UCD team presenting COVend research results at the European Congress of Immunology 2024

Unlocking the Potential of FX06: An Interview with the UCD Systems Biology Ireland Team on Their Research in COVend

Systems Biology Ireland (SBI), University College Dublin (UCD), focuses on developing new therapeutic approaches to disease on a systematic level – through a combination of traditional wet-lab experimental analyses and cutting-edge computational modelling and simulation. One of our key COVend researchers, Prof Günther Eissner, leader of Work Package 5 (Endothelial cell assessment), supervises a dedicated group of researchers who study the impact of COVID-19-triggered cytokines on human microvascular pulmonary endothelial cells (ECs). By combining wet and dry lab techniques, Günther’s team can gain detailed insights into the immune response, viability, stress signalling and other cellular processes. The researchers have made significant progress in understanding how the drug FX06, an innovative therapeutic candidate for the treatment of acute respiratory distress syndrome (ARDS), can counteract the pro-inflammatory effects of a SARS-CoV2 infection and the underlying mechanisms behind its action. We had the opportunity to speak with Günther and his team to learn more about their research.

What are you and your team currently working on under the EU-funded project COVend?

Günther: Firstly, Zhiran Wang, a PhD student and wet lab researcher, along with the assistance of a former member Dr. Simeng Li, conducted a thorough review of the literature to identify cytokines and chemokines present in the serum of critically ill COVID-19 patients. The so-called severe cytokine cocktail is applied to a human microvascular endothelial cell line in vitro [1]. Through our research, we aim to determine the potential effects of these cytokines on cell death, phenotypical changes, and immunological activation, as well as the migration of immunological effector cells through the endothelial monolayer – all indicators of inflammation. To further investigate the protective properties of the promising drug FX06, it was co-administered to the ECs. By using state-of-the-art functional assays and systems biology analyses, our team hopes to gain insights into the signal transduction pathways involved in cytokine signalling and how FX06 may counteract their effects.

[1] research done in a laboratory dish or test tube

Tell us about what is each team member in charge of?

Zhiran: Dr. Simeng Li provided me with 9-month training on various methodologies and data analysis techniques. Mr. Lorcan Doyle was in charge of cytotoxicity assays, and he discovered that FX06 can effectively decrease the cytotoxic effects of CD8+ T cells. Our current Master student, Mr. Kevin Wu, found that the protective effects of FX06 involve the RhoA-pROCK1 signalling pathway. Our current Research Scientist, Mr. Sudharshan Rao, in charge of western blot analysis, is investigating the signalling pathways and further insight into the mechanism of action of FX06.

What does a typical day at the lab look like for COVend’s researcher at SBI?

Zhiran: My daily schedule is quite flexible – some days, I start working early in the morning, while other days, the experiments are scheduled for the afternoon. On average, I schedule 2-4 different experiments per week. Our assays to e.g. assess transendothelial migration of immunological effector cells across the blood vessel wall usually run from Monday to Friday, but they do not take up my entire day, allowing me to work on other tasks in between. While sample preparation for flow cytometry experiments can be tedious, the actual experiments run very quickly. Imaging can be quite relaxing once all the setups are ready, but staying in the dark room for long can be tiring. To ensure a cohesive analysis of the data obtained, I leave the data analysis for the end of each week. This also gives me the opportunity to take a break from experiments and do some literature reading, allowing me to organise our data to further explain it and generate new ideas.

Seeding cells for a particular experiment

Assessing the quality of tissue cultures with a microscope

I have access to multiple machines (flow cytometer, variable types of microscopes, data analysis software) while working on the COVend project. They are located at the different departments of UCD, which means I have an opportunity to also work with people outside SBI. Prof Alfonso Blanco from Flow Cytometry Core at Conway Institute and Prof Jeremy Simpson from the College of Science offered valuable support, which contributed to the success of my research results. It is exciting to have an opportunity to work with those state-of-the-art tools, as my supervisor, Prof Günther Eissner, is very open-minded and willing to try new methodologies. I get to learn how to use new machines and even those fancy microscopes that very few people are working with. Training new lab members and students takes me some extra time but, in the end, I gain valuable supervisory experience which is beneficial for my personal career development.

Can you share with us the major breakthroughs that your team has achieved so far in your research on endothelial protection mechanisms?

Günther: Our research has revealed that FX06 has the potential to protect ECs from transendothelial migration induced by COVID-19-triggered cytokines. This is accomplished by restoring the integrity of the vascular lining and by restructuring stress fibres in ECs. Importantly, this is achieved through the inhibition of the RhoA-ROCK-1 signal transduction pathway and the elimination of pro-inflammatory/pro-angiogenic signals from ECs. In simple words, it means that FX06 can remove danger signals from ECs by blocking very specific signal transduction pathways and thus resolve inflammation.

How does FX06 work? How does it treat acute respiratory distress syndrome (ARDS)?

Günther: We have only generated data from laboratory experiments so far. Therefore, it is too early to say if and how exactly FX06 could treat ARDS in patients. However, our findings suggest that FX06 may have the potential to protect ECs from inflammation caused by COVID-19-triggered cytokines. Although ARDS of different aetiologies has common clinical symptoms, it can be very different at the molecular level and the individual response is also heterogeneous from patient to patient. Nevertheless, our data offer unprecedented insights into the mechanism of EC protection and may have significant relevance for the treatment of severe respiratory diseases and beyond.

What are your expectations for the study’s results and how it may impact our current understanding of treating ARDS, COVID-19 pathology, and other diseases?

Günther: We will further explore the mechanism of action of the FX06 drug to gain a better understanding of its effects. We are confident that our data and findings will support the ethical approval and marketing authorization of FX06 for treating not only COVID-19, but potentially other non-infectious forms of ARDS once the relevant clinical trials are successfully completed.

Are there any future research plans to build upon the findings of your study?

Günther: Our group has a long-standing expertise in the research on endothelium protective drugs. As part of our research, we have discovered that mesenchymal stem/stromal cells (MSCs) and their extracellular vesicles (MSC-EVs) have a synergistic effect with one of these drugs. We are interested in investigating whether MSCs and MSC-EVs can also enhance the effects of FX06. This exciting topic is currently being studied in our group as part of a Research-Master program.

How do you plan to disseminate your research findings and make an impact in the medical community?

Günther: To date, we have successfully shared our research findings with a diverse range of audiences through presentations at four conferences and workshops, both nationally and internationally. In addition, we have leveraged blog entries to further promote our research and garner even more interest. Our team is now in the process of compiling an original manuscript that will be submitted to a high-ranked peer-reviewed immunology journal, which wraps up all the wet and dry lab activities we performed thus far.