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[Display omitted] • The expression of cMET was predominantly observed in cholangiocarcinoma (CCA) cell lines which supported the cMET as a target for CCA. • In this study, we screened and characterized novel single chain variable fragment (ScFv) targeted cMET and generated an anti-cMET CAR-NK cells to treat CCA. • The anti-cMET CAR-NK cells exhibited specificity and favorable cytotoxicity against high-cMET CCA cell line in in vitro model. Cholangiocarcinoma (CCA) presents a significant clinical challenge which is often identified in advanced stages, therby restricting the effectiveness of surgical interventions for most patients. The high incidence of cancer recurrence and resistance to chemotherapy further contribute to a bleak prognosis and low survival rates. To address this pressing need for effective therapeutic strategies, our study focuses on the development of an innovative cellular immunotherapy, specifically utilizing chimeric antigen receptor (CAR)-engineered natural killer (NK) cells designed to target the cMET receptor tyrosine kinase. In this investigation, we initiated the screening of a phage library displaying human single-chain variable fragment (ScFv) to identify novel ScFv molecules with specificity for cMET. Remarkably, ScFv11, ScFv72, and ScFv114 demonstrated exceptional binding affinity, confirmed by molecular docking analysis. These selected ScFvs, in addition to the well-established anti-cMET ScFvA, were integrated into a CAR cassette harboring CD28 transmembrane region-41BB-CD3ζ domains. The resulting anti-cMET CAR constructs were transduced into NK-92 cells, generating potent anti-cMET CAR-NK-92 cells. To assess the specificity and efficacy of these engineered cells, we employed KKU213A cells with high cMET expression and KKU055 cells with low cMET levels. Notably, co-culture of anti-cMET CAR-NK-92 cells with KKU213A cells resulted in significantly increased cell death, whereas no such effect was observed with KKU055 cells. In summary, our study identified cMET as a promising therapeutic target for CCA. The NK-92 cells, armed with the anti-cMET CAR molecule, have shown strong ability to kill cancer cells specifically, indicating their potential as a promising treatment for CCA in the future. [ABSTRACT FROM AUTHOR]

[Display omitted] • The regulatory effects of IL-10 on NK cells during liver fibrosis were evaluated. • IL-10 enhanced the immune functions of NK cells. • IL-10 promoted circulating NK cell migration to the fibrotic liver. • The IL-10–NK cell axis may be a promising treatment strategy for liver fibrosis. Interleukin 10 (IL-10) and natural killer (NK) cells have the potential to combat liver fibrosis. However, whether NK cells play an important role in the anti-fibrotic effects of IL-10 is not sufficiently elucidated. In this study, we investigated the regulatory effects of IL-10 on NK cells during liver fibrosis. Fibrotic mice induced with carbon tetrachloride were treated with or without IL-10 in the presence or absence of NK cells. Liver damage and fibrosis were assessed using hematoxylin and eosin and Sirius Red staining and serum transaminase and liver hydroxyproline assays, respectively. NK cell distribution, quantity, activation, cytotoxicity, development, and origin were analyzed using immunohistochemistry, immunofluorescence, and flow cytometry. Enzyme-linked immunosorbent assay was used to determine chemokine levels. In the presence of NK cells, IL-10 gene intervention improved liver fibrosis and enhanced NK cell accumulation and function in the liver, as evidenced by increased NKG2D, interferon-γ, and CD107a expression. Furthermore, IL-10 promoted the migration of circulating NK cells to the fibrotic liver and elevated C-C motif ligand 5 levels. However, depletion of NK cells exacerbated liver fibrosis and impaired the anti-fibrotic effect of IL-10. The anti-fibrotic effect of IL-10 relies on the enhancement of NK cell immune function, including activation, cytotoxicity, development, and migration. These results provide valuable insights into the mechanisms through which IL-10 regulates NK cells to limit the progression of liver fibrosis. [ABSTRACT FROM AUTHOR]

• NK92 cells expressing novaferon possessed enhanced cytotoxicity. • NK92-nova possessed activated JAK/STAT signaling pathway. • The cytotoxicity of NK92-nova was partially dependent on the NKG2D/NKG2DL pathway. With significant developments in chimeric antigen receptor T-cell therapy, adoptive immunotherapy has unlocked new levels of treatment for malignancies. Natural killer (NK) cells are promising alternative immune effector cells for this strategy. Multiple anti-tumor therapies are largely dependent on type I interferon (IFN) signaling. Type I IFNs enhance NK cell cytotoxicity. Novaferon (nova) is an unnatural, novel IFN-like protein produced by gene shuffling of IFN-α with strong biological activity. To augment the antitumor activity of NK cells, we generated NK92-nova cells that stably express nova. We found that NK92-nova cells mediated enhanced pan-cancer antitumor activity compared to NK92-vec cells. The increased antitumor activity was associated with the enhanced secretion of cytokines, such as IFN-γ, perforin, and granzyme B. Meanwhile, most of the activating receptors were upregulated in the NK92-nova cells. After co-culture with NK92-nova cells, the expression of NKG2D ligands on the HepG2 cells increased, resulting in an enhanced susceptibility of HepG2 cells to NK92 cell-mediated cytolysis. NK92-nova cells significantly inhibited HepG2 tumor growth in a xenograft model without systemic toxicity. Therefore, NK92-nova cells are a novel and safe strategy for cancer immunotherapy. [ABSTRACT FROM AUTHOR]

• Activation of NK cells occurs through interaction between TLR-2 expressed on NK cells and LPG of Leishmania parasites. • NK cells are able to increase the activation of DCs that are infected with Leishmania parasites. • NK cells can be more effective than CD8+ T lymphocytes in parasite eradication. • Leishmania LPG can limit NK cell activation by inhibiting IL-12 synthesis in macrophages. • Leishmania GP63 directly binds to NK cells and modulates NK cell phenotype. NK cells are known as frontline responders that are efficient in combating several maladies as well as leishmaniasis caused by Leishmania spp. As such they are being investigated to be used for adoptive transfer therapy and vaccine. In spite of the lack of antigen-specific receptors at their surface, NK cells can selectively recognize pathogens, accomplished by the activation of the receptors on the NK cell surface and also as the result of their effector functions. Activation of NK cells can occur through interaction between TLR-2 expressed on NK cells and. LPG of Leishmania parasites. In addition, NK cell activation can occur by cytokines (e.g., IFN-γ and IL-12) that also lead to producing cytokines and chemokines and lysis of target cells. This review summarizes several evidences that support NK cells activation for controlling leishmaniasis and the potentially lucrative roles of NK cells during leishmaniasis. Furthermore, we discuss strategies of Leishmania parasites in inhibiting NK cell functions. Leishmania LPG can utilizes TLR2 to evade host-immune responses. Also, Leishmania GP63 can directly binds to NK cells and modulates NK cell phenotype. Finally, this review analyzes the potentialities to harness NK cells effectiveness in therapy regimens and vaccinations. [ABSTRACT FROM AUTHOR]

• The metabolic reprogramming of NK cells plays an important role in liver cancer. • Immune and metabolic checkpoints are believed to interact in the functioning of the NK cells. • Targeting NK cell immunometabolism will be the promising treatment strategy. Natural killer (NK) cells are being used effectively as a potential candidate in tumor immunotherapy. However, the migration and transport of NK cells to solid tumors is inadequate. NK cell dysfunction, tumor invasiveness, and metastasis are associated with altered metabolism of NK cells in the liver cancer microenvironment. However, in liver cancers, metabolic impairment of NK cells is still not understood fully. Evidence from various sources has shown that the interaction of NK cell's immune checkpoints with its metabolic checkpoints is responsible for the regulation of the development and function of these cells. How immune checkpoints contribute to metabolic programming is still not fully understood, and how this can be beneficial needs a better understanding, but they are emerging to be incredibly compelling to rebuilding the function of NK cells in the tumor. It is expected to represent a potential aim that focuses on improving the efficacy of therapies based on NK cells for treating liver cancer. Here, the recent advancements made to understand the NK cell's metabolic reprogramming in liver cancer have been summarized, along with the possible interplay between the immune and the metabolic checkpoints in NK cell function. Finally, an overview of some potential metabolic-related targets that can be used for liver cancer therapy treatment has been presented. [ABSTRACT FROM AUTHOR]

• This review outlines recent advances of CAR NK cell therapy in hematological malignancies and solid tumors. • We reviewed the supremacy of CAR NK cell therapy over CAR T cell therapy. • We concentrate on the current manufacturing procedure of CAR NK cell and its potential challenges. • We focused on innovative and contemporary policies that surmount the limitations of CAR NK cell therapy in solid tumors. Adoptive cell treatment (ACT) utilizing chimeric antigen receptors (CAR) diverts the specificity of safe cells against a target-specific antigen and portrays exceptional potential for cancer treatment. While CAR T cell treatment has risen as a breakthrough with unprecedented results within the therapeutic procedures of human malignancies, different deficiencies including challenging and costly generation processes, strict patient qualification criteria, and undesirable toxicity have ruined its application. Unlike T cells, the application of natural killer (NK) cells has attracted consideration as a reasonable alternative owing to the major histocompatibility complex (MHC)-independency, shorter life expectancy, the potential to create an off-the-shelf immune product, and potent antitumor properties. In this article, we provide an updated review of the differences between CAR T and CAR NK cells, current enhancements in CAR NK design, the available sources for collecting NK cells, and strategies for the transduction step of the CARs to NK cells. Furthermore, we focus on the published and ongoing preclinical and clinical studies of CAR NK treatment strategies both in hematologic malignancies and solid tumors. We also discuss limitations and plausible solutions to improve the perseverance, function, safety, and efficacy of CAR NK cells with a special focus on solid tumors. [ABSTRACT FROM AUTHOR]

Myeloid-derived suppressor cell (MDSC) -mediated immune suppression, and natural killer (NK) and/or T cell-mediated immune responses play important roles in Chronic myeloid leukemia (CML). However, detailed regulation mechanisms of these immune cells in CML have not been fully elucidated. The proportion of MDSCs and effector NK cells in newly diagnosed CML patients, patients during TKI treatment, and healthy donors (HD) were detected using flow cytometry. Serum levels and mRNA expression of Arg1 and iNOS in newly diagnosed CML patients, patients during TKI treatment, and HD were measured by ELISA and qPCR, respectively. Effect of CML serum or peripheral blood mononuclear cells (PBMCs) on HD derived CD3+ T cell proliferation was evaluated by CFSE-labeled HD CD3+ T cells co-cultured with PBMCs and serum from CML patients. Effect of CML serum on NK cells killing activity was evaluated via detecting apoptosis of K562 cells. Proportion of Gr-MDSCs and the serum levels of Arg1 and iNOS were significantly increased in patients at diagnosis, and reduced following TKI treatment. However, the proportion of effector NK cells were decreased in patients at diagnosis, and increased following TKI treatment. Serum and PBMC from CML patients suppressed HD derived T cell proliferation in vitro. Additionally, serum from CML patients enhanced HD derived NK cell killing activity in vitro , while the addition of Arg1 inhibitor to CML serum suppressed this phenomenon. Gr-MDSCs and effector NK cells play an important role in the pathogenesis of CML, inhibiting the function of MDSC and restoring the function of NK cells is expected to be a therapeutic strategy for CML. [ABSTRACT FROM AUTHOR]

• Ginsenoside Rg3 increasd NK cell cytolytic activity. • Ginsenoside Rg3 activated NK cells through the MAPK/ERK signaling pathway. • Ginsenoside Rg3 enhanced the functional maturation of CB-derived NK cells. • Ginsenoside 20(R)-Rg3 was effective at activating NK cells and was activated through the MAPK/ERK signaling pathway. Ginseng is one of the most widely used herbal remedies for various diseases worldwide. Ginsenoside Rg3 (G-Rg3), the main component of ginseng, possesses several pharmacological properties, including anti-inflammatory, anti-tumor, antioxidant, anti-obesity, and immunomodulatory activities. However, the effect of G-Rg3 on natural killer (NK) cells in humans is not fully understood. Here, we investigated the effect of G-Rg3 on NK cell function and differentiation and elucidated the underlying mechanism. G-Rg3 increased NK cell cytotoxicity and simultaneously increased the expression of NK-activating receptors, NKp44, NKp46, and NKp30. Additionally, G-Rg3 increased the mRNA expression of NK cytolytic molecules, granzyme B and perforin. The expression of CD107a, a marker of NK cell degranulation, also increased in G-Rg3–treated NK cells. We therefore proceeded to identify which MAPK signaling pathway was involved in G-Rg3–mediated cytolytic activity. Treatment with G-Rg3 increased the phosphorylation levels of extracellular signal–regulated kinase (ERK), whereas ERK inhibition eliminated G-Rg3–induced NK cell cytotoxicity, suggesting the involvement of the ERK pathway. G-Rg3 did not affect the rate of differentiation of human cord-blood–derived NK cells; however, it increased the functional maturation of differentiated NK cells and promoted their cytotoxicity. The G-Rg3 isomer, 20(R)-Rg3, effectively activated NK cells via the extracellular signal-regulated kinase (ERK) signaling pathway, whereas 20(S)-Rg3 had no effect on NK cell activity. Altogether, the results demonstrated that 20(R)-Rg3 promoted NK cell activity via activation of the MAPK/ERK pathway, suggesting that 20(R)-Rg3 may be used as an activator of NK cell cytotoxicity for the treatment of diverse types of cancers. [ABSTRACT FROM AUTHOR]

• Bufalin could enhance the killing efficacy of NK cells to suppress hepatocellular carcinoma. • The molecular mechanism analysis showed that bufalin could inhibit MICA shedding by reducing ADAM9 expression. • Bufalin may be a potential agent in treatment of hepatocellular carcinoma. Bufalin, as a Chinese traditional anti-tumor agent, has been studied about inhibiting proliferation and promoting apoptosis of liver cancer, however, there are few reports on immune modulating function. We used the human liver cancer cell lines along with 91 pathologically-verified postoperative hepatocellular carcinoma (HCC) specimens to assess immune modulating function of bufalin. We found that bufalin directly balances stimulatory and inhibitory receptors on the surface of NK cells and indirectly activates natural killer (NK) cells by inhibiting MICA shedding, which prevented immune escape and indirectly enhanced NKG2D-dependent immune surveillance. This study showed that bufalin can directly or indirectly regulate the immune response, which provides a new theoretical basis for the clinical application of "Huachansu injection". [ABSTRACT FROM AUTHOR]

• Major immune effector cells like CD8+ T lymphocytes and natural killer cells are able to kill or inhibit Hepatocellular carcinoma (HCC) cells and their cytotoxic ability can be enhanced or down-regulated by certain cytokines or drugs. • HCC cells can impair the function of major immune effector cells through a variety of mechanisms. • Organoids or direct contact cell co-culture especially with primary HCC cells and tumor infiltrating lymphocytes provide better understanding of cell–cell interactions between major immune effector cells and HCC cells. • Immunotherapy like checkpoint inhibitors and adoptive cell transfer showed notable effectiveness and safety in preventing HCC recurrence as adjuvant therapy. Hepatocellular carcinoma (HCC) is the most common liver neoplasm with high morbidity and mortality. Tumor immunotherapy might be promising adjuvant therapy for HCC after surgery. To better develop HCC immunotherapy, comprehensive understanding of cell–cell interactions between immune effector cells and HCC cells remains crucial. To review the existing studies to summarize the cell–cell interactions between major immune effector cells and HCC cells providing new data for HCC immunotherapy. A systematic review was conducted by searching PubMed database covering all papers published in recent five years up to January 2020. The guidelines of the preferred reporting items for systematic reviews were firmly followed. There are 9 studies researching the interactions between CD8+ T lymphocytes and HCC cells and 22 studies researching that between natural killer (NK) cells and HCC cells. Among the 9 studies, 6 studies reported that CD8+ T lymphocytes showed cytotoxicity towards HCC cells while 3 studies found CD8+ T lymphocytes were impaired by HCC cells. Among the 22 studies, 20 studies presented that NK cells could inhibit HCC cells. Two studies were found to report NK cell dysfunction in HCC. Based on the systematic analysis, we concluded that CD8+ T lymphocytes and NK cells can inhibit HCC cells. While in turn, HCC cells can also result in the dysfunction of those effector cells through various mechanisms. Organoids and direct contact cell co-culture with primary HCC cells and TILs should be the most innovative way to investigate the interactions and develop novel immunotherapy. [ABSTRACT FROM AUTHOR]

• NK cells were declined in the peripheral blood of AML patients. • NK cells displayed altered profile of functional receptors in AML patients. • Siglec-7 expression were reduced in NK cells in AML patients. • Siglec-7+ NK cells displayed higher effector function. Immune dysfunction is an established risk factor in acute myeloid leukemia (AML). The cytotoxicity of natural killer (NK) cells is greatly impaired in AML, and the profile of NK cell receptors is markedly altered in AML; however, this is not yet well characterized. In this study, we found the downregulation of Siglec-7 could be utilized as a potential marker of NK cell dysfunction in AML. The absolute numbers and percentages of NK cells were declined in the peripheral blood of patients with AML, and the levels of activating receptors NKG2D, NKp46, and NKp30 were reduced in NK cells from patients with AML compared with healthy controls. In contrast, the levels of inhibitory receptors TIM-3, ILT-4, ILT-5, and PD-1 were increased in NK cells from patients with AML. Of note, the level of Siglec-7 in NK cells from patients with AML was significantly lower than that in NK cells from healthy controls, and Siglec-7+ NK cells displayed higher levels of activating receptors and stronger cytotoxicity when compared with Siglec-7– NK cells. Our data indicate that decreased Siglec-7 level may predict NK cell dysfunction in AML, and NK cells may be promising targets of immunotherapy for AML. [ABSTRACT FROM AUTHOR]