Project 3: Mapping the Evolution of Chronic Transplant Injury in the Context of CMV Infection

项目3：绘制CMV感染背景下慢性移植损伤的演变 总结/摘要 CMV感染在器官移植的背景下驱动炎症，并且与慢性炎症相关。 同种异体移植物中的排斥反应，这会加速同种异体移植物丢失和移植失败。的级联 在CMV感染的情况下触发移植物炎症的分子机制知之甚少。 器官背景下CMV感染和疾病的全基因组、蛋白质组和免疫组分析 移植可以产生一个全面的全球数据库，以了解CMV感染和再激活 在免疫功能低下的宿主中，并帮助设计新的策略，以防止病毒再激活和抑制抗- 捐献者免疫。在项目3中，我们建议研究免疫反应的演变， 肾移植受者的排斥反应、慢性排斥反应和巨细胞病毒感染 移植物转录干扰（Aim 1）、外周T细胞和B细胞受体（TCR和 BCR）通过下一个基因表达测序（NGS），以及体液反应的变化，以重新激活 通过高通量抗体谱分析（Aim 2）确定抗原表位。此外，我们建议生成 通过测量供体来源的细胞游离的变化用于非侵入性监测慢性移植物损伤的生物标志物 通过大规模多重PCR（mmPCR）和高度精细化基因集（kSORT和uCRM）检测DNA（dd-cfDNA） 从以前的高通量微阵列研究中挖掘出来（Aim 3）。我们建议使用一组肾脏 来自美国两个最大的多器官移植项目（UCSF和UCLA）的移植患者。

PROJECT 3: Mapping the Evolution of Chronic Transplant Injury in the Context of CMV Infection SUMMARY/ABSTRACT CMV infection drives inflammation in the context of organ transplantation, and is associated with chronic rejection in the allograft, which drives accelerated allograft loss and transplant failure. The cascade of molecular mechanisms that trigger graft inflammation in the context of CMV infection are poorly understood. Whole genome, proteome, and immunome profiling of CMV infection and disease in the context of organ transplantation can generate a comprehensive global database to understand CMV infection and reactivation in an immunocompromised host and help design novel strategies to prevent viral reactivation and dampen anti- donor immunity. In Project 3, we propose to study the evolution of immune responses that drive acute rejection, chronic rejection, and CMV virus infection in kidney transplant recipients by serial assessments of graft transcriptional perturbations (Aim 1), clonal expansion of peripheral T cell and B cell receptors (TCRs and BCRs) through next gene expression sequencing (NGS), and variations in humoral responses to reactivation of antigenic epitopes by high-throughput antibody profiling (Aim 2). In addition, we propose to generate biomarkers for non-invasive monitoring of chronic graft injury by measuring changes in donor derived cell free DNA (dd-cfDNA) by massively multiplexed PCR (mmPCR) and highly refined gene-sets (kSORT and uCRM) mined from previous high-throughput microarray studies (Aim 3). We propose to use a cohort of kidney transplant patients from two of the largest multi-organ transplant programs in the US (UCSF and UCLA).