#Fulbright – Blog about neuroblastoma research

#JournalClub with Alysia: How exosomes can promote cancer spread?

Hi, it’s Alysia, here to give you some insight on exosomes’ impact on tumor metastatic niche formation! I found an interesting paper related to my work that I want to share with you: Tumour-derived exosomes drive immunosuppressive macrophages in a pre-metastatic niche through glycolytic dominant metabolic reprogramming.

Exosomes have been a groundbreaking field of research due to their hypothesized ability to create a pre-metastatic niche through non-cancerous cell manipulation. They achieve this by relaying oncogenes and proteins from cancer cells in their molecular “cargo”. In this study, Morrissey et al. examined the role of tumor-derived exosomes (TDEs) in increasing programmed death ligand-1 (PDL-1) expression in macrophages. In pre-metastatic tumour microenvironments, there is an increase in inflammation and an immunosuppressive response, thus promoting tumorigenesis. PDL-1 is increased by TDEs manipulating the signalling of toll-like receptor 2 (TLR2) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB), which increases lactate production and glucose uptake in an event called the “Warburg Effect”, which is common in tumour microenvironments. TDEs taken from lung cancer samples were injected into mice, and PDL-1 expression levels of macrophages were higher than in healthy lung samples.

To accomplish this, Lewis lung carcinoma tumour cells (derived from mice) labelled with a green fluorescent protein (LLC-GFP+) were injected into mice with tumours; seven days later, they were injected with either non-GFP LLC exosomes or control mouse lung epithelial cell exosomes (MLE-12). The mice injected with non-GFP LLC exosomes had no metastatic difference compared to the control mice. However, the mice injected with LLC-GFP+ exosomes had an increase in micro-metastases, which was confirmed using fluorescent microscopy to observe LLC-GFP+ occurrence. With confocal microscopy, they observed an increase in PDL-1 expression in lung tissue compared to the control mice. These findings suggest that PDL-1 expression is preferentially increased in lung tumour tissue by injecting LLC-GFP⁺exosomes.

Figure A. Schematic of experimental design. Figure B. Micro-metastatic burden quantified by LLC-GFP% in the lungs of s.c. GFP-LLC tumour-bearing mice treated intravenously (i.v.) with MLE-12 or LLC exosomes. Representative dot plots of GFP expression in the lungs and summarized data of LLC-GFP% are shown. Each dot represents data from one mouse. *p < 0.05, one-way ANOVA with multiple comparisons. Figure C. Confocal microscopic analysis of lung GFP+ micro-metastases from primary s.c. LLC tumor-bearing mice. Figure D. Confocal microscopic analysis of PD-L1 expression in LLC Exo- or LLC Exo+ -treated lungs. *Adapted from Morrissey et al.*

Researchers also examined the clinical significance of human TDEs and their manipulation of macrophages in the lymph nodes of non-small-cell lung cancer (NSCLC) patients. They found that there was increased expression of PDL-1 in the lymph nodes of these patients. To further study this, the researchers treated macrophages with either MLE-12 or LLC exosomes and then put them in a co-culture with OVA transgenic T-cells (a type of immune cell). The T-cells had a decrease in cell proliferation, which was later reversed by adding neutralizing α-PD-1 to the co-culture. This led them to think that PDL-1 had an inhibitory effect on T-cell function through macrophage stimulation. In summary, this study found that TDEs have an impact on PDL-1 expression found in tumour tissue and pre-metastatic tissue, resulting in creating an immunosuppressed environment for cancer cell growth. In my project, I’m interested in what cargo exosomes can carry from parental tumour cells to non-cancerous cells when creating a pre-metastatic niche. Knowing that there has been documented manipulation of the cellular makeup by exosomes encourages me to continue my research using neuroblastoma cells to elucidate the oncogenic markers relayed by exosomes. This research is very exciting, and I hope to shed some light on exosome impact in neuroblastoma tumour microenvironments with my own work!

Written by Alysia Scott

Women in Science: Mary Golda Ross

A woman from a background of adversity cannot be shaken when confronted with resistance. Mary Golda Ross was, “the kind of person who would walk through a door and stick in her foot to make sure that it stayed open for others”. She followed her passion and held her own to become the first Native American female engineer in the 1940s. Ross was a pioneer of rocket science and was an integral part of, still to this day, classified research on interplanetary space travel.

Mary Golda Ross was the great-great-granddaughter of John Ross, the chief of the Cherokee nation. About 70 years before Mary was born, John was tirelessly resisting the seizure and erasure of Cherokee lands and culture by the U.S. Government. In 1838, due to the Indian Removal Act, tens of thousands of Cherokee were forcibly removed from their homes and forced to march over 1,200 miles (about 1,930 kilometers) into present-day Oklahoma. Thousands died during the journey. Mary was born in Park Hill, Oklahoma in 1908. As a student in the Cherokee Nation, Mary was very bright and caught on to mathematics quickly. During her university classes, Mary was ostracized for her interest in STEM as she found herself alone on one side of the classroom with the men on the other. As was the case with many women in science at the time, she both learned and excelled academically with a massive lack of support.

Courtesy of the Smithsonian Institution, USA.

After receiving her bachelor’s degree, Mary held teaching positions and became a statistical clerk at the Bureau of Indian Affairs. While working, she took astronomy classes at the present-day University of Northern Colorado and earned her master’s degree in mathematics in 1938. A full 100 years after the Trail of Tears. Soon after, World War II had started and Mary moved to California to help with the war efforts, as 1 in 4 women worked outside the home during this period. She was hired as a mathematician in 1942 by Lockheed Aircraft Corporation, an aerospace engineering company. Her first project was analyzing the effects of pressure on the design of the P-38 Lightning, the fastest fighter jet in the world at the time, reaching speeds of 400 mph (640 km/h) in a level flight. Mary’s ambition did not stop there as she began to wonder how to become involved in space travel.

When the war ended, Lockheed saw her brilliance and sent her to UCLA where she earned a professional certification in engineering in 1949. Carrying on her engineering work in 1952, Mary helped found Lockheed’s Advanced Development Program, otherwise known as Skunk Works. She was the only woman engineer on this team, and most of the research from this program is still classified. However, it is known that Mary developed technology for space exploration and orbiting satellites. Mary’s work also helped develop the Agena spacecraft used in the Gemini and Apollo space missions. In 1966, Mary was a primary author for NASA’s Planetary Flight Handbook Vol. III, a still relevant source for space travel. Even suggesting the possibility of travelling to Mars and Venus.

Courtesy of San Diego Air and Space Museum, USA.

Mary retired in 1973 and was influential in both the American Indian Science and Engineering Society and the Society of Women Engineers. She worked hard to encourage increased participation of Native American youth in STEM fields. As of 2021, there are less than 1% of Native Americans that comprise the STEM workforce; only 2% of the U.S. population is Native American. She advocated for more resources offered to Native Americans interested in STEM and increased knowledge of her ancestors’ history. Mary attended the 2004 opening of the Smithsonian National Museum of the American Indian in Washington, D.C., wearing a traditional Cherokee dress. Upon her death in 2008, Mary endowed the museum with $400,000. Mary had told the Los Altos Town Crier, “The museum will tell the true story of the Indian – not just the story of the past, but an ongoing story”. Mary had dreams that reached the stars, and she never stopped chasing them. She trailblazed a path for those who followed in her footsteps to find a way, no matter what barriers were present.

Written by Alysia Scott.

Hey there, this is Alysia!

Hey there, this is Alysia! I recently joined the Cancer Bioengineering team as a first-year PhD student. At the beginning of 2022, I was selected as a Fulbright-RCSI StAR Programme student. I spent the summer in anticipation of what to expect from my new home. I had never travelled outside of America before! However, moving to Dublin has been a great new adventure with tons of amazing experiences and new friendships. One of the first things I did when I got here was going to cafes all over town to get to know the area. Also, to indulge in the foodie scene of Dublin. I also did some great walking tours, joined a tag rugby team, got cosy with the pub culture, and found some unique thrifting shops. Dublin has a lot of charm and hidden gems wedged in between the cobblestone streets. As an American in Dublin, I thought I would have a really hard time adjusting. However, everyone has been so friendly and helpful! I’ve been able to catch on to some of the local dialogue and sometimes catch myself about to say, “it’ll be grand!”

One of my favourite things I’ve done since being in Dublin is exploring the surrounding area through hiking. I’m from Colorado in the U.S., so the mountains here are quite a bit different. However, I can greatly appreciate the hiking culture that everyone seems to enjoy. I’ve been able to go on hikes with colleagues and friends alike in new stunning locations. My first introduction to Irish hiking was along the Howth cliff walk. Not only was the wind strong enough to fly me back to America, but the rain was “lashing”, and the temperature was absolutely, “Baltic”. I was soaked to the bone but could not be happier about getting outside and finding a social hobby that would help me adjust to my new home.

It’s only been a few months into my four-year journey in Ireland, but I’m looking forward to so many opportunities. Being so close to the rest of Europe, I’ve already travelled to Paris, London, and Barcelona. I was even in Brussels and Luxembourg for a Fulbright Seminar visiting the EU and NATO! I plan on travelling around Europe some more, exploring the coasts of Ireland, and of course, sampling all the new cuisine! I really lucked out with a great team here in the Bioengineering Group. We have conferences here in Ireland as well as internationally, which helps me explore. I can’t believe I’m here in Dublin doing cancer research. Being selected for Fulbright is an incredible opportunity for me to fall in love with Dublin while doing a PhD program.

Alysia, Ronja and Lin, Barcelona, February 2023

Written by Alysia Scott

How neuroblastoma cells spread?

Since I joined neuroblastoma research, I have been puzzled by the fact that half of the children with neuroblastoma have the disease spread at the time of diagnosis. It is still a puzzle whether cells spread and primary tumour growth happen simultaneously or more adventurous cancer cells escape the primary tumour location later.

At a cancer conference, I met Prof Ewald who studies this process in breast cancer. I was fascinated by the approach and started to look for opportunities to join his lab. To tell the truth, very few exist for mid-stage career scientists! One of them is the Fulbright program.

One day, I opened my email saying that I received a Fulbright-HRB Health Impact Scholar Award to travel to Johns Hopkins University and adapt their 3D models to learn how neuroblastoma spreads. It was a life-changing experience both personally and professionally. The amount of experimental data collected over 4 months of work did not fit a 1TB memory stick! Indeed, this short journey was just a start of a new research inquiry.

On my return home, the greatest task that remained was to make sense of every single experiment. Cian Gavin took over and spent almost a year systematising, characterising it, and placing it into a context. It was meticulous work with very little known about invasion strategies in neuroblastoma. Now, we are happy to share our findings published on Cancers.

Where do we go now? Well, our next step is to understand the cellular players behind neuroblastoma invasion and how we can target them to stop neuroblastoma spread. It won’t be a short and sweet journey, but we are ready for it!

This fantastic and rewarding work was supported by Fulbright Commission Ireland, National Children’s Research Centre, Health Research Board, Science Foundation Ireland, the National Institutes of Health/National Cancer Institute (Prof Ewald), Alex’s Lemonade Stand Foundation for the COG Childhood Cancer Repository (Prof Reynolds) and the National Institutes of Health/National Cancer Institute (Prof Reynolds).

Home, sweet home…

Checking the calendar, it has been a month since my last post! That month was a transition time. Collecting all that accumulated, shipping self and stuff home, settling back and reinventing the life. Learning to drive on the left side, getting used to the narrow roads and ‘snow-flake’ junctions.

I never had a problem with driving on the right side in the US. Though, my entire driving experience as a driver has happened in Ireland driving on the left side of the road. Apparently, something that you absorb as a toddler, youngster, teenager and young adult never leaves you. It is written in your ‘hard-drive and operation system’ as a default option. So, now I have always to pay attention particularly when roads are empty…

What is my other big discovery? I was adopted by the big-big family and happily accepted this fact! I met 3 sisters and 3 brothers-in-law and a grandpa. Now, I miss our Sunday dinners and chit-chats…

2018 Raleigh Fulbright Enrichment Seminar

The theme of the seminar was American Security: Integrating Multiple Perspectives. With the great support from North Carolina University, we stepped out our comfort zones and explored security topics through lenses of our cultural backgrounds and life experiences. Food Security, Energy Security and Environmental Security. Eighty-four Fulbrighters from almost 40 countries were sharing their stories on how these issues are dealt with in their home countries! We learnt to talk through being open-minded, find things in common and come to a balanced solution. This is not a-one-size-fits-all solution. We have more in common than we have thought. We have become friends and partners who build bridges and connect people and countries. The Fulbright Programme helped us to realise it!

A part of the activities was a site visit. I picked the Food Bank for the very simple reason – I have never heard about such a concept. My imagination fueled by perceptions drew a warehouse full of canned and dried food for a ‘rainy day’. How big was my surprise when I saw an absolutely different picture! Many dedicated people with the huge help of volunteers collect, sort, pack and distribute all type of food from vegetables to meat for people who can’t afford to buy it themselves. They collect fresh vegetables that do not meet perfection standards (called also number 2) from farmers. Giant sweet potatoes, ‘ugly’ squash or oranges – they all have the same nutrition value as their glamour looking brothers and sisters. Why #2 should be left in the field? So the Food Bank takes them in. The Bank also educates people on how to cook healthy meals from raw products.

Happy Thanksgiving 2018!

On November 22, almost all Americans and visitors celebrated Thanksgiving. Roads and airports were overcrowded a day before. Turkeys, potatoes, corn and pumpkin tarts were consumed in astronomical amounts. The New York Times cherry picked 50 recipes from the 50 states. Yamee!!!! Have a look – I am voting for Alaska’s one!

So, how was my experience? As a native American – together with the family! I drove through big and small towns, beautiful autumn sunflower fields and forests covering a mile after a mile. When I thought that I was nearly there GPS updated my route with instructions to drive another 5-10 miles. So after an hour, I joined the big gathering in Grandpa’s house in northern Harford County, Maryland.

Many generations sat at the family table making it noisy, warm and live. Grandpa said the prayer. Turkey was served with many tasty sides. Some food combinations were entirely new for me… Having turkey with sweet strawberry jello was unusual. Or with cinnamon-sprinkled baked apples. Both were delicious on their own right! Two recipes I pencilled down for my family – a broccoli salad and a strawberry jello though will use them differently. If the broccoli salad perfectly fits into my vision of appetisers, the jello is for the desserts collection.

After the main men were watching sports, children were surfing the Internet, and the women cleaned the dishes and put cakes, tarts and torts on the table. Chitchat spiced with jokes and old days funny stories were all day long. What else would you wish for?

Sharing this day with this American family recalled childhood memories when I was stepping into my Granny’s arms in a warm kitchen smelled with baked pies and pastries from chilly and wet outdoors.

Americans see Thanksgiving as a day of feasting, football and family. Why not? Traditions are not bad at all. In the modern busy life, family dinners link us together letting stop the moment, smile at each other and thank for being here and now. By the end of the day, the family matters more than any anything else. Isn’t it?

Sightseeing discoveries in Baltimore

Coming to live and work for a short period ignites opposite feelings. Should I see all the places recommended by Tripadvisor or do it at my pace as it happens? What if I miss something worthy? Perhaps, your own intuition balances somehow both.

I have discovered and loved two buildings in Baltimore. One is the building of Barnes & Noble bookstore in Inner Harbour and the other is Mount Vernon Place Church. Both have a great history behind and give a very warm feeling when you are inside.

Barnes & Noble bookstore is located in the former Power Plant. The features are easily spotted. From outside, the building looks like a Plant for modern social activities. Ugly slightly, isn’t it? Though, it is a different feeling when you enter the bookstore. The Plant scaffolds, chimneys and pipes are nicely crafted into a warm welcoming environment. Even lights are dimmed as back then. Rambling through the bookshelves and feeling the magic of the place and unread stories on them. You can pick up a book, sit where you are and enjoy the reading. Maybe it is the feeling of my childhood full of books and hours of reading?

Mount Vernon Place Church is next to the Washington Monument and recalls old Catholic Cathedrals in a Victorian Gothic Style. Built in 1872, the Church is actually the United Methodist Church from the very beginning. It is beautiful inside as well as outside. The stained glass and carvings have Nature themes. A big rose window in the rear of the church was inspired by the one in Notre Dame. It has an interesting history which you can read here. I discovered this place by chance and am glad that this chance came from my host family. I saw the interior not only the Church but also Asbury House. The mansion was a home to the first bishop of the Methodist Church in America – Francis Asbury (1745-1816). Have a look at the pictures, the furniture is well preserved, the staircase is similar to the one you can see in Slane Castle. Its light design makes it looks flying. Indeed, not everyone could use it at that times. Servants had a plain version at the back of the house, so none could have seen them moving in and out. Lots of blue and carved wood in the rooms. Incredibly beautiful.

Mount Vernon Place Church & Asbury House

Taste of Guinness in Baltimore

What is a must-see in Ireland? Right: visit Guinness brewery at St. James’s Gate and have a pint of right Guinness. It is one of the most recognised and famous beer brands. Rumours say Dublin is the only place where Guinness tastes Guinness. Traditions, traditions and traditions. Though the one we like most Guinness Draught is a relatively new addition – it was introduced almost 200 years after the brewery establishment in 1759 by Arthur Guinness.

So, what if you like tastes ‘outside the box’? Then Guinness brewery in Baltimore is for you. Respecting the Father, they do completely different stuff. Imagine, 16 different tastes, including the one we know! You can have a guided tour of the brewery, hear the great story and do a beer tasting. Have you tried one? Not, the one in the pub or with the friends at BBQ. It is a special way to feel the bouquet of flavours and taste the difference. There is a difference between the beer drunk straight from the bottle and from the glass. Because you can smell it as simple as that.

Taste of Guinness, the Guinness Brewery opened in Baltimore, MD, the USA in 2017

During our guided tour, we rambled inside the experimental brewery, learn the basics of beer production, tasted 4 types of beer: Guinness Blonde, Oatmeal Pale Ale, Guinness Draught and Guinness Milk Stout. Three were absolutely new for me. I liked Oatmeal Pale Ale, found Milk Stout a bit dessert style, Blonde – too citrusy. Do not forget, another 12 you can taste at the bar in a special set! However, to enjoy the most you have to bring your friends. All is much better with the right company!

Beer tasting comrades at the Fri night out

Halfway through

Can you control the time? I can’t and know none who can. It flies, when things around you work out, and drags on when not. The time flies for me here in Baltimore. It feels so intense sometimes and then slightly slows down. I take pictures almost of everything: the path’s signposts when rambling in the network of Johns Hopkins Buildings, the first frosty morning, joyful deer at the backyard of my host family house, outdated clothes in the shop…

In the past, I had a similar journey to Ireland. It was 3 months research placement during my PhD. Did I like it – oh, yes I did! I travelled a lot, felt romantic and changed my life on my return home. But I did not run a diary or tag my way on Facebook. I have learnt the lesson: do it even more intense as you can’t travel back in time and write down your experience. It may be funny or educational to read in a couple of years! I become addicted to it though not always have time to do it.

I like the people who I am working with. They are a fantastic bunch of self-motivators and open-minded personalities. They are workaholics either naturally like me or because of the exciting projects they do like I do. Who knows, but very likely because of both. Isn’t it a dream to have an exciting project and great people around you? The luck like this gives you wings.

The American enthusiast studying Russian and my Mum

The host family – is my other great luck! This luck was crafted as a parallel story when none knew how the Fulbright application and an American enthusiast learning Russian may intersect. You would not believe, but parallel lines can be non-parallel sometimes! His journey to my home city in Russia paved the way to the opportunity to stay at his aunt’s house.

Every day 50 min drive to and from Hopkins opens up the other side of the local lifestyle and infrastructure. What are the rush hours? How many drivers are signalling before taking a turn? How do they call the shopping trolley? How parking system works?

Experiencing life as an American working in Baltimore.