Summer Research 2019: Neuroimmune Dysfunction in Chronic Fatigue Syndrome

Throughout the summer of 2019, I have worked on a research project at the Institute of Behavioral Medicine Research with a focus on evaluating the contribution of EBV dUTPase to the neuroimmune dysfunction associated with Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS). The main component of my contribution was conducting gene expression analysis on brains of wild-type or TLR2 knock-out mice injected with EBV dUTPase through qRT-PCR, with additional parts of the research project being continued in the fall semester.

Prior to this project, I had previously spent three years participating in research, however, it was typically limited to preparing materials for an experiment or assisting another lab technician with an experiment. During that time, I learned some of the basic scientific concepts that outlined the research, but once in charge of my own independent project, I was able to gain a much deeper understanding of the science behind the experiments. Most of my courses have centered on the sciences as a pre-medical student, but they have been limited to mainly biology, chemistry, and biochemistry. This project not only improved my knowledge in these areas, but also expanded it to much deeper concepts such as immunology and virology.

The transformation within myself, though, came directly from managing the project. I have previously participated in group projects, as well as completed my own, independent projects for courses at Ohio State, yet none have compared to the skills that are needed to complete an independent research project. This project improved my time management skills, attention to detail, problem solving skills, and ability to communicate ideas such that I believe I have become a better leader and team-member, as well as a better student and scientist.

When conducting the experiments, time management was absolutely necessary in order to conduct the experiments correctly, but also to complete the project in an adequate amount of time. Several steps required incubation periods, however, it was always important to maximize this time in order to prepare other components of the experiment so that the experiment would flow quickly and efficiently. Additionally, on a larger scale, I needed to plan out the timeline of the experiments over several weeks in order to stay on track to complete the gene expression analysis prior to classes beginning once again. Gene expression analysis began with isolating the RNA from the left and right hemispheres of brain samples from nearly twenty mice, then developing the cDNA and conducting qRT-PCR on each sample. This process would take several days, with some overnight wait periods in between, so it was critical that I planned each day out to maximize my time within the week.

Throughout every single step of each experiment, attention to detail was very important to ensure that I was adding the correct amounts of each substrate and buffer at very small quantities, sometimes as small as 1.0 uL, so that the reaction would proceed properly. Attention to detail was especially important when creating the plates for qRT-PCR, which involved ninety-six well plates with each well containing only 20 uL of solution that included less than 10 uL of cDNA. Each well needed an accurate ratio of cDNA to the various buffers, primers, and enzymes in order for DNA amplification to occur.

Even though my attention to detail did greatly improve, mistakes were also made at several points throughout the experiments, and this provided an opportunity for me to work on my problem-solving skills as well as develop better communication skills. When something did go wrong with an experiment, it was important that I communicated these issues with my Principal Investigator as well as the other members of the lab so that we could all work on solving the problem together. When the RNA wasn’t precipitating out correctly during isolation, I communicated this, and we were able to adjust the protocol to include overnight incubations at -20C to increase pellet size. When designing the plates, we were able to configure layouts that allowed for multiple samples to be run at once to improve cost efficiency. When one plate didn’t amplify correctly, I was able to problem solve with the lab technician to determine the mistakes made previously and make adjustments for the next run. Not all communication improvements came out of mistakes, however, as I was able to practice reporting my data during lab meetings each week and receiving feedback on how to improve for the week following.

Following my undergraduate degree, I plan to continue my education in medical school and pursue a career as a physician. On the surface level, the skills I learned in performing academic research were good practice for continuing research throughout my training to become a physician. The soft skills, such as improved time management and effective communication, will also be very beneficial to being a physician and a scientist when communicating with patients or managing my schedule.

More immediately, these skills will help me to become a better student. Improving my problem-solving skills has made me a more curious student and has created a desire to pursue further research and more challenging academic courses so I can continue to develop and apply these skills. This project has also encouraged me to pursue a thesis for research distinction prior to graduation so I can continue my research, as well as improve my abilities to analyze data and draw conclusions from my work.