Thesis: Understanding why your veggies are good for you: The role of hydrogen peroxide in electrophile-induced Nrf2/ARE activation
Majors: Biology & Honors
Concentration: Ethics and Health Care
Postgraduate Plans: I plan on attending Columbia University Mailman School of Public Health to pursue a Master’s degree in Public Health: Sociomedical Sciences.
Briefly explain your topic and what you were looking to find.
The majority of healthcare spending in the United States goes towards chronic disease treatment. Activating the Nrf2 transcription factor can prevent chronic diseases such as diabetes, cancer, heart disease, and neurodegenerative diseases. Through binding to the antioxidant response element (ARE), Nrf2 upregulates genes that encode cytoprotective (cell-protecting) proteins, including AKR1C1 and HO-1.
You can find Nrf2 activators in vegetables and other natural plant products such as kale, ginger, broccoli sprouts, and green tea. Studies conflict on whether or not reactive oxygen species (ROS) produced by Nrf2 activators play a meaningful role in Nrf2/ARE activation. We hypothesized that while not significantly activating ARE-regulated gene expression on their own, ROS (hydrogen peroxide, in particular) will significantly enhance the activation of those genes by an electrophile, namely sulforaphane extracted from broccoli sprouts. Human keratinocyte cells were treated with sulforaphane and varying concentrations of hydrogen peroxide (H2O2, one type of ROS) and dtBHQ (which produces many different types of ROS) and Western blot analysis was conducted to examine the effects of varying kinds of reactive oxygen species.
What motivated you to conduct this research?
The clinical relevance of Nrf2 is astounding and extremely diverse! Sulphoraphane is currently being used in clinical trials for a myriad of conditions, including diabetes, asthma, and cystic fibrosis. The possibility of discovering a way to make these activators even more powerful in its disease-mitigating capacity, a “super-sulforaphane”, if you will, is compelling and exhilarating. I have been a part of the Eggler Lab since my freshman year, and it is truly like a family! Studying the specific topic of hydrogen peroxide within a larger narrative of the Nrf2 pathway allowed me to work closely and collaboratively with my mentor and my labmates/friends.
What is the most interesting thing you learned while conducting your research?
My results show that H2O2 can enhance the activation of HO-1 by an electrophile (sulphoraphane), yet has little to no effect on the induction of another ARE-regulated antioxidant protein, AKR1C1. This is interesting, since these two proteins both play protective antioxidant roles within the cell and are regulated by the same sequence, yet respond completely differently after treatment with a single stressor, namely hydrogen peroxide. This showed me how complex each biochemical structure and process truly is. Even two proteins that have responded similarly in other experiments can yield different results under another set of conditions.
How do you plan on using the information that your thesis uncovered? What kind of attention do you hope your research triggers?
I think the most unexpected thing I discovered while performing research is that there are so many confounding variables to consider. I expected to come across at least some difficulty because science tends to do whatever it wants. However, my mentor, my colleagues, and I came across one factor after another that tweaked the results just a little bit. Thus, I discovered that even the soundly-designed experiments require consistent and continuous optimization. Since we must try to account for everything that may be skewing our data – the cell density while the cells are growing and undergoing treatment, the humidity of the chamber they are growing in, the age and condition of the media that they are kept in, and even the types of sponges we use during transfer – can affect the results drastically. Troubleshooting every step of the way is key!
How do you plan on using the information your research/writing has uncovered, and/or, what kind of change or attention do you hope that your research/writing triggers?
My mentor, Dr. Eggler, is preparing a manuscript that will likely include some of the research that my labmates and I have done on this project! It is important, once good research is performed, to relay these findings to the public. I hope that my project can help to contribute to the existing conversation about the role that reactive oxygen species play in antioxidant responses. I hope that it triggers more questions and thus more exploration into every minute inner-working of Nrf2’s wonderfully complex pathway. The more we know about this influential transcription factor, the better! When I learn and read about health disparities, many discussions include the topic of nutrition and its effect on the prevalence of certain diseases. Thus, this research not only has relevance clinically, but also socially. Most importantly, I have gained invaluable critical thinking and quantitative analysis skills from this entire thesis-writing process.