63 Research Reflection by Nia Brooks

Faculty Mentor: Tracey Lamb (Pathology, University of Utah)

My love for research developed from an aptitude for problem-solving and enjoyment of using self-logic to reason and justify the unknown. In January 2022, I joined the University of Utah’s pathology department and began working under the direction of Dr. Tracey Lamb and PhD candidate Marshall Roedel. My first project sought to determine primary sources of interferon-gamma cytokine production in the brain during Experimental Cerebral Malaria (ECM)pathogenesis. This project introduced me to several novel methods of cell analysis such as flowcytometry, single-cell RNA sequencing, and in-vitro assays. My mentor and I were able to conclude that out of the various cell types assessed (e.g., microglia, astrocytes, etc.), CD8+ T-cells proved to be an important source of interferon-gamma in the brain, which allows for Blood Brain Barrier (BBB) disruption. This lab experience allowed me to explore the use of murine models, practice tissue extraction, and become familiar with the immune system. In January 2022, I began working with Dr. Elliot Asare and Daniel Harkness on an analytical research study to gauge Black medical students’ interests in surgical careers. As a result of the study, I transitioned into aiding the development of the newly established Utah Summer Training Academy for Rising Surgeons (USTARS) program. For the past three summers, I have produced and distributed surveys using Qualtrics XM software to program participants. Once completed, I compiled data to analyze and develop into presentable reports. Working with the USTARS program provided a way for me to exercise my interest in medicine alongside my enthusiasm for involvement in diversity initiatives. These separate experiences made me wonder about the intersectionality between biomedical research and diversity studies, especially the mutual impacts of both fields. The 2022 Yale BioMed SURF Summer Program introduced me to dermatology research and what a career as a physician-scientist could look like. Under the direction and mentorship of, Dr. Keith Choate, Dr. Jing Zhou, and MS2 Mitra Mani, I investigated the effects of two different mutations on the expression of Cyclin G-Associated Kinase (GAK) in a patient presenting Acanthosis nigricans. The patient did not present underlying conditions that would characteristically warrant this condition. This patient did, however, have an inherited constitution splice mutation and a spontaneous insertion mutation in GAK, the latter of which results in a premature termination of protein translation. The two goals of the study were to explore how the GAK insertion mutation alters the GAK signaling pathway and how the constitutional splice mutation might alter RNA splicing. A key moment of this experience was attending the Foundation for Ichthyosis & Related Skin Types National Conference, where people worldwide came to hear the latest advances and professional counsel on these conditions. Through engaging in casual conversation, the sheer number of different stories each patient had to share astounded me. I saw and spoke to patients that represented a wide range of gender, ethnicity, nationality, and age. However, there was a particular interaction that instilled a new sense of purpose and inspiration as to why I chose this career path. I rejoined the Choate Lab the following summer as an AMGEN Scholar. Alongside Dr. Khadim Shah, I worked on the detection of DNA strand breaks in Ichthyosis with Confetti (IWC). IWC is caused by a frameshift mutation in the KRT10 gene that causes its mislocalization. Displaced keratin disrupts the normal keratin intermediate filament network and leads to nuclear instability and DNA damage. Some of this damage is repaired, leading to patches of revertant skin. In order to investigate the type of DNA damage occurring, we cultured U-2 OS cells transduced with KRT10 (both the mutant and normal). I plated harvested cells combined with agarose to perform two comet assays, which visualize DNA damage. An alkaline comet assay visualized single-strand DNA breaks while neutral comet assays visualized double-strand breaks. A greater difference in DNA Tail percentage between the mutant and wild-type in the alkaline comet assay implicated that single-strand breaks were the prominent form of damage. This conclusion can aid in narrowing what DNA repair mechanisms may be involved in IWC. As I prepare to graduate, I conclude my journey in the Lamb lab with the completion of my honors thesis. My thesis will cover the localization and regulation of beta-catenin during experimental cerebral malaria pathogenesis in hopes of investigating its effects on endothelial cell integrity. As grateful as I am for the research experiences I have had in my undergraduate career, I am even more excited to continue research in dermatopathology and apply it to improving patient care and education in underserved communities.