistically, we did not find changes in traditional modulators of goblet cell effectors such as the NOTCH pathway or specific cytokines. We found MMP17 expression in smooth muscle cells as well as lamina propria cells such as macrophages. Together, our data suggest that MMP17 extrinsically alters goblet cell maturation which is sufficient to alter clearance in a helminth infection model

Synergistic Interferon-Alpha-Based Combinations for Treatment of SARS-CoV-2 and Other Viral Infections

Background: There is an urgent need for new antivirals with powerful therapeutic potential and tolerable side effects. Methods: Here, we tested the antiviral properties of interferons (IFNs), alone and with other drugs in vitro. Results: While IFNs alone were insufficient to completely abolish replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), IFNα, in combination with remdesivir, EIDD-2801, camostat, cycloheximide, or convalescent serum, proved to be more effective. Transcriptome and metabolomic analyses revealed that the IFNα–remdesivir combination suppressed SARS-CoV-2-mediated changes in Calu-3 cells and lung organoids, although it altered the homeostasis of uninfected cells and organoids. We also demonstrated that IFNα combinations with sofosbuvir, telaprevir, NITD008, ribavirin, pimodivir, or lamivudine were effective against HCV, HEV, FLuAV, or HIV at lower concentrations, compared to monotherapies. Conclusions: Altogether, our results indicated that IFNα can be combined with drugs that affect viral RNA transcription, protein synthesis, and processing to make synergistic combinations that can be attractive targets for further pre-clinical and clinical development against emerging and re-emerging viral infections

INSL6 Protective Effects in Heart Failure

Background The insulin/insulin‐like growth factor/relaxin family represents a group of structurally related but functionally diverse proteins. The family member relaxin‐2 has been evaluated in clinical trials for its efficacy in the treatment of acute heart failure. In this study, we assessed the role of insulin‐like peptide 6 (INSL6), another member of this protein family, in murine heart failure models using genetic loss‐of‐function and protein delivery methods. Methods and Results Insl6‐deficient and wild‐type (C57BL/6N) mice were administered angiotensin II or isoproterenol via continuous infusion with an osmotic pump or via intraperitoneal injection once a day, respectively, for 2 weeks. In both models, Insl6‐knockout mice exhibited greater cardiac systolic dysfunction and left ventricular dilatation. Cardiac dysfunction in the Insl6‐knockout mice was associated with more extensive cardiac fibrosis and greater expression of fibrosis‐associated genes. The continuous infusion of chemically synthesized INSL6 significantly attenuated left ventricular systolic dysfunction and cardiac fibrosis induced by isoproterenol infusion. Gene expression profiling suggests liver X receptor/retinoid X receptor signaling is activated in the isoproterenol‐challenged hearts treated with INSL6 protein. Conclusions Endogenous Insl6 protein inhibits cardiac systolic dysfunction and cardiac fibrosis in angiotensin II– and isoproterenol‐induced cardiac stress models. The administration of recombinant INSL6 protein could have utility for the treatment of heart failure and cardiac fibrosis

The G9.62+0.19-F Hot Molecular Core - The infrared view on very young massive stars

(abridged) We present the results of an extensive infrared study of the massive star-forming region G9.62+0.19. The data cover information from broad- and narrow-band filters in the wavelength range from 1 to 19 micrometer and are obtained with ESO's infrared cameras ISAAC and TIMMI2 and with SpectroCam-10 (Mt. Palomar). The high sensitivity and resolution provided by these facilities revealed intriguing new details of this star-forming region and especially about the embedded hot molecular core (HMC) - component F. We analyse the newly found infrared sub-structure of four objects in this HMC region. While one of these objects (F2) is probably a foreground field star, the nature of the brightest object in the near-infrared there (F1) remains somewhat enigmatic. Our new astrometry proves that this object is not coincident with the peak of the molecular line emission of the HMC, but displaced by 1.7 arcsecs (nearly 10000 AU on a linear scale). We estimate this object to be an additional embedded object with a dense dust shell. Very near the HMC location we find L' band emission which strongly rises in flux towards longer wavelengths. We presume that this emission (F4) arises from the envelope of the HMC which is known to be associated with a molecular outflow roughly aligned along the line of sight. Thus, the clearing effect of this outflow causes strong deviations from spherical symmetry which might allow infrared emission from the HMC to escape through the outflow cavities. This presents the first direct detection of an HMC at a wavelength as short as 3.8 micron. At 11.7 and 18.75 micron, the HMC counterpart F4 ultimately proves to be the most luminous IR source within the G9.62+0.19-F region.Comment: 39 pages, 10 figures, accepted for publication in Astronomy & Astrophysics. The paper contains several colour images. It is highly advisable to go to the following website to get a high-resolution version of this preprint: http://www.tls-tautenburg.de/research/tls-research/papers/linz/G9.62.htm

Discovery of Broad-Spectrum Antiviral Agents by Drug Repurposing for Rapid Response to Emergent Viral Pandemics: Building Digital Tools to Advance Translational Research

The ongoing COVID-19 pandemic caused by the emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in 2019 has highlighted urgency for a more efficient and comprehensive paradigm for antiviral development. The majority of antiviral development in the past has relied on the ‘one drug, one virus’ model, which focuses on a method of drug discovery wherein one drug exploits the biological mechanisms of one specific species of virus. However, this is an inefficient method for drug development which often dissuades many pharmaceutical companies from investing resources towards antiviral discovery. Broad spectrum antivirals agents (BSAAs) are compounds that have antiviral activity against a broad range of viruses. They have become the subject of increasing attention in antiviral development because of the advantages they confer in potentially treating multiple viral infections with one drug. Drug repurposing is also an increasingly utilized technique in antiviral development that relies on the discovery of new indications for previously developed drugs, and offers the advantage of saving time and money in the drug development process. These two approaches to antiviral drug discovery can be combined to drastically cut down on time, money, and other resource requirements of traditional drug development. If these techniques are successful, they could yield a broad arsenal of antiviral drugs that will not only help combat current viral epidemics, but also increase global preparedness for emerging viruses in the future. In this master’s thesis, I intend to describe my efforts that contributed to the building of a web-based database, DrugVirus.info, that aggregates information to promote discovery and development of novel BSAA candidates. Additionally, I will present SARS-Coronavirus-2.info, a highly accessible, multilingual website which served as a method for rapid dissemination of information in the early days of the COVID-19 pandemic. Finally, using information gathered from work on both resources, I will discuss the impact of each in the context of the emergence of SARS-CoV-2, as well as how the pharmaceutical response to COVID-19 will impact the future of BSAA development

Nafamostat-Interferon-alpha Combination Suppresses SARS-CoV-2 Infection In Vitro and In Vivo by Cooperatively Targeting Host TMPRSS2

SARS-CoV-2 and its vaccine/immune-escaping variants continue to pose a serious threat to public health due to a paucity of effective, rapidly deployable, and widely available treatments. Here, we address these challenges by combining Pegasys (IFN alpha) and nafamostat to effectively suppress SARS-CoV-2 infection in cell culture and hamsters. Our results indicate that Serpin E1 is an important mediator of the antiviral activity of IFN alpha and that both Serpin E1 and nafamostat can target the same cellular factor TMPRSS2, which plays a critical role in viral replication. The low doses of the drugs in combination may have several clinical advantages, including fewer adverse events and improved patient outcome. Thus, our study may provide a proactive solution for the ongoing pandemic and potential future coronavirus outbreaks, which is still urgently required in many parts of the world.Peer reviewe