Date of Award
12-2025
Document Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Genetics and Biochemistry
Committee Chair/Advisor
James Morris
Committee Member
Meredith Morris
Committee Member
Cheryl Ingram-Smith
Committee Member
Lesly Temesvari
Abstract
Trypanosoma brucei is a protozoan parasite which causes human African trypanosomiasis and the livestock wasting disease, nagana. These parasites have highly specialized peroxisomes, called glycosomes, which house a variety of essential metabolic pathways, including glycolysis. Glycolysis is the sole means of energy production in the infective life stage of T. brucei making glycosomes and the glycolytic pathway promising targets for therapeutic strategies. In this work, we have studied the glycosome as a therapeutic target from multiple perspectives. The first portion of this work addresses drug trafficking. The efficacy of small-molecule inhibitors of glycosomal enzymes is restricted by the rate of trafficking to the glycosomal compartment; this challenge can be mitigated by the appendage of a type one peroxisomal targeting sequence (PTS1). With the goal of enhancing this targeted drug delivery system, we quantified the uptake of a panel of FITC conjugated PTS1 tripeptides and found that specific tripeptides exhibit variable rates of uptake. The second portion of this work focuses on drug discovery and the study of glycosome biology. We established a cell line and protocols for the use of the pHluorin2 fluorescent pH biosensor in T. brucei. This cell line, as well as cell lines expressing glucose and ATP biosensors, has allowed for the development of a high throughput drug discovery strategy for identifying molecules that disrupt the glycolytic pathway. In addition to drug screening, we have used the pH and glucose biosensors to characterize the knockdown phenotypes of two proteins that we identified as putative sodium proton exchangers. Together, these data advance our understanding of the potential of targeting glycosomes as a therapeutic modality.
Recommended Citation
Pizarro, Sabrina Sutton, "Exploring Glycosomes as Therapeutic Targets for Trypanosoma brucei" (2025). All Dissertations. 4138.
https://open.clemson.edu/all_dissertations/4138