High Throughput Screening: Bacterial Mechanosensitive Channels as Drug Targets

描述（由申请人提供）：医疗保健行业的主要问题之一是多重耐药感染。医院中诸如金黄色葡萄球菌等耐药细菌的继发性感染以及多重耐药结核分枝杆菌已成为全球性的卫生问题。这项提议的目的是分离出潜在的广谱抗生素，这可能会彻底改变我们治疗常见和多重耐药细菌感染的方式，甚至可能是生物恐怖主义的受害者。我们以前发现，微小的结构变化会导致大孔细菌机械敏感通道错门，严重影响细菌细胞的生长和活力。有许多特征使这些通道成为药理学靶点的理想候选者。这些通道：(1)在细菌生长的所有阶段表达，这表明激动剂甚至可以攻击那些倾向于长时间保持休眠状态的微生物，比如结核分枝杆菌。(2)基本上存在于每一种细菌中，但为微生物所特有的；在任何哺乳动物中都没有发现家族成员。(3)高度保守，因此药物可能是广谱的。(4)最后，我们为该项目提供了许多工具，包括零菌株，表达构建，以及校园内使用高通量筛选（HTS）设施的批准。总之，本提案的目的是开发该检测方法，并筛选HTS文库中的200,000个化合物，以分离针对机械敏感通道的潜在抗生素药物化合物。公共卫生相关性：我们建议分离新一代抗生素，这将是治疗多重耐药和其他难以治疗的感染的理想选择。分离出的抗生素也可能对紧急情况有影响，包括治疗生物恐怖主义行为的受害者。

DESCRIPTION (provided by applicant): One of the major problems in the health care industry is multi-drug resistant infections. Secondary infections of drug-resistant bacteria such as S. aureus in hospitals, and the multidrug resistant forms of M. tuberculosis, have become global health problems. The aim of this proposal is to isolate potential broad-spectrum antibiotic agents that could revolutionize how we treat common and multi-drug-resistant bacterial infections, and possibly even victims of bio-terrorism. We previously found that small structural changes can cause large- pore bacterial mechanosensitive channels to mis-gate, severely compromising the growth and viability of the bacterial cell. There are many features that make these channels ideal candidates for pharmacological targets. The channels are: (1) expressed in all phases of bacterial growth, suggesting that agonists would attack even microbes that have a tendency to remain dormant for long periods of time, such as M. tuberculosis. (2) found in essentially every bacterial species, but are unique to microbes; no family members are found in any mammal. (3) highly conserved, so drugs are likely be broad spectrum. (4) Finally, we have numerous tools for the project including null strains, expression constructs, and the approval for the use of the High Throughput Screening (HTS) facility on campus. In sum, the aim of this proposal is to develop the assay, and screen the 200,000 compounds in the HTS library to isolate compounds that are potential antibiotic drugs that target mechanosensitive channels. PUBLIC HEALTH RELEVANCE: We propose to isolate a new generation of antibiotics that would be ideal for treating multi-drug resistant and otherwise difficult to treat infections. The antibiotics isolated could also have implications in emergency situations including treating victims of acts of bioterrorism.