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.

项目摘要 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，最可能是因为其异质性。 的综合征。描绘器官衰竭的MODS内在型和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 战略布局然而，很难从危重患者中获得临床标本， 对器官特异性机制变化的研究。细胞外囊泡是一种球形微粒 被双层磷脂膜包围。外泌体或小EV（sEV）是EV的一种亚型，其通过以下方式形成： 内体生物发生小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通讯器， 由远处的细胞引起，这会影响炎症的特征。我们的初步数据显示，从 脓毒症儿童患者的血清与无脓毒症儿童的sEV相比具有明显的促炎特征， 脓毒症和来自脓毒症患者的体外sEV可以诱导免疫细胞中的非典型炎症反应。以来 循环sEV表现出起源细胞的特征，它们已被用作液体活检。因此，sEV 保持作为器官特异性变化有用生物标志物的潜力。没有可用的sEV生物储存库， 儿科败血症研究，部分原因是缺乏标准化的sEV分离方法。整体 我们建议的目标是建立标准化程序，为sEV生物标志物建立可靠的生物储存库 重症脓毒症患者的研究。这一建议将证明假设，质量和一致性 血浆和血清样品的分离和纯化方案，能够建立可靠的生物储存库， 脓毒症中sEV的未来研究。我们将利用两个基于重症监护室的大型存储库 它有儿科重症感染性和非感染性研究的生物样本。R21阶段目标1是 开发用于以高产率和纯度收集和分离sEV的方法，目标2是 证明了库存sEV对于RNA货物谱的高通量分析的适用性。一次里程碑， 我们将进入R33阶段，以回顾性地表征sEV内型， 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特征进行分类。这些调查的结果将使小说 在儿科败血症中开发sEV的生物储存库。这个生物储存库将使研究人员能够探索 器官特异性分子标记用于sEV的机制研究。