T-BET and EOMES sustain mature human NK cell identity and antitumor function
Pamela Wong, Washington University School of Medicine in St. Louis
Jennifer A. Foltz, Washington University School of Medicine in St. Louis
Lily Chang, Washington University School of Medicine in St. Louis
Carly C. Neal, Washington University School of Medicine in St. Louis
Tony Yao, Washington University School of Medicine in St. Louis
Celia C. Cubitt, Washington University School of Medicine in St. Louis
Jennifer Tran, Washington University School of Medicine in St. Louis
Samantha Kersting-Schadek, Washington University School of Medicine in St. Louis
Sathvik Palakurty, Washington University School of Medicine in St. Louis
Natalia Jaeger, Washington University School of Medicine in St. Louis
David A Russler-Germain, Washington University School of Medicine in St. Louis
Nancy D. Marin, Washington University School of Medicine in St. Louis
Margery Gang, Washington University School of Medicine in St. Louis
Julia A Wagner, Washington University School of Medicine in St. Louis
Alice Y. Zhou, Washington University School of Medicine in St. Louis
Miriam T. Jacobs, Washington University School of Medicine in St. Louis
Mark Foster, Washington University School of Medicine in St. Louis
Timothy Schappe, Washington University School of Medicine in St. Louis
Lynne Marsala, Washington University School of Medicine in St. Louis
Ethan McClain, Washington University School of Medicine in St. Louis
Patrick Pence, Washington University School of Medicine in St. Louis
Michelle Becker-Hapak, Washington University School of Medicine in St. Louis
Bryan Fisk, Washington University School of Medicine in St. Louis
Allegra A. Petti, Washington University School of Medicine in St. Louis
Obi L. Griffith, Washington University School of Medicine in St. Louis
Malachi Griffith, Washington University School of Medicine in St. Louis
Melissa M. Berrien-Elliott, Washington University School of Medicine in St. Louis
Todd A. Fehniger, Washington University School of Medicine in St. Louis
Abstract
Since the T-box transcription factors (TFs) T-BET and EOMES are necessary for initiation of NK cell development, their ongoing requirement for mature NK cell homeostasis, function, and molecular programming remains unclear. To address this, T-BET and EOMES were deleted in unexpanded primary human NK cells using CRISPR/Cas9. Deleting these TFs compromised in vivo antitumor response of human NK cells. Mechanistically, T-BET and EOMES were required for normal NK cell proliferation and persistence in vivo. NK cells lacking T-BET and EOMES also exhibited defective responses to cytokine stimulation. Single-cell RNA-Seq revealed a specific T-box transcriptional program in human NK cells, which was rapidly lost following T-BET and EOMES deletion. Further, T-BET- and EOMES-deleted CD56bright NK cells acquired an innate lymphoid cell precursor-like (ILCP-like) profile with increased expression of the ILC-3-associated TFs RORC and AHR, revealing a role for T-box TFs in maintaining mature NK cell phenotypes and an unexpected role of suppressing alternative ILC lineages. Our study reveals the critical importance of sustained EOMES and T-BET expression to orchestrate mature NK cell function and identity.
