Understanding MPOX transmission

Susana Guerra is a Professor of Microbiology at the Autonomous University of Madrid Medical School. After working with pox viruses for 24 years, her research work took an unexpected turn in 2022. Today, she is one of Europe’s leading MPOX researchers (formerly known as Monkeypox), on a mission to understand why the 2022 MPOX virus (MPXV) strain is so much more contagious than previous strains. Her quest brought her to Euro-BioImaging’s EMBL Node, along with her PhD student, Joseph McGrail. Their imaging project, supported by ISIDORe funding, employs confocal fluorescence microscopy and electron microscopy (TEM) to better understand the viral transmission mechanisms in three different strains of MPOX in murine models. 

It was June 2022. Prof. Susana Guerra was at the Mt Sinai hospital in New York City on a Fulbright scholarship when the MPOX outbreak began. In response to this emergency, Susana’s work on Vaccinia Virus was put on hold. Her research focus shifted immediately to investigating MPXV. Over the course of her three-month stay at Mt Sinai, Susana worked with patients, dermatologists and other clinicians, to diagnose and characterize the MPXV. During this time, she contributed to three publications: on MPOX diagnostics (PCR), genomics analysis, and ultrastructural characterisation. All this work has shaped  her curiosity to understand the monkeypox virus infection mechanisms and evolution. 

“ISIDORe has really opened the door for us to integrate several scientific services and expertise across Europe,  and has been a facilitator for the overall project implementation. Thanks to ISIDORe and Euro-BioImaging, we could have free access to state-of-the-art imaging expertise and technologies offered at EMBL.” 

• Susana Guerra, Professor of Microbiology, Autonomous University of Madrid Medical School

Shedding light on MPOX transmission

In her research, Susana aims to compare the virus transmission of different MPOX strains in murine cells and other organisms, and investigate which factors lead to variation in transmission efficiency - using proteomics, light microscopy, electron microscopy, virus titration and western blotting. Gaining knowledge on commonalities can facilitate research on targeted development of effective vaccines, therapeutics and to increase protection against different variants of the virus.

“When I returned to Spain, the biggest challenge was to get access to the required  portfolio of imaging technologies to dive deeper into the virus’s transmission mechanisms,” says Susana. “At my home institute, there is only one confocal microscope, and it is hardly available. Pursuing research on MPOX also requires access to the BSL-3 facility. I need to simultaneously negotiate and coordinate agreements with several institutions and sometimes wait for a long time for individual slots where we can perform experiments.” 

“ISIDORe has really opened the door for us to integrate several scientific services and expertise across Europe,  and has been a facilitator for the overall project implementation. Thanks to ISIDORe and Euro-BioImaging, we could have free access to state-of-the-art imaging expertise and technologies offered at EMBL.” 

Susana Guerra and her PhD student Joseph McGrail explore MPXV with fluorescence microscopy at EMBL's ALMF, part of Euro-BioImaging's EMBL Node.

Addressing some of the challenges of infectious disease research

Through ISIDORe, the Euro-BioImaging team connected Susana and Joseph with Instruct ERIC for proteomics analysis by mass spectrometry at the University of Helsinki, and to the European Virus Archive (EVAg) for accessing biological resources for their project. 

They were able to make use of all of these services and expertise free of charge thanks to the ISIDORe Monkeypox Emergency Call and the TNA program.

In addition, the ISIDORe award also helped Susana to negotiate a timely access to the BSL-3 facility at Instituto de Salud Carlos III (ISCIII) which is a partner of ERINHA, the European Research Infrastructure on Highly Pathogenic Agent and coordinator of ISIDORe.

“Susana’s and Joseph’s user project is an interesting example for highlighting missions of ISIDORe, which stands for Integrated Services for Infectious Diseases Outbreak research,” explains Arina Rybina, Euro-BioImaging’s Scientific Project Manager in ISIDORe and Advanced Imaging Specialist. “The ISIDORe consortium has facilitated a spectrum of complex research projects from basic research to vaccine development - by providing scientists free access to multiple European Life Science Research Infrastructures (LS-RIs) and Networks.”

“These partners work together in the consortium and also develop innovative cross-disciplinary services pipelines,” Arina continues. “When a researcher from academia or industry approaches the consortium and asks for help, the partners are promptly pulling strings to investigate and respond to this request. The RI project managers and facility experts provide consultation, find the most suitable expertise and technology and support the researchers during the project application, implementation and dissemination phases.”

The benefit of cutting-edge microscopy

Coming to EMBL has been a very good experience for both Joseph and Susana. In a short time, they were able to acquire many datasets, thanks in part to the work and preparations for the on site experiments that were carried out by EMBL staff. 

Stefan Terjung is the Operational Manager of the Advanced Light Microscopy Facility at EMBL. He helped Joseph to set up the fluorescence microscopy imaging pipeline and taught him how to use the microscope in order to speed up the data acquisition process. “With light microscopy, thanks to Stefan Terjung, we are able to quickly acquire many images of the infected cells, map the subcellular distribution of viral proteins, and study particular features of fluorescently labeled  intracellular organelles, such as actin filaments,” explains Joseph.

Viola Oorschot is an expert in Electron Microscopy at EMBL’s Electron Microscopy Core Facility (EMCF) in Heidelberg. She’s been working with Joseph and Susana to explore the samples of murine cells infected with different MPOX strains by Transmission Electron Microscopy. “It’s a very interesting research project, and very relevant. In fluorescence microscopy, you can highlight and see what you label with markers. Using electron microscopy, we can observe and characterize  what is happening in and around cells and the virus particles - in the ultrastructural context and at much higher resolution,” explains Viola. 

Left: Rachel Mellwig and Viola Oorschot, on the Electron Microscope at EMBL EMCF, and right, Arina Rybina talks with Susana Guerra and Joseph McGrail about their imaging project at EMBL.

“I’ve seen lots of different viruses on a microscope over the course of my career,” remarks Viola, “But this is my first time imaging the MPXV. Virus structures are beautiful in general, but at the same time alarming  considering their impact on health, on society.”

“Through the electron microscopy images obtained, quantification of the different viral forms in MPXV infection is being undertaken to better understand the differences in viral morphogenesis between the 2022 outbreak MPXV and older MPXV strains. This would not have been possible without ISIDORe´s support and help from professionals such as Viola and Rachel,” remarks Susana.

Towards concrete scientific results

Shortly after completing their visit, the first results of their comparative study on MPOX contagiousness integrating the proteomics analysis part of their project was accepted for publication, "Comparative Analysis of 2022 Outbreak MPXV and Previous Clade II MPXV" (doi: 10.1002/jmv.70023). A second publication featuring results of the EM and LM imaging work done at EMBL is currently in preparation along with the analysis of obtained data.

Joseph and Susana are also looking forward to taking advantage of the FAIR data consultation offered through Euro-BioImaging for bringing their imaging data sets to open access repositories and sharing those with scientific communities.

“This is one of the greatest rewards of open, funded access to cross-disciplinary scientific services like through the Horizon Europe funded project ISIDORe,” concludes Arina. “It is truly inspiring to facilitate research and knowledge acquisition, and to watch research collaborations develop across Europe and beyond. While the ISIDORe project will be coming to an end soon, we hope for similar initiatives and funding opportunities to sustain such a framework in near future. We must build on all the hard work and procedures we established in ISIDORe, in support of research on pathogens of concern and effective response to arising emergencies and pandemic preparedness.”