(4dw) Continuous Improvement in Gene Therapies Facilitated By Electrically Mediated Processes

Gene therapy has undergone major advancements since the first FDA approved therapy for B-cell leukemia in 2017. Now, there are over 25 different gene therapies approved to treat diseases like leukemia, melanoma, spinal muscular atrophy, sickle cell anemia, and more. However, some of these formulations remain individualized and expensive. My lab will have three major objectives: 1) Utilize and improve viral gene therapy methods to assist with these therapies, as well as combine with other physical methods of gene delivery, including gene electrotherapy, 2) Investigate the possibility of electrically mediated purification methods for viruses, and 3) Utilize atomic force microscopy to analyze cancerous cells that do not have DNA to code for the common surface proteins that are utilized in chimeric antigen receptor (CAR) T cell therapies.

Research Experience:

Postdoctoral Research:

I am currently conducting my postdoctoral research with Dr. Caryn L. Heldt in the Heldt Bioseparations Laboratory at Michigan Technological University. One major problem in viral vaccine and gene therapy manufacturing is the purification of full viral capsids. My project is to find structural and chemical differences between gene-of-interest-containing (full) capsids versus the “empty” capsids. These minor differences can be exploited to further purify viral capsids. I am utilizing atomic force microscopy (AFM) and chemical force microscopy (CFM) to determine these changes at an atomic level. I am characterizing two serotypes of adeno associated virus (AAV) to see the changes between full and empty capsids and how these change between serotypes. I am also working on a side project to create a small-scale purification and characterization method for AAVs. This would allow our lab to create our own samples of empty and full AAV capsids from a stock we receive from a collaborator rather than purchasing expensive pre-purified stock from a company.

In addition to my research, I mentor 6 graduate students and multiple undergraduate students in the Heldt lab. I have brought in my previous flow cytometry experience to assist in a graduate student’s project as well as assisting all students in their writing and presentations skills. I have also gained grant writing experience by assisting Dr. Heldt in putting together a National Institute of Health (NIH) S10 Shared Equipment Grant (SIG) for a bio-specific AFM for the campus.

PhD Research:

In December 2023, I obtained my PhD from the University of South Florida with a dissertation entitled “Exploration of Corona Charge as a Novel in Vitro Human T Cell DNA Transfection Method.” My research focused on optimizing a non-contact form of gene electrotransfer. This method utilized corona charge, a plasma created in atmospheric air, to deliver plasmid DNA to cells. Two cell lines were utilized: Jurkat cells, an immortalized T-cell line (semi-attached cells), and B16-F10 cells, a murine melanoma line (attached cells). I designed a new dish to perform high voltage treatments and to ensure the direction of the electric field and a set of parameters to best deliver green fluorescent protein plasmid DNA (GFP pDNA) or Luciferase to Jurkat and B16F10 cells. These parameters include current, number of ions in cell solution, and temperature of cell solution during treatment. Success was analyzed via flow cytometry, immunohistochemistry, and luminescence plate reading.

With this work, I was able to limit the variable field. Voltage changes did not cause a statistically significant difference in uptake from the cells, but current changes did. I determined stable regions of current created by the corona to ensure more consistent delivery. Additionally, there was a trend where cells with pDNA present had a higher viability than cells without additional pDNA present. This work proved the potential for a non-contact form for gene elctrotransfer to mammalian cells.

Graduate Research:

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.

M.S. 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.” I conducted this research in the Gene and Drug Delivery Lab at the University of South Florida. 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 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. This research served as the starting point for my PhD research.

Teaching Interests:

Previous Teaching Experience:

During my graduate program, I was a teaching assistant (TA) for multiple classes: Chemical Engineering Unit Operations Lab I and II, Engineering of Biological Systems, and Research Design, Methods, and Interpretation. My longest appointment was to the Unit Operations Lab series, where I served as a TA for seven semesters. I served as the lead TA for four semesters, which included setting up the Canvas course, assisting in mentoring new TAs, and hosting seminar sessions when the course instructor was traveling or on medical leave. In 2020, I received my department’s Outstanding TA Award. During the COVID-19 pandemic, I assisted in moving the lab course to an online and remote mode. This included creating a take-home kit for students to explore concepts like fluid flow and pump curves. This project won the 2021 University of South Florida Student Success Academic Excellence Award and was presented at the 2022 AIChE annual meeting in the Education Division. During my time as a TA for Engineering of Biological Systems, I lead the transition from Scantron examinations to Zipgrade, which works in a similar way but has more features like a multiple answer option and the ability to grade using a smartphone. During my time as a TA, I created a workshop entitled “TA Survival Guide” which assists teaching assistants and faculty on how to make a mutually beneficial relationship to ensure student (both undergraduate and graduate) success.

Teaching Interests and Goals:

One of the benefits of teaching a laboratory course as a graduate student is staying current on concepts taught in a variety of core chemical engineering courses. I would feel most comfortable teaching separations, reactions, and material balance courses, but I am confident I can teach any core ChemE course and multiple electives. With my specific research and knowledge, I would also be able to take on more bio-specific courses. I plan to integrate real world examples from a variety of industries, including pharma, food, paper, petroleum, and chemical processing. I also plan to give students the opportunity to work on their scientific communication through a variety of mediums, including oral presentations and short executive summaries. I plan to create an inclusive environment and evolve my teachings semester-to-semester with student feedback and teaching trends by attending conferences and AIChE Education Division sessions.