Bacillus anthracis Targets Involved in Chemokine-Mediated Antimicrobial Activity

炭疽杆菌靶标参与趋化因子介导的抗菌活性

• 批准号: 9029273
• 负责人: MOLLY A HUGHES
• 金额: $ 39.5万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-05 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9029273
• 关键词: A/J Mouse; ATP-Binding Cassette Transporters; Anthrax disease; Anti-Bacterial Agents; Antibiotics; Antimicrobial Cationic Peptides; Antimicrobial Effect; Bacillus (bacterium); Bacillus anthracis; Bacillus anthracis spore; Bacteria; Bacterial Proteins; C57BL/6 Mouse; CXC Chemokines; CXC chemokine receptor 3; CXCL10 gene; CXCL11 gene; CXCL9 gene; CXCR3 gene; Cell division; Cells; Charge; Chemicals; Co-Immunoprecipitations; Data; Development; Electron Microscopy; Escherichia coli; Exhibits; Foundations; Future; Genes; Germination; Gram-Negative Bacteria; Health; Host Defense; Human; Immune response; Immune system; Immunofluorescence Microscopy; Immunoprecipitation; In Vitro; Infection; Infectious Agent; Integral Membrane Protein; Interferons; Invaded; Lead; Leukocytes; Libraries; Listeria monocytogenes; Lung; LytA enzyme; Mediating; Multi-Drug Resistance; Organism; Outcome; Phenotype; Plasmids; Play; Predisposition; Proteins; Publishing; Recombinants; Recruitment Activity; Reporting; Reproduction spores; Resistance; Role; Site; Site-Directed Mutagenesis; Staging; Testing; Therapeutic; Therapeutic Agents; Work; antimicrobial; base; chemokine; constriction; crosslink; drug resistant microorganism; fighting; genetic approach; in vivo; innovation; insight; killings; macromolecule; microorganism; migration; mouse model; mutant; novel; pathogen; receptor; response; small molecule

DESCRIPTION (provided by applicant): Bacillus anthracis Targets Involved in Chemokine-Mediated Antimicrobial Activity Chemokines are chemotactic cytokines that function in host defense by orchestrating leukocyte migration to sites of infection. However, a number of chemokines have also been found to directly kill a range of pathogenic microorganisms through an as yet undefined mechanism. We previously reported that the interferon-inducible CXC chemokines, CXCL9, CXCL10, and CXCL11, block Bacillus anthracis spore germination, reduce spore viability, and kill vegetative cells, with CXCL10 being the most effective. We also reported that C57BL/6 mice, which are resistant to pulmonary B. anthracis Sterne strain infection, produced significant levels of CXCL9, CXCL10, and CXCL11 in their lungs following spore challenge; whereas, highly susceptible A/J mice did not generate significant levels of these chemokines during pulmonary infection. In vivo neutralization of CXCL9, CXCL9/CXCL10, or CXCL9/CXCL10/CXCL11 rendered C57BL/6 mice susceptible to pulmonary anthrax whereas neutralization of their shared receptor CXCR3, which is the receptor expressed on leukocytes recruited to the site of infection by CXCL9, CXCL10, CXCL11, had no impact on survival. These findings support that CXCL9, CXCL10, and CXCL11 have direct antimicrobial effects against B. anthracis both in vitro and in vivo. To identify the vegetative cell target(s) of CXCL10, we screened a B. anthracis transposon mutant library and found that disruption of ftsX, which encodes the transmembrane protein of a widely conserved prokaryotic ABC transporter, resulted in a CXCL10-resistant phenotype. Deletion of the ftsX gene in B. anthracis (i.e., DftsX) resulted in resistance of vegetative cells to CXCL10, and complementation of ftsX restored CXCL10 susceptibility. In contrast, DftsX spores remained susceptible to CXCL10, suggesting that spores have a different CXCL10 target. To further investigate the role of FtsX, as well as other B. anthracis targets of CXCL10, we propose three Specific Aims: 1) Determine the role of FtsX in susceptibility of vegetative cells to CXCL10; 2) Identify spore target(s) of CXCL10; and 3) Determine the role of spore and vegetative bacterial targets of CXCL10 during in vivo infection. These studies will provide a key foundation for the development of innovative therapeutic strategies for treating infections caused by not only B. anthracis but also a range of pathogenic, potentially multi-drug resistant microorganisms.

描述（由申请人提供）：参与趋化因子介导的抗菌活性的炭疽芽孢杆菌靶标趋化因子是趋化细胞因子，通过协调白细胞迁移至感染部位来在宿主防御中发挥作用。然而，还发现许多趋化因子可以通过尚未明确的机制直接杀死一系列病原微生物。我们之前报道过，干扰素诱导的 CXC 趋化因子 CXCL9、CXCL10 和 CXCL11 可阻止炭疽芽孢杆菌孢子萌发、降低孢子活力并杀死营养细胞，其中 CXCL10 最有效。我们还报道，对肺炭疽杆菌 Sterne 菌株感染具有抵抗力的 C57BL/6 小鼠在孢子攻击后，其肺部产生了显着水平的 CXCL9、CXCL10 和 CXCL11；然而，高度易感的 A/J 小鼠在肺部感染期间并未产生显着水平的这些趋化因子。 CXCL9、CXCL9/CXCL10 或 CXCL9/CXCL10/CXCL11 的体内中和使 C57BL/6 小鼠易患肺炭疽，而中和其共享受体 CXCR3（CXCR3 是 CXCL9、CXCL10 募集到感染部位的白细胞上表达的受体） ，CXCL11，对生存没有影响。这些发现支持 CXCL9、CXCL10 和 CXCL11 在体外和体内对炭疽芽孢杆菌具有直接抗菌作用。为了鉴定 CXCL10 的营养细胞靶标，我们筛选了炭疽芽孢杆菌转座子突变体文库，发现编码广泛保守的原核 ABC 转运蛋白跨膜蛋白的 ftsX 的破坏导致了 CXCL10 抗性表型。炭疽杆菌中 ftsX 基因（即 DftsX）的缺失导致营养细胞对 CXCL10 产生抗性，而 ftsX 的补充则恢复了 CXCL10 的敏感性。相反，DftsX 孢子仍然对 CXCL10 敏感，表明孢子具有不同的 CXCL10 靶标。为了进一步研究 FtsX 以及 CXCL10 的其他炭疽杆菌靶标的作用，我们提出了三个具体目标： 1) 确定 FtsX 在营养细胞对 CXCL10 敏感性中的作用； 2) 鉴定CXCL10的孢子靶标； 3)确定CXCL10的孢子和营养细菌靶标在体内感染过程中的作用。这些研究将为开发创新治疗策略提供关键基础，这些策略不仅可以治疗炭疽杆菌，还可以治疗一系列致病性、潜在的多重耐药微生物引起的感染。

项目成果

Mechanistic insights and therapeutic opportunities of antimicrobial chemokines.

抗菌趋化因子的机制见解和治疗机会。

• 发表时间: 2019
• 影响因子: 7.3
• 作者: Crawford, Matthew A;Margulieux, Katie R;Singh, Arpita;Nakamoto, Robert K;Hughes, Molly A
• 通讯作者: Hughes, Molly A

Genome Sequences of Multidrug-Resistant, Colistin-Susceptible and -Resistant Klebsiella pneumoniae Clinical Isolates from Pakistan.

来自巴基斯坦的多重耐药、粘菌素敏感和耐药肺炎克雷伯菌临床分离株的基因组序列。

• 发表时间: 2016-12-15
• 作者: Crawford, Matthew A;Timme, Ruth;Lomonaco, Sara;Lascols, Christine;Fisher, Debra J;Sharma, Shashi K;Strain, Errol;Allard, Marc W;Brown, Eric W;McFarland, Melinda A;Croley, Tim;Hammack, Thomas S;Weigel, Linda M;Anderson, Kevin;Hodge, David R
• 通讯作者: Hodge, David R

CXC Chemokines Exhibit Bactericidal Activity against Multidrug-Resistant Gram-Negative Pathogens.

CXC 趋化因子对多重耐药革兰氏阴性病原体具有杀菌活性。

• 发表时间: 2017
• 影响因子: 6.4
• 作者: Crawford, Matthew A;Fisher, Debra J;Leung, Lisa M;Lomonaco, Sara;Lascols, Christine;Cannatelli, Antonio;Giani, Tommaso;Rossolini, Gian Maria;Doi, Yohei;Goodlett, David R;Allard, Marc W;Sharma, Shashi K;Khan, Erum;Ernst, Robert K;Hughes, Moll
• 通讯作者: Hughes, Moll

Resistome of carbapenem- and colistin-resistant Klebsiella pneumoniae clinical isolates.

耐碳青霉烯和粘菌素的肺炎克雷伯菌临床分离株的耐药性。

• 发表时间: 2018
• 影响因子: 3.7
• 作者: Lomonaco, Sara;Crawford, Matthew A;Lascols, Christine;Timme, Ruth E;Anderson, Kevin;Hodge, David R;Fisher, Debra J;Pillai, Segaran P;Morse, Stephen A;Khan, Erum;Hughes, Molly A;Allard, Marc W;Sharma, Shashi K
• 通讯作者: Sharma, Shashi K