Otoupal Peter B, Cordell William T, Bachu Vismaya, Sitton Madeleine J, Chatterjee Anushree
Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, CO, 80303, USA.
BioFrontiers Institute, University of Colorado at Boulder, Boulder, CO, 80303, USA.
Commun Biol. 2018 Sep 3;1:129. doi: 10.1038/s42003-018-0135-2. eCollection 2018.
The ever-increasing threat of multi-drug resistant bacteria, a shrinking antibiotic pipeline, and the innate ability of microorganisms to adapt necessitates long-term strategies to slow the evolution of antibiotic resistance. Here we develop an approach, dubbed Controlled Hindrance of Adaptation of OrganismS or CHAOS, involving induction of epistasis between gene perturbations to deter adaption. We construct a combinatorial library of multiplexed, deactivated CRISPR-Cas9 devices to systematically perturb gene expression in . While individual perturbations improved fitness during antibiotic exposure, multiplexed perturbations caused large fitness loss in a significant epistatic fashion. Strains exhibiting epistasis adapted significantly more slowly over three to fourteen days, and loss in adaptive potential was shown to be sustainable. Finally, we show that multiplexed peptide nucleic acids increase the antibiotic susceptibility of clinically isolated Carbapenem-resistant in an epistatic fashion. Together, these results suggest a new therapeutic strategy for restricting the evolution of antibiotic resistance.
多重耐药细菌构成的威胁日益增加、抗生素研发渠道不断萎缩，以及微生物与生俱来的适应能力，都使得我们需要制定长期策略来减缓抗生素耐药性的演变。在此，我们开发了一种方法，称为“生物体适应的可控阻碍”（CHAOS），该方法涉及诱导基因扰动之间的上位性以阻止适应。我们构建了一个多重失活的CRISPR-Cas9装置的组合文库，以系统地扰动基因表达。虽然单个扰动在抗生素暴露期间提高了适应性，但多重扰动以显著的上位性方式导致了较大的适应性损失。表现出上位性的菌株在三到十四天内适应得明显更慢，并且适应性潜力的损失被证明是可持续的。最后，我们表明多重肽核酸以上位性方式增加了临床分离的耐碳青霉烯类细菌的抗生素敏感性。总之，这些结果提示了一种限制抗生素耐药性演变的新治疗策略。
