Combating Multidrug-Resistant Bacteria by Integrating a Novel Target Site Penetration and Receptor Binding Assay Platform Into Translational Modeling.
Combating Multidrug-Resistant Bacteria by Integrating a Novel Target Site Penetration and Receptor Binding Assay Platform Into Translational Modeling.
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DOI：
10.1002/cpt.2205
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发表时间：
2021-04
影响因子：
6.7
通讯作者：
Bulitta, Juergen B.
中科院分区：
文献类型：
--
作者：
Lang, Yinzhi;Shah, Nirav R.;Tao, Xun;Reeve, Stephanie M.;Zhou, Jieqiang;Moya, Bartolome;Sayed, Alaa R. M.;Dharuman, Suresh;Oyer, Jeremiah L.;Copik, Alicja J.;Fleischer, Brett A.;Shin, Eunjeong;Werkman, Carolin;Basso, Kari B.;Deveson Lucas, Deanna;Sutaria, Dhruvitkumar S.;Megroz, Marianne;Kim, Tae Hwan;Loudon-Hossler, Victoria;Wright, Amy;Jimenez-Nieves, Rossie H.;Wallace, Miranda J.;Cadet, Keisha C.;Jiao, Yuanyuan;Boyce, John D.;LoVullo, Eric D.;Schweizer, Herbert P.;Bonomo, Robert A.;Bharatham, Nagakumar;Tsuji, Brian T.;Landersdorfer, Cornelia B.;Norris, Michael H.;Soo Shin, Beom;Louie, Arnold;Balasubramanian, Venkataraman;Lee, Richard E.;Drusano, George L.;Bulitta, Juergen B.
多重耐药细菌正在造成严重的全球健康危机。在过去几十年中，制药行业在抗生素发现和开发方面的投资急剧下降，减缓了新技术的采用。我们必须基于最新技术创造新的机制见解，并使用转化策略来优化患者治疗。尽管药物开发依赖于最小抑菌浓度测试和建立的体外和小鼠感染模型，但对革兰氏阴性菌外膜渗透性的有限理解提出了重大挑战。我们的团队已经开发了一个平台，使用最新的技术来表征完整细菌中的靶位点渗透和受体结合，从而为翻译建模提供信息并指导新的发现。增强的测定可以使用多重液相色谱串联质谱法定量β-内酰胺抗生素和β-内酰胺酶抑制剂的外膜渗透性。虽然已知β-内酰胺抗生素与多种不同的青霉素结合蛋白（PBP）结合，但几乎总是在裂解细菌中研究其结合特征。开发了完整细菌周质中PBP结合的新测定法，并通过蛋白质组学鉴定了蛋白质。为了表征细菌的形态变化响应PBP结合，高通量流式细胞术和延时共聚焦显微镜与荧光探针提供了前所未有的机制的见解。此外，定量胞质受体结合和胞内药物浓度的新测定法告知靶位点占有率。通过定量和系统药理学建模整合这些机制数据，以最大限度地杀灭细菌，并最大限度地减少体外和小鼠感染模型中的耐药性。这种转化方法有望为严重感染患者确定抗生素联合给药策略。
Multidrug-resistant bacteria are causing a serious global health crisis. A dramatic decline in antibiotic discovery and development investment by pharmaceutical industry over the last decades has slowed the adoption of new technologies. It is imperative that we create new mechanistic insights based on latest technologies, and use translational strategies to optimize patient therapy. Although drug development has relied on minimal inhibitory concentration testing and established in vitro and mouse infection models, the limited understanding of outer membrane permeability in Gram-negative bacteria presents major challenges. Our team has developed a platform using the latest technologies to characterize target site penetration and receptor binding in intact bacteria that inform translational modeling and guide new discovery. Enhanced assays can quantify the outer membrane permeability of β-lactam antibiotics and β-lactamase inhibitors using multiplex liquid chromatography tandem mass spectrometry. While β-lactam antibiotics are known to bind to multiple different penicillin-binding proteins (PBPs), their binding profiles are almost always studied in lysed bacteria. Novel assays for PBP binding in the periplasm of intact bacteria were developed and proteins identified via proteomics. To characterize bacterial morphology changes in response to PBP binding, high-throughput flow cytometry and time-lapse confocal microscopy with fluorescent probes provide unprecedented mechanistic insights. Moreover, novel assays to quantify cytosolic receptor binding and intracellular drug concentrations inform target site occupancy. These mechanistic data are integrated by quantitative and systems pharmacology modeling to maximize bacterial killing and minimize resistance in in vitro and mouse infection models. This translational approach holds promise to identify antibiotic combination dosing strategies for patients with serious infections.
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影响因子：
4.9
作者：
Bulitta, Juergen B.;Ly, Neang S.;Tsuji, Brian T.
通讯作者：
Tsuji, Brian T.
影响因子：
4.9
作者：
BILELLO, JA;BAUER, G;DRUSANO, GL
通讯作者：
DRUSANO, GL
影响因子：
5.2
作者：
Appelbaum, Peter C.
通讯作者：
Appelbaum, Peter C.
影响因子：
3.2
作者：
Dougherty, TJ;Kennedy, K;Pucci, MJ
通讯作者：
Pucci, MJ
影响因子：
6.4
作者：
Bulman ZP;Chen L;Walsh TJ;Satlin MJ;Qian Y;Bulitta JB;Peloquin CA;Holden PN;Nation RL;Li J;Kreiswirth BN;Tsuji BT
通讯作者：
Tsuji BT