Combatting antibiotic resistance with synthetic biology technologies

项目摘要/摘要 抗菌素耐药性的出现是对我们全体人口最严重的健康威胁之一。 来自耐药细菌的感染现在太普遍了，一些病原体已经对 多个抗生素类别。疾病控制和预防中心(CDC)最近估计， 在美国，耐药细菌每年导致200多万人患病和23,000多人死亡。 随着抗药性感染率的上升，迫切需要新的诊断方法来迅速 确定对感染最有效的治疗方法。不幸的是，目前的执行方法 抗生素敏感性试验(AST)包括从临床样本中培养微生物并确定 他们通过细胞生长对抗生素的敏感性。这种“黄金标准”技术非常耗时。 (至少48-72小时)，并可能导致适当治疗的显著延误、长期患病、 死亡的风险、不适当的抗生素使用和增加的耐药性传播。对于一些感染，比如 淋病、门冬氨酸氨基转移酶甚至不在临床上进行，而是根据治疗失败来推断。简而言之，它 迫切需要开发新的策略来快速诊断和防止药物放大 抵抗。抗生素暴露可在敏感微生物中触发一组标志性mRNAs的表达 在短短几分钟内，提高了使用RNA检测-而不是细胞生长-作为 快速、基于表型的AST的新方法。我们将开发创新的RNA传感器技术， 在一个临床相关、低成本和易于使用的诊断平台中评估这些分子特征。 为了实现这一目标，我们将使用合成生物学方法来设计高度敏感的基因传感器 MRNA.这些传感器将部署在可排列和冷冻干燥的无细胞表达系统中 到低成本的固态基板上，比如纸。其结果将是一种新的抗生素诊断方法 理想的性能、存储和分发特性。RNA传感器技术将得到开发和应用 通过高优先级细菌生物体进行验证。值得注意的是，我们将首次定义 淋球菌的易感性，疾控中心最近将其提升为美国关注的主要原因。 其AST功能目前在临床环境中还不存在。这项工作将迎来一个新的 快速诊断抗生素耐药性的技术，有可能改变对 当今日益严重的抗菌素耐药性问题。

PROJECT SUMMARY / ABSTRACT The emergence of antimicrobial resistance is one of the most serious health threats to our entire population. Infections from resistant bacteria are now too common, and some pathogens have become resistant to multiple antibiotic classes. The Centers for Disease Control and Prevention (CDC) recently estimated that drug-resistant bacteria account for more than 2 million illnesses and over 23,000 deaths every year in the U.S. With rising rates of drug-resistant infections, there is pressing need for new diagnostic methods that can rapidly determine the most effective therapy for an infection. Unfortunately, the current method for performing antibiotic susceptibility testing (AST) involves growing microorganisms from clinical samples and determining their sensitivity to antibiotics through cell growth. This “gold standard” technique is extremely time-consuming (minimum 48-72 hours) and can result in significant delays in appropriate therapy, prolonged illness, greater risk of death, inappropriate antibiotic use, and increased spread of resistance. For some infections like gonorrhea, AST is not even performed in the clinic and instead inferred based on treatment failure. In short, it is imperative that new strategies are developed to rapidly diagnose and prevent the amplification of drug resistance. Antibiotic exposure can trigger the expression of a signature set of mRNAs in susceptible microbes in as rapidly as a few minutes, raising the exciting possibility of using RNA detection – not cell growth – as a new means for rapid, phenotype-based AST. We will develop innovative RNA sensor technology that evaluates these molecular signatures within a clinically-relevant, low-cost, and easy-to-use diagnostic platform. To achieve this, we will use synthetic biology approaches to engineer highly-sensitive genetic sensors of mRNA. These sensors will be deployed in cell-free expression systems that can be arrayed and freeze-dried onto low-cost, solid-state substrates like paper. The result will be a new class of antibiotic diagnostics with ideal performance, storage, and distribution characteristics. The RNA sensor technology will be developed and validated with high priority bacterial organisms. Notably, we will, for the first time, define RNA signatures of susceptibility for N. gonorrhoeae, which the CDC recently elevated as a major cause for concern in the U.S. and for which AST capabilities do not currently existing in the clinical setting. This work will usher in a new technology for rapidly diagnosing antibiotic resistance, with the potential to transform the management of today's growing antimicrobial resistance problem.