Browsing by Author "Crawford, Jeremy Chase"

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    Caspase-8 and FADD prevent spontaneous ZBP1 expression and necroptosis
    (National Academy of Sciences, 2022) Rodriguez, Diego A.; Quarato, Giovanni; Liedmann, Swantje; Tummers, Bart; Zhang, Ting; Guy, Cliff; Crawford, Jeremy Chase; Palacios, Gustavo; Pelletier, Stephane; Kalkavan, Halime; Shaw, Jeremy J. P.; Fitzgerald, Patrick; Chen, Mark J.; Balachandran, Siddharth; Green, Douglas R.; Medical and Molecular Genetics, School of Medicine
    Caspase-8 and Fas-associated death domain (FADD) play key roles in the regulation of cell death by necroptosis. The absence of either protein results in early embryonic lethality due to the activation of the kinase receptor interacting protein kinase-3 (RIPK3) and its phosphorylation of the necroptosis executioner, mixed-lineage kinase-like (MLKL). We genetically engineered and characterized a mouse model to monitor MLKL phosphorylation in the absence of necroptosis in vivo. Ablation of caspase-8 or FADD resulted in the transcriptional upregulation in several tissues of Z-DNA binding protein-1 (ZBP1), a cytosolic nucleic acid sensor capable of activating RIPK3, and ZBP1 was required for spontaneous phosphorylation of MLKL. Our findings provide a mechanism by which the FADD/Caspase-8 complex prevents necroptosis.
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    Neuroblastoma Formation Requires Unconventional CD4 T Cells and Arginase-1–Dependent Myeloid Cells
    (American Association for Cancer Research, 2021) Van de Velde, Lee-Ann; Allen, E. Kaitlynn; Crawford, Jeremy Chase; Wilson, Taylor L.; Guy, Clifford S.; Russier, Marion; Zeitler, Leonie; Bahrami, Armita; Finkelstein, David; Pelletier, Stephane; Schultz-Cherry, Stacey; Thomas, Paul G.; Murray, Peter J.; Medicine, School of Medicine
    Immune cells regulate tumor growth by mirroring their function as tissue repair organizers in normal tissues. To understand the different facets of immune-tumor collaboration through genetics, spatial transcriptomics, and immunologic manipulation with noninvasive, longitudinal imaging, we generated a penetrant double oncogene-driven autochthonous model of neuroblastoma. Spatial transcriptomic analysis showed that CD4+ and myeloid populations colocalized within the tumor parenchyma, while CD8+ T cells and B cells were peripherally dispersed. Depletion of CD4+ T cells or CCR2+ macrophages, but not B cells, CD8+ T cells, or natural killer (NK) cells, prevented tumor formation. Tumor CD4+ T cells displayed unconventional phenotypes and were clonotypically diverse and antigen independent. Within the myeloid fraction, tumor growth required myeloid cells expressing arginase-1. Overall, these results demonstrate how arginine-metabolizing myeloid cells conspire with pathogenic CD4+ T cells to create permissive conditions for tumor formation, suggesting that these protumorigenic pathways could be disabled by targeting myeloid arginine metabolism. SIGNIFICANCE: A new model of human neuroblastoma provides ways to track tumor formation and expansion in living animals, allowing identification of CD4+ T-cell and macrophage functions required for oncogenesis.
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    The interaction between RIPK1 and FADD controls perinatal lethality and inflammation
    (Elsevier, 2024) Rodriguez, Diego A.; Tummers, Bart; Shaw, Jeremy J. P.; Quarato, Giovanni; Weinlich, Ricardo; Cripps, James; Fitzgerald, Patrick; Janke, Laura J.; Pelletier, Stephane; Crawford, Jeremy Chase; Green, Douglas R.; Medical and Molecular Genetics, School of Medicine
    Perturbation of the apoptosis and necroptosis pathways critically influences embryogenesis. Receptor-associated protein kinase-1 (RIPK1) interacts with Fas-associated via death domain (FADD)-caspase-8-cellular Flice-like inhibitory protein long (cFLIPL) to regulate both extrinsic apoptosis and necroptosis. Here, we describe Ripk1-mutant animals (Ripk1R588E [RE]) in which the interaction between FADD and RIPK1 is disrupted, leading to embryonic lethality. This lethality is not prevented by further removal of the kinase activity of Ripk1 (Ripk1R588E K45A [REKA]). Both Ripk1RE and Ripk1REKA animals survive to adulthood upon ablation of Ripk3. While embryonic lethality of Ripk1RE mice is prevented by ablation of the necroptosis effector mixed lineage kinase-like (MLKL), animals succumb to inflammation after birth. In contrast, Mlkl ablation does not prevent the death of Ripk1REKA embryos, but animals reach adulthood when both MLKL and caspase-8 are removed. Ablation of the nucleic acid sensor Zbp1 largely prevents lethality in both Ripk1RE and Ripk1REKA embryos. Thus, the RIPK1-FADD interaction prevents Z-DNA binding protein-1 (ZBP1)-induced, RIPK3-caspase-8-mediated embryonic lethality, affected by the kinase activity of RIPK1.