A comprehensive library of human transcription factors for cell fate engineering (scRNA-Seq)

Human pluripotent stem cell (hPSC)-derived cells offer an unprecedented opportunity to model diverse human tissue functions. However, the field lacks a generalized approach to rapidly and efficiently generate individual cell types of interest and to incorporate them into complex but defined tissues. To systematically explore the transcription factor (TF)-mediated hPSC programming landscape, we constructed the Human TFome: a comprehensive human TF library containing 1,564 TF (1,732 splice-isoforms). We discovered 241 previously unreported factors that individually converted hPSCs into diverse lineages with up to 99% efficiency in four days without alteration of external factors. Among these, we programmed neurons, fibroblasts, oligodendrocytes and endothelial cells that have molecular, transcriptomic and functional similarity to primary cells. Splice-isoform-specificity doubled endothelial conversion efficiency into in vivo-functional cells. Our cell-autonomous approach enabled parallel and orthogonal hPSCs programming into multiple cell types simultaneously without altering any microenvironmental cues. We generated in vivo-engraftable induced oligodendrocytes that expedited myelination within synthetically accelerated cerebral organoids.

人类多能干细胞（human pluripotent stem cell, hPSC）衍生细胞为建模多样的人类组织功能提供了前所未有的机遇。然而，该领域目前缺乏一种通用方法，能够快速高效地生成目标特异性细胞类型，并将其整合至结构复杂但定义明确的组织中。为系统性探究转录因子（transcription factor, TF）介导的人类多能干细胞重编程全景图谱，本研究构建了人类转录因子组库（Human TFome）：一套涵盖1564种转录因子、包含1732种剪接异构体（splice-isoform）的综合性转录因子文库。本研究发现了241种此前未被报道的转录因子，它们可单独将人类多能干细胞定向转化为多种细胞谱系，四天内转化效率最高可达99%，且无需改变外部培养条件。其中，我们成功诱导生成了神经元、成纤维细胞、少突胶质细胞（oligodendrocytes）与内皮细胞（endothelial cells），这些细胞在分子特征、转录组谱与功能特性上均与原代细胞高度相似。剪接异构体特异性可使内皮细胞向具有体内功能的细胞的转化效率提升一倍。本研究采用的细胞自主性策略，可在不改变任何微环境信号的前提下，同时将人类多能干细胞并行且正交地定向重编程为多种不同细胞类型。我们还生成了可体内移植的诱导型少突胶质细胞，其可在人工加速发育的大脑类器官中促进髓鞘形成进程。

搜集汇总

背景与挑战

背景概述

该数据集基于单细胞RNA测序（scRNA-Seq），提供了一个全面的人类转录因子库（Human TFome），包含1,564个转录因子（1,732个剪接异构体），用于高效编程人类多能干细胞（hPSCs）生成多种细胞谱系（如神经元、成纤维细胞等），在4天内转换效率高达99%，且无需外部因素干预。其特点包括剪接异构体特异性增强细胞功能、细胞自主编程支持并行生成多种细胞类型，以及应用于加速脑类器官髓鞘形成等组织工程。