(2fk) A Novel in Vitro Cell Transfection Method: Optimization of Corona Charge Instrumentation and Parameters

I have been a part of five research projects from high school to my dissertation work, including nanoparticle engineering, Alzheimer’s and virus research, and gene and drug delivery methods. Through these research projects, I have had the opportunity to learn multiple chemical, biological, and engineering techniques and multiple mediums for presenting my research to the public.

High School and Undergraduate Research (Chemical Engineering):

The summer after my sophomore year of high school, I was exposed to the research environment at the Particle Engineering Lab at the University of Florida. There, I synthesized and purified polyhydroxy fullerenes. After purification, I employed different characteristic techniques to study the molecules, including Fourier-transform infrared (FTIR) spectroscopy, inductively coupled plasma mass spectrometry, and nuclear magnetic resonance spectroscopy. These molecules would later be used in experiments for MRI contrasts as well as a potential tumor treatment that would produce fewer free radicals.

While completing my undergraduate studies in chemical engineering, I joined Dr. Norma Alcantar’s lab at the University of South Florida (USF) where I became a part of 2 major projects: directed drug delivery using hydrogels and Alzheimer’s research. I analyzed the degradation rate of hydrogels in varying pH mediums using FTIR. I later determined the medication release rate of hydrogels in different environments. For the second project, I tested the effects of mucilage of the Opuntia Ficus-Indica on amyloids, a fiber that is involved in neurodegenerative diseases, like Alzheimer’s. The use was hypothesized to disrupt the formation of fibers, which could prevent the progression of neurodegenerative diseases. I analyzed different concentrations of mucilage using an FTIR and the program Omnic to determine the amount of monomers, oligomers, and fibers at the different concentrations and monitor the degradation of the fibers over time. I was one of thirty undergraduate students honored with the 2017 USF Excellence In Undergraduate Research Award for my work. I presented this project at the Society of Women Engineers (SWE) WE17 Conference in Austin, TX as a National Finalist for the Graduate Poster Competition.

International Research Experience for Students (National University of Singapore):

In the summer of 2018, I was selected as one of five students to participate in the NSF-funded program, the International Research Experience for Students (IRES) at the National University of Singapore. Through this program, I was invited to work under Dr. Ganesh Anand (now a professor at Penn State) to determine how osmolytes impacted the structures of viruses to help create more effective vaccines. Viruses change in different environments and by testing the structure in each environment, the weak points can be determined for viruses within the human body to create better vaccinations. I transformed and cultured bacteria cells to obtain a purified virus for experiments. Using Chymotrypsin assays and Hydrogen-Deuterium Exchange Mass Spectrometry, I determined the structural changes of the Turnip Crinkle Virus in the presence of aggregating and stabilizing osmolytes, such as sucrose. I was able to determine that as the concentration of sucrose increased, the virus became more stable in certain areas. Knowing the areas that become more stable in sucrose allows for vaccines to be designed with that in mind.

There were two main goals for this program. The first was to expose students to a different culture, both inside and outside of the lab. The second was to effectively communicate research through different mediums, including a traditional scientific poster and through a weekly Twitter post. This exposed me to a new way of distributing scientific knowledge and taught me how to describe my research to different levels of expertise. As shown by the coverage of the COVID-19 pandemic, it is imperative that scientists and engineers are able to explain their research not only to experts in our field but to the general public who will be the ones consuming our products. I have presented this work at SWE’s WE18 Conference in Minneapolis, MN as a National Finalist for the Graduate Rapid-Fire Competition. I received First Place in the Collegiate Competition at SWE’s WELocal Tampa 2019.

Master’s Thesis (Biomedical Engineering):

In December 2019, I successfully defended my Master’s Thesis entitled “The Combined Effect of Heat and Corona Charge on Molecular Delivery to a T Cell Line In vitro” to obtain an M.S. in Biomedical Engineering from the University of South Florida. I conducted this research under Dr. Mark Jaroszeski in the Gene and Drug Delivery Lab. This project involved me designing instrumentation and other custom pieces for the apparatus to deliver small molecules to a cell line using corona charge. Corona charge is a non-contact form of electrogenetherapy (previously known as electroporation). My goal was to use the custom apparatus to tune corona charge parameters to deliver the small molecule Sytox^TM into an immortalized human T cell line (Jurkats). This included varying parameters such as voltage, current, and temperature to increase molecular delivery. I found parameter combinations that induced statistically significant delivery compared to controls while still maintaining viability for at least 48 hours after treatment. A paper discussing this work is currently being edited for submission.

Ph.D. Dissertation (Chemical Engineering):

My dissertation work is a continuation of my thesis under Dr. Mark Jaroszeski in the Gene and Drug Delivery Lab. I established candidacy in Spring 2021 and plan to graduate in Summer 2023 with a Ph.D. in Chemical Engineering from the University of South Florida. Using what I learned through my Master’s Thesis, I was able to create a novel transfection dish that will direct the electric field through the cell solution. The previous dish design was not able to be insulated for high voltage and involved too many sharp points to ensure the direction of the electric field, so this new dish design mitigated that problem. With this new dish, I am working on designing a set of parameters to best deliver green fluorescent protein plasmid DNA (GFP pDNA) to Jurkat cells. These parameters include current, amount of ions in cell solution, and temperature of cell solution during treatment. One paper on related work has been submitted to the American Vacuum Society and there are plans to produce another paper with the results of the ongoing experiments. This work, as well as my thesis work, has been presented at multiple colloquiums and conferences, including at the American Institute of Chemical Engineers and Biomedical Engineering Society Conferences.

Research Interests:

My future research goals involve me working with a cross-disciplinary group of researchers working to develop novel cancer therapies that are targeted, translational, and less expensive than ever before. Immunotherapies are one of the current trends in cancer research and I would like to get more involved in that approach. I currently work on the delivery aspect and I would like to get more involved in the protein engineering, testing, and optimization of immunotherapies. While this is my primary goal, I know that trends in how we treat cancer can change quickly and my goal is to gain the knowledge to allow me to stay up to date on those trends to eventually create a long-lasting and prosperous research lab.