通过单细胞 RNA-Seq 解析体细胞重编程过程中的细胞命运决定。

Resolving Cell Fate Decisions during Somatic Cell Reprogramming by Single-Cell RNA-Seq.

机构信息

CAS Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou 510530, China; Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou Medical University, Guangzhou 511436, China; Guangzhou Regenerative Medicine and Health GuangDong Laboratory (GRMH-GDL), Guangzhou 510005, China.

出版信息

Mol Cell. 2019 Feb 21;73(4):815-829.e7. doi: 10.1016/j.molcel.2019.01.042. Epub 2019 Feb 13.

DOI:10.1016/j.molcel.2019.01.042

PMID:30772174

Abstract

Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs), which is a highly heterogeneous process. Here we report the cell fate continuum during somatic cell reprogramming at single-cell resolution. We first develop SOT to analyze cell fate continuum from Oct4/Sox2/Klf4- or OSK-mediated reprogramming and show that cells bifurcate into two categories, reprogramming potential (RP) or non-reprogramming (NR). We further show that Klf4 contributes to Cd34+/Fxyd5+/Psca+ keratinocyte-like NR fate and that IFN-γ impedes the final transition to chimera-competent pluripotency along the RP cells. We analyze more than 150,000 single cells from both OSK and chemical reprograming and identify additional NR/RP bifurcation points. Our work reveals a generic bifurcation model for cell fate decisions during somatic cell reprogramming that may be applicable to other systems and inspire further improvements for reprogramming.

摘要

体细胞可被重编程为诱导多能干细胞（iPSCs），这是一个高度异质的过程。在这里，我们以单细胞分辨率报告体细胞重编程过程中的细胞命运连续体。我们首先开发了 SOT 来分析由 Oct4/Sox2/Klf4 或 OSK 介导的重编程过程中的细胞命运连续体，并表明细胞分为两类，即重编程潜能（RP）或非重编程（NR）。我们进一步表明，Klf4 有助于 Cd34+/Fxyd5+/Psca+角质细胞样 NR 命运，而 IFN-γ 会阻碍 RP 细胞向嵌合体相容的多能性的最终转变。我们分析了来自 OSK 和化学重编程的超过 150,000 个单细胞，并鉴定了其他 NR/RP 分岔点。我们的工作揭示了体细胞重编程过程中细胞命运决策的通用分岔模型，该模型可能适用于其他系统，并为重编程提供进一步的改进。