Angel Zavala Custodio

Angel Zavala Custodio is a rising senior at the University of Texas at Austin studying Neuroscience and Kinesiology Exercise Science, optimized high through put screening of Na_v1.2 to identify pathological variants associated with schizophrenia and develop better therapeutic strategies.

Voltage-gated sodium channels (Na_v) are responsible for initiating an action potential within most electrically excitable cells. Na_v1.2, encoded by the SCN2A gene, is an essential sodium channel within the central nervous system, and its associated missense mutations have led to diverse neurodevelopmental and psychiatric disorders. However, how these mutations impact Na_v1.2 function is not fully understood. Upon reflecting on my experiences at ӳ��ý, I am reminded of the talented scientists I had the privilege to work with. My mentors not only taught me how to perform various lab techniques, but how to think like a scientist. Each day was a journey filled with adventure and challenge that allowed me to grow as a researcher and think of high-throughput and novel solutions to problems. Additionally, I was surrounded by extraordinary, intelligent, and gifted peers who always motivated me to exceed my goals and were always there to support me. I am so grateful for the team I was able to work with as it showed me how to think critically about a problem but also how to help people from diverse backgrounds through research.Thus, the various effects produced by these missense variants suggest the need to systematically characterize their specific functional consequences. We utilized saturated mutagenesis to create a library of 10,000 SCN2A gene variants. Additionally, to investigate functional disparities within variants and categorize loss-of-function (LOF) and gain-of-function (GOF) variants from wild type, we utilized a previously established cellular assay based on the cytotoxicity resulting from the intracellular accumulation of Na+ ions. Through using lentivirus vectors and puromycin selection, we were able to efficiently deliver our test library into HEK293 cells. Furthermore, our results indicated a combination of a sodium influx promoter (Veratridine) and a sodium efflux inhibitor (Ouabain) successfully categorized LOF and GOF SCN2A variants within HEK293 cells. We were further able to validate the biophysical properties of Na_v1.2 by performing large-scale precise electrophysiology protocols in an automated and standardized manner. Our results provide the potential to utilize high-throughput drug screening to identify channel variants that exhibit significant functional alterations. Comprehending the effects of these mutations on SCN2A provides insight into understanding the molecular and biophysical properties of the channel required to develop an efficient therapeutic strategy.