Aminoglycoside antibiotics (AGAs) are a mainstay in clinical management of life-threatening Gram-negative bacterial infections. AGAs are widely available and highly economical;however, their use is limited due to ototoxicity and nephrotoxicity. We sought to identify agents that would block aminoglycoside ototoxicity without blocking its antimicrobial activity. Using an in vivo zebrafish (Danio rerio) neuromast assay, we identified two structurally related compounds (PROTO1 and 2) from a library of diverse, drug-like compounds that protect zebrafish mechanosensory neuromast cells from neomycin-induced cell death. The protective effect of PROTO1/2 is dose-dependent and is not due to inhibition of AGA uptake into zebrafish neuromasts. Furthermore, neither PROTO1 nor PROTO2 inhibit AGA antibacterial activity in E. coli. PROTO1 is non-toxic in animals and confers AGA resistance in explanted mouse utricles and human kidney cells. In this application we propose four Specific Aims to carry out pre-clinical optimization of the lead compounds, to identify the molecular targets of the protective drugs and determine the molecular pathways that are involved in both AGA-induced ototoxicity and its prevention by PROTO1/2. Specifically, in Aim 1, we will synthesize and evaluate the hair cell protective effects of PROTO1/2 analogues with two goals: 1). to increase the potency of the protective activity, and;2). to develop a structure activity relationship (SAR) that would allow us to modify PROTO1/2 for target identification studies (Aim 3).
In Aim 2, we will develop AGA-induced toxicity assays using human kidney cells, zebrafish neuromast hair cells and explanted mouse utricles.
In Aim 3, we will carry out in vivo pre-clinical evaluation of PROTO1/2-derived agents.
In Aim 4, we will use SAR data to design and synthesize an affinity capture reagent based on PROTO1/2 and will carry out target identification studies. These convergent and complementary studies will aid in the development of target-based hair cell protective agents and expand the molecular understanding of AGA-induced cell damage. The use of many drugs is limited by serious side effects. We have discovered compounds that may help reduce hearing loss and damage to kidneys caused by an important class of antibiotics. Studies in this proposal will develop more potent protective compounds, identify how these compounds work and provide preliminary data for their development as drugs.

Public Health Relevance

Relevance The use of many drugs is limited by serious side effects. We have discovered compounds that may help reduce hearing loss and damage to kidneys caused by an important class of antibiotics. Studies in this proposal will develop more potent protective compounds, identify how these compounds work and provide preliminary data for their development as drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
5R01DC009807-04
Application #
8288091
Study Section
Auditory System Study Section (AUD)
Program Officer
Freeman, Nancy
Project Start
2009-08-17
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$346,194
Indirect Cost
$108,339
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
Chowdhury, Sarwat; Owens, Kelly N; Herr, R Jason et al. (2018) Phenotypic Optimization of Urea-Thiophene Carboxamides To Yield Potent, Well Tolerated, and Orally Active Protective Agents against Aminoglycoside-Induced Hearing Loss. J Med Chem 61:84-97
Stawicki, Tamara M; Esterberg, Robert; Hailey, Dale W et al. (2015) Using the zebrafish lateral line to uncover novel mechanisms of action and prevention in drug-induced hair cell death. Front Cell Neurosci 9:46
Thomas, Andrew J; Wu, Patricia; Raible, David W et al. (2015) Identification of small molecule inhibitors of cisplatin-induced hair cell death: results of a 10,000 compound screen in the zebrafish lateral line. Otol Neurotol 36:519-25
Thomas, Andrew J; Hailey, Dale W; Stawicki, Tamara M et al. (2013) Functional mechanotransduction is required for cisplatin-induced hair cell death in the zebrafish lateral line. J Neurosci 33:4405-14
Esterberg, Robert; Coffin, Allison B; Ou, Henry et al. (2013) Fish in a Dish: Drug Discovery for Hearing Habilitation. Drug Discov Today Dis Models 10:
Vlasits, Anna L; Simon, Julian A; Raible, David W et al. (2012) Screen of FDA-approved drug library reveals compounds that protect hair cells from aminoglycosides and cisplatin. Hear Res 294:153-65
Ou, Henry; Simon, Julian A; Rubel, Edwin W et al. (2012) Screening for chemicals that affect hair cell death and survival in the zebrafish lateral line. Hear Res 288:58-66
Ou, Henry C; Keating, Sarah; Wu, Patricia et al. (2012) Quinoline ring derivatives protect against aminoglycoside-induced hair cell death in the zebrafish lateral line. J Assoc Res Otolaryngol 13:759-70
Hirose, Yoshinobu; Simon, Julian A; Ou, Henry C (2011) Hair cell toxicity in anti-cancer drugs: evaluating an anti-cancer drug library for independent and synergistic toxic effects on hair cells using the zebrafish lateral line. J Assoc Res Otolaryngol 12:719-28
Ou, Henry C; Santos, Felipe; Raible, David W et al. (2010) Drug screening for hearing loss: using the zebrafish lateral line to screen for drugs that prevent and cause hearing loss. Drug Discov Today 15:265-71

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