Under-oil open microfluidic system (UOMS) for studying systemic fungal infection

用于研究全身真菌感染的油下开放式微流体系统 (UOMS)

基本信息

批准号：
10552700
负责人：
David J Beebe
金额：
$ 77.08万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10552700
关键词：
Address Antibiotics Bacterial Infections Biological Assay Blood Circulation Candida Candida albicans Candidiasis Catheters Cell Count Cells Clinical Collecting Cell Critical Illness Devices Disease Drug Targeting Drug resistance Effectiveness Environment Foundations Generations Genetic Genetic Determinism Growth Health Care Costs Healthcare Heterogeneity Human body Immunocompromised Host In Vitro Incidence Individual Infection Intervention Libraries Life Liquid substance Measurement Measures Methods Microbial Biofilms Microfluidic Microchips Microfluidics Morbidity - disease rate Mycoses Neoplasm Metastasis Oils Patients Pharmaceutical Preparations Phenotype Physiological Prevention Process Property Sepsis Site Solid Surface System Technology Therapeutic Virulence Virulence Factors Wettability Work Yeasts attributable mortality candida biofilm candidemia fungus in vitro Assay in vivo innovation instrumentation mortality mutant novel operation pathogenic fungus screening tool

项目摘要

Project Summary/Abstract Opportunistic fungal pathogens are a leading cause of healthcare associated bloodstream infections. Candida yeasts, specifically, cause 80-90% of biofilm-associated invasive fungal infections and mortality rates can approach 50%. Furthermore, the incidence of Candida infections is rising with the increased use of catheter and other device-based interventions. To date, the majority of work related to fungal infections and potential treatments has focused on biofilms and their prevention. However, recent evidence suggests that the dispersion of yeast cells from the biofilm (into the bloodstream) and the persistence of these dispersed cells are perhaps more important virulence factors and represent significant but underutilized treatment targets. Further, existing assays and instrumentation are not amenable to measuring dispersion and phenotyping dispersed cells. Nor do they recapitulate in vivo conditions (e.g. flow, interaction with host cells). Thus, here we will develop a new type of under-oil open microfluidic system (UOMS) to quantify the dispersive capacity of biofilms and assess the phenotype of dispersed cells in Candida mutants and clinical isolates. The UOMS platform is built on the foundation of a newly observed phenomena called Exclusive Liquid Repellency (ELR). ELR provides a unique environment where liquid is completely repelled from a solid surface to eliminate biofouling. Additionally, ELR expands the capabilities of simple open microfluidic devices allowing us to overcome the limitations of current methods and provide a system capable of quantitatively studying fungal dispersion. We will first (Aim 1) develop an under-oil microchannel device to measure the dispersive capacity of Candida biofilms and virulence phenotypes. Second (Aim 2) we will automate the UOMS and develop a single- cell distribution assay to measure the phenotype of individual dispersed cells. And finally (Aim 3), we will use the UOMS platform to develop dispersion phenotype profiles for Candida mutant libraries and clinical isolates. We will measure the dispersive capacity and phenotype of dispersed cells from hundreds of Candida albicans clinical isolates and mutants available in existing libraries, providing clues to the genetic determinants of dispersion.

项目概要/摘要机会性真菌病原体是医疗保健相关血液感染的主要原因。念珠菌属具体来说，酵母菌会导致 80-90% 的生物膜相关侵袭性真菌感染，并且死亡率可接近50%。此外，随着导管和导管使用的增加，念珠菌感染的发生率也在上升。其他基于设备的干预措施。迄今为止，大部分工作与真菌感染和潜在的治疗的重点是生物膜及其预防。然而，最近的证据表明，分散酵母细胞从生物膜（进入血液）的数量以及这些分散细胞的持久性也许是更重要的毒力因素，代表着重要但未充分利用的治疗目标。此外，现有的测定和仪器不适合测量分散性和表型分析分散的细胞。它们也没有概括体内条件（例如流动、与宿主细胞的相互作用）。因此，在这里我们将开发一种新型油下开放式微流体系统（UOMS）来量化生物膜并评估念珠菌突变体和临床分离株中分散细胞的表型。伦敦大学管理学院平台建立在一种新观察到的现象的基础上，称为排斥液体（ELR）。 ELR 提供了一个独特的环境，液体与固体表面完全排斥，以消除生物污垢。此外，ELR 扩展了简单开放式微流体设备的功能，使我们能够克服现有方法的局限性，提供一种能够定量研究真菌的系统分散。我们将首先（目标 1）开发油下微通道装置来测量油下微通道的分散能力念珠菌生物膜和毒力表型。第二（目标 2）我们将使 UOMS 自动化并开发一个单一的细胞分布测定来测量单个分散细胞的表型。最后（目标 3），我们将使用 UOMS 平台用于开发念珠菌突变体文库和临床分离株的分散表型谱。我们将测量数百个临床白色念珠菌的分散细胞的分散能力和表型现有文库中提供的分离株和突变体，为分散的遗传决定因素提供了线索。