Gene expression in control and DOCK8 CRISPR KHYG1 NK cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE101467 | GEO | 2017/09/15
SECONDARY ACCESSION(S): PRJNA394516
REPOSITORIES: GEO
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Project description:NK cells are a promising cellular therapy for cancer, with challenges in the field including persistence, functional activity, and tumor recognition. Briefly priming blood NK cells with rhIL-12, rhIL-15, and rhIL-18 (12/15/18) results in memory-like NK cell differentiation and enhanced responses against cancer. We developed a novel platform centered upon an inert tissue factor scaffold for production of heteromeric fusion protein complexes (HFPC). The first use of this platform combined IL-12, IL-15 and IL-18 receptor engagement (HCW9201) and the second adds CD16 engagement (HCW9207). HCW9201 and HCW9207 stimulated activation and proliferation signals in NK cells, but HCW9207 had decreased IL-18 receptor signaling. RNAseq and multidimensional mass cytometry revealed strong parallels between HCW9201 and 12/15/18. Moreover, HCW9201 stimulation improved NK cell metabolic fitness, and resulted in the DNA methylation remodeling characteristic of memory-like differentiation. HCW9201- and 12/15/18-primed similar increases in short-term and memory-like NK cell cytotoxicity and IFN-g production against leukemia targets, as well as equivalent control of leukemia in NSG mice. Thus, HFPCs represent a protein engineering approach that solves many problems associated with multi-signal receptor engagement on immune cells, and HCW9201-primed NK cells will be advanced as an ideal approach for clinical GMP-grade memory-like NK cell production for cancer therapy.erm and memory-like NK cell cytotoxicity and IFN-g production against leukemia targets, as well as equivalent control of leukemia in NSG mice. Thus, HFPCs represent a protein engineering approach that solves many problems associated with multi-signal receptor engagement on immune cells, and HCW9201-primed NK cells will be advanced as an ideal approach for clinical GMP-grade memory-like NK cell production for cancer therapy.
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Project description:is involved in the formation of immune signaling complexes. To date only limited and moreover 35 conflicting data exist regarding the role of ADAP in NK cells. To extend existing knowledge we 36 investigated ADAP-dependency of NK cells in the context of in vivo infection with the intracellular 37 pathogen Listeria monocytogenes (Lm). Ex vivo analysis of infection-primed NK cells revealed 38 impaired expression of IFN-γ and chemokines as well as impaired cytotoxic capacity in NK cells 39 lacking ADAP as indicated by reduced CD107a surface expression and inefficient perforin 40 production. However, ADAP-deficiency had no global effect on NK cell morphology or intracellular 41 distribution of CD107a-containing vesicles. Proteomic definition of ADAPko and wild type NK cells 42 did not uncover obvious differences in protein composition during the steady state and moreover, 43 similar early response patterns were induced in NK cells upon infection independent of the genotype. 44 In line with protein network analyses that suggested an altered migration phenotype in naïve 45 ADAPko NK cells, in vitro migration assays uncovered significantly reduced migration of both naïve 46 as well as infection-primed ADAPko NK cells compared to wild type NK cells. We propose that this 47 migration defect might account at least in part for the fact that during in vivo infection significantly 48 lower numbers of ADAPko NK cells accumulate in the spleen i.e. the site of infection. In conclusion, 49 we show here that during systemic Lm infection in mice ADAP is essential for efficient cytokine and 50 chemokine production, cytotoxic capacity and migration of NK cells.
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Project description:Natural killer (NK) cells play a critical role in early host defense to infected and transformed cells. Here we show that mice deficient in Eri1, a conserved 3’-to-5’ exoribonuclease that represses RNA interference, have a cell-intrinsic defect in the homeostasis and terminal maturation of NK cells. Eri1–/– NK cells show delayed acquisition of Ly49 receptors in the bone marrow and a selective reduction in Ly49D and Ly49H activating receptors in the periphery. Furthermore, antigen-specific Ly49H+ NK cells deficient in Eri1 fail to expand efficiently in mouse cytomegalovirus (MCMV) infection. Consequently, we find that Eri1 is required for immune-mediated control of MCMV titers. In contrast to NK cells, T cells deficient in Eri1 differentiate into effectors that produce high levels of Th2 cytokines. Here we identify miRNAs as the major endogenous small RNA target of Eri1 in mouse lymphocytes. NK and T cells deficient in Eri1 display an approximately two-fold increase in all miRNAs. In Eri1-deficient T cells, ectopic expression of wildtype Eri1 is sufficient to rescue defects in both miRNA homeostasis and Th2 cytokine production. Thus mouse Eri1 negatively regulates miRNA homeostasis and is required to promote normal lymphocyte effector functions.
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