Open Access

Show Abstract ▼

Abstract: Malignant tumors often escape anticancer immune surveillance by suppressing the cytotoxic functions of T lymphocytes. While many of these immune evasion networks include checkpoint proteins, small molecular weight compounds, such as the amino acid L-kynurenine (LKU), could also substantially contribute to the suppression of anti-cancer immunity. However, the biochemical mechanisms underlying the suppressive effects of LKU on T-cells remain unclear. Here, we report for the first time that LKU suppresses T cell function as an aryl hydrocarbon receptor (AhR) ligand. The presence of LKU in T cells is associated with AhR activation, which results in competition between AhR and hypoxia-inducible factor 1 alpha (HIF-1α) for the AhR nuclear translocator, ARNT, leading to T cell exhaustion. The expression of indoleamine 2,3-dioxygenase 1 (IDO1, the enzyme that leads to LKU generation) is induced by the TGF-β-Smad-3 pathway. We also show that IDO-negative cancers utilize an alternative route for LKU production via the endogenous inflammatory mediator, the high mobility group box 1 (HMGB-1)-interferon-gamma (IFN-γ) axis. In addition, other IDO-negative tumors (like T-cell lymphomas) trigger IDO1 activation in eosinophils present in the tumor microenvironment (TME). These mechanisms suppress cytotoxic T cell function, and thus support the tumor immune evasion machinery.

Open Access

Show Abstract ▼

Abstract: Galectin-9 is one of the key proteins employed by a variety of human malignancies to suppress anti-cancer activities of cytotoxic lymphoid cells and thus escape immune surveillance. Human cancer cells in most cases express higher levels of galectin-9 compared to non-transformed cells. However, the biochemical mechanisms underlying this phenomenon remain unclear. Here we report for the first time that in human cancer as well as embryonic cells, the transcription factors hypoxia-inducible factor 1 (HIF-1) and activator protein 1 (AP-1) are involved in upregulation of transforming growth factor beta 1 (TGF-β1) expression, leading to activation of the transcription factor Smad3 through autocrine action. This process triggers upregulation of galectin-9 expression in both malignant (mainly in breast and colorectal cancer as well as acute myeloid leukaemia (AML)) and embryonic cells. The effect, however, was not observed in mature non-transformed human cells. TGF-β1-activated Smad3 therefore displays differential behaviour in human cancer and embryonic vs non-malignant cells. This study uncovered a self-supporting biochemical mechanism underlying high levels of galectin-9 expression operated by the human cancer and embryonic cells employed in our investigations. Our results suggest the possibility of using the TGF-β1 signalling pathway as a potential highly efficient target for cancer immunotherapy.

Open Access

Show Abstract ▼

Abstract: Human cancer cells operate a variety of effective molecular and signaling mechanisms which allow them to escape host immune surveillance and thus progress the disease. We have recently reported that the immune receptor Tim-3 and its natural ligand galectin-9 are involved in the immune escape of human acute myeloid leukemia (AML) cells. These cells use the neuronal receptor latrophilin 1 (LPHN1) and its ligand fibronectin leucine rich transmembrane protein 3 (FLRT3, and possibly other ligands) to trigger the pathway. We hypothesized that the Tim-3-galectin-9 pathway may be involved in the immune escape of cancer cells of different origins. We found that studied breast tumors expressed significantly higher levels of both galectin-9 and Tim-3 compared to healthy breast tissues of the same patients and that these proteins were co-localized. Increased levels of LPHN2 and expressions of LPHN3 as well as FLRT3 were also detected in breast tumor cells. Activation of this pathway facilitated the translocation of galectin-9 onto the tumor cell surface, however no secretion of galectin-9 by tumor cells was observed. Surface-based galectin-9 was able to protect breast carcinoma cells against cytotoxic T cell-induced death. Furthermore, we found that cell lines from brain, colorectal, kidney, blood/mast cell, liver, prostate, lung, and skin cancers expressed detectable amounts of both Tim-3 and galectin-9 proteins. The majority of cell lines expressed one of the LPHN isoforms and FLRT3. We conclude that the Tim-3-galectin-9 pathway is operated by a wide range of human cancer cells and is possibly involved in prevention of anti-tumor immunity.

Show Abstract ▼

Abstract: The progression of acute myeloid leukemia (AML)—the most severe blood/bone marrow cancer—is determined by the ability of malignant cells to escape host immune surveillance. However, the systemic regulatory mechanisms underlying this phenomenon remain largely unknown. In this study, we discovered a fundamental systemic biochemical strategy that allows AML cells to employ physiological systems within the body to survive and escape immune attack. We found that AML cells use a crucial human adrenal cortex hormone (cortisol) to induce the expression of neuronal receptor latrophilin 1 (LPHN1), which facilitates exocytosis. This receptor interacts with the blood plasma protein fibronectin leucine rich transmembrane protein 3 (FLRT3) to cause secretion of the immune suppressor galectin-9, which impairs the anticancer activities of cytotoxic lymphoid cells. AML is a cancer of the blood and bone marrow that originates from self-renewing malignant immature myeloid cells and rapidly progresses into a systemic, and very often fatal, malignancy.1 AML cells employ physiological systems in the body to produce factors required for proliferation/disease progression.2,3 This includes the hijacking of stem cell factor (SCF), a major hematopoietic growth factor that controls AML progression and thus can become highly oncogenic.2,3 The expression and release of SCF can be triggered by AML cells via cytokines (e.g., interleukin-1β).2 Recent evidence clearly demonstrated that AML cells are also capable of impairing the activities of cytotoxic lymphoid cells (e.g., natural killer (NK) cells and cytotoxic T cells).4 One of the biochemical mechanisms underlying this phenomenon lies in the ability of AML cells to secrete the protein galectin-9. This tandem-type galectin binds the immune receptor Tim-3 and induces a variety of intracellular and cell-to-cell signaling events leading to the inactivation of NK cells, as well as the death of cytotoxic T cells.4,5 We recently reported that the process of galectin-9 secretion in AML cells is stimulated by the unique G protein-coupled receptor LPHN1, which normally functions in neurons to facilitate exocytosis.4,6 LPHN1 is also found in hematopoietic stem cells (HSCs), but its expression disappears at the early stages of their maturation.4,7 However, upon malignant transformation, AML cells preserve their abilities to express LPHN1 and to produce high levels of galectin-9 and Tim-3, in which the latter is involved in trafficking galectin-9 during the secretion process (HSCs express neither galectin-9 nor Tim-34). It is currently unknown which molecular mechanisms trigger elevated levels of LPHN1 expression in primary human AML cells, and in general, the mechanisms of upregulation of LPHN1 expression at the genomic level remain unclear. It is also unknown whether FLRT3, a natural LPHN1 ligand,4,8 is present in human blood plasma and in other tissues associated with AML. Unraveling these mechanisms is crucial to understanding the pathways that control the ability of AML cells to protect themselves against cytotoxic lymphoid cells and, thus, was the aim of the present study.

Open Access

Show Abstract ▼

Abstract: In this study we used 5 nm gold nanoparticles as delivery platforms to target cancer cells expressing the immune receptor Tim-3 using single chain antibodies. Gold surfaces were also covered with the cytotoxic drug rapamycin which was immobilised using a glutathione linker. These nanoconjugates allowed highly specific and efficient delivery of cytotoxic rapamycin into human malignant blood cells.