Britt Coenen

Hard work and highlights!

01/09/2023

My journey as a PhD student started one year ago, joining a multi-disciplinary research project aiming to develop a fast and sensitive method to detect sepsis by measuring radical production in immune cells. In my previous blog post I already gave you some background information about my project, and my first milestone in measuring free radical production in neutrophils. Today, I’m excited to update you on my recent endeavours and discoveries.

First of all, I’ve been gathering a lot of results on the effects different isolation methods can have on monocytes. I did this together with my first master student Petri de Jager, therefore marking my debut as a mentor. Our commitment to gathering results led us to early morning starts at 07:00 and late-night sessions extending until 01:00. But it was all worth it, since I won a poster prize during the annual GBB symposium while presenting our findings! Perhaps I’ll delve deeper into these results in a future blog post, or you might even find them in a published paper soon.

However, as anyone who has embarked on a PhD journey knows, it involves more than just the primary project. In my case, I’ve been helping out with several experiments from colleagues, performing review experiments for submitted papers and working in the weekends for check-ups on mice. Next to that, I also took on a role in the young NCOH board at the start of 2023. This opportunity has allowed me to connect with researchers from various Dutch research institutes who share a common objective: exploring and prioritizing research gaps through a One Health perspective.

Currently I’m preparing for the upcoming Society for Free Radical Biology and Medicine (SFRBM) conference, scheduled to take place in Uruguay this November. This international gathering of researchers and experts in the field of free radicals and antioxidants presents a golden opportunity to share my research on nanoscale MRI and ROS measurement in immune cells. I am eager to connect with other scientists in the field, exchange ideas, and gather insights that will further advance my research.

Some background and first successes

01/04/2023

To start, a little bit of background: My PhD project is a multi-disciplinary project involving three research groups within the University of Groningen and the UMCG (department of Molecular Immunology and the department of Biomedical Engineering) and the emergency department at the UMCG (including Acutelines, a biobank for acute medicine). The research aims to develop a fast and sensitive method to detect sepsis by measuring radical production in immune cells. In this blog post, I’ll share some general information and updates on my project.

I’ll start with a brief introduction on sepsis. Sepsis is a life-threatening dysregulated host response to infection, leading to organ dysfunction. The global burden is astonishingly high: 20% of all deaths worldwide are due to sepsis. Every year, more people in the Netherlands die of sepsis than of any type of cancer, myocardial infarction or traffic accidents. Early recognition of sepsis is essential for timely initiation of adequate care and lower morbidity and mortality. However, sepsis recognition in the early phase and prediction of the clinical course are difficult as signs and symptoms may be nonspecific and vary among individuals. At this point, the golden standard in recognition is based on a scoring system assessing the performance of multiple organ systems. And the specificity of this scoring system? An “astonishing” 47%, meaning that you might as well flip a coin to diagnose a patient.

Next to this scoring system, inflammatory markers that can be measured in the blood are used in sepsis diagnosis. The production of radicals (reactive oxygen species [ROS] to be specific) by immune cells is one of the earliest responses in inflammation, and precedes the production of these inflammatory markers by multiple hours. In sepsis, the function of immune cells is disturbed leading to increased ROS production, eventually resulting in organ damage. So why not look at this early response molecule to diagnose sepsis? Well, ROS are challenging to detect due to their high reactivity and low abundance. Fortunately, this challenge has recently been overcome by the development of nanoscale MRI, which allows fast measurement of ROS using fluorescent defects in Nano diamonds. The goal of my project is therefore to develop a fast and sensitive diagnostic test for the early detection of sepsis using this nanoscale MRI method.

In the past six months, I’ve been busy reading a lot of literature and gathering my first results in the lab. Moreover, I developed an optimized method for the isolation of immune cells that are mainly responsible for ROS production, the monocytes and neutrophils. This is important because I need to be able to isolate these immune cells from blood without them getting activated in the process. Next to that, I also had a first try in using the nanoscale MRI method to detect free radical production in freshly isolated neutrophils. It was the first time ever that we tried to do this, and I’m happy to share with you that it worked! I was able to do a “baseline” measurement of these neutrophils, after which I stimulated them with a microbial stimulus to start producing ROS. Over time (20 minutes to be precise), a clear increase in ROS production was measured every 2 minutes, eventually resulting in the orchestrated death of the neutrophils. This is also known as NETosis, but I will not go into detail about that for now. In the attached picture (special thanks to Alina Sigaeva) you can see the outline of a single neutrophil (purple) with the characteristic lobular nucleus (darker areas in the middle), and a diamond (bright yellow/orange spot).

Overall, I’m excited about the potential implications of my research and the positive impact it could have on sepsis diagnosis. I’m looking forward to continuing my work in the upcoming years and exploring the mechanisms behind ROS in sepsis.

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