基于单细胞解析和类器官模拟的人类成体子宫细胞图谱与内膜分化调控机制研究

The human uterus is a highly dynamic tissue that undergoes repeated damage repair and regeneration. However, it's still difficult to repair and reconstruct large scale, full thickness uterine tissues. Organoid technology is promising approach to solve these problems, but the key cell subset of uterine organoid and the regulatory mechanism remains unclear: Endometrial stem progenitor cells were shown to be involved in the regeneration of the damaged functional layer during the menstrual cycle and continuous growth upon embryo implantation. However, the precise identify, function and regulatory mechanisms of the uterus epithelial stem progenitors during the period of adult uterine cyclical repair and regeneration in vivo remains unclear. This project aims to build cell atlas of normal human uterine use single-cell sequencing analysis and reconstruct the interaction network of uterine stem progenitor cell subsets and their microenvironment. Furthermore, techniques of organoid culture is used to mimic and regulate the interaction network of uterine stem progenitor cells. The ultimate goal of this project is the utility of uterus organoid in the uterine repair and reconstruction. The study will systematically reveal the rules of uterine stem progenitor cells, the microenvironment and their interactions in uterine periodic regeneration, and build a new uterine repair and reconstruction technology based on human uterine organoids.

人类子宫组织具有周期性完美再生能力和高度可塑性，但大面积、全层性子宫组织修复重建仍面临巨大挑战。类器官技术有望解决该临床难题，而子宫类器官构建的关键人成体子宫干祖细胞亚群特征及与子宫再生微环境互作调控机制尚不明确。基于前期研究本项目拟通过单细胞测序分析技术解析正常人子宫组织细胞图谱，构建子宫干祖细胞亚群及周期性再生微环境互作网络；进一步通过类器官培养技术体外模拟、调控子宫干祖细胞与微环境互作；最终研究体外构建子宫类器官在子宫修复重建的效用。本研究的开展将系统揭示子宫组织干祖细胞微环境及其互作在子宫周期性再生中的规律，形成基于成体人子宫类器官的子宫修复重建新技术。

子宫组织细胞微环境成分、关系复杂，生理情况下可实现子宫组织的完美修复和再生，而大面积、全层性子宫修复重建困难的背景。且目前的研究局限主要包括：缺乏对子宫上皮干/祖细胞标志物及其体内分化调控机制的研究；缺乏对子宫周期性再生修复过程微环境细胞亚群、标志物、分子特征动态变化等的系统解析；缺乏对微环境细胞亚群间互作网络图谱解析；缺乏可模拟体内复杂微环境的的子宫类器官构建技术。本项目首先利用单细胞RNA-seq技术绘制了人体子宫组织在增生期和分泌期中细胞亚群图谱以及细胞间相互作用图谱。该图谱强调了人子宫月经周期中细胞亚群的时间动态变化和谱系结构，并且提供了月经周期中细胞亚群的通信和调控网络。其次我们利用分离的上皮干细胞构建子宫内膜上皮类器官库，并通过共培养验证了可以通过与SFRP4+基质细胞共培养来提高子宫内膜上皮类器官的构建效率；最后通过动物实验我们明确了关键的SFRP4+基质细胞亚群就能够有效促进子宫内膜损伤修复和重建。本研究提供了迄今为止最详细的子宫组织动态的细胞亚群图谱之一，可为子宫生物学提供新知识，为子宫的再生研究提供细胞亚群结构的参考。同时我们的研究从解析增生期子宫组织出发，鉴定了子宫内膜再生过程中的关键细胞亚群及其功能，可为子宫内膜损伤修复再生提供有力的指导。