#JournalClub: Anti-Cancer Immunotherapy

Hi there, Federica here! In the fast-paced world of scientific research, staying informed about the latest studies and breakthroughs is crucial. It enables researchers to build upon existing knowledge, avoid redundant efforts, and discover new directions for their work. That’s why we’ve started a new series of blog posts highlighting recent papers and explaining their significance for our research.

Recently, a fascinating study explored an innovative method to boost the effectiveness of cancer immunotherapy: “A combination of a TLR7/8 agonist and an epigenetic inhibitor suppresses triple-negative breast cancer through triggering anti-tumour immune“.

The researchers investigated a combination of immune checkpoint blockade (ICB) and other drugs to turn “immune-cold” tumours (which evade the immune system) into “immune-hot” tumours (which the immune system can attack). They developed a special delivery system using nanoparticles called metal-organic frameworks (MOFs). These nanoparticles were loaded with two types of drugs—a TLR7/8 agonist and an epigenetic inhibitor (BRD4 inhibitor). To make the nanoparticles even more effective, they were coated with vesicles from the cancer cells themselves. This coating helps the nanoparticles specifically target cancer cells.

But how does it work?

The nanoparticles are designed to find and enter triple-negative breast cancer (TNBC) cells. Once inside, the drugs prompt the cancer cells to break apart and release signals that alert the immune system. These signals attract dendritic cells, which then activate CD8+ T cells—the body’s natural cancer fighters. The TLR7/8 agonist further enhances this immune response, making the treatment more powerful.

In both laboratory tests and animal models, this method showed significant promise. It not only slowed down tumour growth but also improved the body’s immune response to cancer. Importantly, the study found that this approach could remodel the tumour environment, making it more hostile to cancer cells. For example, they wanted to verify that their combined delivery system could really boost the body’s ability to fight tumours. They focused on a protein called calreticulin (CRT) that, when it shows up on the surface of tumour cells, helps the immune system spot and remove them. They found that when they used their special delivery system (CM@UN and MCM@UN), the levels of CRT on the surface of tumour cells went way up. This was especially true for the MCM@UN group, showing just how powerful their method was in getting the immune system to attack the tumours.

The original image was published in J Nanobiotechnology. 2024; 22: 296.

So, why is this study important for my work?

The principles of enhancing the immune system’s ability to fight cancer are central to both the research in the study and in my project. Like the nanoparticles in the study, mRNA vaccines can be designed to specifically target cancer cells, ensuring that the treatment reaches its intended destination. Another similarity is how the drugs activate the immune system, which parallels how mRNA vaccines work—by training the immune system to recognise and attack cancer cells.

I find this study really interesting as it sheds light on innovative strategies for cancer treatment and provides valuable insights that can inform and inspire our research on developing mRNA vaccines for childhood neuroblastoma!

Written by Federica Cottone

What Can VHI Women’s Mini Marathon Do For You?

I have never been a runner. In my head, the word ‘marathon’ was linked to professional athletes and the Olympics or Athletics competitions. I could not imagine anyone doing a race as long as one gets guts. However, once I found the courage and motivation to explore my body’s limits. It was probably our team fundraising ‘Hell&Back’ challenge in 2019 that ignited that courage. We did many! Virtual VHI MiniMarathon, the Dublin Mountain Way in a Day, Cyclothone, to name a few.

Eventually, I decided to do it on my own in 2024. I booked the VHI Mini Marathon 2024 entry and started my training. Having many fitness apps at my disposal let me pick the right training plan. 10K sounded manageable. The fitness watch kept all my training records, so I got a very good understanding of my body potential and limits to do 10K.

On that sunny day, hundreds and hundreds of women were getting to the starting line. Each set their own ambition and target. I had three: 1) jogging from start to finish, 2) finishing within 1hr and 20 min, and 3) supporting the Conor Foley Neuroblastoma Cancer Research Foundation.

So, June 2nd was fast approaching, and I injured my calf during the Wicklow hike. However, giving up was not an option, but healing was required. So, I decided to stop running for a week and gradually get back on track.

The atmosphere was cheering and empowering, and the sun was bright and hot. The green ‘wave’ began their race at about 12:45. Running in a big company was easy. It is less doubtful if you are in a group of allies. Distance and time flew. I was sinking into the diversity and variety of running women and supporters along the road, enjoying every minute. Some took over me, and I took over some. I stuck to my training pace to ensure my power and energy were not draining quickly under the sun. My fitness watch counted the distance, heart bit and steps, showing that I was slightly slower on average than during my training. I decided not to break the limits and kept running, listening to my body. Somewhere at the back of my head, I hoped to speed up at the end, if any. Unfortunately for me, the final 1-2 km were up the hill, so the slop ate my efforts.

Being a researcher puts every experience in perspective. We tend to analyse the flow of any information, sometimes unconsciously, by asking questions and reflecting. One of the running advantages is that you see an accomplishment right now at the finish line. Your fitness watch provides all the data to plan and complete a given challenge with very good accuracy. It is not about luck. It is about your trust in your body and mind. You are in the driving seat! Delighted with my accomplishment!

So, what is next on my to-do list? Definitely another race, very likely Run in the Dark.

Written by Olga Piskareva

Class 2024: Congratulations to Ciara, Ellen and Rabia!

Massive congratulations on the official moulding of PhD and MSc by Research to our promising young scientists: Rabia Saleem, Dr Ciara Gallagher and Dr Ellen King! Great accomplishments!

Three different journeys, with two through the COVID-19 pandemic. The full range of ups and downs. Who said that the PhD is a straight line? It has never been. It is more like the Irish weather: some days are sunny and bright, and some have scattered showers, gale winds and stormy snow, with sunshine developing elsewhere. The journey was spiced up with publications, conferences, travels, days out and fundraising events with the team.

It is a proud moment for me as well. 🙂 Three PhD and one MSc by Research students graduated within the last 12 months.

Of note, Ellen was behind our Twitter activities in the past, making our team visible!

Wish you all the best of luck on your new adventure!

Olga Piskareva

Walking the Wicklow Way, 87/129 km

Little did I know about hikers when I moved to Ireland in 2004. Who they are and how they get around. My knowledge was limited to Rosalind Franklin’s love of hiking. I could not even imagine that one day I’d try their shoes.

However, things have changed since then! Spiced by the COVID-19 pandemic and various fundraising activities inspired by my team, my daily walking transformed into regular one-day hiking here and there. Luckily, my spouse shares the same attitude. So, we decided to explore longer walks one day.

The first go was the Dublin Mountain Way (42 km) in a Day. We started in Glensmole-Tallagh on a dry and sunny morning and finished in Shankill in the dark and pouring rain with a short recharge at Johnny Foxes. We were delighted with ourselves and raised the bar.

So, last week, we attempted the Wicklow Way. After studying the route, accommodation options and our fitness, we agreed on three days of walking in the north-to-south direction and 2 nights of sleep in B&Bs. We also monitored the weather forecast to make the most of this adventure. So, May 10-12 were the best. However, accommodation became a quest. Nevertheless, luck was on our side, and we found two nice places: one was near Roundwood, and the second was in Glenmalure.

Early morning of May 10th, we cheerfully started our journey in Marley Park. The day was fab; the topics for a chat were endless. We were walking away from Dublin. My fitness watch counted the steps and kilometres. During the walk, we got a confirmation that our accommodation in Glenmaluer had been upgraded to a room with a shower. Happy days!

37 km later, we arrived at our first B&B. It was actually a fancy hostel where all the guests walked in socks, parking their heavy boots in the lobby. It was the night of aurora borealis, but we did not know about it. We were tired and fell asleep before 11 pm. The next morning, none of the guests shared any pics or insights. Apparently, everyone was knocked down by the long day in Wicklow.

May 11th. Fueled with a tasty Irish breakfast, we said “Goodbye” to our hosts and headed further. While walking slowly to warm up for a long day ahead, I noticed that my calf was strained and walking downhill had become unpleasant. Where did it happen? I had no idea. We reached Glendalough around noon.

The day was warm, the car parks were full, and everyone enjoyed the beauty of Glendalough and the sunshine. We stretched our legs and backs on greens. During our light lunch, we discussed our options: 1) evacuation home or 2) continuing and hoping for the best. I did not give up. But our walking pace considerably slowed down.

We covered another ~15 km from Glendalough to Glenmalure and landed in the Glenmalure Lodge – the healing station for all hikers, cyclers, and bikers alike. People gathered outside, and strangers had cheat-chat sharing their tricks and tips for a better hike. Something adventurous was in the air.

Our friendly host picked us up at the Lodge and drove to their B&B. We stayed late and hoped to catch a glimpse of aurora borealis. The sky was cloudy. We saw nothing. While disappointed, our bodies cried for long rest, and we did not resist.

May 12 was the last leg of our journey. My calf did not improve. We took the shortest option to finish in Aughrim. I doubled the dose of painkillers. Then, we moved tirelessly, enjoying the sunshine and the forest.

This part of the Wicklow Way was mostly foresty. The forest was magical and a bit surreal. We agreed that it is perfect for various fantasy and horror movies. My fitness watch signposted that its battery was low, but it continued to count the steps and kilometres.

Overall, our hike had a fair amount of ups and downs. Some climbs and descents were quite steep before Glendalough. Then, they became more gradual, working well for my injured calf.

The luck was again on our side in Aughrim. We saw a taxi – a rarity in this area. The cheerful driver dropped us at the bus stop in Arklow. In 15 min, we were on the way to Dublin, relaxing and enjoying the peaceful countryside from the bus seat.

Our tally was 87 km in 3 days, fully recharged mind but worn body. Would I do it again? Absolutely!

How things work in science: Gene editing technology

Few advancements in biomedical sciences hold as much promise for revolutionising cancer research as CRISPR-Cas9. This ground-breaking gene-editing tool has sparked a wave of innovation, offering precision and efficiency in manipulating the human genome in the fight against cancer.

Now, what is it? CRISPR is basically an acronym for a very long name Clustered Regularly Interspaced Short Palindromic Repeats Associated Protein 9 or CRISPR-Cas9 for short. It was found in simple organisms such as archaea and bacteria. Interestingly, this is a component of bacterial immune systems that can cut DNA. So, this feature was proposed for use as a gene editing tool, a kind of precise pair of molecular scissors that can cut a target DNA sequence. So, the CRISPR-Cas9 scissors allow us to precisely edit the DNA sequence of living organisms by adding in (knock-in) or removing (knockout) a gene of interest.

For cancer research, for example, the CRISPR-Cas9 scissors can be used to introduce therapeutic genes or correct mutations associated with cancer predisposition syndromes. Meanwhile, those scissors can also disrupt genes involved in treatment resistance, sensitising cancer cells to existing therapies.

Jennifer Doudna and Emmanuelle Charpentier have won the 2020 Nobel Prize in Chemistry “for the development of a method for genome editing.”. A nice accompanying piece was published in The Conversation, highlighting the history of these scissors and the politics behind it.

Jennifer Doudna explains this revolutionary genetic engineering tool in a TED lecture. However, she warns:

“All of us have a huge responsibility to consider carefully both the unintended consequences as well as the intended impacts of a scientific breakthrough.”

I hope you enjoyed it!

Witten by Rabia Saleem

Congratulations to a new Dr in the house: Dr Ellen King

Huge congrats to a newly minted Dr Ellen King!  She passed her PhD viva on April 9. This is a testimony to your dedication, strong will and hard work. May this PhD be the beginning of many more successful endeavours, Ellen!

We thank examiners Prof Sally-Ann Cryan (RCSI) and Prof Joanne Lysaght (TCD) for the time and expertise they provided.

We also thank the RCSI PhD Programme for their generous support!

From left to right: Prof Joanne Lysaght, Dr Ellen King, Dr Olga Piskareva & Prof Sally-Ann Cryan

How things work in science: targeting cell components.

How do researchers study cells? How do we get the nitty gritty?

We use many methods to tag and chase various cell components. One of my favourites is fluorescent microscopy. It allows the use of nearly all spectrum of colours from blue to purple in one go. However, we prefer to narrow it down to 2-3 colours and avoid their overlap.

How does it work? First, we use DAPI or Hoescht, which are blue fluorescent dyes used to stain DNA. This way, we tag the nucleus of the cell. Then, we tag a protein of interest. In our case, it was MYCN, a protein that acts as a transcription factor. MYCN amplification is associated with poor prognosis in neuroblastoma. As a transcription factor, it binds to genomic DNA and is located in the nucleus. We used a specific antibody that was labelled with a green fluorescent dye. Look at the image below. The green colour pattern overlaps with the blue colour. Then, we tagged the cytoskeleton, a complex of various proteins that hold the cell architecture and dynamics. We used phalloidin with red fluorescence. It is a highly selective bicyclic peptide and a popular choice for staining actin filaments.

Neuroblastoma organoids stained with DAPI, Phalloidin and anti-MYCN antibody. This work was done during the Fulbright journey to Ewald’s Lab at Johns Hopkins

Now, we can enjoy visualising cells and test different research questions. For example, how do cells respond to a drug? Or how do neuroblastoma cells spread?

Written by Olga Piskareva

How things work in science: Scaffolding

At the Cancer Bioengineering Group, we use different types of scaffolds to mimic the 3D structure of tumours outside the body. We use these scaffolds to test new therapeutics and understand the tumour microenvironment. But I bet you didn’t think we had this in common with spiders?

Spiders make their webs by producing silk from specialized glands in their abdomen. They release the silk through spinnerets located at the back of their abdomen, then use their legs to manipulate the silk strands into intricate patterns, depending on the species and purpose of the web.

The process of web building begins with a scaffold. The specialized glands that spiders use are called spinnerets, and they produce liquid silk proteins that solidify into a thread when they come into contact with air. Using their many legs, spiders can manipulate the threads by changing the speed and tension they enforce on the silk, thus controlling thickness, stickiness and strength. They first lay a framework of non-sticky threads, known as scaffolding. And layer by layer, different species of spiders will add their own artistic sticky silk design to the scaffold depending on their aim. Take the deadly redback spider for example, these guys have a utilitarian approach to web building relying on their webs mainly for shelter and capturing prey. As such, they don’t put much effort into producing irregular and messy homes. In comparison, the orb-weaving spider produces “Mona Lisa”-like designs, with complex geometric patterns and intricate designs. The differences in effort seem to come from the environments in which the webs are located, with the redbacks choosing more sheltered environments and thus not needing much strength to their webs. Whereas orb-weaving spiders are more adapted to a range of environments, from forests to grasslands to urban gardens. So, while the redback gets a lot of attention for their neurotoxic venom, they need to step up their artistic skills to match that of their orb-weaving colleagues.

The redback spider and its webs are reminiscent of an aggressive tumour, which is erratic, dangerous, and unpredictable. We want to find the “anti-venom” for such tumours so we can wipe them out for good.

Watch this amazing web-building timelapse by BBC Earth.

Written by Ellen King

How things work in Science: Tìr na nÒg

In humans, NANOG, SOX2, and OCT4 are transcription factors that maintain the undifferentiated state of embryonic stem cells (ESCs). NANOG was first discovered in 2003 by Chambers et al. and Mitsui et al. as a transcription factor in ESCs responsible for cellular self-renewal. More importantly, it enables continuous self-renewal of cancer stem cells, leading to metastasis when the regulatory genes involved do not function normally. These have been identified as cancer stem cells, with NANOG being a marker of “stemness”. In multiple cancer types, NANOG has various effects, including cellular expression of mesenchymal phenotype, cellular invasion/migration, repressed apoptosis, drug resistance, and increased angiogenesis. In pathways, NANOG either promotes or represses the expression of other genes that lead to cancer-favoured cellular behaviour. Overall, a higher expression level of NANOG is usually indicated in cases of poor prognosis.

NANOG is even more interesting due to its eponym, which comes from Tìr na nÒg. A Celtic myth of the Land of Youth, where the Tuath Dé resided in a supernatural land of paradise. This land offered beauty, health, joy, and everlasting youth to the inhabitants. As the myth goes, the Tuath Dé were gods of the land, and the god that ruled, Manannán mac Lir, was the first ancestor of humans. In various Celtic legends, humans are invited by the gods to visit Tìr na nÒg on great adventures.

However, time passes much slower in Tìr na nÒg, making it precarious for humans to return to their own world. As is the fateful tale of Oisín, who fell in love with the Tìr na nÒg goddess, Niamh. He travelled with her to Tìr na nÒg, where they lived happily in paradise. Upon a visit back to Ireland, Oisín realized that all his family had died over the years. When Oisín found a group of men who were struggling to move a giant rock, he stopped to lend them a hand while on his horse. However, the weight of the rock caused his saddle strap to snap. He fell from his horse, and when he touched the ground, he suddenly aged 300 years all at once.

Written by Alysia Scott

Sources:

Gawlik-Rzemieniewska, Natalia, and Ilona Bednarek. “The role of NANOG transcriptional factor in the development of malignant phenotype of cancer cells.” Cancer biology & therapy vol. 17,1 (2016): 1-10.

“The Story of Tír Na NÓg.” Celtic Titles, 10 Feb. 2022

Congratulations to Dr Ciara Gallagher!


Huge congrats to a newly minted Dr Ciara Gallagher!  She defended her PhD on March 8 – International Women’s Day. Your enthusiasm and perseverance are truly fascinating! May this be the stepping stone towards a brighter future, Ciara!

We thank examiners Dr Marie McIlroy (RCSI) and Prof Jan Škoda (Masaryk Uni) for the time and expertise they provided.

We also thank the Irish Research Council for their generous support!

Dr Ciara Murphy (Chair), Dr Olga Piskareva (Supervisor), Dr Ciara Gallagher, Prof Jan Skoda (examiner), Dr Marie McIlroy (Examiner)