Fas/APO-1 gene transfer for human malignant glioma

• Published in: Cancer research (Chicago, Ill.) Vol. 55; no. 13; pp. 2936 - 2944
• Main Authors: WELLER, M, MALIPIERO, U, RENSING-EHL, A, BARR, P. J, FONTANA, A
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 01-07-1995
• Subjects: Antibodies - administration & dosage; Antigens, Surface - administration & dosage; Antigens, Surface - genetics; Antineoplastic agents; Apoptosis; Base Sequence; Biological and medical sciences; Cycloheximide - pharmacology; Dexamethasone - pharmacology; DNA Primers - chemistry; fas Receptor; Gene Transfer Techniques; Genetic Therapy; Glioma - genetics; Glioma - therapy; Humans; Immunotherapy; In Vitro Techniques; Interferon-gamma - pharmacology; Medical sciences; Membrane Glycoproteins - metabolism; Molecular Sequence Data; Pharmacology. Drug treatments; Tumor Cells, Cultured; Human; Intracranial; Nervous system diseases; Cytotoxicity; Monoclonal antibody; Malignant tumor; In vitro; Mechanism; Genetic transfer; Resistance; Glioma; Cell death; Central nervous system disease; Established cell line; Reversibility; Gene therapy; Tumor cell
• ISSN: 0008-5472; 1538-7445

Human malignant glioma cells are susceptible to apoptosis induced by antibodies to Fas/APO-1, a cytokine receptor protein of the nerve growth factor/tumor necrosis factor receptor superfamily. Here we show that a critical level of cell surface expression of Fas/APO-1 is a prerequisite for induction of glioma cell apoptosis via Fas/APO-1. Although Fas/APO-1 mRNA was expressed in three Fas/APO-1 antibody-resistant glioma cell lines, these cells expressed either little Fas/APO-1 protein (LN-319 and LN-405) or an abnormal Fas/APO-1 protein that was not translocated to the cell membrane and therefore functionally inactive (LN-308). Although all glioma cell lines expressed mRNA for Fas/APO-1-delta TM, a soluble form of Fas/APO-1 lacking the transmembrane domain, none of the cell lines released detectable amounts of soluble Fas/APO-1, a potential endogenous antagonist of Fas/APO-1-mediated glioma cell apoptosis. Stable transfection of three resistant glioma cell lines with a human Fas/APO-1 cDNA expression vector dramatically enhanced cell surface expression of Fas/APO-1 and induced susceptibility to Fas/APO-1 antibody-mediated apoptosis. These data indicate that malignant glioma cells, unlike other tumor cells, uniformly harbor the intracellular cascade required for Fas/APO-1-mediated apoptosis. Low level of Fas/APO-1 expression results from inefficient transcription and translation of the Fas/APO-1 gene or the synthesis of mutant Fas/APO-1 proteins. gamma-Interferon, tumor necrosis factor-alpha, and interleukin 1 beta augmented Fas/APO-1-mediated apoptosis of Fas/APO-1-transfected glioma cells by acting on the subcellular suicidal cascade triggered by Fas/APO-1 activation. Dexamethasone attenuated Fas/APO-1 antibody-induced apoptosis, not only of constitutively Fas/APO-1-positive glioma cells, but also of Fas/APO-1-transfected glioma cells. The antiapoptotic effect of dexamethasone could be overcome by preexposure of the glioma cells to gamma-interferon or by coexposure to Fas/APO-1 antibodies and cycloheximide. Thus, Fas/APO-1 gene transfer and combined immunotherapy using Fas/APO-1 antibodies and cytokines may overcome Fas/APO-1 antibody resistance of Fas/APO-1-negative human malignant glioma cells, which may represent subpopulations within single gliomas or form a separate subgroup of human malignant gliomas.