Defining cellular mechanisms of chronic graft failure in transplanted hearts with single cell multi-omics

项目摘要/摘要 心脏移植是终末期心力衰竭患者的最终治疗选择。虽然短期内 存活率有了很大的改善，长期结果受到慢性移植物衰竭的限制。心脏 同种异体移植物肥厚(CAH)是一种临床现象，指的是左心室质量逐渐增加。 和心脏移植物的壁厚。CAH最早可在移植后2个月发病， 常导致细胞环境进行性改变，包括心肌细胞肥大和纤维化 并长期积累导致移植物失败。移植患者细胞系统的完整图景 代表着知识上的重大鸿沟。单细胞rna测序(scrna-seq)已经成为一种强有力的工具。 分析复杂结构中不同的细胞类型和细胞状态以及单个细胞的伪时间拓扑 然而，器官缺乏推断细胞谱系的能力，而细胞谱系对于确定疾病的起源是重要的 致病细胞。在这个项目中，我们将开发一种新的单细胞多组学测序技术，并 几种计算工具，可以同时检测细胞谱系标记(即DNA)和 来自相同细胞的细胞命运标记(即RNA)。我们还将区分所有细胞的供体和宿主身份 研究它们在CAH中的不同作用，这些作用在很大程度上是未知的。我们假设CAH是由特定的 免疫细胞亚型与内皮细胞向间充质细胞转化的驱动 心肌细胞。在目标1中，我们将开发新的技术来定义统一的细胞谱系和细胞命运 心脏细胞的路线图。目标2将剖析移植心脏细胞成分的时间变化 在CAH早期发病期间。目标3是用时间序列单细胞追踪CAH进展过程中的细胞谱系 多元组学。该项目的成功完成将导致供体和宿主来源的细胞的鉴定 作为新的治疗靶点来延长移植患者的生命。

Project Summary/Abstract Heart transplantation is a definitive treatment option for patients with end-stage heart failure. While short-term survival has made substantial improvements, long-term outcomes are limited by chronic graft failure. Cardiac allograft hypertrophy (CAH) is a clinical phenomenon referring to the gradual increase of left ventricular mass and wall thickness of cardiac grafts. CAH onset can occur as early as 2 months after transplantation, which often lead to progressive changes in cellular environment including cardiomyocytes hypertrophy and fibrosis and accumulate to graft failure in long term. The complete landscape of cellular system in transplant patients represents a major gap in knowledge. Single cell RNA sequencing (scRNA-seq) has emerged as powerful tool to analyze diverse cell types and cell states as well as pseudo-time topologies of single cells in complex organs, which however lacks the power to infer cell lineages that is important for identify the origins of pathogenic cells. In this project, we will develop a novel single cell multi-omics sequencing technology and several computational tools, which will enable simultaneous detection of cell lineage markers (i.e. DNAs) and cell fate markers (i.e. RNAs) from same cells. We will also distinguish donor and host identities for all cells to study their distinct roles in CAH that are largely unknown. We hypothesized that CAH is initiated by specific immune cell subtypes and driven by endothelial to mesenchymal transition that is fueled by enlarged cardiomyocytes. In aim 1, we will develop novel technology to define a unifying cell lineage and cell fate roadmap for cardiac cells. Aim 2 will dissect temporal changes of cellular components in transplanted hearts during CAH early onset. Aim 3 is to trace cell lineages during CAH progression with time-series single cell multi-omics. Successful completion of this project will lead to the identification of donor and host originated cell populations as novel therapeutic targets to extend the life of transplant patients.