Biology subjects

Munir, M.

Publications and source records attributed to Munir, M..

6 recordsLinked to original sources

bioRxiv2023

Structural and Mechanistic Usage and Preference of Human, Dog, and Bat Receptors by Rabies Virus

Rabies is a lethal zoonotic viral disease causing approximately 59,000 human deaths annually. Recently, several cellular receptors for rabies virus (RABV) entry and internalization have been identified. However, none of these receptors have been demonstrated to be indispensable for RABV entry. Here we describe the RABV receptor preference in vivo, utilizing a replication-competent vesicular stomatitis virus (VSV), in which the VSV surface glycoprotein was replaced with rabies virus glycoprotein. To investigate the specific role of RABV receptors in promoting RABV entry in non-permissive cell line, HaCaT cells were used as a cellular model refractory for RABV infection. Employing virus binding and quantification studies, we demonstrated that ITGB1 and mGluR2 are potential receptors for RABV entry and replication. Consequently, knockout (KO) cell lines corresponding to each of the ITGB1 and mGluR2 receptors were generated using CRISPR/Cas9 mediated knockout. Surprisingly, RABV was still able to enter and replicate in the generated KO cell lines, yet the replication and entry of RABV in KO cells lacking mGluR2 and ITGB1 were significantly reduced; respectively. These findings suggest that RABV utilize these receptors in series rather than sequentially. To test whether RABV utilizes similar receptor preference among human, dog, and bats, the A549, Pa-Br and MDCK cell lines that overexpress receptor orthologs from their respective species were infected with rVSV-dG-RABV-G-GFP and quantified for virus binding and released virus progeny. Our findings revealed that in human cells, ITGB1 increased virus entry, while nAChR enhanced virus replication. In bat cells, ectopic expression of nAChR allowed enhanced virus entry and internalization. While MDCK cells overexpressing ITGB1 enhanced the levels of virus entry and replication. Conclusively, our study, reveals the RABV distinct receptor preference, influenced by the underlying pathways that occur during the interaction between the virus and receptor in different cell lines. Additionally, it emphasizes the significance of host-specific factors in virus entry and replication. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=146 SRC="FIGDIR/small/564510v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@1f02ccaorg.highwire.dtl.DTLVardef@10cb0edorg.highwire.dtl.DTLVardef@33d3a4org.highwire.dtl.DTLVardef@8bbe58_HPS_FORMAT_FIGEXP M_FIG C_FIG Author SummaryRabies is a fatal neurological disease, characterized by broad host range and tissue tropism. In accordance with the global goal of eliminating dog-mediated human rabies by 2030, studying the underlying mechanism of RABV entry across distinct species would enable adjustment of RABV control strategies. Owing to RABV wide tropism, multiple cellular receptors have been identified for RABV entry into host cells. Previous studies have proposed that some of RABV receptors could serve as promising candidates for development of antiviral drugs (1). From this perspective, we focused on elucidating RABV receptor preference for viral entry in human, dog, and bat cells. In addition to determining whether RABV utilizes these receptors in parallel or in series which would indicate the potential of the identified RABV cellular receptors as targets for antiviral drugs against rabies. Our results demonstrated varying receptor preference of RABV across species. In addition to revealing that none of RABV receptors solely, govern the broad host range of rabies, suggesting that RABV antiviral drugs targeting host cellular factors may not effectively inhibit RABV entry into cells, while antiviral drugs targeting virus glycoprotein may exhibit greater efficacy. Collectively, our study, contribute to providing mechanistic model for RABV entry in different species.

microbiology↗

bioRxiv2023

Genome-wide CRISPR activation screen identifies JADE3 as an antiviral activator of NF-kB

The innate immune system features a web of interacting pathways that require exquisite regulation. To identify novel nodes in this immune landscape we conducted a gain of function, genome-wide CRISPR activation screen with influenza A virus. We identified both appreciated and novel antiviral genes, including JADE3 a protein involved in directing the histone acetyltransferase HBO1 complex to modify chromatin and regulate transcription. JADE3 is both necessary and sufficient to restrict influenza A virus infection. Interestingly, expression of the closely related paralogues JADE1 and JADE2 are unable to restrict influenza A virus infection, suggesting a distinct function of JADE3. We identify both shared and unique transcriptional signatures between uninfected cells expressing JADE3 and JADE2. These data provide a framework for understanding the overlapping and distinct functions of the JADE family of paralogues. Specifically, we find that JADE3 expression activates the NF-kB signaling pathway, consistent with an antiviral function. Therefore, we propose JADE3, but not JADE1 or JADE2, activates an antiviral genetic program involving the NF-kB pathway to restrict influenza A virus infection.

microbiology↗

bioRxiv2021

Mapping Molecular Gene Signatures Among Respiratory Viruses Based on Large-Scale and Genome-wide Transcriptomics Analysis

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emerging RNA virus causing COVID-19 disease across the globe. SARS-CoV-2 infected patients exhibit acute respiratory distress syndrome which can be compounded by endemic respiratory viruses and thus highlighting the need to understand the genetic bases of clinical outcome under multiple respiratory infections. In this study, 42 individual datasets and a multi-parametric based selected list of over 12,000 genes against five medically important respiratory viruses (SARS-CoV-2, SARS-CoV-1, influenza A, respiratory syncytial virus (RSV) and rhinovirus were collected and analysed in an attempt to understand differentially regulated gene patterns and to cast genetic markers of individual and multiple co-infections. While a certain cohort of virus-specific genes were regulated (negatively and positively), notably results revealed a greatest correlation among gene regulation by SARS-CoV-2 and RSV. Furthermore, out of analysed genes, the MAP2K5 and NFKBIL1 were specifically and highly upregulated in SARS-CoV-2 infection in vivo or in vitro. In contrast, several genes including GPBAR1 and SC5DL were specifically downregulated in SARS-CoV-2 datasets. Additionally, we catalogued a set of genes that were conserved or differentially regulated across all the respiratory viruses. These finding provide foundational and genome-wide data to gauge the markers of respiratory viral infections individually and under co-infection.

immunology↗

bioRxiv2021

Immunogenicity and Protective Efficacy of an Intranasal Live-attenuated Vaccine Against SARS-CoV-2 in Preclinical Animal Models

The global deployment of an effective and safe vaccine is currently a public health priority to curtail the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we evaluated a Newcastle disease virus (NDV)-based intranasal vectored-vaccine in mice and hamsters for its immunogenicity, safety and protective efficacy in challenge studies with SARS-CoV-2. The recombinant (r)NDV-S vaccine expressing spike (S) protein of SARS-CoV-2 administrated via intranasal route in mice induced high levels of SARS-CoV-2-specific neutralizing immunoglobulin A (IgA) and IgG2a antibodies and T cell-mediated immunity. Hamsters vaccinated with two doses of vaccine showed complete protection from clinical disease including lung infection, inflammation, and pathological lesions after SARS-CoV-2 challenge. Importantly, a single or double dose of intranasal rNDV-S vaccine completely blocked SARS-CoV-2 shedding in nasal turbinate and lungs within 4 days of vaccine administration in hamsters. Taken together, intranasal administration of rNDV-S has the potential to control infection at the site of inoculation, which should prevent both the clinical disease and transmission to halt the spread of the COVID-19 pandemic.

immunology↗

bioRxiv2020

A Scalable Topical Vectored Vaccine Candidate Against SARS-CoV-2

The severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) caused an ongoing unprecedented global public health crises of coronavirus disease in 2019 (CoVID-19). The precipitously increased death rates, its impact on livelihood and trembling economies warrant the urgent development of SARS-CoV-2 vaccine which would be safe, efficacious and scalable. Owing to unavailability of the vaccine, we propose a de novo synthesised avian orthoavulavirus 1 (AOaV-1)-based topical respiratory vaccine candidate against CoVID-19. Avirulent strain of Newcastle disease virus, proto-type virus of AOaV-1, was engineered to express full length spike (S) glycoprotein which is highly neutralizing and major protective antigen of the SARS-CoV-2. Broad-scale in vitro characterization of recombinant vaccine candidate demonstrated efficient co-expression of the hemagglutinin-neuraminidase (HN) of AOaV-1 and S protein of SARS-CoV-2, and comparable replication kinetics were observed in cell culture model. The recombinant vaccine candidate virus actively replicated and spread within cells independently of exogenous trypsin. Interestingly, incorporation of S protein of SARS-CoV-2 into the recombinant AOaV-1 particles attributed the sensitivity to anti-SARS-CoV-2 antiserum and more prominently to anti-AOaV-1 antiserum. Finally, our results demonstrated that the recombinant vaccine vector stably expressed S protein after multiple propagation in chicken embryonated eggs, and this expression did not significantly impact the in vitro growth characteristics of the recombinant. Taken together, the presented respiratory vaccine candidate is highly attenuated in primates per se, safe and lacking pre-existing immunity in human, and carries the potential for accelerated vaccine development against CoVID-19 for clinical studies.

microbiology↗

bioRxiv2019

Genome-Wide Classification of Type I, Type II and Type III Interferon-Stimulated Genes in Chicken Fibroblasts

Interferons (IFNs) play central roles in establishing innate immunity and mediating adaptive immunity against multiple pathogens. Three known types of IFNs identify their cognate receptors, initiate cascades of signalling events and eventually result in the induction of myriad of IFN-stimulated genes (ISGs). These ISGs perform multitude of functions and cumulatively corroborate a bespoke antiviral state to safeguard hosts against invading viruses. Owing to unique nature of chickens immune system and lack of foundational profiling information on the nature and dynamic expression of IFN-specific ISGs at the genome scale, we performed a systematic and extensive analysis of type I, II and III IFN-induced genes in chicken. Employing pan-IFN responsive chicken fibroblasts coupled with transcriptomics we observed an overwhelming over-representation of up-regulated ISGs by all types of IFNs. Intriguingly, prediction of IFN-stimulated response element (ISRE) and gamma-IFN activation sequence (GAS) revealed a substantial number of GAS motifs in selective significantly induced ISGs in chicken. Extensive comparative, genome-wide and differential expression analysis of ISGs under equivalent signalling input catalogue a set of genes that were either IFN-specific or independent of types of IFNs used to prime fibroblasts. These comprehensive datasets, first of their kinds in chicken, will establish foundations to elucidate the mechanisms of actions and breadth of antiviral action of ISGs which may propose alternative avenues for the targeted antiviral therapy against viruses of poultry and public health importance.

immunology↗