Characterizing the protective effects of caffeine and other natural products in a

描述（由申请人提供）：在美国，数百万人患有由内耳感觉毛细胞永久性损伤引起的听力损失。毛细胞损伤通常是由于暴露于过量的声音或特定的合法药物，如氨基糖苷类抗生素。这项拟议研究的总体目标是发现新的药物，防止氨基糖苷类药物诱导的毛细胞死亡，并保护需要这些救命抗生素的患者的听力损失。在这项工作中，我们将使用斑马鱼侧线，这是一个强大的体内模型，用于研究毛细胞死亡和保护机制。我们的初步数据表明，咖啡因显着保护毛细胞免受氨基糖苷类损伤。这项研究的具体目标是确定咖啡因提供保护的机制，并确定其他天然产品，防止氨基糖苷类药物诱导的毛细胞死亡。该项目有两个具体目标：1）测试咖啡因耳保护是通过抑制p53和腺苷信号传导介导的假设，2）测试天然产物是预防氨基糖苷类耳毒性的新药物的假设。我们以前的研究表明，氨基糖苷类激活受损毛细胞中的p53。咖啡因是一种已知的p53抑制剂，部分通过其与腺苷信号的相互作用。对于目标1，我们将在氨基糖苷类药物治疗期间使用药理学和遗传操作来调节p53和腺苷受体。咖啡因是一种植物生物碱和其他天然产物，包括植物和微生物提取物，可能是一种尚未开发的新型耳保护剂。在目标2中，我们将进行一系列药物发现实验，以确定作为毛细胞保护剂的天然产物，并表征其作用机制。这些研究将为听觉药物的开发提供新的先导化合物。通过专门针对咖啡因，一种广泛使用的化合物，具有良好的安全性，我们增加了我们的发现可以迅速转化为临床应用的可能性。学生研究人员是这个项目的各个方面不可或缺。学生将学习尖端的细胞生物学和药物发现技术，这将加强他们的生物医学职业机会，同时积极促进发现 预防药物性听力损失的新化合物。

DESCRIPTION (provided by applicant): Millions of people in the U.S. suffer from hearing loss caused by permanent damage to sensory hair cells of the inner ear. Hair cell damage often results from exposure to excessive sound or particular licit drugs, such as the aminoglycoside antibiotics. The overall goal of this proposed research is to discover novel drugs that prevent aminoglycoside-induced hair cell death and preserve hearing loss in patients requiring these life-saving antibiotics. For this work we will use the zebrafish lateral line, which is a powerful n vivo model for studying mechanisms of hair cell death and protection. Our preliminary data indicate that caffeine significantly protects hair cells from aminoglycoside damage. The specific goal of this research is to determine the mechanism by which caffeine confers protection, and to identify additional natural products that prevent aminoglycoside-induced hair cell death. This project has two specific aims: 1) Test the hypothesis that caffeine otoprotection is mediated by inhibition of p53 and adenosine signaling, and 2) Test the hypothesis that natural products are novel agents for preventing aminoglycoside ototoxicity. Our previous research demonstrates that aminoglycosides activate p53 in damaged hair cells. Caffeine is a known p53 inhibitor, in part via its interaction with adenosine signaling. For Aim 1, we will use pharmacologic and genetic manipulation to modulate p53 and adenosine receptors during aminoglycoside treatment. Caffeine is a plant alkaloid and other natural products, including plant and microbial extracts, likely represent an untapped source of novel otoprotective agents. In Aim 2, we will conduct a series of drug discovery experiments to identify natural products that act as hair cell protectants and to characterize their mechanisms of action. These studies will provide new lead compounds for auditory drug development. By specifically targeting caffeine, a widely used compound with a well-characterized safety profile, we increase the likelihood that our findings may be quickly translated into clinical application. Student researchers are integral to all aspect of this project. Students will learn cutting- edge cell biology and drug discovery techniques that will bolster their biomedical career opportunities, while actively contributing to the discovery of new compounds that prevent drug-induced hearing loss.