In vivo CRISPR screens reveal a HIF-1α-mTOR-network regulates T follicular helper versus Th1 cells

• Published in: Nature communications 2022-02, Vol.13 (1), p.805-805, Article 805
• Main Authors: Huang, Bonnie, Phelan, James D., Preite, Silvia, Gomez-Rodriguez, Julio, Johansen, Kristoffer H., Shibata, Hirofumi, Shaffer, Arthur L., Xu, Qin, Jeffrey, Brendan, Kirby, Martha, Anderson, Stacie, Yang, Yandan, Gossa, Selamawit, McGavern, Dorian B., Staudt, Louis M., Schwartzberg, Pamela L.
• Format: Article
• Language: English
• Subjects: 38/39; 45/41; 49/31; 49/47; 631/208/4041/3196; 631/250/1619/554/1898; 631/250/2152/2153/1982; 631/553/1833; 64/60; 96/95; Animals; Antibodies; Antibody Formation; Autophagy; CD4 antigen; Cell differentiation; Cell Differentiation - immunology; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR; Gene Expression; Gene Knockout Techniques; Genetic analysis; Germinal Center - immunology; Glycolysis; Humanities and Social Sciences; Hypoxia-Inducible Factor 1, alpha Subunit - genetics; Hypoxia-inducible factor 1a; Immunity, Humoral - immunology; Lymphocytes; Lymphocytes T; Mechanistic Target of Rapamycin Complex 1 - metabolism; Mice; Microenvironments; multidisciplinary; Myc protein; Nutrient cycles; Science; Science (multidisciplinary); T-Lymphocytes, Helper-Inducer - immunology; T-Lymphocytes, Helper-Inducer - metabolism; Th1 Cells - immunology; Th1 Cells - metabolism; TOR protein; TOR Serine-Threonine Kinases - genetics; TOR Serine-Threonine Kinases - metabolism; Transcriptomics; VHL protein; Virus Diseases - immunology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-28378-6

T follicular helper (Tfh) cells provide signals to initiate and maintain the germinal center (GC) reaction and are crucial for the generation of robust, long-lived antibody responses, but how the GC microenvironment affects Tfh cells is not well understood. Here we develop an in vivo T cell-intrinsic CRISPR-knockout screen to evaluate Tfh and Th1 cells in an acute viral infection model to identify regulators of Tfh cells in their physiological setting. Using a screen of druggable-targets, alongside genetic, transcriptomic and cellular analyses, we identify a function of HIF-1α in suppressing mTORC1-mediated and Myc-related pathways, and provide evidence that VHL-mediated degradation of HIF-1α is required for Tfh development; an expanded in vivo CRISPR screen reveals multiple components of these pathways that regulate Tfh versus Th1 cells, including signaling molecules, cell-cycle regulators, nutrient transporters, metabolic enzymes and autophagy mediators. Collectively, our data serve as a resource for studying Tfh versus Th1 decisions, and implicate the VHL-HIF-1α axis in fine-tuning Tfh generation. T follicular helper (Tfh) and T help type 1 (Th1) cells both arise from naïve CD4 T cells, but detailed knowledge of their differentiation remains incomplete. Here the authors pursue an in vivo CRISPR screen to identify genes, focusing on druggable targets, regulating Tfh versus Th1 to provide a resource for related studies, while also implicating HIF-1α and VHL in this regulation.