Back in the Lab
June 16, 2024
Hello again! Allow me to reintroduce myself. I’m Eleanor Helm and I am now a rising senior at the University of Delaware where I study cellular and molecular biology. It has been a year since I last worked at Fox Chase, and I’m so excited to be back for a second summer. At the end of the program last year, I couldn’t believe it was over so fast. Now, I’m extremely grateful to have the chance to return and keep learning more this summer, this time in a new lab. Last year I was in the Balachandran lab, learning about Z-RNA and necroptosis, but this summer I will be studying the role of β-catenin in liver cancer as part of the Font-Burgada lab. I’m so excited to get this new experience, learn additional techniques, and get a different perspective on cancer research.
Another thing I am particularly looking forward to is the chance to help and support the new students in the Empower fellowship this year. I loved doing research last summer, but there were also a lot of challenges. I hope that I will not only be able to give advice to the incoming fellows and reassure them that they aren’t the only ones who might be struggling to learn all of these new concepts, but also to be able to share in the excitement and successes they will have. Already, I’m so impressed with the other students this year, and I’m glad to be able to spend the summer learning from them as well.
My personal goal for this summer is to work more independently and collect some real data. Now that I have some lab experience, I feel like I’m over the learning curve that I faced last year and will be able to get started on my project much more quickly. From all that I learned last year, I now have a much better understanding of how to read research papers and protocols, as well as how to do basic lab techniques like pipetting, immunofluorescence staining, and making solutions. Already, I feel more confident in doing experiments. For example, this week, I learned how to use a cryostat to make slices of a liver to put on a microscope slide. Then I followed an immunofluorescence staining protocol to stain the slides for particular proteins in the liver cells. Last year, I was just learning how to properly use a micropipette!
All in all, I am very excited to see where this summer takes me. I feel very lucky to have met so many great people so far–the other Empower fellows, my mentor Abhinav Illendula, and my PI Dr. Joan Font-Burgada–and I can’t wait to keep learning and growing with them over the course of these 11 weeks.
Halfway There
July 14, 2024
The past five weeks have gone by so quickly now that I’ve started working on my project. This summer I’m investigating the effects of a mutation in an important transcription factor called β-catenin. My lab has observed that this β-catenin mutation leads to liver tumors in mice with diseased livers but not in healthy mice. In order to determine why the β-catenin mutants behave differently in healthy versus diseased liver tissue, we are working to identify differences in the expression of certain pathway proteins by using immunofluorescence staining. This method allows us to visualize increases or decreases of protein levels in these pathways, and if we do see a difference, this may explain why the mutant cells generate tumors in diseased livers but not healthy ones.
Each week, I have been working to improve our method of staining these key proteins. Some proteins appear more clearly when you pressure cook the samples, so I have been experimenting with pressure cooking them to get rid of background fluorescence. I’m excited to finally get a clear signal and begin to compare the level of protein expression between healthy and diseased livers so I can answer our research question of why mutant cells behave differently in healthy and diseased tissue.
After staining so many liver samples, I definitely feel confident in following this protocol. But even though I’m confident now, it took a lot of trial and error to get to this point. First, I struggled to learn how to use a machine called a cryostat to make slices of the livers to put on the microscope slides. Then there was an issue with the slides themselves which caused the liver tissue to completely fall off. I also had to learn how to use the microscope to look at my samples which has probably been my favorite thing to learn so far. The immunofluorescence staining protocol takes two days to complete, so when you can finally sit down at the microscope and look at your slides, it is such an immediate payoff to see your proteins light up under the microscope’s lasers.
Even though there have been challenges with getting a clear staining, I think I’m close to being able to see my proteins. It has been a fascinating experience to see how my mentor and PI work through challenges with my project, and being a part of their conversations has allowed me to see how scientists think through these types of problems and brainstorm solutions. I appreciate that they involve me with their discussions and are helping me learn how to problem solve like them and get to the next step in the research.
I’m really excited to get to a point where I can begin quantifying my data, and I look forward to sharing my data at the end of the summer. I’m also really excited to hear what the other fellowship students have been working on. I’ve enjoyed getting a preview of their projects in the past few weeks as we have started to put together our presentations and abstracts, but I’m sure the final product will be even better. I can’t wait to see where our research ends up in another 5 weeks.
A Bittersweet Goodbye
August 26, 2024
It has been a week since the end of the program, and I already miss working in the Font-Burgada lab. Finishing my project took so much concentration and hard work that I didn’t have time to really consider that the summer was almost over. But, as I look back on those short few months, I’ve been able to think about how much I truly loved being a part of the research at Fox Chase and how wonderful my experience there was. I am so proud of the work that I was able to do this summer. 
I not only learned how to carry out new protocols like immunofluorescence staining and genotyping, but I also got to see how a different lab worked and pick up on the habits of a lot of really talented scientists. I especially liked being able to sit in on lab meetings where I got to hear about all of the different projects everyone had going on, see how the lab worked through problems together, and get an idea of how all of the little pieces came together to tell a story about their research.
I’m also so thankful for my mentor, Abhinav, who taught me so much this summer and always found a way to include me in the lab’s research. It is rare to have a mentor who invests so much time and attention in helping you grow, and I will always be thankful for his guidance and friendship. Especially in the final week before the fellowship’s symposium, he was still helping me polish my presentation and put together something I was proud to show to my friends, family, and the scientists in the audience. As the day of the final symposium got closer and I was frantically circling and counting cells on my immunofluorescence slide scans, he was a calming presence, helping me tie everything together so that on the day of my presentation, I knew the research inside and out.
We began the summer by asking why constitutively active β-catenin leads to liver tumors in mice with diseased livers but not in healthy mice. Specifically, we wanted to know how mutated constitutively active β-catenin results in the elimination of the mutant cells from healthy livers. This led us to investigate if there was a change in the level of protein expression related to endoplasmic reticulum (ER) stress or oxidative stress in these mutant cells. We used immunofluorescence staining to check for levels of two proteins – BiP and CHOP – which are both involved in ER stress, and we found that there were increased levels of both when β-catenin is constitutively active, indicating that there was more ER stress in the mutant cells. Based on the lab’s past research, they also observed that these β-catenin mutant cells were shrinking over time as they were eliminated in the healthy liver, so we wanted to investigate why that was happening. The lab suspected that this was the result of oxidative stress occurring in the mutant cells, so they treated mice with an antioxidant to see if this would avoid the shrinking they saw in the mutant cells. Then, they collected the mice livers, and I stained the cells to observe how large the cells were. For this, I used a computer program to measure the area of cells in both antioxidant-treated mice and control mice, and we found that the treated mice did avoid the mutant cell shrinkage unlike the control. 
This shows that there is oxidative stress occurring in the β-catenin mutant cells. Both of these forms of stress may contribute to the decreased cellular fitness we see in the mutant cells, resulting in their elimination in the healthy liver. With this research, the lab can now compare this model to a diseased liver where the same mutation leads to cancer.
Now that I am leaving Fox Chase after two amazing summers, I realize that I wouldn’t be the person or scientist I am today without the support and encouragement of all the incredible people I’ve met there. I’m so lucky to have had the chance to really contribute something this summer as well as benefit from all of the opportunities I was given. Coming back and getting more experience in the lab has just shown me again how much I love science and research, and I’m so glad to have been able to be a part of this incredible community of scientists. Thank you so much for everything, Fox Chase.