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.

项目总结

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