Single-cell transcriptome reveals the novel role of T-bet in suppressing the immature NK gene signature
Chao Yang,
Jason R Siebert,
Robert Burns,
Yongwei Zheng,
Ao Mei,
Benedetta Bonacci,
Demin Wang,
Raul A Urrutia,
Matthew J Riese,
Sridhar Rao,
Karen-Sue Carlson,
Monica S Thakar,
Subramaniam Malarkannan
Affiliations
Chao Yang
Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States; Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, United States
Jason R Siebert
Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States; Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, United States
Robert Burns
Bioinfomatics Core, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States
Yongwei Zheng
Laboratory of B-Cell Lymphopoiesis, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States
Ao Mei
Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States; Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, United States
Benedetta Bonacci
Flow Cytometry Core, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States
Demin Wang
Laboratory of B-Cell Lymphopoiesis, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States
Raul A Urrutia
Department of Surgery, Medical College of Wisconsin, Milwaukee, United States
Matthew J Riese
Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, United States; Laboratory of Lymphocyte Biology, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States; Department of Medicine, Medical College of Wisconsin, Milwaukee, United States
Sridhar Rao
Laboratory of Stem Cell Transcriptional Regulation, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States
Karen-Sue Carlson
Department of Medicine, Medical College of Wisconsin, Milwaukee, United States; Laboratory of Coagulation Biology, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States
Monica S Thakar
Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States; Department of Pediatrics, Medical College of Wisconsin, Milwaukee, United States
Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Blood Center of Wisconsin, Milwaukee, United States; Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, United States; Department of Medicine, Medical College of Wisconsin, Milwaukee, United States; Department of Pediatrics, Medical College of Wisconsin, Milwaukee, United States
The transcriptional activation and repression during NK cell ontology are poorly understood. Here, using single-cell RNA-sequencing, we reveal a novel role for T-bet in suppressing the immature gene signature during murine NK cell development. Based on transcriptome, we identified five distinct NK cell clusters and define their relative developmental maturity in the bone marrow. Transcriptome-based machine-learning classifiers revealed that half of the mTORC2-deficient NK cells belongs to the least mature NK cluster. Mechanistically, loss of mTORC2 results in an increased expression of signature genes representing immature NK cells. Since mTORC2 regulates the expression of T-bet through AktS473-FoxO1 axis, we further characterized the T-bet-deficient NK cells and found an augmented immature transcriptomic signature. Moreover, deletion of Foxo1 restores the expression of T-bet and corrects the abnormal expression of immature NK genes. Collectively, our study reveals a novel role for mTORC2-AktS473-FoxO1-T-bet axis in suppressing the transcriptional signature of immature NK cells.
