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

基本信息

批准号：
9184554
负责人：
Joanne Hsu
金额：
$ 6.1万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
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/9184554
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 Clinical 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

项目摘要

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（由申请人提供）：氨基糖苷抗生素（AGA）广泛用于治疗多种难治性细菌感染。 AGA的最重要的副作用和治疗局限性是内耳损伤（耳毒性），可导致不可逆的听力损失和肾脏损伤（肾毒性），这可能导致肾衰竭。预防AGA诱导的耳毒性和肾毒性一直是生物医学研究中的主要治疗目标。朱利安·西蒙（Julian Simon）博士实验室的一个主要目标是确定眼球保护原型类似物（Proto）的生物学靶标，以确定其在防止AGA诱导的内耳毛细胞死亡方面的作用机理。为了实现这一目标，实验室最近通过光性下拉与质谱法结合了斑马鱼中的几个候选原始目标。在该奖学金提案中，三个具体目标旨在检验以下假设：这些质谱法鉴定的原始候选目标之一在控制毛细胞死亡和对agas的Surviva中起着至关重要的作用：1）验证原始质谱和最有希望的质谱质量光谱群体识别的候选目标，使用蛋白质斑点和竞争PULLEDS和竞争Pulllds和竞争; 2）使用表面等离子体共振（SPR）生物传感器对原始靶标之间的相互作用进行动力学分析； 3）研究了使用哺乳动物细胞系和有条件的基因敲除小鼠研究原始靶标对AGA诱导的毛细胞毒性的影响。预计生物学原始靶标的鉴定将提高对调节毛细胞死亡和生存的细胞途径的理解，并促进新的治疗策略的发展，以防止听力和平衡疾病。奖学金提案将在听力保护研究和临床前药物开发方面提供宝贵的实践经验和综合知识，为独立研究职业铺平了稳固的基础，作为听觉和生物医学研究领域的研究者/科学家。