Biobank of small extracellular vesicles for pediatric sepsis

小儿脓毒症小细胞外囊泡生物库

• 批准号: 10740537
• 负责人: Jennifer Melissa Kaplan
• 金额: $ 21.93万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-17 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10740537
• 关键词: Affect; Biochemical; Biogenesis; Biological; Biological Markers; Blood Cells; Blood specimen; Body Fluids; Categories; Cell Communication; Cells; Characteristics; Child; Childhood; Classification; Clinical; Clinical Trials; Collection; Complex; Critical Care; Critical Illness; Data; Data Set; Development; Diagnostics Research; Disease; Distant; Endosomes; Endothelial Cells; Freezing; Functional disorder; Funding; Future; Genomics; Goals; Harvest; Health; Heart; Hepatocyte; Heterogeneity; Human; Immune; Immune response; In Vitro; Infection; Inflammatory; Inflammatory Response; Investigation; Kidney; Lipids; Liquid substance; Liver; Lung; Membrane; Methodology; MicroRNAs; Mind; Molecular; Molecular Profiling; Morbidity - disease rate; Multiple Organ Failure; National Institute of General Medical Sciences; Nucleic Acids; Organ; Organ failure; Outcome; Patients; Pediatric cohort; Phase; Phased Innovation Awards; Phenotype; Phospholipids; Plasma; Population Heterogeneity; Precision therapeutics; Procedures; Process; Proteins; Proteomics; Protocols documentation; Quality Control; RNA; RNA analysis; Reproducibility; Research; Research Personnel; Sampling; Sepsis; Septic Shock; Serum; Source; Specimen; Standardization; Syndrome; Therapeutic Research; Time; Tissues; beta-Lactams; biobank; biomarker identification; biophysical properties; cell type; cohort; effective therapy; exosome; extracellular vesicles; high throughput analysis; improved; individual variation; lipidomics; liquid biopsy; metabolomics; mortality; nano-string; novel; organ injury; particle; patient subsets; pediatric patients; pediatric sepsis; personalized medicine; pharmacometrics; potential biomarker; prospective; repository; sample collection; septic; septic patients; tool; trait; transcriptome sequencing; transcriptomics; treatment strategy

PROJECT SUMMARY Sepsis is a dysregulated host response to an infection and can lead to multiple organ dysfunction syndrome (MODS). There are no effective treatments for sepsis-induced MODS most likely because of the heterogeneity of the syndrome. Delineating MODS endotypes and molecular signatures of organ failures may lead to a better understanding of the mechanisms involved in sepsis heterogeneity and allow for personalized treatment strategies. It is difficult, however, to obtain clinical specimens from critically ill patients that would enable investigations into organ-specific mechanistic changes. Extracellular vesicles (EVs) are spherical microparticles enclosed by bilayer phospholipid membranes. Exosomes or small EVs (sEVs) are a subtype of EV formed by endosomal biogenesis. Small EVs can be released from almost any cell type into a variety of bodily fluids and contain many cellular components. The cell-specific cargo can serve as cell-to-cell communicators and be taken up by distant cells which can affect the inflammatory profile. Our preliminary data show that sEVs harvested from serum of pediatric patients with sepsis have a distinct pro-inflammatory trait compared to sEVs of children without sepsis and in vitro sEVs from septic patients can induce atypical inflammatory responses in immune cells. Since circulating sEVs manifest characteristics of the cell of origin, they have been used as liquid biopsy. Thus, sEVs hold potential as useful biomarker for organ-specific changes. There are no available biorepositories of sEVs for pediatric sepsis research, in part, because of the lack of standardized methodology for sEV isolation. The overall goal of our proposal is to establish standardized procedures for reliable biorepositories for sEV biomarker research in critically ill patients with sepsis. This proposal will prove the hypothesis that quality and consistency of isolation and purification protocols of plasma and serum samples enable setting-up reliable biorepositories for future research on sEVs in sepsis. We will take advantage of two large critical care-division based repositories which has biospecimens from pediatric critically ill septic and non-septic studies. The R21 phase Aim 1 is to develop a methodology for sample collection and isolation of sEVs with high yield and purity and Aim 2 is to demonstrate suitability of banked sEVs for high throughput analyses of RNA cargo profile. Once milestones for the R21 phase are met we will proceed to the R33 phase to retrospectively characterize sEV endotypes in critically ill patients with specific organ injuries (Aim 3) and then prospectively determine whether patients can be classified based on their sEV characteristics. Results from these investigations will allow for the novel development of a biorepository of sEVs in pediatric sepsis. This biorepository will enable investigators to explore organ-specific molecular signatures for mechanistic studies of sEVs.

项目概要 脓毒症是宿主对感染的反应失调，可导致多器官功能障碍综合征 （MODS）。对于脓毒症引起的 MODS 没有有效的治疗方法，很可能是因为存在异质性 综合症的。描绘 MODS 内型和器官衰竭的分子特征可能会带来更好的结果 了解脓毒症异质性的机制并允许个性化治疗 策略。然而，从危重病人身上获取临床标本是很困难的。 研究器官特异性机制变化。细胞外囊泡 (EV) 是球形微粒 被双层磷脂膜包围。外泌体或小型 EV（sEV）是 EV 的一个亚型，由 内体生物发生。小型 EV 可以从几乎任何细胞类型释放到各种体液中 含有许多细胞成分。细胞特异性货物可以作为细胞间的通讯器并被采取 远处的细胞可以影响炎症状况。我们的初步数据显示，sEV 是从 与没有脓毒症的儿童的 sEV 相比，脓毒症儿童患者的血清具有明显的促炎特征。 脓毒症和脓毒症患者的体外 sEV 可以诱导免疫细胞非典型炎症反应。自从 循环的 sEV 表现出起源细胞的特征，它们已被用作液体活检。因此，sEV 具有作为器官特异性变化的有用生物标志物的潜力。没有可用的 sEV 生物储存库 儿科脓毒症研究的部分原因是缺乏 sEV 分离的标准化方法。整体 我们提案的目标是为 sEV 生物标记物建立可靠的生物样本库的标准化程序 脓毒症危重患者的研究。该提案将证明质量和一致性的假设 血浆和血清样品的分离和纯化方案使得能够建立可靠的生物样本库 败血症中 sEV 的未来研究。我们将利用两个基于重症监护部门的大型存储库 其中有来自儿科重症脓毒症和非脓毒症研究的生物样本。 R21 阶段目标 1 是 开发一种高产量和高纯度的 sEV 样品采集和分离方法，目标 2 是 证明存储的 sEV 适用于 RNA 货物概况的高通量分析。一旦达到里程碑 满足 R21 阶段后，我们将进入 R33 阶段，以回顾性地表征 sEV 内型 患有特定器官损伤的危重患者（目标 3），然后前瞻性地确定患者是否可以 根据其 sEV 特性进行分类。这些调查的结果将允许小说 开发儿童脓毒症 sEV 生物储存库。该生物样本库将使研究人员能够探索 用于 sEV 机制研究的器官特异性分子特征。