Research Title: Determining if Radioprotection Can Be Enhanced Through the Use of Deinococcus radiodurans MDP and Tardigrada Dsup Protein: Implications for Space Exploration

 

Abstract: Ionizing radiation is the number one risk to astronaut health beyond low Earth orbit. This type of radiation, consisting of subatomic particles, X-rays, and gamma rays, can strip electrons from atoms, causing severe DNA damage, often leading to cancer. Some organisms, known as extremophiles, can

survive substantial exposure to ionizing radiation. The Tardigrada, uses Dsup, its damage suppressor protein, to protect its DNA during stress. Deinococcus radiodurans protects its repair proteins against radiation using manganese-decapeptides (MDP). The purpose of this study was to determine if radioresistance can be enhanced using both Dsup and MDP, a multimodal therapy that protects both cellular repair proteins and DNA. It was hypothesized that cells expressing both Dsup and MDP would exhibit enhanced radioprotection, resulting in less DNA damage and higher survivability and proliferation. Cells expressing both Dsup and MDP, or Dsup or MDP alone, were irradiated at either 0 Gy or 100 Gy of X-ray radiation or 0.5 Gy of HZE radiation, and DNA damage, viability, and proliferation were assessed. Results showed that irradiated cells expressing either Dsup or MDP alone showed enhanced radioprotection, as previous studies found; however, when the proteins were combined, radioprotection was weaker than in control HEK293 cells, indicating there might be a negative interaction between the two. Subsequent research will use RNA sequencing and a cell cycle assay to measure expression levels and explore the interaction between Dsup and MDP.

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What initially inspired you to choose your research topic. How did your research evolve over time?   As an aspiring astronaut, I knew I wanted to study space. When I started my research, I was particularly captivated by how inhospitable space is for humans, and how little progress we have made to change this in spite of flying rockets and spaceships almost daily now. I studied the many risks to astronaut health in space, which led me to ionizing radiation (subatomic particles, gamma rays, X-rays), the most immediate risk to astronaut health. I initially looked at the effects of radiation on the body, and uncovered that some microorganisms on Earth, known as extremophiles, can withstand radiation in their own way through specific repair mechanisms. It gave me the idea to try to combine the defense systems of these extremophiles with human cells in order to enhance our resistance to radiation. I used two extremophiles, Deinococcus radiodurans, which developed the ability to protect its repair proteins from radiation, and Tardigrada, which can shield its DNA. My goal was to engineer a two-way repair system in human cells using both mechanisms.
 

Did you work with a mentor outside of HHS? How did the mentorship experience impact you? My two mentors were outside of HHS, one working for NASA and the other working at the Mason Lab at Weill Cornell Medicine, the lab where I conducted my research. Being mentored taught me a lot about the power of collaboration and how exchanges are needed to take fresh perspectives on challenges faced when going from theory to practice. Beyond the mentorship, the opportunity to work in a real research lab allowed me to understand the scientific world on a deeper level. Of course, our teacher, Ms. Blunt, was another one of my mentors, who pushed me to discover what real science looks like.
 

What skills, scientific or other, have you developed through the program? I learned to think like a scientist and how to navigate through issues with experiments, constantly redesigning my methodology. I learned how to analyze results and draw new hypotheses or conclusions, especially when the outcome was not as expected. I gained practical skills through hands-on experiments at the lab, but also in data science, as I independently needed to quantify and validate my results. The program as a whole revealed the importance of being able to communicate with very different audiences and levels of expertise, through presentations in lab meetings or in class, through posters or slides at conferences, and writing papers in an academic style. These three years of research certainly taught me resilience and commitment.
 

What has been the most rewarding aspect of your research experience? I faced many setbacks during my research experience, but being able to test and achieve what I had envisioned on a topic that seemed so much out of reach has been incredibly rewarding. The long hours, efforts, and stress were definitely worth it. 

 

How has the science research experience shaped your academic or career goals?
Science research has confirmed my passion for uncovering and solving challenges. I intend to continue in this scientific field, but I also know that I want my research to be practical, and not theoretical. I really want it to be about creating solutions. In addition, this experience has not only strengthened my passion for space but also widened my perspective, pushing me to discover more fields and topics and to be curious about everything.

 

What advice would you give to future students entering the Science Research Program? My main advice would be to choose something you are deeply passionate about. Research is not easy; it requires a true commitment, and failure is totally part of the process. If you research something you really love, your project is not going to be just a research project, but it will become yours, and you will deeply own it. My other advice: never give up, even when you are struggling, there is always another route. It is all worth it!

 

Research Title: The Effect of ɑ-tocopherol on Locomotor Ability of the Drosophila melanogaster Mutant for Glutathione S-Transferase Theta 3 (GstT3)

 

Abstract: Over 200,000 children in the United States live with Juvenile Arthritis (JA), an autoimmune condition that causes chronic inflammation as well as muscle and tissue atrophy. The human GSTT1 (hGSTT1) gene, part of the Glutathione S-Transferase Theta series, has been linked to this muscle erosion causing locomotor deficiencies. Since current JA treatments can be neurotoxic, it is necessary to identify safer and less toxic treatments. Ɑ-tocopherol (Vitamin E) is an antioxidant known to protect and repair muscle tissue. This study examined whether ɑ-tocopherol supplementation could rescue the locomotor deficiency associated with flies expressing UAS-RNAi against GstT3, the Drosophila ortholog of hGSTT1, and whether there is functional conservation between the human and fly gene by conducting a genetic cross to rescue the motor delay by overexpressing human GSTT1. Flies knocked down for GstT3 in muscle tissue were raised on 0, 20, 200, 1,000, and 2,000µM ɑ-tocopherol diets and tested using a negative geotaxis assay. Performance of the knockdowns increased significantly as the ɑ-tocopherol increased (p<.0001), but plateaued between 1,000 and 2,000µM, suggesting a saturation effect. The overexpressed hGSTT1 flies performed significantly better than knockdowns and indistinguishably from wild type at all ɑ-tocopherol concentrations (p>.05), demonstrating functional conservation between hGSTT1 and GstT3. Since functional conservation was established, future research should assay muscle breakdown in ɑ-tocopherol treated versus untreated flies and investigate how GstT3 interacts with immune pathways to better treat JA.

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What initially inspired you to choose your research topic. How did your research evolve over time? I chose my research topic because I have always been interested in the “why” behind Juvenile Arthritis (JA), something that I have been struggling with since I was two. Over time, I realized that I wanted to focus on finding a better treatment for JA since the majority of the current treatments have many harmful side-effects. Reading through the existing literature, I found ɑ-tocopherol (Vitamin E) and all the the incredible benefits it has on the human body, particularly in tissue development. My research took off from there when I began to use Drosophila melanogaster to test if Vitamin E really could be a safer treatment. 

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Did you work with a mentor outside of HHS? How did the mentorship experience impact you? At the end of my sophomore year, I was introduced to Dr. Cale Whitworth, a scientist at the Bloomington Drosophila Stock Center at Indiana University. Dr. Whitworth helped me to figure out which gene I should use in the Drosophila melanogaster to best simulate the mechanisms that occur in Juvenile Arthritis. Additionally, Ms. Blunt helped me to learn everything I needed to know about taking care of the fruit flies so that I could get my data with minimal interruptions. 

What skills, scientific or other, have you developed through the program? Throughout the program, I have seen both my critical thinking and communication skills improve significantly. Going into the program, I knew very little about the science behind my study and had to build my knowledge from the ground up. Additionally, this class requires many presentations and I have learned how to communicate effectively with both peers and experts. 

 

What has been the most rewarding aspect of your research experience? The most rewarding aspect of my research experience is seeing that my study could have real benefits in the world. Knowing that there is so much more research that could be done about Vitamin E and Juvenile Arthritis is very fulfilling. 

 

How has the science research experience shaped your academic or career goals? The science research experience has shaped my career goals by allowing me to solidify that I want to work on the business side of science. I plan to major in finance in college and I know that the skills that science research taught me will help me be successful. 

 

My advice to students applying next year is to research a topic that you truly care about. You will spend more time than you think working on it, and having genuine interest makes challenges easier to overcome.