Dissertation
Identification and characterization of Inhibitory compounds for S. neurona
Doctor of Philosophy (PhD), Washington State University
01/2020
Handle:
https://hdl.handle.net/2376/111398
Abstract
The apicomplexan parasite Sarcocystis neurona is the causative agent of debilitating neurologic disease in horses, marine mammals, and several other domestic and wild animals. A new therapy to treat infected animals is needed. To address this critical need, we developed, validated, and implemented a high-throughput screen to test repurposed FDA-approved drugs and tartrolon E (trtE), a promising anti-apicomplexan natural product. We identified 19 new inhibitory compounds of S. neurona, including trtE. The lead inhibitory compound, trtE, inhibited S. neurona parasites in vitro at nM concentrations. To investigate the mechanism of action of trtE, we examined the changes in the transcriptome of trtE-treated Toxoplasma gondii parasites. We identified the conserved gene TgME49_272370 to be significantly upregulated in a rapid dose-dependent manner, which indicated a role for the gene in response to treatment. Using a combination of informatics and proteomics, we predicted the protein product of TgME49_272370 to have multiple transmembrane domains, localize to the plasma membrane, and contain multiple phosphorylation sites. Deletion of the TgME49_272370 gene by CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) resulted in a 68% increase in parasite resistance to trtE treatment. In further uncovering the mechanism by which trtE inhibits apicomplexan parasites, we may identify a shared pathway critical to parasite survival and advance the search for new treatments.
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Details
- Title
- Identification and characterization of Inhibitory compounds for S. neurona
- Creators
- Gregory David Bowden
- Contributors
- Roberta M. O'Connor (Advisor)Kelly A. Brayton (Committee Member)James A. Brozik (Committee Member)Carlos E. Suarez (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Veterinary Medicine, College of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Number of pages
- 132
- Identifiers
- 99900581703001842
- Language
- English
- Resource Type
- Dissertation