Biological Activity of Novel Azolium-Based Therapeutics against TDP-43-Mediated-Toxicity in Transfected Normal Neuronal and Carcinoma Cell Lines
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Session: 1
Room: Eastern Market
Alisha R. Kennedy - Columbus State University
Co-Author(s): Monica Frazier, Columbus State University, Columbus, GA; Kerri Taylor, Columbus State University, Columbus, GA; Dr. Alexandra Taraboletti, University of the District of Columbia, Washington, DC.
Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig’s disease, is a progressive neurodegenerative disease that damages motor neurons that run from the brain into muscles via the spinal cord. When damaged, these neurons strip patients of voluntary movement and the progression is fatal. Despite the variation in the mean survival time for ALS patients, the average unfortunately is low, ranging from a few months to up to 10 years. A hallmark feature in ALS and other neurodegenerative diseases is the presence and over-expression of the TDP-43 protein. Knowing that the TDP-43 protein is a signature feature in neurogenerative disease, specifically frontotemporal dementia (FTD) and ALS, allows it to be a promising target for exploring new therapeutic interventions to try to minimize or completely halt the progression of such diseases. ALS currently does not have a cure, but there are treatment options to help manage the symptoms of the disease. The most prescribed treatment option, Riluzole, is one of two FDA-approved oral drugs for ALS. Riluzole is a drug in the benzothiazole family that works to block the release of glutamate. However, Riluzole and other ALS treatments on the market fail to meaningfully extend the lives of ALS patients and effectively manage patients’ symptoms to increase their quality of life. We have synthesized, characterized, and tested the biological activity of a series of novel azolium-based salts (benzothiazole and triazole, designated BS and T3, respectively) against WI-38 and H1299 cells. This project will explore the effects of these novel salts against TDP-43 transfected HEK cells and SH-SY5Y cells through the analysis of cellular metabolism on increasing levels of TDP-43 mediated-toxicity and its alterations of the cell cycle. Additionally, this project will explore possible drug-binding sites for the downregulation or complete inhibition of TDP-43 production. To achieve these goals, the cell lines will all be transfected with the protein using a Lipofectamine kit. To ensure transfection success, western blots will be used to test for the target protein. If successful, the cell lines will be drugged with the most promising of the novel drugs for cell viability assays, cell cycle analysis, analysis of reactive oxygen species (ROS) levels, and the analysis of mitochondrial dysfunction. Riluzole and/or Cisplatin, a common cancer drug, will be used as controls for the drug testing. Currently, the cytotoxicity assays of some BS and most T3 drugs on WI38 and H1299 cells show a dose-dependent decrease in cell proliferation in cancer cells and dose-dependent toxicity in normal cells. The next step is to take the most effective drugs, i.e., those that selectively decrease cancer cell populations, from the cytotoxicity assays and apply them to the transfected cell lines to see how they perform to reduce or inhibit TDP-43 mediated toxicity.
Funder Acknowledgement(s): CSU Interdisciplinary Initiative Grants – Center of Experiential Learning;CSU Student Research and Creative Endeavors Grants;Louis Stokes Alliance for Minority Participation (LSAMP) National Science Foundation Grant No. HRD-1817519
Faculty Advisor: Dr. Monica Frazier, frazier_monica@columbusstate.edu
Role: For this research project, I synthesized the benzothiazole (BS) salts, cultured each cell line, solubilized the drugs, and drugged the cells. I also ran the MTT assays for each cytotoxicity assay. I will also be transfecting the cell lines with the target protein and conduct all necessary tests and assays, including western blot, cell viability and MTT assays, cell cycle analysis, ROS analysis, mitochondrial dysfunction analysis.