TNFSF9 Is a Prognostic Biomarker and Correlated with Immune Infiltrates in Pancreatic Cancer.

BACKGROUND: TNFSF9 gene has been found to play an anti-tumor role and regulate the function of immune cells. However, the prognostic role of TNFSF9 in pancreatic cancer and its relationship with immune cell infiltration have not been studied. METHODS: We used Oncomine, UALCAN, and GEPIA databases to analyze the expression of TNFSF9 in pancreatic cancer. We used Kaplan-Meier plotters, GEPIA, and UALCAN to evaluate the effect of TNFSF9 on clinical prognosis. We further used TIMER to study the correlation between TNFSF9 and cancer immune infiltrate cells. In addition, we used GEPIA to analyze the correlation between TNFSF9 expression and gene markers of immune infiltrate cells. RESULTS: TNFSF9 mRNA expression level was remarkably increased in pancreatic cancer than that in normal tissues (both P < 0.05). In addition, high TNFSF9 expression was significantly related to poor overall survival (OS) and relapse-free survival (RFS) in pancreatic cancer (OS HR = 2.02, P = 0.0012; RFS HR = 2.63, P = 0.022). Moreover, high TNFSF9 expression in pancreatic cancer patients was associated with worse OS in stage 1 to 2 but not stage 3 and stage 4. Specifically, TNFSF9 expression and CD8+ T cell infiltration of pancreatic cancer was negatively correlated. TNFSF9 expression showed strong correlations with M1 macrophages in pancreatic cancer. CONCLUSIONS: Our results suggest that TNFSF9 is associated with prognosis and CD8+ T cell infiltration levels in patients with pancreatic cancer. Further, TNFSF9 expression potentially contributes to the modulation of M1 polarization of macrophages. These findings indicate that TNFSF9 can be serves as a prognostic biomarker in determining the prognosis of pancreatic cancer and is associated with different types of phenotypes of immune cell infiltration.

Aberrant CD137 ligand expression induced by GATA6 overexpression promotes tumor progression in cutaneous T-cell lymphoma.

CD137 and its ligand, CD137L, are expressed on activated T cells and antigen-presenting cells, respectively. Recent studies have shown that CD137L and CD137 are aberrantly expressed by tumor cells, especially in some hematopoietic malignancies, and interactions between these molecules on tumor cells promote tumor growth. In this study, we investigated the roles of CD137L and CD137 in cutaneous T-cell lymphoma (CTCL), represented by mycosis fungoides and Sézary syndrome. Flow cytometric analysis showed that primary Sézary cells and CTCL cell lines (Hut78, MyLa, HH, SeAx, and MJ) aberrantly expressed CD137L. CD137L expression by tumor cells in CTCL was also confirmed by immunohistochemistry. Anti-CD137L-neutralizing antibody inhibited proliferation, survival, CXCR4-mediated migration, and in vivo growth in CTCL cell lines through inhibition of phosphorylation of AKT, extracellular signal-regulated kinase 1/2, p38 MAPK, and JNK. Moreover, suppression of CD137L signaling decreased antiapoptotic proteins Bcl-2 and phosphorylated Bad. We also explored the transcription factor regulating CD137L expression. Because GATA6 has been proposed as an oncogene in many types of tumors with aberrant CD137L expression, we examined GATA6 expression and the involvement of GATA6 in CD137L expression in CTCL. DNA hypomethylation and histone acetylation induced GATA6 overexpression in CTCL cells. Furthermore, chromatin immunoprecipitation, luciferase reporter assay, and knockdown by short hairpin RNA showed that GATA6 directly upregulated CD137L expression. Inhibition of GATA6 resulted in decreased survival and in vivo growth in CTCL cells. Collectively, our findings prompt a novel therapeutic approach to CTCL based on the discovery that the GATA6/CD137L axis plays an important role in the tumorigenesis of CTCL.

Enhanced antitumor activity mediated by human 4-1BB-engineered T cells.

4-1BB (CD137) is a costimulatory molecule transiently expressed on the T-cell surface after TCR engagement, whereas its ligand 4-1BBL can be found on professional antigen-presenting cells, but more importantly, also on tumor cells. As the role of the 4-1BB/4-1BBL pathway has emerged central to CD8(+) T-cell responses and survival, we sought to test its relevance in the context of genetically modified human T cells. To that end, T cells purified from healthy donors and from vaccinated-melanoma patients were transduced to express high levels of constitutive 4-1BB. 4-1BB-transduced T cells were cocultured with melanoma tumor lines and exhibited enhanced cytokine secretion, upregulation of activation markers as well as increased cytotoxicity in a chick-chorioallantoic membrane model of human melanoma tumors. In addition, these cells expanded and proliferated at a higher rate, expressed heightened levels of the antiapoptotic molecule Bcl(XL) and were also relatively insensitive to immunosuppression mediated by transforming growth factor-ß, compared to control cells. We also show that 4-1BBL expression on the target cell is essential to 4-1BB-mediated functional improvement. Overall, we conclude that the modification of human T cells with 4-1BB yields enhanced antitumor function which may have important applications in therapies based on the genetic modification of patient lymphocytes.

Determination of the activity of CD134 (OX-40) and CD137 (4-1BB) adhesive molecules by means of flow cytometry in patients with colorectal cancer metastases to the liver.

UNLABELLED: Colorectal carcinoma (CRC) is one of the most common reasons of mortality in patients diagnosed with neoplasms. In nearly 20% of patients with colorectal carcinoma metastatic lesions are diagnosed. In general, survival of patients with metastatic lesions to the liver and other organs is poor. Conventional therapy of colorectal carcinoma is based on the surgical excision of the tumor, chemotherapy, and radiotherapy. THE AIM OF THE STUDY: was to determine the expression of CD134 and CD137 molecules inside the tumor, at the border of the tumor, in the healthy tissue, and peripheral blood, considering patients with colorectal carcinoma metastases to the liver. MATERIAL AND METHODS: The study group comprised 39 patients subject to surgical treatment at the Department of General and Gastroenterological Surgery, due to colorectal carcinoma with liver metastases. CD134 and CD137 adhesive molecule levels were determined inside the tumor, at the border of the tumor, and in the healthy margins of the surgical incision. Additionally, the authors evaluated the peripheral blood level of the above-mentioned molecules on the day of the surgical procedure, and 10 days, thereafter. RESULTS: The mean CD134 levels were the highest inside the tumor, significantly decreasing towards the direction of healthy tissues. The average peripheral blood molecule levels were four-fold higher on the day of the surgical procedure, as compared to values obtained on the tenth postoperative day. This dependency also concerned the remaining statistical measures.The mean CD137 levels showed no significant difference, regardless their location. The authors observed significant, peripheral blood, CD137 level differences, considering the day of the surgical procedure and tenth postoperative period. The mean CD137 peripheral blood level was several times higher on the day of the surgical procedure, as compared to the postoperative period. CONCLUSIONS: The determination of the activity of CD134 and CD137 molecules might create opportunities to plan treatment and predict prognosis in case of colorectal carcinoma. Proper immuno-therapeutic management which is based on the expression of the above-mentioned molecules might help determine the risk of metastases, preventing from their development. In advanced cases treatment of liver metastases might be possible.

Triple expression of B7-1, B7-2 and 4-1BBL enhanced antitumor immune response against mouse H22 hepatocellular carcinoma.

OBJECTIVES: Costimulatory signals are essential for T-cell activation and hence play a very important role in antitumor immunity. B7 and 4-1BBL which belongs to tumor necrosis factor (TNF) family provide costimulatory interaction for T-cell activation and function. This study investigated the role of B7 and 4-1BBL in the amplification of tumor immunity by transduction of the B7-1, B7-2 and 4-1BBL into mouse hepatocellular carcinoma cell line H22. METHODS: The tumorigenicity of H22 variants expressing either B7-1, B7-2 (H22/B7-1/B7-2) or 4-1BBL was compared with an H22 variant expressing B7-1, B7-2 and 4-1BBL (H22/B7-1/B7-2/4-1BBL). The study next investigated whether the combination of B7-1/B7-2 and 4-1BBL cell injection induced cytotoxic T lymphocyte (CTL) response and IL-2/IFN-γ secretion. The immune mechanisms underlying this combination treatment were then analyzed. RESULTS: Syngeneic BALB/c mice injected with H22/B7-1/B7-2/4-1BBL cells that expressed elevated levels of B7-1, B7-2 and 4-1BBL showed a tumor development frequency of 50% compared with 100% in mice injected with the H22 parental line, H22/neo, H22/B7-1/B7-2 and H22/4-1BBL. Mice inoculated with H22 tumor cells expressing B7-1, B7-2 and 4-1BBL developed a strong cytotoxic T lymphocyte response and long-term immunity against wild-type tumor, suggesting a synergistic effect between the B7 and 4-1BBL costimulatory pathways. Results showed that H22/B7-1/B7-2/4-1BBL tumor vaccines probably protect the infiltrating lymphocytes from apoptosis and induce NF-κB activation to improve T-cell-mediated antitumor response. CONCLUSIONS: In this study, the antitumor consequences of using B7-1, B7-2 and 4-1BBL gene transfer have demonstrated the therapeutic potential of gene therapy approach for hepatocellular carcinoma.

The role of soluble and cell-surface expressed 4-1BB ligand in patients with malignant hemopoietic disorders.

The TNFR family member 4-1BB and its ligand 4-1BBL are involved in the costimulation of T-cells and tumor-derived soluble (s)4-1BBL may influence the interaction of malignant cells with the immune system. Here, we report that cell-surface-expressed (c)4-1BBL can be expressed on mononuclear blood cells from patients with acute myeloid leukemia (AML) (n = 35), myelodysplasia (n = 5) or non-Hodgkin lymphoma (n = 11) and can be coexpressed on varying proportions of lymphoid or myeloid malignant cells and on dendritic cells differentiated from AML-blasts. Direct correlations between c- and s4-1BBL were not found in the investigated cases. Up to now expression of 4-1BBL has not been described on primary myeloid malignant cells, but only on malignant cells of lymphoid or solid tumor origin or on tumor cell lines. With our work we further contribute to the understanding of the potential role of c/s4-1BBL in immune reactions and its influence on the interaction of tumor and immunoreactive cells.

Suppression of neointimal hyperplasia after vascular injury by blocking 4-1BB/4-1BB ligand pathway.

AIM: T cell-mediated immunity is involved in the pathogenesis of atherosclerosis and arteriosclerosis. This study examined whether the 4-1BB pathway affects the development of arteriosclerosis after vascular injury. METHODS AND RESULTS: The left or right femoral arteries of adult male mice weighing 22 to 25 g were injured with a straight spring wire. The injured artery was excised 28 days later. Confocal microscopy revealed intense expression of both 4-1BB and 4-1BBL in the developing neointima and media. Similar results were obtained on immunoblotting analysis of lysates of the injured arteries. We gave mice an injection of 100 microg or 200 microg 4-1BB-fused with human immunoglobulin (Ig) every other day over the course of 5 days. As compared with untreated controls (intima/media ratio, 2.13 +/- 0.37, n=10), the intima/media ratio was smaller in mice treated with 200 microg of 4-1BBIg (1.20 +/- 0.30, n=6, p<0.05), but not in mice treated with 100 microg of 4-1BBIg (1.56 +/- 0.27, n=9). CONCLUSIONS: 4-1BB inhibits neointimal hyperplasia after vascular injury. Our findings suggest that 4-1BB is involved in injury-induced neointimal hyperplasia and may be an effective target for the treatment of neointimal hyperplasia.