Published in NAR: formation of the SARS-CoV-2 methylation complex

The CARE partner Jagiellonian University (JU) deciphered the interaction between three viral proteins that are necessary to viral replication. To be able to replicate, coronaviruses modified their single-stranded RNA genome by adding a methylated cap to mimic the host cell mRNAs and thus highjack the host enzymes that will then replicate the virus RNA.

Three viral non-structural proteins – nsp14, nsp10 and nsp16 – have previously been shown to be involved in the implementation of the methylation cap, with nsp14 and nsp16 performing the methylation while nsp10 acts as a co-factor to both. However, structural information available suggested that the interaction between the three proteins would be impossible considering their conformation. To solve this mystery, JU applied orthogonal methodologies, i.e., used different techniques to evaluate the interactions between the three proteins.

The results shows that nsp14, nsp16 and nsp10 are able to form a trimer complex, built around nsp10. This trimer brings together two consecutive activities required for RNA cap formation, the one led by nsp14 and the one led by nsp16, likely contributing to the RNA capping mechanism.

These findings contribute to the current understanding of the intricate interplay among nsp14, nsp10 and nsp16 during the replication process, particularly in the context of RNA cap methylation in the coronaviral genome. This research may have implications for the development of novel targeted therapeutic strategies for the control of coronaviral infections.

To learn more, click here: Despite the odds: formation of the SARS-CoV-2 methylation complex

Published in Virology: a new cell line permissive to human coronavirus 229E

The CARE partner Jagiellonian University (JU) developed a new cell line that can be infected by the human coronavirus 229E (HCoV-229E), a member of the alphacoronavirus family. HCoV-229E is one of the coronaviruses that poorly propagate in cell lines and require extensive cell culture adaptation leading to changes in the virus phenotype. New cell lines are thus necessary to study in vitro HCoV-229E more easily. 

To create this new cell line, JU co-transduced CD13 and transmembrane serine protease 2 (TMPRSS2) in A549 cell line (lung-derived cell line) using lentiviral vectors. HCoV-229E can use several pathways to enter a permissive cell, one using CD13, an aminopeptidase N while another one is using TMPRSS2 to proteolytically prime the viral spike, leading to membrane fusion. Cells overexpressing CD13 and TMPRSS2 (A549++ cells) and infected by HCoV-229 showed viral RNA replication and a significant rise on virus titer, confirming their ability to produce infectious HCoV-229E progeny.  

Moreover, JU evaluated the possible entry pathways of HCoV-229E in A549++ cells by inhibiting the TMPRSS2-mediated membrane fusion entry pathway and/or the endocytic pathway. The results indicate that although HCoV-229E prefers TMPRSS2-mediated entry, it can also readily use the endocytic pathway in the absence of TMPRSS2. 

JU has thus developed a robust and physiologically relevant cell line model that is permissive to HCoV-229E clinical isolate replication. The data provide further insight into the potential of lentiviral transduction in developing permissive cell models for viral infection studies. Moreover, this line constitutes a uniform platform for studies on multiple members of the coronaviridae family. 

To learn more, click here: An engineered A549 cell line expressing CD13 and TMPRSS2 is permissive to clinical isolate of human coronavirus 229E 

Published in American Chemical Society Pharmacology & Translational Science: Effective, but Safe? Physiologically Based Pharmacokinetic (PBPK)-Modeling-Based Dosing Study of Molnupiravir for Risk Assessment in Pediatric Subpopulations

Recognising that treatment options for vulnerable paediatric populations with COVID-19 are limited to intravenous remdisivir, the CARE team at Helmholtz Centre for Infection Research (HZI, Germany) wanted to validate the assumption that oral molnupiravir should not be prescribed to under 18-year-olds, due to the potential for cartilage and bone toxicity, which would put normal paediatric development at risk.

Through the application of physiologically based pharmacokinetic (PBPK) modelling, the HZI team was able to show that to match the efficacy of oral molnupiravir seen in adults, a proportionately much higher dose would be needed in paediatric populations, substantially increasing the toxicity risk.

To learn more, click here: Effective, but Safe? Physiologically Based Pharmacokinetic (PBPK)-Modeling-Based Dosing Study of Molnupiravir for Risk Assessment in Pediatric Subpopulations

Introducing Scifeon – a CARE SME organisation

Scifeon was set up in 2016 by Thomas P. Boesen (PhD) to help scientists have better, more flexible, adaptable and agile informatics support that can take them and their research to the next level.

Dr. Boesen’s vision was to provide scientists with the next generation of research data management software to help them in their research.

Scifeon is a Software as a Service (SaaS) platform. This comprises the Electronic Laboratory Notebook (ELN), Laboratory Information Management System (LIMS) and Scientific Data Management System (SDMS). These systems collect, organize and securely store valuable research data for easier processing, better collaboration and faster results – giving scientists more time for science.

Scifeon provides scientists with better informatics on a day-to-day basis. It is the next generation of research data management software specifically developed for scientists by scientists.

Why did Scifeon choose to get involved in CARE?

Scifeon recognised CARE’s need for a flexible data management solution that could be adapted to its highly variable research effort. This involved the creation of CARE’s data management plan as well as creating and operating the data management cloud platform, CARE-4-DATA. The platform houses protocols for several biochemical and cellular assays and allows researchers from different institutions to collaborate on testing compounds in these assays and sharing the results.  It also contains a registry of virus strains and plasmids that are present at several of the CARE partner organisations.

In addition to Dr. Boesen, the Scifeon team includes

What has Scifeon delivered for CARE?

The CARE-4-DATA platform houses protocols for several biochemical and cellular assays and allows researchers from different institutions to collaborate on testing compounds in these assays and sharing the results.  It also contains a registry of virus strains and plasmids that are present at several of the CARE partner organisations.

What benefits has Scifeon enjoyed through participating in CARE? 

Scifeon has seen its software platform grow and improve through incorporating feedback from CARE scientists.

Want to know more about Scifeon?  

https://www.scifeon.com/

https://www.scifeon.com/contact/

CARE External Newsletter – December 2024

The new issue of our biannual newsletter is out. In this edition we learn about an impressive app developed by AbbVie to determine genetic factors related to COVID-19 risk, plus we share news of an exciting partnership between University of Dundee and Novartis. We also introduce CARE partner Scifeon and the important role they play in managing CARE’s laboratory informatics.

Read the Newsletter here: CARE External Newsletter – December 2024