Using nanopocket membranes to capture bacterial outer membrane vesicles from biofluids

使用纳米袋膜从生物液中捕获细菌外膜囊泡

• 批准号: 10425443
• 负责人: THOMAS R GABORSKI
• 金额: $ 18.44万
• 依托单位: ROCHESTER INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-09 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10425443
• 关键词: APACHE II; Age; Albumins; Antibiotics; Antibodies; Antigens; Bacteria; Benchmarking; Biological; Biological Markers; Cause of Death; Cells; Chronic; Clinical; Clinical Data; Complex; Critical Care; Culture Media; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Early treatment; Enterobacteriaceae; Escherichia coli; Filtration; Focal Infection; Future; Health Status; Hospitals; Hour; Human; Immune response; Infection; Inflammatory; Liquid substance; Measures; Membrane; Molecular Diagnostic Testing; Organism; Outcome; Parents; Patients; Phosphate Buffer; Physicians; Physiological; Plasma; Proteins; Radial; Retrospective Studies; Saline; Sampling; Scientist; Sepsis; Severity of illness; System; Techniques; Technology; Testing; Tissues; Ultracentrifugation; Variant; Vesicle; Work; detection method; diagnostic assay; diagnostic biomarker; extracellular vesicles; molecular diagnostics; molecular marker; mortality; novel; point of care; point-of-care diagnostics; potential biomarker; pressure; rapid diagnosis; septic patients; success; tool

PROJECT SUMMARY Sepsis is not only the most expensive condition treated in US hospitals, but also a leading cause of death. Sepsis occurs when host pro-inflammatory immune responses become abnormally elevated due to a dysregulated or aberrant host-response to infection. Diagnostic methods for sepsis can vary between hospitals, but often involve scoring systems (e.g. APACHE II and SOFA) that grade the severity of illness in patients. Many of the altered physiological parameters measured by these scoring systems are not necessarily specific to sepsis, which makes it difficult to diagnose sepsis in early stages. Timing of a patient’s sepsis diagnosis is often predictive of their clinical outcome, underlining the need for a more definitive molecular diagnostic test. However, a recent study found that in the majority (70.1%) of sepsis cases, a specific causal organism could not be determined, likely due to aggressive antibiotics or localized infections. Bacterial outer membrane vesicles (OMVs) are attractive diagnostic biomarkers because of: A) their abundance and ability to circulate throughout the body and pass tissue barriers more easily than bacteria themselves; B) their robustness - unlike their bacterial cell “parent,” OMVs can withstand the inundation of broad-spectrum antibiotics; and C) their unique features that could allow for differentiation between bacterial species. The objective of this proposal is two-fold: Identify whether bacterial OMVs could be a molecular diagnostic biomarker for sepsis and develop a rapid approach to isolate them from patient plasma. We seek to develop a straightforward high-purity and rapid separation technology that effectively isolates and purifies OMVs from biofluids, including plasma. We will implement a modified tangential flow filtration approach, similar to those used by biopharma for high-capacity and high- yield purification, with a new nanopocket membrane that effectively captures and releases bacterial OMVs with minimal loss. Success of these aims will pave the way for the future development of a one-step, point-of-care diagnostic test for sepsis using bacterial OMVs as the molecular biomarker, as well a more complex diagnostic test that more accurately quantifies OMV levels in the patient’s biofluids to help direct early treatment and reduce mortality. This project will be led by an interdisciplinary team of experts in identifying potential biomarkers in E. coli; membranes, materials, and bioseparations; and a critical care physician scientist.

项目摘要 败血症不仅是美国医院治疗的最昂贵的病情，而且是主要原因 死亡。当宿主促炎性免疫反应异常变得异常时，会发生败血症 由于对感染的宿主反应失调或异常，因此升高。诊断方法 败血症在医院之间可能有所不同，但通常涉及评分系统（例如Apache II和沙发） 那个患者的疾病严重程度。许多改变的生理参数 通过这些评分系统衡量不一定特定于败血症，这使得很难 在早期诊断败血症。患者败血症诊断的时间通常是预测的 他们的临床结果，强调了对更明确的分子诊断测试的需求。 但是，最近的一项研究发现，在大多数（70.1％）的败血症病例中，这是一个特定原因 无法确定生物体，可能是由于侵袭性抗生素或局部感染引起的。 细菌外膜蔬菜（OMV）是有吸引力的诊断生物标志物，因为：a） 它们的丰度和能够在整个身体中循环并更容易通过组织屏障 比细菌本身； b）它们的鲁棒性 - 与细菌细胞“父”不同，OMV可以 承受广谱抗生素的淹没； c）它们的独特功能可以 允许细菌之间的区分。 该提案的目的是两个倍：确定细菌OMV是否可以是分子 败血症的诊断生物标志物，并开发出一种将它们与患者血浆分离的快速方法。 我们试图开发一种直接的高纯度和快速分离技术 分离株并从包括血浆在内的生物流体中净化OMV。我们将实施一个修改的 切向流滤波器方法，类似于生物武器用于高容量和高位的方法 产生纯化，并有一种新的纳米材料膜，可有效捕获并发行 细菌OMV损失最小。这些目标的成功将为未来铺平道路 开发使用细菌OMV作为败血症的一步，即时诊断测试 分子生物标志物以及更复杂的诊断测试，更准确地量化OMV 患者生物流体的水平有助于指导早期治疗并降低死亡率。这个项目将 由识别大肠杆菌潜在生物标志物的专家跨学科团队领导； 膜，材料和生物序列；和重症监护物理科学家。

项目成果

The Effect of Antibiotics on Outer Membrane Vesicle Production.

抗生素对外膜囊泡产生的影响。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Dinu,MilenaA;Michel,Lea;Gaborski,Thomas;Perdue,Janai;Kasper,Anna;Morehouse,Ryan
• 通讯作者: Morehouse,Ryan

Triggering toxicity: How antibiotics can enhance the release of outer membrane vesicles from Escherichia coli during sepsis.

引发毒性：抗生素如何在败血症期间增强大肠杆菌外膜囊泡的释放。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Michel,LeaV;Konovalova,Anna;Hellman,Judith;Gaborski,Thomas
• 通讯作者: Gaborski,Thomas

Optimizing the production, purification, characterization of E. coli OMVs.

优化大肠杆菌 OMV 的生产、纯化和表征。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Kasper,Anna;Henretta,Sarah;Dinu,Milena;Smith,Callum;Videva,Martina;Crawford,Jamie;Shanahan,Caitlin;Gaborski,Thomas;Michel,LeaV
• 通讯作者: Michel,LeaV

Investigating the Application of Size-Exclusion Chromatography to Purify Bacterial Extracellular Vesicles.

研究尺寸排阻色谱法在纯化细菌细胞外囊泡中的应用。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Gaborski,Thomas;Guernsey,Cara;Henretta,Sara;Hosseinifakhr,Atiyeh;Torabian,Pantea;Kasper,Anna;Michel,Lea
• 通讯作者: Michel,Lea

Outer membrane vesicles as molecular biomarkers for Gram-negative sepsis: Taking advantage of nature's perfect packages.

• DOI: 10.1016/j.jbc.2022.102483
• 发表时间: 2022-10
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Michel, Lea Vacca;Gaborski, Thomas
• 通讯作者: Gaborski, Thomas