Investigating the PROTO Targets in Regulating Drug-Induced Hair Cell Death

研究调节药物诱导的毛细胞死亡的 PROTO 靶点

• 批准号: 8967107
• 负责人: Joanne Hsu
• 金额: $ 5.8万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967107
• 关键词: Address; Adverse effects; Affinity; Aminoglycoside Antibiotics; Antibiotics; Attenuated; Auditory; Bacterial Infections; Binding; Binding Proteins; Biological; Biomedical Research; Biosensor; Cell Death; Cell Line; Cell Survival; Cells; Cessation of life; Coupled; Development; Disease; Fellowship; Foundations; Goals; Gram-Positive Bacteria; Hair Cells; Healthcare; Hearing; Hearing problem; Homologous Protein; Human; In Vitro; Infection; Investigation; Kidney; Kidney Failure; Kinetics; Knockout Mice; Knowledge; Laboratories; Labyrinth; Lead; Life; Light; Mammalian Cell; Mass Spectrum Analysis; Measurement; Measures; Molecular; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Preclinical Drug Development; Preclinical Drug Evaluation; Prevention; Prevention strategy; Proteins; Rattus; Refractory; Research; Research Personnel; Research Proposals; Role; Scientist; Sensory Hair; Solid; Surface Plasmon Resonance; Testing; Therapeutic; Toxic effect; Validation; Vertigo; Western Blotting; Zebrafish; analog; career; cell injury; design; equilibration disorder; experience; hearing impairment; in vivo; insight; lateral line; nephrotoxicity; novel; novel therapeutic intervention; ototoxicity; prevent; public health relevance; response; small molecule

DESCRIPTION (provided by applicant): Aminoglycoside antibiotics (AGAs) are widely used in the treatment of a variety of refractory bacterial infections. The most important side effects and therapeutic limitations of AGAs are inner ear damage (ototoxicity) which can produce irreversible hearing loss and kidney damage (nephrotoxicity) which can lead to kidney failure. Prevention of AGA-induced ototoxicity and nephrotoxicity has been a major therapeutic objective in biomedical research. One major goal at the laboratory of Dr. Julian Simon is to identify the biological target of the otoprotective PROTO1 analogs (PROTO) to determine their mechanism of action in preventing AGA-induced inner ear hair cell death. To achieve this goal, the laboratory recently identified several candidate PROTO targets in zebrafish through photoaffinity pulldowns coupled with mass spectrometry. In this fellowship proposal, the three specific aims are designed to test the hypothesis that one of these mass spectrometry- identified PROTO candidate targets plays a critical role in controlling hair cell death and surviva in response to AGAs: 1) Validation of the interactions between PROTO and the most promising mass spectrometry-identified candidate targets using western blotting and competition pulldowns; 2) kinetic analysis of interactions between PROTO and its targets using a surface plasmon resonance (SPR) biosensor; 3) investigation of the effects of the PROTO targets on AGA-induced hair cell toxicity using mammalian cell lines and conditional knockout mice. Identification of the biological PROTO target is expected to advance the understanding of the cellular pathways that regulate hair cell death and survival and promote the development of new therapeutic strategies to prevent hearing and balance disorders. The fellowship proposal will provide an invaluable hands-on experience and comprehensive knowledge in hearing protection research and preclinical drug development paving a solid foundation towards an independent research career as an investigator/scientist in the field of auditory and biomedical research.

描述（申请人提供）：氨基糖苷类抗生素（AGA）广泛用于治疗多种难治性细菌感染。 AGA 最重要的副作用和治疗限制是内耳损伤（耳毒性），它会导致不可逆的听力损失；以及肾脏损伤（肾毒性），可能导致肾衰竭。预防 AGA 引起的耳毒性和肾毒性一直是生物医学研究的主要治疗目标。 Julian Simon 博士实验室的一个主要目标是确定耳保护剂 PROTO1 类似物 (PROTO) 的生物靶点，以确定其预防 AGA 诱导的内耳毛细胞死亡的作用机制。为了实现这一目标，该实验室最近通过光亲和下拉结合质谱鉴定了斑马鱼中的几个候选 PROTO 靶点。在该奖学金提案中，三个具体目标旨在检验以下假设：这些质谱鉴定的 PROTO 候选靶点之一在控制毛细胞死亡和响应 AGA 的存活方面发挥着关键作用：1) 使用蛋白质印迹和竞争下拉验证 PROTO 与最有希望的质谱鉴定的候选靶点之间的相互作用； 2）使用表面等离子共振（SPR）生物传感器对PROTO与其靶标之间的相互作用进行动力学分析； 3) 使用哺乳动物细胞系和条件敲除小鼠研究 PROTO 靶点对 AGA 诱导的毛细胞毒性的影响。生物 PROTO 靶标的鉴定有望促进对调节毛细胞死亡和存活的细胞途径的理解，并促进预防听力和平衡障碍的新治疗策略的开发。该奖学金提案将为听力保护研究和临床前药物开发提供宝贵的实践经验和全面的知识，为作为听觉和生物医学研究领域的调查员/科学家的独立研究生涯奠定坚实的基础。