Donor specific extracellular vesicle characterization for monitoring lung transplant rejection

标题 供体组织特异性细胞外囊泡特性在肺移植排斥反应监测中的应用 移植 项目摘要（摘要） 肺移植仍然是治疗终末期呼吸衰竭患者的唯一方法。 然而，肺移植患者的发病率和死亡率仍然是所有实体器官中最高的 移植，并直接与移植肺排斥反应和免疫抑制并发症。 然而到目前为止，还没有可靠的生物标志物平台来监测移植肺的状态。一个准确的，非- 侵入性、时间敏感的生物标记物平台将对患者存活具有显著的转化影响， 生活质量，因为它将使早期干预和免疫抑制滴定。 细胞外囊泡（EV），包括外泌体，由许多组织释放到外周血中， 流通EV代表了稳定的和组织特异性的蛋白质组和RNA特征谱，反映了 它们的起源组织的条件状态。组织特异性EV谱和它们的RNA特征在组织中是不同的。 健康状况与损伤/病理学。因此，了解来自身体的组织特异性EV谱， 液体有希望作为其来源组织状态的“液体活组织检查”。但是没有一个小组研究过 移植中的组织特异性EV谱。根据我们实验室过去三年所做的工作，我们 发现移植的组织/器官释放一个稳定的和可检测的供体特异性EV池到组织中。 接受者血液/体液。此外，移植组织的免疫排斥导致免疫系统的明显变化。 供体EV量及其蛋白质组学和microRNA谱。这些发现现已得到验证 在临床环境中。在这个指导研究项目中，我们建议调查供体肺特异性的作用， 来自受体血浆的EV蛋白质组学和RNA特征作为肺领域的新型生物标志物平台 移植诊断学 由于迄今为止还没有研究表征移植肺特异性和受体免疫细胞EV池， 我们建议在两种体内移植模型中研究这种新的概念：（1）建立的小鼠模型， 在急性排斥与接受的受控设置下的原位肺移植，和（2）人 肺移植在这两种情况下，我们将表征移植肺特异性和受体T细胞特异性 EV池，它们在健康条件下的蛋白质组和RNA特征与排斥反应，以发现不同的 与移植排斥相关的特征成功完成这一项目提案将奠定 为未来开发一种非侵入性的、基于EV的新型生物标志物检测方法以监测肺部疾病奠定了基础 移植排斥反应

Title Donor tissue specific extracellular vesicle characterization for monitoring rejection in lung transplantation Project Summary (Abstract) Lung transplantation remains the only curative therapy in patients with end stage respiratory failure. However, morbidity and mortality in lung transplant patients remains the highest amongst all solid organ transplants, and is directly associated with transplant lung rejection and complications of immunosuppression. Yet to this date, there is no reliable biomarker platform to monitor the transplant lung status. An accurate, non- invasive, time-sensitive biomarker platform would have significant translational impact on patient survival and quality of life as it would enable early intervention and immunosuppression titration. Extracellular vesicles (EVs), including exosomes, are released by many tissues into peripheral circulation. EVs represent stable and tissue-specific proteomic and RNA signature profiles that reflect the conditional state of their tissue of origin. Tissue specific EV profiles and their RNA signatures are distinct in conditions of health versus injury/ pathology. Therefore, understanding tissue specific EV profiles from bodily fluids has promise to serve as a “liquid biopsy” of the status of their tissue of origin. But no group has studied tissue specific EV profiles in transplantation. From work done over the past three years in our laboratory, we found that transplanted tissues/ organs release a stable and detectable pool of donor-specific EVs into the recipient blood/ bodily fluids. Further, immune rejection of the transplanted tissue leads to distinct changes in the donor EV quantity and in their proteomic and microRNA profiles. These findings have now been validated in the clinical setting. In this mentored research project we propose to investigate the role of donor lung specific EV proteomic and RNA signatures from recipient plasma as a novel biomarker platform in the field of lung transplantation diagnostics. Since no study to date has characterized transplant lung specific and recipient immune cell EV pools, we propose to investigate this novel concept in two in vivo transplant models: (1) established mouse model of orthotopic lung transplantation under controlled settings of acute rejection versus acceptance, and; (2) human lung transplantation. In both settings we will characterize transplant lung specific and recipient T cell specific EV pools, their proteomic and RNA signatures in conditions of health versus rejection to discover distinct profiles associated with transplant rejection. Successful completion of this project proposal would lay the foundation for future development of a non-invasive, EV-based novel biomarker assay to monitor lung transplant rejection.