Feinstein Institute for Medical Research, Northwell Health, Manhasset, NY
This summer I had the wonderful opportunity to be a research intern at the Feinstein Institute for Medical Research, a research facility that aims to be the number one disease-related research facility in the country. My lab group focuses on pediatric sepsis and was under the advisement of Dr. Clifford Deutschman, a world-renowned sepsis expert. Sepsis is defined by the American Medical Association as a life threatening, organ-dysfunction caused by a dysregulated host response to infection. It is the leading cause of death in critically ill patients, accounting for more than 300,000 deaths in the United States each year. The pathobiology is poorly understood and there are no specific therapeutic approaches. It has been proposed that mitochondrial dysfunction plays a role in sepsis pathology. Along with my mentor, Dr. Ariel Brandwein, we looked at the role of mitochondria in sepsis, specifically the mitochondrial complex II in liver cells of mice.
Our lab performs a procedure called Cecal Ligation and Puncture (CLP) in which an incision is made in the abdomen of the mice, exposing the cecum. The cecum was ligated (meaning a surgical thread is tied around the tip of the intestines), punctured with a needle, and then placed back into the abdomen. The purpose of this surgery is to mimic the effects of sepsis in mice. This summer, we euthanized mice at baseline, 24, and 48 hours post-CLP. We used a machine called the Seahorse XF Analyzer in order to measure the oxygen consumption rate during mitochondrial respiration. After euthanizing the mice, we would remove their livers and isolate the mitochondria. Using these samples, we would place them in the Seahorse machine to measure the oxygen consumed after injecting a substrate that stimulates Complex II activity. By isolating the complex II pathway, we were able to see whether the complex was functioning properly in the mitochondria.
We also performed a Complex II Enzyme Activity Assay that is a colorimetric assay using a microplate with wells coated in an anti-complex II monoclonal antibody to purify the enzyme. Complex II (succinate dehydrogenase) is unique because it is the only complex that participates in both the citric acid cycle and the Electron Transport Chain. It consists of two enzyme reactions: the oxidation of succinate to fumarate and the reduction of ubiquinone to ubiquinol. After the purification of the enzyme, the production of ubiquinol is coupled to the reduction of DCPIP in the assay in order to measure the production rate. Through this experiment, we were able to measure the complex II enzyme activity.
At the beginning of the summer, we had hypothesized that oxygen consumption rate (OCR) and Complex II enzyme activity would decrease in hepatic (liver) mitochondria from baseline to 24 to 48 hours post-CLP. However, over the course of the experiments, there was little to no change from baseline to 24 and 48 hours post-procedure seen in the resulting Seahorse data. This could indicate that the liver’s mitochondria are not compromised during sepsis as much as in other organs. We also theorized that we had not done enough experiments waiting long enough after surgery and that the mice were not sick enough to show the debilitating effects in the liver mitochondria. If my internship had lasted longer, we would have performed another Seahorse experiment at 72 hours post-CLP in order to see whether the mice’s livers were not functioning as well at a later time point.
This is my second summer doing pre-clinical research, and while I have enjoyed both experiences, they have solidified for me that research is not what I want to do for the rest of my life. While I still love science and biochemistry, I have always known that I want to be a physician, but I do not think an MD/Ph.D. is in my future. This experience did have many positive outcomes as it is useful to have the skills to do lab benchwork and to be able to analyze results. We also had weekly lab meetings where I was able to practice my presentation skills and sum up our findings each week. The head of the lab, Dr. Deutschman, also gave us a chance to test our knowledge by throwing quick questions at us while we presented. This made us think on our toes and apply hypotheses to our findings. While it could have been embarrassing to stumble over our thoughts or not be able to provide an answer, his method taught me so much and reiterated concepts in my head until I understood them even better. I value the overall process of research and admire anyone who decides to continue with the job as it is a very long, slow process with many frustrations along the way, but that is how progress is made and lives are saved down the line. One big take-away I have from Dr. Deutschman is that scientific discoveries should never be hidden. Research takes a long time and the only way anyone is going to make progress is through sharing their findings with others. Science is all about collaboration.
I am thankful to have been placed with the physician I was with because I also had the opportunity to shadow him in the Pediatric Intensive Care Unit. I have not had experience in pediatrics and this gave me a chance to expand my knowledge of the medical field. It was interesting to see the different cases that pediatric physicians deal with versus those concerning adults. My mentor was a wonderful teacher, both at the hospital and in the lab. On rounds, he was simultaneously quizzing his residents while trying to make everything understandable for me. With his research, he taught me so much about metabolism and the mitochondria, making me feel even more prepared for my upcoming biochemistry classes at Williams. He gave me the chance to run experiments on my own and over the course of the summer my techniques improved and resulted in better and better results. It was interesting to work in a lab full of MD’s, MS’s, and Ph.D.’s all with varying backgrounds and different focuses in molecular biology. I enjoyed having the chance to learn from them all and hear about their lives.
One of my favorite aspects of the internship was the weekly “Lunch and Learn” where all the high school and college interns would gather to hear from different researchers at the Feinstein Institute. We listened to lectures from highly skilled individuals with interesting topics such as “Decoding neural signals to hack the nervous system” or “Using lasers to fight disease.” While I did not have much background in these areas, the lecturers did a great job of speaking at a level that was comprehensible for college science students. I thoroughly enjoyed learning about all the other amazing work this Institute was doing, as it is pretty incredible, groundbreaking research.
This opportunity was only possible because of the Alumni Sponsored Internship Program and for that I am grateful. Thank you to the Class of 1972 and the ’68 Center for Career Exploration for providing me with this opportunity. It has given me many useful skills and will allow me to further pursue my interests in the medical field.
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