Anti-GPC3-CAR T cells suppress the growth of tumor cells in patient-derived xenografts of hepatocellular carcinoma

© 2017 Jiang, Jiang, Chen, Lai, Wei, Li, Lin, Wang, Wu, Liang, Liu, Peng, Yu, Weng, Du, Pei, Liu, Yao, Xue and Li. Background: The lack of a general clinic-relevant model for human cancer is a major impediment to the acceleration of novel therapeutic approaches for clinical use. We propose to establish and characterize primary human hepatocellular carcinoma (HCC) xenografts that can be used to evaluate the cytotoxicity of adoptive chimeric antigen receptor (CAR) T cells and accelerate the clinical translation of CAR T cells used in HCC. Methods: Primary HCCs were used to establish the xenografts. The morphology, immunological markers, and gene expression characteristics of xenografts were detected and compared to those of the corresponding primary tumors. CAR T cells were adoptively transplanted into patient-derived xenograft (PDX) models of HCC. The cytotoxicity of CAR T cells in vivo was evaluated. Results: PDX1, PDX2, and PDX3 were established using primary tumors from three individual HCC patients. All three PDXs maintained original tumor characteristics in their morphology, immunological markers, and gene expression. Tumors in PDX1 grew relatively slower than that in PDX2 and PDX3. Glypican 3 (GPC3)-CAR T cells efficiently suppressed tumor growth in PDX3 and impressively eradicated tumor cells from PDX1 and PDX2, in which GPC3 proteins were highly expressed. Conclusion: GPC3-CAR T cells were capable of effectively eliminating tumors in PDX model of HCC. Therefore, GPC3-CAR T cell therapy is a promising candidate for HCC treatment.Link_to_subscribed_fulltex

Targeted calcium influx boosts cytotoxic T lymphocyte function in the tumour microenvironment

Adoptive cell transfer utilizing tumour-targeting cytotoxic T lymphocytes (CTLs) is one of the most effective immunotherapies against haematological malignancies, but significant clinical success has not yet been achieved in solid tumours due in part to the strong immunosuppressive tumour microenvironment. Here, we show that suppression of CTL killing by CD4+CD25+Foxp+ regulatory T cell (Treg) is in part mediated by TGFβ-induced inhibition of inositol trisphosphate (IP3) production, leading to a decrease in T cell receptor (TCR)-dependent intracellular Ca2+ response. Highly selective optical control of Ca2+ signalling in adoptively transferred CTLs enhances T cell activation and IFN-γ production in vitro, leading to a significant reduction in tumour growth in mice. Altogether, our findings indicate that the targeted optogenetic stimulation of intracellular Ca2+ signal allows for the remote control of cytotoxic effector functions of adoptively transferred T cells with outstanding spatial resolution by boosting T cell immune responses at the tumour sites

Peroxiredoxin 3 Is a Redox-Dependent Target of Thiostrepton in Malignant Mesothelioma Cells

Thiostrepton (TS) is a thiazole antibiotic that inhibits expression of FOXM1, an oncogenic transcription factor required for cell cycle progression and resistance to oncogene-induced oxidative stress. The mechanism of action of TS is unclear and strategies that enhance TS activity will improve its therapeutic potential. Analysis of human tumor specimens showed FOXM1 is broadly expressed in malignant mesothelioma (MM), an intractable tumor associated with asbestos exposure. The mechanism of action of TS was investigated in a cell culture model of human MM. As for other tumor cell types, TS inhibited expression of FOXM1 in MM cells in a dose-dependent manner. Suppression of FOXM1 expression and coincidental activation of ERK1/2 by TS were abrogated by pre-incubation of cells with the antioxidant N-acetyl-L-cysteine (NAC), indicating its mechanism of action in MM cells is redox-dependent. Examination of the mitochondrial thioredoxin reductase 2 (TR2)-thioredoxin 2 (TRX2)-peroxiredoxin 3 (PRX3) antioxidant network revealed that TS modifies the electrophoretic mobility of PRX3. Incubation of recombinant human PRX3 with TS in vitro also resulted in PRX3 with altered electrophoretic mobility. The cellular and recombinant species of modified PRX3 were resistant to dithiothreitol and SDS and suppressed by NAC, indicating that TS covalently adducts cysteine residues in PRX3. Reduction of endogenous mitochondrial TRX2 levels by the cationic triphenylmethane gentian violet (GV) promoted modification of PRX3 by TS and significantly enhanced its cytotoxic activity. Our results indicate TS covalently adducts PRX3, thereby disabling a major mitochondrial antioxidant network that counters chronic mitochondrial oxidative stress. Redox-active compounds like GV that modify the TR2/TRX2 network may significantly enhance the efficacy of TS, thereby providing a combinatorial approach for exploiting redox-dependent perturbations in mitochondrial function as a therapeutic approach in mesothelioma

A conceptual analysis of the experience of aloneness as a necessary condition for the making of art

This Thesis tries to show how the experience of aloneness comes in\ud many forms for the artist working in the fine arts. It is argued in\ud the chapter Alone with the Idea Part 1, which concentrates on an\ud analysis of my creative process as the writer of the Thesis, that the\ud experience of being intimate with the idea to the exclusion of all else,\ud is the essential experience of aloneness that artists seek to attain,\ud whether they are children, adolescents or adults. The chapters Solitude\ud and Alone Together explore the ways in which the physical and social\ud dimensions of aloneness are important in facilitating the experience of\ud being alone with the idea. It is suggested that only when a territory\ud has been established and, for the artist working in the company of\ud others, greeting rituals have been enacted can the artist turn his\ud attention to the making of art. The chapter Alone with the Idea Part 2\ud describes how the emotional demands of art have to be reconciled with\ud the emotional demands in an artist's private life. The chapter on\ud Autonomy concludes that the autonomous artist shows an independence of\ud thought and spirit in his art and a capacity to penetrate the depths\ud of his being in order to realise that which is uniquely his own. The\ud chapters Loneliness and Alienation propose that such feeling states\ud can provide the subject matter for art forms; that for those whose life\ud experiences have made them lonely or alienated the making of art can\ud be regenerative; and that art for the dissident adolescent, is a potent\ud form for the needful expression of his condition. The Conclusion\ud describes the art teacher whose capacity to be alone in a general life\ud sense allows students the life—space in which to be alone with their\ud ideas. Verbatim transcripts of recorded conversations held with\ud seven American and Canadian artists in Canada in 1975-76 are\ud included as Appendices

Health: Education, promotion and application in the primary school setting

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Christian principles for urban policy

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