My honor’s Capstone experience was a little different from other honors students. This was because I graduated a semester early, and therefore had to complete my Capstone project during the Fall semester only. Thankfully, I already had a head start on my project because of the work I had completed during the previous summer as an Honors Scholar. The first step of my Senior Capstone project this Fall was to conduct further analysis of the expression level of each amino acid transporter gene of interest. This was accomplished by comparing the graphs of the expression levels for each gene I had constructed during my summer research. I also normalized the data to show the fold-change of each gene’s expression in cancerous versus normal tissue. This allowed a more intuitive understanding of the relative expression of amino acid transport proteins between mouse and human liver tissue, both normal and cancerous.
Our work this past summer also allowed us to determine the exact amino acid transporters to study for comparison between mouse and human genomes. By utilizing bioinformatic “in silico” analysis rather than actual cells, we were able to better generate testable hypotheses and insure that we were not expending time and resources studying solute carriers that are not expressed in liver cancer.
Our work during the fall suggested there were significant differences between mouse and human solute carrier gene expression. On average, the amino acid transporter ASCT2 was suppressed in both human and mouse liver cancer; however, LAT1 was enhanced in humans and suppressed in mice. Among the mouse datasets we studied, there was also significant variation between certain treatments. For instance, ASCT2 was suppressed in all of the samples except for the DENA treatment. Diethylnitrosamine treatment, or DENA, is a chemically induced cirrhosis of the liver and we therefore hypothesized the disparity could be due to the significant scar tissue found in cirrhosis of the liver. We concluded this because scarring in the liver would mean there was an abundance of stromal cells (which are the surrounding support cells) instead of hepatocytes (which are the liver cells) compared to the other samples. We saw similar results in the mouse DENA treatment for LAT1. The DENA treatment was the only sample with a significant elevation in the expression of the gene for LAT1 in mice.
Further research was necessary however to determine if the results from our bioinformatics data analysis was confirmed in actual cells. Therefore, the second step in my project was to shadow one of the PhD candidates in Dr. Bode’s lab in order to learn the proper protocols to grow, maintain and harvest a stock of cancerous liver cells. I also learned how to perform several laboratory analysis procedures such as: isolating RNA, making cDNA and performing RT-qPCR analysis in order to determine the quantitative level of RNA expression in liver cells.
I utilized the skills I had learned in order to help grow a stock of cancerous mouse liver cells for use in future screenings. I then harvested the cells and performed the necessary RNA level analysis. I met with my mentor to determine if the data was sufficient or if any of the experiments needed to be repeated. Unfortunately, we ran into some issues while attempting to get the RT-qPCR analysis to work, and the experiments had to be repeated.
The final step in my Capstone project was to compare the data from the bioinformatics analysis to the levels of RNA and protein expression measured in the lab. This analysis was designed to allow us to either validate or adjust our preliminary conclusions based on the “in silico” bioinformatics analysis this past summer. At this point in our project, there are still a few other tasks that need to be completed before we can finalize our conclusions. My project has served as a starting point for several other lab members who have taken on the project for continued study. We are planning on publishing our data in a scientific journal once the project is fully completed.
I am very grateful for the opportunities that being an Honors Scholar provided me with this past year. One of the aspects of being an Honors Scholar I enjoyed the most was the opportunity for individualized, hands-on training in the lab. My mentors were amazing and really provided me with a unique insight into scientific research and collaboration that is not available in a classroom setting. They were also very understanding of my ever-changing travel schedule due to my medical school interviews throughout this past semester. Over the course of the Fall semester, I traveled to 11 medical schools in several mid-western states for interviews. I have been placed on several wait-lists/accepted alternate lists and have received full acceptances to three medical schools thus far.
I was fortunate to also receive a generous merit scholarship to the University of Illinois – College of Medicine. I am currently planning on attending medical school at U of I, unless I get accepted at Cleveland Clinic from the accepted alternate list which I am currently on. At this point, it is my goal to specialize in surgical oncology. It is my sincere hope that I will continue learning about novel treatments and interventions for patients suffering from cancer through hands-on bench-work in a lab during medical school.
I am confident that the initial foundation of knowledge and experiences I have gained at NIU will prepare me well for my continued cancer research throughout medical school. I believe my experiences in the lab will also prepare me well for my future career as a physician, especially if I specialize in removing tumors through a career in surgical oncology. Although the path of my future education is still somewhat uncertain, this I do know: my experience as an Honors Scholar was one of the best experiences I had while at NIU. My work in the lab has had a profound impact on my personal development as well as my future career planning, and for that I am very grateful!