Surprising Efficacy of Discounted Antibiotics vs. MDR Gram-Negative Pathogens Occurring Through Innate Immune Sensitization

折扣抗生素对抗通过先天免疫敏化产生的耐多药革兰氏阴性病原体的惊人功效

• 批准号: 10665716
• 负责人: Monika Kumaraswamy
• 金额: $ 47.4万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10665716
• 关键词: Acinetobacter baumannii; Adopted; Animals; Antibiotic Resistance; Antibiotic susceptibility; Antibiotics; Antimicrobial Cationic Peptides; Antimicrobial susceptibility; Attention; Azithromycin; Bacteremia; Bacterial Physiology; Bacteriology; Biological Assay; Blood Platelets; Case Series; Cellular Assay; Clinical; Clonal Expansion; Complement; Cytokine Signaling; Cytology; Data; Defect; Development; Diagnostic; Enterobacter cloacae; Enzymes; Escherichia coli; Evolution; FDA approved; Face; Genetic; Host Defense; Human; Immune; Immune signaling; Immune system; Immunologic Factors; Immunologic Sensitization; Immunologics; Infection; Innate Immune System; Integration Host Factors; Interferons; Investigation; Kinetics; Klebsiella pneumoniae; Laboratories; Letters; Macrophage; Medicine; Metabolic; Methods; Modeling; Modern Medicine; Morbidity - disease rate; Multi-Drug Resistance; Mus; Natural Immunity; Nosocomial Infections; Pathway interactions; Patients; Phagocytes; Phagocytosis; Pharmaceutical Preparations; Pharmacologic Substance; Physicians; Production; Proteus mirabilis; Pseudomonas aeruginosa; Public Health; Publishing; Research; Resistance; Respiratory Burst; Rod; Sepsis; Series; Serum; Shapes; Signal Transduction; Standardization; Superbug; Tazobactam; Testing; Therapeutic; Thinking; Translating; Transmission Electron Microscopy; Variant; Whole Blood; Work; antimicrobial; antimicrobial drug; antimicrobial peptide; bactericide; beta-Lactamase; beta-Lactams; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; cathelicidin; cathelicidin antimicrobial peptide; catheter associated UTI; cell killing; effectiveness analysis; extracellular; forgetting; genome sequencing; high risk; human neutrophil peptide 1; immune clearance; immunoregulation; in vivo; in vivo Model; inhibitor; methicillin resistant Staphylococcus aureus; mortality; mouse model; multi-drug resistant pathogen; neutrophil; novel therapeutics; pathogen; pathogenic bacteria; pharmacologic; pneumonia model; receptor; resistant strain; synergism; therapeutically effective; translational impact; treatment guidelines; triphenylmethylphosphonium; whole genome

The burgeoning antibiotic resistance crisis is an imminent threat to the practice of modern medicine and global public health. In particular, nosocomial infections caused by multi-drug resistant (MDR) Gram- negative rods (GNRs) such as carbapenem-resistant strains of Klebsiella pneumoniae (Kp), Escherichia coli (Eco), Enterobacter cloacae (Ecl), Proteus mirabilis (Pm), Acinetobacter baumannii (Ab) and Pseudomonas aeruginosa (Pa) are spreading through horizontal transfer and clonal expansion with limited therapeutic options and high attendant morbidity and mortality. We are challenging the current standard MIC/MBC testing in bacteriologic media, which is totally agnostic to the host immune system. Even before the first diagnostic encounter with a physician, a patient’s infection is already being combatted by numerous endogenous immune effectors, including antimicrobial peptides, serum complement and phagocytic cells. While tremendous effort has been extended to identify and exploit synergy between different classes of pharmaceutical antibiotics, very little work has been conducted to study the way pharmaceutical antibiotics interact with endogenous antimicrobial defenses. We have documented unexpected activity of azithromycin (AZM), the most commonly prescribed antibiotic in the US, to sensitize highly MDR strains of Kp, Ab and Pa to innate immune killing – even though this drug is not even included in the AST panel when such isolates are encountered. Second, we have documented unexpected activity of β-lactamase inhibitors (BLIs) such as tazobactam (TAZ) and avibactam (AVI) to sensitize highly MDR strains of Kp and Ab to innate immune killing – even though these agents are never contemplated to have direct action beyond blocking β- lactamase enzymes. In the present proposal, we will identify synergistic interactions wherein AZM or BLIs sensitize MDR GNRs to killing by host AMPs, serum complement or phagocytic cells using checkerboard and kinetic killing assays, assays of bacterial physiology including bacterial cytological profiling (BCP), and studies of sensitization to complement, platelets, and phagocytes. Then using Ab and Kp as model MDR GNRs, we will translate innate immune sensitization by AZM and BLIs to effective therapeutic options in murine models of pneumonia, sepsis and UTI, using pharmacologically or genetically targeted mice to have defects in innate immunity (cathelicidin, complement, neutrophils) to identify key synergistic host factors. Careful attention is paid to provide alternative methods for mechanism of action determination (e.g. metabolic precursor incorporation, transmission electron microscopy, whole genome sequencing of induced resistant variants) and to assess other potential immunomodulatory effects of AZM and BLIs on cytokine signaling. As our earlier work with MDR Gram+ pathogens (MRSA, VRE) and β-lactam sensitization to innate immunity was corroborated in clinical series and influenced treatment guidelines, we foresee immediate translational impact of the current investigations on MDR GNR pathogens.

日益严重的抗生素耐药性危机对现代医学实践和全球公共卫生构成迫在眉睫的威胁，特别是由多重耐药（MDR）革兰氏阴性杆菌（GNR）（例如肺炎克雷伯菌的碳青霉烯类耐药菌株）引起的医院感染。 (Kp)、大肠杆菌 (Eco)、阴沟肠杆菌 (Ecl)、奇异变形杆菌 (Pm)、不动杆菌鲍曼不动杆菌 (Ab) 和铜绿假单胞菌 (Pa) 通过水平转移和克隆扩增进行传播，治疗选择有限，并且伴随的发病率和死亡率很高，我们正在挑战目前细菌学介质中的标准 MIC/MBC 测试，该测试对于宿主来说是完全不可知的。甚至在第一次与医生进行诊断之前，患者的感染就已经被许多内源性免疫效应物所对抗，包括抗菌肽、血清补体和免疫系统。虽然人们付出了巨大的努力来识别和利用不同类别的药物抗生素之间的协同作用，但对药物抗生素与内源性抗菌防御相互作用的方式进行的研究却很少，我们已经记录了阿奇霉素（AZM）的意外活性。美国最常用的抗生素，使 Kp、Ab 和 Pa 的高度 MDR 菌株对先天免疫杀伤敏感——尽管这种药物甚至不包括在 AST 组中，而当这些分离株被其次，我们已经充分记录了 β-内酰胺酶抑制剂 (BLI)，例如他唑巴坦 (TAZ) 和阿维巴坦 (AVI) 的意想不到的活性，可以使 Kp 和 Ab 的 MDR 菌株对先天免疫杀伤敏感，尽管这些药物从未被认为具有直接的杀伤作用。在本提案中，我们将确定 AZM 或 BLI 是否使 MDR 敏感的协同相互作用。使用棋盘和动力学杀伤测定、细菌生理学测定（包括细菌细胞学分析 (BCP)）以及对补体、血小板和吞噬细胞的敏化研究，然后使用 Ab 和 Kp 作为模型，研究 GNR 对宿主 AMP、血清补体或吞噬细胞的杀伤作用。 MDR GNR，我们将把 AZM 和 BLI 的先天免疫敏化转化为肺炎、脓毒症和 UTI 小鼠模型的有效治疗方案，使用具有先天免疫缺陷（组织素、补体、中性粒细胞）的药理学或基因靶向小鼠来识别关键的协同宿主因子，并特别注意为作用机制的确定提供替代方法（例如代谢前体掺入、透射电子显微镜、整体）。诱导耐药变异的基因组测序）并评估 AZM 和 BLI 对细胞因子信号传导的其他潜在免疫调节作用。 VRE）和β-内酰胺对先天免疫的敏感性在临床系列中得到证实，并影响了治疗指南，我们预计当前对 MDR GNR 病原体的研究将产生直接转化影响。

项目成果

Potent Activity of Ertapenem Plus Cefazolin Within Staphylococcal Biofilms: A Contributing Factor in the Treatment of Methicillin-Susceptible Staphylococcus aureus Endocarditis.

厄他培南加头孢唑林在葡萄球菌生物膜内的有效活性：治疗甲氧西林敏感金黄色葡萄球菌心内膜炎的影响因素。

• 发表时间: 2022-05
• 影响因子: 4.2
• 作者: Gilbertie, Jessica;Ulloa, Erlinda R;Daiker, Jennifer C;Nguyen, Khanh;Smelter, Dan;Rose, Warren;Geriak, Matthew;Schnabel, Lauren V;Nizet, Victor;Sakoulas, George
• 通讯作者: Sakoulas, George

Evaluating Organism-Wide Changes in the Metabolome and Microbiome following a Single Dose of Antibiotic.

评估单剂量抗生素后整个生物体代谢组和微生物组的变化。

• 发表时间: 2020-10-06
• 影响因子: 6.4
• 作者: Vrbanac, Alison;Patras, Kathryn A;Jarmusch, Alan K;Mills, Robert H;Shing, Samuel R;Quinn, Robert A;Vargas, Fernando;Gonzalez, David J;Dorrestein, Pieter C;Knight, Rob;Nizet, Victor
• 通讯作者: Nizet, Victor

Current Paradigms of Combination Therapy in Methicillin-Resistant Staphylococcus aureus (MRSA) Bacteremia: Does it Work, Which Combination, and For Which Patients?

目前治疗耐甲氧西林金黄色葡萄球菌 (MRSA) 菌血症的联合治疗范例：是否有效、哪种组合以及适用于哪些患者？

• 发表时间: 2021
• 作者: Rose, Warren;Fantl, Michael;Geriak, Matthew;Nizet, Victor;Sakoulas, George
• 通讯作者: Sakoulas, George

Comparative molecular profiling of multidrug-resistant Pseudomonas aeruginosa identifies novel mutations in regional clinical isolates from South India.

多重耐药铜绿假单胞菌的比较分子分析鉴定了印度南部地区临床分离株的新突变。

• 发表时间: 2024-02
• 作者: Menon, Nitasha D;Somanath, Priyanka;Jossart, Jennifer;Vijayakumar, Gayathri;Shetty, Kavya;Baswe, Manasi;Chatterjee, Meghna;Hari, Malavika B;Nair, Samitha;Kumar, V Anil;Nair, Bipin G;Nizet, Victor;Perry, J Jefferson P;Kumar, Geetha B
• 通讯作者: Kumar, Geetha B

Machine Learning of Bacterial Transcriptomes Reveals Responses Underlying Differential Antibiotic Susceptibility.

细菌转录组的机器学习揭示了差异抗生素敏感性背后的反应。

• 发表时间: 2021
• 影响因子: 4.8
• 作者: Sastry, Anand V;Dillon, Nicholas;Anand, Amitesh;Poudel, Saugat;Hefner, Ying;Xu, Sibei;Szubin, Richard;Feist, Adam M;Nizet, Victor;Palsson, Bernhard
• 通讯作者: Palsson, Bernhard