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The µSiM-hNVU - a human BBB platform for the study of brain injury mechanisms during systemic infection

µSiM-hNVU - 用于研究全身感染期间脑损伤机制的人类 BBB 平台

基本信息

批准号：
10518863
负责人：
Britta Engelhardt
金额：
$ 59.95万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-03 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10518863
关键词：
Address Affect Alleles Animal Model Animals Apolipoprotein E Astrocytes Biological Assay Biological Models Blood Blood - brain barrier anatomy Brain Brain Injuries Cell Culture System Cell Line Cells Complex Critical Care Critical Illness Development Devices Diagnostic Diffuse Diffusion Disease Encephalitis Encephalopathies Endothelium Engineering Environment Enzyme-Linked Immunosorbent Assay Exhibits Exposure to Extravasation Fluorescence Foundations Functional disorder Gene Expression Profiling Genetic Transcription Glass Glycocalyx Goals Hematological Disease Hematology Hemoglobin Heparin Lyase Human Impaired cognition In Vitro Infiltration Inflammation Inflammatory Intervention Invaded Leukocytes Lipoproteins Matrix Metalloproteinases Measurement Membrane Microfabrication Microglia Microscopic Microscopy Modeling Molecular Monitor Myeloid Cells Nuclear Optics Outcome Pathway interactions Pattern Pericytes Permeability Pharmacology Phase Physiological Play Population Property Proteins Reporter Reporting Resolution Risk Risk Factors Role S100A8 gene Sepsis Side Signal Transduction Silicon Survivors System Systemic infection Testing Thick Tissue Microarray Validation Work antibody inhibitor apolipoprotein E-3 base blood-brain barrier function blood-brain barrier permeabilization brain endothelial cell cerebrovascular design digital experience in vivo induced pluripotent stem cell innovation live cell imaging microphysiology system monocyte multidisciplinary mutant nanomembrane neuroinflammation neurovascular unit neutrophil pre-clinical preclinical study prevent protein expression response sensor technology septic patients solute stem cells systemic inflammatory response therapy development tool trafficking

项目摘要

Abstract Long-term cognitive impairment affects more than 70% of sepsis survivors, but the underlying mechanisms remain unknown. Though widely hypothesized, evidence of blood-brain barrier (BBB) dysfunction in septic patients is limited by practical barriers to diagnostic studies in critically ill subjects. While BBB breakdown and cognitive impairment are seen in animal models of sepsis, the complexity of sepsis in vivo and differences between animal and human responses means that animal models cannot unambiguously identify the circulating factors that cause brain injury in human sepsis. Therefore, we propose to develop the µSiM-hNVU as an `on-chip' platform featuring a human iPSC-derived neurovascular unit (NVU; brain microvascular endothelial cells, pericytes and astrocytes). The `blood side' will allow the flow-based introduction of blood- borne cells and molecules with known or hypothesized roles in sepsis related brain injury, and the `brain side' will feature iPSC-derived microglial cells serving as a reporter of the brain inflammatory status. The human NVU will be built on a device platform – the µSiM – featuring ultrathin silicon nanomembranes that provide for unhindered solute exchange between `blood' and `brain' compartments and glass-like optical quality for live cell imaging and high-resolution microscopy. In the R61 phase, the device platform will be advanced for ease-of- use including `plug-and-play' modules for flow and barrier measurements (TEER, diffusion), and compatibility with a small-volume, digital-ELISA assay for secreted proteins. The µSiM-hNVU will be validated with functional assays of blood-brain barrier (BBB) function, protein expression studies, and transcriptional analysis. We will also build a iPSC NVU in which each cellular component of the NVU carries the ApoE4 allele. The expression of the ApoE4 lipoprotein drives BBB dysfunction by a known pathway and increases the risk of cognitive impairment in humans and animals experiencing brain inflammation. We will use the ApoE4-NVU as a `diseased BBB on a chip” which we hypothesize will show enhanced vulnerabilities to candidate mechanisms of brain injury identified by our team and others. Specifically, we will test the hypotheses that 1) pre-activated monocytes invade the brain and drive microglial activation; 2) the damage associated molecular pattern (DAMP) complex S100A8/A9 drive BBB breakdown to promote leukocyte infiltration and neuroinflammation; and 3) circulating factors that degrade endothelial glycocaylx (e.g., heparinase) or contribute to systemic inflammation (cell-free hemoglobin) promote CNS infiltration of leukocytes and subsequent neuroinflammation.

摘要