Humanized mice reveal a macrophage-enriched gene signature defining human lung tissue protection during SARS-CoV-2 infection. Kenney et al.

Summary of the study: The human immunological mechanisms defining the clinical outcome of SARS-CoV-2 infection remain elusive. This knowledge gap is mostly driven by the lack of appropriate experimental platforms recapitulating human immune responses in a controlled human lung environment. Here, we report a mouse model (i.e. HNFL mice) co-engrafted with human fetal lung xenografts (fLX) and a myeloid-enhanced human immune system to identify cellular and molecular correlates of lung protection during SARS-CoV-2 infection. Unlike mice solely engrafted with human fLX, HNFL mice are protected against infection, severe inflammation, and histopathological phenotypes. Lung tissue protection from infection and severe histopathology associate with macrophage infiltration and differentiation, and the upregulation of a macrophage-enriched signature composed of eleven specific genes mainly associated with the type I interferon signaling pathway. Our work highlights the HNFL model as a transformative platform to investigate, in controlled experimental setting, human myeloid immune mechanisms governing lung tissue protection during SARS-CoV-2 infection. This Mendeley dataset contains all of the Supplemental Items associated with this study. This includes the following:- Table S1. Related to Figure 1,4 and 6. Single-cell RNA sequencing gene-defining clusters and cluster annotation in naïve NRGL-LX and HNFL-LX, and in inoculated HNFL-LX (Excel file). - Table S2. Related to Figure 1 and 5. Proteomic analysis Matrix: Naive NRGL-LX vs. HNFL-LX, Naïve HNFL-LX vs. 2DPI HNFL0LX, Naïve NRGL-LX vs. 2DPI NRGL-LX (Excel file).- Table S3. Related to Figure 5. Phospho-proteomics analysis Matrix: Naïve HNFL-LX vs. 2DPI HNFL0LX, Naïve NRGL-LX vs. 2DPI NRGL-LX (Excel file).- Table S4. Related to Figure 5. List of differentially expressed genes and IPA scores from bulk RNA sequencing analysis of naïve and inoculated NRGL-LX (Excel file).- Table S5. Related to all figures. List of the mice and fetal donor ID used in this study (Excel file).- Movie S1. Related to Figure 2. Time-lapse (2DPI, 4DPI, 6DPI and 12DPI) 3D in vivo imaging of SARS-CoV-2 infection in NRGL mice (mp4 file).THIS DATASET IS ARCHIVED AT DANS/EASY, BUT NOT ACCESSIBLE HERE. TO VIEW A LIST OF FILES AND ACCESS THE FILES IN THIS DATASET CLICK ON THE DOI-LINK ABOV

Humanized mice reveal a macrophage-enriched gene signature defining human lung tissue protection during SARS-CoV-2 infection. Kenney et al.

Summary of the study: The human immunological mechanisms defining the clinical outcome of SARS-CoV-2 infection remain elusive. This knowledge gap is mostly driven by the lack of appropriate experimental platforms recapitulating human immune responses in a controlled human lung environment. Here, we report a mouse model (i.e. HNFL mice) co-engrafted with human fetal lung xenografts (fLX) and a myeloid-enhanced human immune system to identify cellular and molecular correlates of lung protection during SARS-CoV-2 infection. Unlike mice solely engrafted with human fLX, HNFL mice are protected against infection, severe inflammation, and histopathological phenotypes. Lung tissue protection from infection and severe histopathology associate with macrophage infiltration and differentiation, and the upregulation of a macrophage-enriched signature composed of eleven specific genes mainly associated with the type I interferon signaling pathway. Our work highlights the HNFL model as a transformative platform to investigate, in controlled experimental setting, human myeloid immune mechanisms governing lung tissue protection during SARS-CoV-2 infection. This Mendeley dataset contains all of the Supplemental Items associated with this study. This includes the following:- Spreadsheet S1. Related to Figure 1,4 and 6. Single-cell RNA sequencing gene-defining clusters and cluster annotation in naïve NRGL-LX and HNFL-LX, and in inoculated HNFL-LX (Excel file). - Spreadsheet S2. Related to Figure 1 and 5. Proteomic analysis Matrix: Naive NRGL-LX vs. HNFL-LX, Naïve HNFL-LX vs. 2DPI HNFL0LX, Naïve NRGL-LX vs. 2DPI NRGL-LX (Excel file).- Spreadsheet S3. Related to Figure 5. Phospho-proteomics analysis Matrix: Naïve HNFL-LX vs. 2DPI HNFL0LX, Naïve NRGL-LX vs. 2DPI NRGL-LX (Excel file).- Spreadsheet S4. Related to Figure 5. List of differentially expressed genes and IPA scores from bulk RNA sequencing analysis of naïve and inoculated NRGL-LX (Excel file).- Spreadsheet S5. Related to all figures. List of the mice and fetal donor ID used in this study (Excel file).- Movie S1. Related to Figure 2. Time-lapse (2DPI, 4DPI, 6DPI and 12DPI) 3D in vivo imaging of SARS-CoV-2 infection in NRGL mice (mp4 file).THIS DATASET IS ARCHIVED AT DANS/EASY, BUT NOT ACCESSIBLE HERE. TO VIEW A LIST OF FILES AND ACCESS THE FILES IN THIS DATASET CLICK ON THE DOI-LINK ABOV

Humanized mice reveal a macrophage-enriched gene signature defining human lung tissue protection during SARS-CoV-2 infection. Kenney et al.

Summary of the study: The human immunological mechanisms defining the clinical outcome of SARS-CoV-2 infection remain elusive. This knowledge gap is mostly driven by the lack of appropriate experimental platforms recapitulating human immune responses in a controlled human lung environment. Here, we report a mouse model (i.e. HNFL mice) co-engrafted with human fetal lung xenografts (fLX) and a myeloid-enhanced human immune system to identify cellular and molecular correlates of lung protection during SARS-CoV-2 infection. Unlike mice solely engrafted with human fLX, HNFL mice are protected against infection, severe inflammation, and histopathological phenotypes. Lung tissue protection from infection and severe histopathology associate with macrophage infiltration and differentiation, and the upregulation of a macrophage-enriched signature composed of eleven specific genes mainly associated with the type I interferon signaling pathway. Our work highlights the HNFL model as a transformative platform to investigate, in controlled experimental setting, human myeloid immune mechanisms governing lung tissue protection during SARS-CoV-2 infection. This Mendeley dataset contains all of the Supplemental Items associated with this study. This includes the following:- Spreadsheet S1. Single-cell RNA sequencing gene-defining clusters and cluster annotation (Excel file).- Spreadsheet S2. Proteomic analysis Matrix; _NRG-L vs. HNFL (Excel file)- Movie S1. Time-lapse (2DPI, 4DPI, 6DPI and 12DPI) 3D in vivo imaging of SARS-CoV-2 infection (mp4 file)- Spreadsheet S3. Phospho-proteomics analysis Matrix; _NRG-L vs. HNFL (Excel file)- Spreadsheet S4. List of differentially expressed genes and IPA scores from bulk RNA sequencing analysis (Excel file).- Spreadsheet S5. List of the mice and fetal donor ID used in this study.THIS DATASET IS ARCHIVED AT DANS/EASY, BUT NOT ACCESSIBLE HERE. TO VIEW A LIST OF FILES AND ACCESS THE FILES IN THIS DATASET CLICK ON THE DOI-LINK ABOV

Critical interaction between E1 and E2 glycoproteins determines binding and fusion properties of hepatitis C virus during cell entry

International audienceHepatitis C virus (HCV) envelope glycoproteins E1 and E2 are important mediators for productive cell entry. However, knowledge about their structure, intra- or intermolecular dialogs, and conformational changes is scarce, limiting the design of therapeutic strategies targeting E1E2. Here we sought to investigate how certain domains of E1 and E2 have coevolved to optimize their interactions to promote efficient HCV entry. For this purpose we generated chimeric E1E2 heterodimers derived from two HCV 1a strains to identify and characterize crosstalk between their domains. We found an E1E2 combination that drastically impaired the infectivity of cell culture-derived HCV particles, whereas the reciprocal E1E2 combination led to increased infectivity. Using HCV pseudoparticle assays, we confirmed the opposing entry phenotypes of these heterodimers. By mutagenesis analysis, we identified a particular crosstalk between three amino acids of E1 and the domain III of E2. Its modulation leads to either a full restoration of the functionality of the suboptimal heterodimer or a destabilization of the functional heterodimer. Interestingly, we found that this crosstalk modulates E1E2 binding to HCV entry receptors SR-BI and CD81. In addition, we found for the first time that E1E2 complexes can interact with the first extracellular loop of Claudin-1, whereas soluble E2 did not. These results highlight the critical role of E1 in the modulation of HCV binding to receptors. Finally, we demonstrated that this crosstalk is involved in membrane fusion. Conclusions: These results reveal a multifunctional and crucial interaction between E1 and E2 for HCV entry into cells. Our study highlights the role of E1 as a modulator of HCV binding to receptors and membrane fusion, underlining its potential as an antiviral target. (Hepatology 2014;59:776-788

Isocotoin suppresses hepatitis E virus replication through inhibition of heat shock protein 90

Hepatitis E virus (HEV) causes 14 million infections and 60,000 deaths per year globally, with immunocompromised persons and pregnant women experiencing severe symptoms. Although ribavirin can be used to treat chronic hepatitis E, toxicity in pregnant patients and the emergence of resistant strains are major concerns. Therefore there is an imminent need for effective HEV antiviral agents. The aims of this study were to develop a drug screening platform and to discover novel approaches to targeting steps within the viral life cycle. We developed a screening platform for molecules inhibiting HEV replication and selected a candidate, isocotoin. Isocotoin inhibits HEV replication through interference with heat shock protein 90 (HSP90), a host factor not previously known to be involved in HEV replication. Additional work is required to understand the compound’s translational potential, however this suggests that HSP90-modulating molecules, which are in clinical development as anti-cancer agents, may be promising therapies against HEV

A protein coevolution method uncovers critical features of the Hepatitis C Virus fusion mechanism.

Amino-acid coevolution can be referred to mutational compensatory patterns preserving the function of a protein. Viral envelope glycoproteins, which mediate entry of enveloped viruses into their host cells, are shaped by coevolution signals that confer to viruses the plasticity to evade neutralizing antibodies without altering viral entry mechanisms. The functions and structures of the two envelope glycoproteins of the Hepatitis C Virus (HCV), E1 and E2, are poorly described. Especially, how these two proteins mediate the HCV fusion process between the viral and the cell membrane remains elusive. Here, as a proof of concept, we aimed to take advantage of an original coevolution method recently developed to shed light on the HCV fusion mechanism. When first applied to the well-characterized Dengue Virus (DENV) envelope glycoproteins, coevolution analysis was able to predict important structural features and rearrangements of these viral protein complexes. When applied to HCV E1E2, computational coevolution analysis predicted that E1 and E2 refold interdependently during fusion through rearrangements of the E2 Back Layer (BL). Consistently, a soluble BL-derived polypeptide inhibited HCV infection of hepatoma cell lines, primary human hepatocytes and humanized liver mice. We showed that this polypeptide specifically inhibited HCV fusogenic rearrangements, hence supporting the critical role of this domain during HCV fusion. By combining coevolution analysis and in vitro assays, we also uncovered functionally-significant coevolving signals between E1 and E2 BL/Stem regions that govern HCV fusion, demonstrating the accuracy of our coevolution predictions. Altogether, our work shed light on important structural features of the HCV fusion mechanism and contributes to advance our functional understanding of this process. This study also provides an important proof of concept that coevolution can be employed to explore viral protein mediated-processes, and can guide the development of innovative translational strategies against challenging human-tropic viruses