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Drug Screening Utilizing the Visual Motor Response of a Zebrafish Model of Retinitis Pigmentosa

thesis
posted on 06.05.2020 by Logan C Ganzen
Retinitis Pigmentosa (RP) is an incurable inherited retinal degeneration affecting approximately 1 in 4,000 individuals globally. The aim of this dissertation was to identify drugs that can help patients suffering from the disease. To accomplish this goal, the zebrafish was utilized as a model for RP to perform in vivo drug screening. The zebrafish RP model expresses a human rhodopsin transgene which contains a premature stop codon at position 344 (Tg(rho:Hsa.RH1_Q344X)). This zebrafish model exhibits significant rod photoreceptor degeneration beginning at 7 days post fertilization (dpf). To assess the visual consequence of this rod degeneration the zebrafish behavior visual motor response (VMR) was assayed under scotopic conditions. The Q344X RP model larvae displayed a deficit in this VMR in response to a scotopic light offset. This deficit in behavior was utilized to perform a drug screen to identify compounds that could ameliorate the deficient Q344X VMR. The ENZO SCREEN-WELL® REDOX library was chosen to be screened since oxidative stress may increase RP progression in a non-specific manner. From this library, a β-blocker, carvedilol, was identified as a compound that improved the Q344X VMR behavior. This drug was also able to increase rod number in the Q344X retina. Carvedilol was shown to be capable of working directly on rods by demonstrating that the drug can signal through the adrenergic pathway in the rod-like human Y79 cell line. Since carvedilol is an FDA-approved drug, this screening paradigm was utilized to screen the Selleckchem FDA-approved library to identify more drugs that can potentially be repurposed to treat RP like carvedilol. Additionally, this scotopic VMR assay was used to demonstrate that it can identify behavioral deficits in the P23H RP model zebrafish (Tg(rho:Hsa.RH1_P23H)). This dissertation work provides a potential FDA-approved drug for RP treatment and sets the foundation for future drug screening to identify more drugs to treat different forms of RP.

History

Degree Type

Doctor of Philosophy

Department

PULSe (Life Sciences)

Campus location

West Lafayette

Advisor/Supervisor/Committee Chair

Yuk Fai Leung

Additional Committee Member 2

Daniel Suter

Additional Committee Member 3

Donald Ready

Additional Committee Member 4

Connie Weaver

Licence

Exports