Biomedical Engineering Pre-Grad Intern Gunjan Singh

Gunjan Singh"If someone were to go look at the last pages of Laurel Mountain Elementary's 5th grade yearbook, they would find the caption "scientist" under a smiling picture of a young Indian girl. It was a random answer that came to me when the yearbook editors went around asking all the graduating 5th graders what they wanted to be. For the next five to six years I would state a great variety of other professions for my dream career. I wanted to be a marine biologist who could thoroughly enjoy the beauty of whales and dolphins in the wild sea. When Mars took up my curiosity, I dreamt of becoming a revolutionary pioneer who would refill Mars' atmosphere with oxygen and slowly aid the migration of people to an unpolluted world. Somehow however, when the time for college applications came I reverted back to my fifth grade instinctual response. I more than anything wanted simply to be a scientist, a researcher, a discoverer of new truths. This semester as an IE Pre-Grad Intern has greatly helped pave this path that I hope to be in professionally.

For the past few months, I have had the privilege to work with my graduate student Subhamoy Das and many other students in Dr. Aaron Baker's Cardiovascular Bioengineering and Therapeutics Lab. This lab was the perfect fit because it combined my interest in biomedical engineering with more medical topics. Our lab group works mainly on therapeutic angiogenesis. Cardiovascular diseases, characterized by partial or full blockage of the blood vessels, are a global medical and economic problem that remains the leading cause of death worldwide. While performing a bypass, angioplasty, or inserting a stent have offered solutions for restoring blood supply to affected tissue, many patients with advanced stages of the disease cannot resort to traditional methods for revascularization. Therapeutic angiogenesis offers an appealing alternative that induces revascularization through the delivery of growth factors or stem cells. Unfortunately, clinical trials with this method have shown little success. This semester, we further studied the angiogenic pathway of a diseased state to determine if extensive damage effects appropriate growth factor signaling. Four groups of mice were fed for 10 weeks: wild type mice with a standard diet, wild type mice with a fatty diet, ob/ob (leptin deficient) mice with the standard diet, and ob/ob mice with a fatty diet. The heart and muscle tissue were then examined using qPCR and western blot to determine the concentrations of certain growth factors receptors, co-receptors and other components of the signal pathway. Analysis of the different concentrations between each group of mice were made to determine which factors may have caused the diseased models to go through symptoms of cardiovascular disease. In addition, each tissue sample was H&E stained and immunostained for specific proteins. The data so far has conclusively pointed toward a broken signaling pathway in both muscle and heart tissue. However they don't show similar effects. As a result, we can safely assume that the disease affects the tissues in different ways. More downstream signaling molecules need to be tested before we can elucidate the overall effects.

When I first started I ever imagined knowing so much about the topic of syndecans in therapeutic angiogenesis. In fact, coming into the lab I was so focused on learning all the steps and procedures correctly that I failed to see the larger picture and the purpose behind our research for many months. I was aware that it aimed to help those suffering from atherosclerosis and ischemia but the tiny molecules such as Syndecan, Neuropilin, VEGFR, FGFR, and many others all became jumbled in my head. One mistake I made in this critical learning process, is not taking more of an initiative to learn more about what exactly I was doing. I was too content with the continuous and slightly tedious process of performing one Western blot and qPCR after another. Midway through the process I decided to learn more in attempt to connect the small pieces of the research I was doing. The readings were at times difficult to follow but I had been expecting that. What I was entirely surprised by was how much I did understand and how everything together made complete sense. Before that moment I thought of the research as many small projects, but I realized that everything actually fit together quite cohesively. I understood that we needed to look at protein and gene expression levels using Western blots and qPCR but I didn't quickly realize that we needed to look at that signal pathway because we needed to determine if a lack of co-receptors were preventing efficacy of the growth factors.

One of the most important things I learned working in this lab is that research can get extremely frustrating when it appears to halt and not move forward for potentially several weeks despite hours and hours of labor. There was a point during the research process when I needed to work with Human Fibroblast cells taken from patients. Every time I needed to handle them, I took great care in not somehow "hurting" or worse contaminating the cells in hopes that they would grow and show meaningful results. However, after approximately 2 months of little growth and perplexing data, my graduate student and I came to the conclusion that the different fibroblasts were aging differently and could not be compared. By that time an array of bacteria had sadly made themselves home in many of our lab's cell cultures' and so they needed to be disposed of quickly.

While this mini-project took around a month, there were many more times when smaller two day procedures would fail to work. It's amazing how small variables can make such a large different and that repetition is one of the best ways to confirm results and prove statistical significance.
Graduate school I've realized is quite different from undergrad. The continuous cycle of movement from class to class, homework, and tests are absent but in their place one has to take the time to set up their own schedule that will allow them to reach their goals. This semester has definitely taught me to manage my time and determine my goals. While I was initially determined to go to medical school, I am increasingly looking at an M.D./PhD instead. The ability to do research and discover new things in addition to treating patients is quite appealing. The IE Pre-Graduate Internship has allowed me to learn many key aspects of becoming a graduate student especially learning how to be a researcher, learn how to deal with failure, and how to manage my time. I hope to continue this project through my senior year and hopefully all that I have learned this semester will aid me in receiving an M.D. and a Ph.D."