Establishment of a multi-center biobank of patient-specific induced pluripotent stem cells for pediatric sepsis research

用于小儿脓毒症研究的患者特异性多能干细胞库的建立。 项目总结： 脓毒症是严重感染患者的最终常见途径，导致儿童死亡的人数多于癌症患者。 每年在美国。然而，除了早期抗生素和抗生素外，还没有治疗儿童败血症的方法。 器官支持。患者、器官和细胞水平的多层次异质性具有显著的 阻碍了科学进步。尽管精准医学方法已经被用来筛选患者- 水平差异，我们从根本上缺乏对异质细胞如何 这些反应参与了脓毒症的发病机制。因此，人类败血症生物储存库迫切需要 解开细胞特异性的机制通路，加速新的脓毒症治疗药物的开发。 我们试图通过开发一种史无前例的患者特异性诱导多能性的生物库来弥合这一差距 来自败血症儿童的干细胞(IPSCs)。人的IPSCs可以用作可再生的和 可扩展的任何细胞类型的来源，并已用于许多疾病状态的精确建模。 然而，目前尚不清楚它们是否可以作为一种急性情况下的脓毒症的体外模型。 具有复杂的基因-环境相互作用，主要由宿主反应驱动。通过这个分阶段的 创新奖，我们寻求建立患者特有的IPSC来源细胞捕获疾病的能力 脓毒症患者循环初级细胞亚群的表型和体内反应概述。 在R21阶段，里程碑驱动的开发活动将侧重于建立高效的 用于IPSC的同时采集和储存外周血单个核细胞的样品富集法 从全血中产生、循环初级白细胞和内皮细胞，以及血清。到时候我们会的 建立质量受控的IPSC系，将其分化为单核细胞和内皮细胞，并执行 患者来源细胞与健康人来源细胞的功能和表型特征 捐赠者。在R33阶段，我们将使用最先进的多组测序来确定患者特定的 风险分层脓毒症血清镜像处理IPSC来源的细胞 各自的循环原代细胞。扩大活动范围将包括跨多个中心招募患者， IPSC产生和分发的标准化协议，以及开发生物信息数据库， 将使我们能够用丰富的临床数据来整理和关联组学分析。通过成功的 执行这项建议，我们寻求改造我们现有的儿科脓毒症生物库，以满足 下一代科学研究，促进人类脓毒症的细胞特异性机制研究。

Establishment of a multi-center biobank of patient-specific pluripotent stem cells for pediatric sepsis research. PROJECT SUMMARY: Sepsis is the final common pathway among those with serious infections and kills more children than cancer in the U.S each year. Yet, there are no therapies available for pediatric sepsis beyond early antibiotics and organ support. Multiple levels of heterogeneity at the patient, organ, and cellular levels have significantly impeded scientific progress. Although precision medicine approaches have been used to sift through patient- level differences, we fundamentally lack a comprehensive understanding of how heterogeneous cellular responses contribute to sepsis pathogenesis. Thus, there is a crucial need for human sepsis biorepositories to unravel cell-specific mechanistic pathways and accelerate the development of novel sepsis therapeutics. We seek to bridge this gap by developing an unprecedented biobank of patient-specific induced pluripotent stem cells (iPSCs) derived from children with sepsis. Human iPSCs can be used as a renewable and expandable source of any cell type and have been used for precision modeling of numerous disease states. However, it remains unknown whether they can serve as robust in vitro models of sepsis – an acute condition with complex gene-environment interactions and driven primarily by the host response. Through this phased innovation award, we seek to establish the ability of patient-specific iPSC derived cells to capture disease phenotypes and recapitulate in vivo responses of circulating primary cell subsets in human sepsis. Milestone-driven developmental activities in the R21 phase will focus on establishment of an efficient sample enrichment process to simultaneously collect and bank peripheral blood mononuclear cells for iPSC generation, circulating primary leukocytes and endothelial cells, and serum from whole blood. We will then generate quality-controlled iPSC lines, differentiate them into monocytes and endothelial cells, and perform functional and phenotypic characterization of patient derived cells relative to those derived from healthy donors. In the R33 phase, we will use state-of-the-art multiomic sequencing to determine if patient-specific iPSC derived cells treated with risk-stratified septic serum mirror transcriptomic and epigenomic responses of respective circulating primary cells. Scale up activities will include patient recruitment across multiple centers, standardized protocols for iPSC generation and distribution, and development of a bioinformatic database that will allow us to collate and contextualize omic analyses with rich clinical data. Through the successful execution of this proposal, we seek to transform our existing pediatric sepsis biobank to meet the needs of the next generation of scientific inquiry and facilitate cell-specific mechanistic research in human sepsis.