NK cells recognize virally infected and malignant cells and respond by direct target cell killing and the production of inflammatory cytokines. In humans, we have shown that adoptive transfer of NK cells can induce remission in patients with refractory acute myelogenous leukemia (AML) and that hematopoietic cell transplantation (HCT) can cure AML. Although clinical success is seen in both settings, NK cell survival and function are often suboptimal after transplant, resulting in cancer relapse. We have shown that AML patients who reactivate latent human cytomegalovirus (HCMV) post-HCT reconstitute more terminally differentiated NK cells with potent effector function relative to those who do not reactivate HCMV or are HCMV seronegative. Several clinical studies have also recently shown that HCMV reactivation early after HCT is associated with reduced relapse risk. The molecular basis for enhanced NK cell differentiation and function in response to HCMV is unknown. We have extensive preliminary data showing that, in healthy HCMV seropositive donors and in transplant recipients that experience HCMV reactivation, populations of NK cells lacking the proximal signaling molecules Fc?R1g, SYK and EAT-2 expand. These novel NK cells have a genome wide epigenetic profile markedly similar to that of effector CD8+ T cells with a striking microRNA signature. They are present at stable frequencies over the course of at least one year, and they exhibit enhanced function in response to signaling through CD16. These cells appear to represent a human equivalent to adaptive or memory NK cells in mice that respond specifically to mouse cytomegalovirus (MCMV). Because of their functional and survival properties, there is considerable interest in being able to utilize them for immunotherapy. In this proposal, we aim to study the role of microRNAs in the differentiation of adaptive human NK cells upon HCMV reactivation in NK cells from our unique cohort of transplant patients using an unbiased, cutting-edge deep-sequencing approach. We will test the hypothesis that signaling through activating receptors is required for the expansion of adaptive NK cells and determine the role of pentameric HCMV protein complexes and high-affinity antibodies to these complexes in triggering expansion. We have preliminary data suggesting that the Notch/TCF-1 and Wnt signaling pathways are selectively induced in HCMV-induced adaptive NK cells, and we will study the role of both pathways in the development and differentiation of adaptive NK cells in depth using our well defined transplant cohorts and in vitro assays. We will also combine our recently developed xenogeneic model for adoptive human NK cell transfer with a new model for systemic HCMV infection in mice to study HCMV-induced adaptive NK cell survival, homing and function in vivo. Our team of leading basic and clinical NK cell biologists, HCMV experts and mouse immunologists are well positioned to perform these studies and translate them to therapies that can prime NK cells for potent function and prolonged survival for clinical use in the treatment of cancer.

Public Health Relevance

Human natural killer (NK) cells are a type of circulating white blood cell that can kill cancer cells and virally infected cells. As these immune cells develop, they acquire the ability to kill damaged or sick cells and to secrete cytokines. Although NK cells were thought to be short lived, new emerging data suggests that viral infection not only activates human NK cells, but also increases their function, lifespan and response to both secondary challenge of the previous virus and cancer targets, suggesting that adaptive NK cell responses may not be antigen specific. The real impact of this grant is to understand how viral infection makes NK cells acquire adaptive properties. Our long-term goal is to translate this basic research into strategies that can prime NK cells with potent function and prolonged in vivo survival for clinical use for patients with solid tumors and leukemia.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
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Special Emphasis Panel (ZRG1-IMM-M (03))
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Di Fronzo, Nancy L
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University of Minnesota Twin Cities
Internal Medicine/Medicine
Schools of Medicine
United States
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de Witte, Moniek A; Sarhan, Dhifaf; Davis, Zachary et al. (2018) Early Reconstitution of NK and ?? T Cells and Its Implication for the Design of Post-Transplant Immunotherapy. Biol Blood Marrow Transplant 24:1152-1162
Cichocki, Frank; Wu, Cheng-Ying; Zhang, Bin et al. (2018) ARID5B regulates metabolic programming in human adaptive NK cells. J Exp Med 215:2379-2395
de Witte, Moniek A; Kuball, J├╝rgen; Miller, Jeffrey S (2017) NK Cells and ??T Cells for Relapse Protection After Allogeneic Hematopoietic Cell Transplantation (HCT). Curr Stem Cell Rep 3:301-311
Cichocki, Frank; Valamehr, Bahram; Bjordahl, Ryan et al. (2017) GSK3 Inhibition Drives Maturation of NK Cells and Enhances Their Antitumor Activity. Cancer Res 77:5664-5675
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Cichocki, F; Cooley, S; Davis, Z et al. (2016) CD56dimCD57+NKG2C+ NK cell expansion is associated with reduced leukemia relapse after reduced intensity HCT. Leukemia 30:456-63
Sarhan, Dhifaf; Cichocki, Frank; Zhang, Bin et al. (2016) Adaptive NK Cells with Low TIGIT Expression Are Inherently Resistant to Myeloid-Derived Suppressor Cells. Cancer Res 76:5696-5706
Davis, Zachary B; Cooley, Sarah A; Cichocki, Frank et al. (2015) Adaptive Natural Killer Cell and Killer Cell Immunoglobulin-Like Receptor-Expressing T Cell Responses are Induced by Cytomegalovirus and Are Associated with Protection against Cytomegalovirus Reactivation after Allogeneic Donor Hematopoietic Cell Transpla Biol Blood Marrow Transplant 21:1653-62

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