Author Archives: Cassie Ingram

Looking into the Future

This summer has been an amazing experience. I have gotten a really good look into what life would be like if I chose to go to grad school. While I still don’t know for sure if this is what I intend to do after school ends, I now know so much more about this option. Before doing BSURF, I didn’t really have any idea of what getting a Ph.D. entailed and what graduate school was really like, but now I have spent the last 8 weeks in a lab full of graduate students and post docs and learned so much more about pursuing a career in academia.

I think the most important thing I got out of this summer was being paired up with Lucas as my mentor. He seems to really care about me and my learning and me getting as much as I can out of this experience. I know that I have learned so much from this summer about BME and immunology and that is all thanks to him.

I have also learned more about science—how science is actually done in the lab, how it is written about, and how it is presented. The morning meetings with Dr. G and Jason really helped with this part, as well as all the faculty talks. I feel like I understand more about what a career in science looks like.

I never realized bird songs could be so interesting…

I have to say, the seminars have been incredibly interesting and a great start to my mornings. I like that we have been able to listen to a wide breadth of science being done a Duke and learn about all different types of fields and research being done. In our labs, we have been getting a very in depth look into one subject, but through the seminars we have been able to learn about all different types of research.

One of my favorite seminars was on something very unexpected: bird songs. Dr. Nowicki studies how birds communicate and how their songs evolve, and then uses birds as a model system to better understand human language and communication. This lab does not relate to anything I am studying and the field work they do is pretty different from the bench work I do in the lab every day. Quite honestly, I really did not think that bird songs could be very interesting. However, I walked out of Dr. Nowicki’s talk suddenly incredibly interested in bird songs! The way that Dr. Nowicki presented his research was engaging and easy to understand, so I felt like I had a good idea about what was going on in his lab without being overwhelmed. Additionally, I was interested in learning more.

All the talks have been incredibly interesting and got me thinking about all the different types of research out there. I do think the most influential parts of the talks, though, were the parts where the faculty told us about their career journeys and how they got to where they are today. Those stories really gave me a good understanding of what my career path may look like if I choose to go to grad school. They also let me know that there are many ways to get to the same place, and let me know that there is still time for me to figure out what I want to do in the future. Doing lab work right now is really awesome for whatever I may choose to do, but I still have time to explore my interests and find my path.

Abstract Draft

Self-assembling peptide nanofibers have been shown to have self-adjuvanting properties—stimulating targeted immune responses in vivo and raising long lasting antibody responses without an accompanying inflammatory response. Despite these novel properties, the mechanisms behind nanofiber induced immune responses are still not known. To investigate these mechanisms, the self assembling Q11 peptide sequence was used. Dendritic cell (DC) activation was measured in vitro using the DC2.4 dendritic cell line, which immunohistochemistry identified nanofiber localization within the lymph node in a murine model of immunization. DC activation in vitro was shown to be dependent on the presence and concentration of the Pan-DR-epitope (PADRE) within the nanofiber. Measurements made by flow cytometry showed a dose dependent upregulation of CD83 and MHCII when PADRE containing nanofibers were added to DC cell culture media. Additionally, immunohistochemical analysis of lymph nodes containing nanofibers conjugated with the fluorescent molecule Tetramethyl-6-Carboxyrhodamine (TAMRA) showed a colocalization of injected nanofibers with both macrophages and DCs in vivo. These results indicate that nanofiber induced immune responses function in both canonical pathways, involving DCs, and non-standard pathways, involving macrophages. This provides a framework to begin to understand the complex immune responses generated by self-assembled peptide nanofiber vaccines.

Research is Really Cool

First of all, I would like to say that everybody did a great job with their Chalk Talks and I really enjoyed this week of morning meetings. I know that we have gone in circles a lot of times telling people our names and our favorite animals and what lab we are working in, but this was the first time we got to talk about our research in depth with everybody in BSURF. I think one of the reasons that BSURF is such a cool program is that there are so many different types of research being done by everybody in the program so we can really get a good idea about what kind of research opportunities there are just within Duke. These talks really showed that there is so much to learn and do in the world of science.

Alie’s talk was particularly interesting to me because she works in a neuroengineering lab, and as a Pratt star of course that would peak my interest. Alie’s lab focuses on using psychiatric neuroengineering to study and provide potential treatments for mental illnesses and Alie’s particular project is looking at the involvement of MeCP2 in anti-depressant treatment of socially defeated mice. Alie is trying to figure out whether MeCP2, which is necessary for the proper function of nerve cells, is phosphorylated when mouse models are given imipramine (an antidepressant drug), because the phosphorylation of MeCP2 is shown to have antidepressant effects. Alie explained that she is using the social defeat method to make mice depressed and then treating mice with different amounts of imipramine to see if they are less depressed and whether their MeCP2 levels increase. I think Alie did a great job with her presentation (as did everybody!!) and I really liked learning about her work.

I also did some of my own googling to learn more about Alie’s lab because I was so interested and the work they do there is pretty damn cool. Her PI, Dr. Dzirasa, has this goal of making a pace maker, which is typically used for hearts, for the brain to treat psychiatric disorders. This is the kind of work that is using engineering to solve medial problems in the work that made me interested in biomedical engineering in the first place.

A Day in the Life

My day usually starts with cells. I am culturing dendritic cells, and they need to be passaged on Monday, Wednesday, and Friday. So when I get into lab, I meet with the Lucas, the graduate student I am working with, and we talk about the plan for the day, and then I usually passage my cells shortly after we meet. On Tuesday and Thursday I just take a look at my cells under the microscope to make sure they are still growing healthily. Then I launch into the other tasks of the day.

For the first couple weeks, a lot of what I was doing was learning procedures and finding my way around the lab. I would watch Lucas do a variety of tests and experiments, trying to absorb everything that was going on. I learned how to passage my cells, do ELISAs, stain histology slides, and more. I am now doing more and more on my own now, becoming more confident around the lab. What I do can vary a lot from day to day depending on where Lucas and I are with our experiments, but it all centers around the same main experiments. I split my cells either just to passage them or to prepare them for an experiment. If an experiment is coming up, I will do other prep work for the cell experiment, and if it’s the day of an experiment I will perform the experiment. I may cut lymph node tissues, stain the tissues, or look at the stained tissues under the microscope. And, if it is the day of a blood draw, I might do an ELISA.

One thing that is a constant every day, though, is that whenever I am not doing anything at the bench, I am reading. I have done a ton of reading in the last month and I have learned so much about immunology that I would have probably never learned any other way. I started off by reading the better part of an immunology textbook, and then moved on to reading articles about biomaterial vaccines and engineering the immune system. I have a whole folder of papers on my computer of papers to read, and am making my way through them one by one in my free time. So far, I am enjoying my time in the lab a lot and I am learning so much every single day.

Engineering the Immune System

The focus of my lab is developing and characterizing self-assembling, self-adjuvanting nanofibers and other materials to induce certain immune responses to provide another possible platform for vaccine design and other medical applications. I have been paired with Lucas Shores, a rising second year graduate student, and I am helping him to run his experiments. The project we are working on builds on past projects by Collier Group in order to better understand these peptide nanofibers. We are trying to figure out exactly how the nanofibers interact with the immune system and induce an immune response, and how to engineer this response to me more catered toward a particular disease (by, for example, engaging B cells and Tfh cells, rather than a Th1/Th2 response if you want the vaccine to activate the humoral side of the adaptive immune system).

Lucas is looking specifically at using nanofibers to target the IL-17 cytokine in order to possibly create another treatment for inflammatory diseases, like autoimmune diseases such as Rheumatoid Arthritis. IL-17 is an inflammatory molecule, and so the theory is that if a vaccine can induce an immune response to destroy or deactivate IL-17, then inflammatory symptoms would be reduced. Lucas has designed and synthesized a few peptide epitopes targeting IL-17 and is testing their effectiveness in creating an immune response in mouse models.

In addition to these mouse experiments, Lucas and I are culturing dendritic cells from a cell line in order to do experiments that investigate antigen uptake by dendritic cells and are doing lymph node immunohistochemistry in attempt to further understand which cells the nanofibers interact with when they enter the mouse.

The Stories Behind the Research Papers

Dr. Joel Collier is the PI of the Collier Group—a biomedical engineering lab here at Duke that is working to design new vaccines, and I have been placed in his lab over the summer. He took some time out of his busy schedule to talk to me earlier this week, and I want to share some of what he told me about his experience of becoming a scientist, his favorite parts of the job, and some advice he gave me, and I am looking at a carrer in scienve.

Dr. Collier’s lab is focused on immunoenginnering, but he has not always been interested in this field. He got his undergraduate degree at Rice University in materials science—biomedical engineering, being a pretty new field, wasn’t even a major at Rice University at that time. Dr. Collier didn’t even go straight to graduate school out of undergrad. Rather, he took a few years to work in industry to satisfy his curiosity, and then returned to school at the University of Chicago. All this has led him to be a PI and a professor at Duke University. However, what I found most interesting is Dr. Collier’s perspective on teaching and running a lab.

In the interview, Dr. Collier explained that he believes labs produce two products: new technology and people, and he believes the more important of the two is the people each project produces. Research papers, Dr. Collier elaborated, contain a lot of science, but they do not really tell the story of how a discovery was made. A graduate student comes into a lab a different person than he or she leaves as, having learned more inside and outside of the lab. Each research paper has a story behind it that has to do with the growth of the person writing the paper. Dr. Collier enjoys having the opportunity to cultivate the learning and experiences of his students.

Finally, Dr. Collier dropped his final piece of advice. As a rising sophomore, I still don’t really know what I am going to do after college. In terms of deciding a career path, Dr. Collier told me to strongly consider how the day to day life of a particular job fits my personality because someone doing research, working as a physician, or working in industry has a vastly different everyday life.

What am I doing here?

Coming into BSURF, I honestly had no idea what to expect. I’ve never worked in a lab before, but I have thought for a while that I would like to work in one. I mean, one of the reasons I chose to go to Duke was because they have so many research opportunities, even for undergraduate students, and I knew I would want to do research in college. It is a little weird, though, that I still didn’t really know much about what doing research even entailed, even when I was contacting my PI for the first time to set up a meeting about working in his lab over the summer. I wanted to do research because that was where new discoveries were being made and new technologies were being developed. I am majoring in biomedical engineering because I want to take my love of science and make something out of it. BME labs are developing the medical technologies that are proving means of dealing with medical issues. I suppose my main goal of the summer was to get some hands-on research experience by diving right in so I could figure out if doing research was something I really wanted to do, and if it was, then learning ways to stay involved for the next three years of my college experience.

Now that I have spent a week in the lab, I have a little more of an idea what working in a lab involves, so I know a little more about what I should expect from the next 7 weeks in the lab. At first, I was very overwhelmed because I felt unprepared and like there was no way I was going to understand what they are doing in the lab with my freshman knowledge of biology, chemistry, and engineering. But through asking questions, I quickly started to learn and now I feel like I sort of know what is going on. This summer, I expect to learn more and more about how to do common lab protocols, such as cell culturing and histology. I expect to become more and more educated in the field of immunoengineering so that reading journal articles in this field will no longer be so confusing. I also expect to make new friends through the BSURF program and learn more about doing research from their experiences as well. Finally, I expect to learn a lot from the faculty seminars during our morning meetings about the many different things going on at Duke. I am very excited to keep learning in the next 7 weeks.