Wang Jie, Adrianto Indra, Subedi Kalpana, Liu Tingting, Wu Xiaojun, Yi Qijun, Loveless Ian, Yin Congcong, Datta Indrani, Sant'Angelo Derek B, Kronenberg Mitchell, Zhou Li, Mi Qing-Sheng
Center for Cutaneous Biology and Immunology Research, Department of Dermatology, Henry Ford Health, Detroit, MI, USA.
Immunology Research Program, Henry Ford Cancer Institute, Henry Ford Health, Detroit, MI, USA.
Cell Discov. 2023 Jun 20;9(1):61. doi: 10.1038/s41421-023-00547-x.
Unlike conventional αβT cells, invariant natural killer T (iNKT) cells complete their terminal differentiation to functional iNKT1/2/17 cells in the thymus. However, underlying molecular programs that guide iNKT subset differentiation remain unclear. Here, we profiled the transcriptomes of over 17,000 iNKT cells and the chromatin accessibility states of over 39,000 iNKT cells across four thymic iNKT developmental stages using single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) to define their developmental trajectories. Our study discovered novel features for iNKT precursors and different iNKT subsets and indicated that iNKT2 and iNKT17 lineage commitment may occur as early as stage 0 (ST0) by two distinct programs, while iNKT1 commitments may occur post ST0. Both iNKT1 and iNKT2 cells exhibit extensive phenotypic and functional heterogeneity, while iNKT17 cells are relatively homogenous. Furthermore, we identified that a novel transcription factor, Cbfβ, was highly expressed in iNKT progenitor commitment checkpoint, which showed a similar expression trajectory with other known transcription factors for iNKT cells development, Zbtb16 and Egr2, and could direct iNKT cells fate and drive their effector phenotype differentiation. Conditional deletion of Cbfβ blocked early iNKT cell development and led to severe impairment of iNKT1/2/17 cell differentiation. Overall, our findings uncovered distinct iNKT developmental programs as well as their cellular heterogeneity, and identified a novel transcription factor Cbfβ as a key regulator for early iNKT cell commitment.
与传统的αβT细胞不同，不变自然杀伤T（iNKT）细胞在胸腺中完成向功能性iNKT1/2/17细胞的终末分化。然而，指导iNKT亚群分化的潜在分子程序仍不清楚。在这里，我们使用单细胞RNA测序（scRNA-seq）和转座酶可及染色质测序的单细胞分析（scATAC-seq），对超过17,000个iNKT细胞的转录组和超过39,000个iNKT细胞在四个胸腺iNKT发育阶段的染色质可及性状态进行了分析，以确定它们的发育轨迹。我们的研究发现了iNKT前体和不同iNKT亚群的新特征，并表明iNKT2和iNKT17谱系的定向分化可能早在0期（ST0）通过两个不同的程序发生，而iNKT1的定向分化可能在ST0之后发生。iNKT1和iNKT2细胞均表现出广泛的表型和功能异质性，而iNKT17细胞相对同质。此外，我们发现一种新的转录因子Cbfβ在iNKT祖细胞定向分化检查点高度表达，其表达轨迹与其他已知的iNKT细胞发育转录因子Zbtb16和Egr2相似，并可指导iNKT细胞命运并驱动其效应表型分化。条件性敲除Cbfβ会阻断早期iNKT细胞发育，并导致iNKT1/2/17细胞分化严重受损。总体而言，我们的研究结果揭示了不同的iNKT发育程序及其细胞异质性，并确定了一种新的转录因子Cbfβ作为早期iNKT细胞定向分化的关键调节因子。
