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Exploring the stage-specific roles of Tcf-1 in T cell development and malignancy at single-cell resolution

Cellular & Molecular Immunology volume 18, pages 644–659 (2021)Cite this article

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Abstract

Tcf-1 (encoded by Tcf7) not only plays critical roles in promoting T cell development and differentiation but also has been identified as a tumor suppressor involved in preventing T cell malignancy. However, the comprehensive mechanisms of Tcf-1 involved in T cell transformation remain poorly understood. In this study, Tcf7fl/fl mice were crossed with Vav-cre, Lck-cre, or Cd4-cre mice to delete Tcf-1 conditionally at the beginning of the HSC, DN2–DN3, or DP stage, respectively. The defective T cell development phenotypes became gradually less severe as the deletion stage became more advanced in distinct mouse models. Interestingly, consistent with Tcf7−/− mice, Tcf7fl/flVav-cre mice developed aggressive T cell lymphoma within 45 weeks, but no tumors were generated in Tcf7fl/flLck-cre or Tcf7fl/flCd4-cre mice. Single-cell RNA-seq (ScRNA-seq) indicated that ablation of Tcf-1 at distinct phases can subdivide DN1 cells into three clusters (C1, C2, and C3) and DN2–DN3 cells into three clusters (C4, C5, and C6). Moreover, Tcf-1 deficiency redirects bifurcation among divergent cell fates, and clusters C1 and C4 exhibit high potential for leukemic transformation. Mechanistically, we found that Tcf-1 directly binds and mediates chromatin accessibility for both typical T cell regulators and proto-oncogenes, including Myb, Mycn, Runx1, and Lyl1 in the DN1 phase and Lef1, Id2, Dtx1, Fyn, Bcl11b, and Zfp36l2 in the DN2–DN3 phase. The aberrant expression of these genes due to Tcf-1 deficiency in very early T cells contributes to subsequent tumorigenesis. Thus, we demonstrated that Tcf-1 plays stage-specific roles in regulating early thymocyte development and transformation, providing new insights and evidence for clinical trials on T-ALL leukemia.

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Acknowledgements

We thank Mr. Biliang Zhang and Dr. Ziding Zhang at the College of Biological Sciences, China Agricultural University, for their constructive comments and assistance with the computational analysis. We also thank Dr. Bing Zhang at the Laboratory Animal Center at China Agricultural University for his assistance with animal care. This work was supported by the National Key R&D Program of China (2017YFA0104401), the National Natural Science Foundation of China (31630038, 31970831, and 31571522), and the Project for Extramural Scientists of State Key Laboratory of Agrobiotechnology from China Agricultural University (2019SKLAB6-6 & 2019SKLAB6-7).

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  1. These authors contributed equally: Fang Wang, Zhihong Qi.

Authors and Affiliations

  1. State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuanmingyuan West Road 2, 100193, Beijing, China

    Fang Wang, Zhihong Qi, Yingpeng Yao, Guotao Yu, Tao Feng, Tianyan Zhao, Yaofeng Zhao & Shuyang Yu

  2. Department of Microbiology and Immunology, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA

    Hai-Hui Xue

  3. Regenerative Biology Laboratory, Morgridge Institute for Research, Madison, WI, 53707, USA

    Peng Jiang

  4. Department Hematology, Beijing Jishuitan Hospital, 100096, Beijing, China

    Li Bao

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Contributions

S.Y. designed and supervised the experiments. F.W. analyzed the high-throughput data under the supervision of P.J.; single cells were isolated and selected by G.Y. and T.F. F.W., Z.Q., Y.Y., and G.Y. performed the general experiments. T.Z., H.-H.X., Y.Z., L.B., and S.Y. assisted with the general experiments and interpreted the data. S.Y. and F.W. wrote the manuscript with the aid of all authors.

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Correspondence to Shuyang Yu.

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Wang, F., Qi, Z., Yao, Y. et al. Exploring the stage-specific roles of Tcf-1 in T cell development and malignancy at single-cell resolution. Cell Mol Immunol 18, 644–659 (2021). https://doi.org/10.1038/s41423-020-00527-1

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