Molecular basis and intervention of Staphylococcus aureus agglutination

金黄色葡萄球菌凝集的分子基础及干预

• 批准号: 9180674
• 负责人: Dominique M. Missiakas
• 金额: $ 38.94万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9180674
• 关键词: Abscess; Adhesions; Agglutination; Antibiotics; Aspartate; Bacteremia; Bacteria; Binding; Blocking Antibodies; Blood; Bypass; Calcium; Cell Wall; Cells; Chelating Agents; Clinical; Coagulation Process; Coenzyme A; Communities; Disease; Drug resistance; Electrons; Encapsulated; Endothelial Cells; Endothelium; Event; Factor XIII; Fibrin; Fibrinogen; Fibronectins; Fluorescein; Genes; Genetic; Genus staphylococcus; Human; Immune; In Vitro; Incubated; Intervention; Label; Lesion; Mediating; Methicillin Resistance; Microscope; Modeling; Modification; Molecular; Monoclonal Antibodies; Multiprotein Complexes; Mus; Organ; Pathogenesis; Pathway interactions; Patients; Phagocytes; Phagocytosis; Phenotype; Plasma; Post-Translational Protein Processing; Prothrombin; Recruitment Activity; Resistance; Sepsis; Serine; Skin; Skin Tissue; Soft Tissue Infections; Splanchnic Circulation; Staphylococcus aureus; Staphylococcus epidermidis; Surface; Survival Rate; Symbiosis; Therapeutic antibodies; Time; Tissues; Transferase; Vascular Endothelium; Virulence; Work; base; crosslink; factor A; gene product; in vivo; killings; loss of function; micrography; mortality; mouse model; pathogen; prevent; public health relevance; reconstitution; release factor; small molecule inhibitor; sortase; von Willebrand Factor

DESCRIPTION (provided by applicant): Staphylococcus aureus, a commensal of the human skin and nares, is also an invasive pathogen and frequent cause of skin and soft tissue infections, bacteremia and sepsis. Many clinical isolates are resistant against commonly used antibiotics and these strains are collectively referred to as methicillin-resistant S. aureus (MRSA). The annual survival rate of patients with MRSA sepsis is less than 50%. S. aureus is the only bacterial pathogen known to coagulate plasma and to agglutinate with fibrin cables in blood. We show here that this agglutination phenotype is based on the secreted products from three genes (coa, vwb, clfA) as well as several non-secreted gene products. We propose a new model whereby S. aureus agglutination involves the formation of staphylothrombin (Coa, vWbp)-assembled fibrin cables, which are capped by clumping factor A (ClfA) on the staphylococcal surface and crosslinked by factor XIII. We hypothesize that S. aureus agglutination provides for escape from phagocytic killing. Further, vWbp-mediated recruitment of von Willebrand Factor (vWF) provides a tether for endothelial cells and supports bacterial traffic out of the vasculature thereby promoting S. aureus dissemination to organ tissues where staphylococci replicate as a bacterial community, protected from immune cells. This key virulence strategy may be perturbed with monoclonal antibodies and small molecule inhibitors to either prevent or treat S. aureus sepsis. Preliminary work demonstrated that S. aureus Coa and vWbp each associate with host prothrombin to form multi-protein complexes in human plasma (with prothrombin, fibrinogen, fibronectin and factor XIII) dedicated to the formation of cross-linked fibrin cables. Staphylococcal agglutination involves the surface display of functional ClfA, which requires several gene products [aggABCD, (staphylococcal agglutination genes)] that appear to modify ClfA prior to sortase A-mediated anchoring in the bacterial cell wall envelope (srtA). In addition to genetic loss-of-function analysis, we will also reconstitute agglutination in vitro from purifie components and in vivo by expressing genes in Staphylococcus epidermidis, an opportunistic pathogen that cannot agglutinate or form discrete abscess lesions. Monoclonal antibodies (mAbs) -isolated against purified Coa, vWbp or ClfA- and small molecule inhibitors are being used to perturb staphylococcal agglutination in vitro and in vivo and examined for the provision of protection or therapy in a mouse model of S. aureus sepsis.

描述（由申请方提供）：金黄色葡萄球菌是人体皮肤和鼻孔的寄生虫，也是一种侵入性病原体，是皮肤和软组织感染、菌血症和败血症的常见原因。许多临床分离株对常用抗生素具有耐药性，这些菌株统称为耐甲氧西林S。金黄色葡萄球菌（MRSA）。MRSA败血症患者的年生存率低于50%。S.金黄色葡萄球菌是唯一已知的凝固血浆并与血液中的纤维蛋白缆线凝集的细菌病原体。我们在这里表明，这种凝集表型是基于三个基因（coa，vaccine，clfA）的分泌产物以及几个非分泌的基因产物。我们提出了一个新的模型，S。金黄色葡萄球菌凝集涉及葡萄球菌凝血酶（CoA，vWbp）组装的纤维蛋白缆的形成，所述纤维蛋白缆被葡萄球菌表面上的凝集因子A（ClfA）封端并被因子XIII交联。我们假设S.金黄色葡萄球菌凝集提供了逃避吞噬细胞杀伤。此外，vWbp介导的血管性血友病因子（vWF）募集为内皮细胞提供了一个系链，并支持细菌运输出血管系统，从而促进S。金黄色葡萄球菌传播到器官组织，在那里葡萄球菌作为细菌群落复制，免受免疫细胞的影响。这种关键的毒力策略可能会受到单克隆抗体和小分子抑制剂的干扰，以预防或治疗S。金黄色脓毒症初步研究表明，S。金黄色葡萄球菌CoA和vWbp各自与宿主凝血酶原结合，在人血浆中形成多蛋白复合物（与凝血酶原、纤维蛋白原、纤连蛋白和因子XIII），专用于形成交联的纤维蛋白缆。葡萄球菌凝集涉及功能性ClfA的表面展示，这需要几种基因产物[aggABCD，（葡萄球菌凝集基因）]，其似乎在分选酶A介导的锚定在细菌细胞壁包膜（srtA）中之前修饰ClfA。除了遗传功能丧失分析，我们还将在体外从纯化组分和在体内通过在表皮葡萄球菌（一种不能凝集或形成离散脓肿病变的机会致病菌）中表达基因来重建凝集。针对纯化的CoA、vWbp或ClfA分离的单克隆抗体（mAb）和小分子抑制剂正在用于干扰葡萄球菌的体外和体内凝集，并在小鼠S.金黄色脓毒症