Thymidine kinase activation of ganciclovir in recurrent malignant gliomas: a gene-marking and neuropathological study

2000 ◽  
Vol 92 (5) ◽  
pp. 804-811 ◽  
Author(s):  
Griffith R. Harsh ◽  
Thomas S. Deisboeck ◽  
David N. Louis ◽  
John Hilton ◽  
Michael Colvin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Enzymatic Activity ◽  
Thymidine Kinase ◽  
Tumor Cells ◽  
Malignant Gliomas ◽  
Retroviral Vectors ◽  
Content Type ◽  
Tumor Bed ◽  
Immunohistochemical Evidence ◽  
Fine Print

Object. The gene therapy paradigm of intratumoral activation of ganciclovir (GCV) following transduction of tumor cells by retroviral vectors bearing the thymidine kinase (tk) gene has produced dramatic remissions of malignant gliomas in animal models. In human trials, although the technique has been deemed safe, little antitumor effect has been demonstrated. To evaluate the basis of this inefficacy in human gliomas, the authors conducted a gene-marking trial involving neuropathological and biochemical studies of treated tumor specimens.Methods. Five patients with malignant recurrent gliomas underwent stereotactic biopsy sampling and intratumoral implantation procedures with three aliquots of 106 vector-producing cells (VPCs) in columns. After 5 days, the tumor was resected and the tumor bed reimplanted with VPCs, and a course of GCV was given. Patients received clinical and radiological follow up for 6 months. Tumor specimens were analyzed neuropathologically and for tk gene expression by anti-TK immunohistochemistry and TK enzymatic activity.Four patients tolerated the treatment well but experienced tumor progression. The other developed an abscess after the second operation and died. Increased TK enzymatic activity was demonstrated in the one tumor specimen analyzed. Immunohistochemical evidence of tk gene expression was limited to VPCs. Transduction of tumor cells was not seen. Viable tumor cells were seen near VPCs containing TK. The lymphocytic immune response was mild.Conclusions. Except for the risk of infection inherent in reoperation, this tk—GCV paradigm was both feasible and safe. Pathological studies indicated that limited dissemination of VPCs and vector from the infusion site and failure to transduce tumor cells with the tk gene are major barriers to efficacy.

Histochemical investigation of nonspecific acid phosphatase in cerebral tumors

1971 ◽  
Vol 34 (6) ◽  
pp. 730-740 ◽  
Author(s):  
Ivanka Lolova ◽  
Ana Ivanova ◽  
Sasho Bojinov
Keyword(s):  
Acid Phosphatase ◽  
Enzymatic Activity ◽  
Tumor Cells ◽  
Enzymatic Reaction ◽  
Histochemical Investigation ◽  
Cerebral Tumor ◽  
Content Type ◽  
Different Types ◽  
Tumor Biopsies ◽  
Fine Print

✓ The activity of acid phosphatase in 144 cerebral tumor biopsies was investigated in an attempt to reveal correlations between enzymatic activity and biological states of the tumor cells. The distribution and intensity of the product of enzymatic reaction in different types of tumors, especially in some rare neoplasms (medulloepithelioma, gangliocytoma), showed evidence of the link between vascularization and the activity of the enzyme.

The effect of thymidine kinase transduction and ganciclovir therapy on tumor vasculature and growth of 9L gliomas in rats

1994 ◽  
Vol 81 (2) ◽  
pp. 256-260 ◽  
Author(s):  
Zvi Ram ◽  
Stuart Walbridge ◽  
Thomas Shawker ◽  
Kenneth W. Culver ◽  
R. Michael Blaese ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Thymidine Kinase ◽  
Tumor Cells ◽  
Tumor Regression ◽  
Tumor Vasculature ◽  
Cell Injection ◽  
Content Type ◽  
Ganciclovir Therapy ◽  
Viral Particles ◽  
Fine Print

✓ Eradication of malignant brain tumors by in situ intratumoral, retrovirally mediated transfer of the herpes simplex virus thymidine kinase (HSVtk) gene, which sensitizes the tumor cells to ganciclovir, has recently been demonstrated in animal models. The observation that tumors studied in vitro and in animals can be completely eliminated despite only partial transduction of the tumor suggests a bystander mechanism that affects nontransduced tumor cells. Such a bystander effect is not completely understood and may represent a combination of several factors that lead to tumor eradication. Endothelial cells of the tumor blood vessels were shown to occasionally integrate the retroviral vector and thus become sensitized to ganciclovir. In the presence of vector-producer cells, which continuously release infectious viral particles, diffuse multifocal hemorrhages occurred during ganciclovir administration. When the tumor was composed of cells that had been transduced with the thymidine kinase gene before inoculation, no infectious viral particles were present within the tumor, no transduction of endothelial cells occurred, and no hemorrhages were observed during ganciclovir therapy. These observations suggest that tumor regression may be due, in part, to destruction of in vivo HSVtk-transduced endothelial cells after exposure to ganciclovir, resulting in tumor ischemia as one possible bystander mechanism. The authors investigated this hypothesis using the subcutaneous 9L gliosarcoma tumor model in Fischer rats. The tumors were evaluated with Doppler color-flow and ultrasound imaging during the various phases of the study. Twenty rats received intratumoral injections of HSVtk retroviral vector-producer cells (6 × 107 cells/ml) 21 days after bilateral flank tumor inoculation. Ten rats were subsequently treated with intraperitoneal ganciclovir (15 mg/kg/ml twice a day) for 14 days starting on Day 7 after producer cell injection; 10 control rats received intraperitoneal saline injections (1 ml twice a day) instead of ganciclovir. Ultrasound and flow images were obtained before cell injection, before and during ganciclovir or saline administration, and after cessation of treatment. The number, location, and ultrasonographic appearance of tumor vessels and the tumor volumes were recorded. The number of blood vessels in the tumors increased over time in both groups before treatment. Intratumoral cell injection without ganciclovir administration did not influence tumor growth or intratumoral vasculature. However, tumor vasculature decreased after initiation of ganciclovir therapy in the HSVtk-transduced tumors (p < 0.05). Early patchy or diffuse necrotic changes associated with ultrasonographic evidence of scattered intratumoral hemorrhage occurred in tumors treated with ganciclovir. Reduction of the tumor blood supply may be an important feature of HSVtk transduction-mediated tumor regression and may, at least partially, account for the degree of tumor destruction that occurs despite the lack of transduction of all tumor cells.

Experimental appraisal of the lack of antitumor natural killer cell—mediated immunosurveillance in response to lymphomas growing in the mouse brain

2003 ◽  
Vol 98 (3) ◽  
pp. 599-606 ◽  
Author(s):  
Toshiki Yamasaki ◽  
Kouzo Moritake ◽  
George Klein
Keyword(s):  
Gene Expression ◽  
Nk Cells ◽  
Cell Line ◽  
Tumor Cells ◽  
Mhc Class I ◽  
Nk Cell ◽  
Class I ◽  
Content Type ◽  
The Brain ◽  
Fine Print

Object. Natural killer (NK) cell—mediated immunosurveillance in the brain is currently obscure, in contrast with the intracerebral immune reaction of cytotoxic T lymphocytes (CTLs) to tumor cells. The goal of this study, in which a controlled tumor model was used, was to investigate a relationship between NK cells and major histocompatibility complex (MHC) Class I gene expression in intracerebral tumor—bearing hosts. Methods. A matched set of two cloned tumor cell lines (lymphoma+ and lymphoma−), which differ only in MHC Class I gene expression, was established from the parental YAC-1 cell line (a target widely accepted as being sensitive to murine NK cells). An in vivo rapid elimination assay (REA) was performed using tumor cells labeled with [125I] 5-iodo-2-deoxyuridine to evaluate intracerebral NK cell—mediated defense immunity. There was no difference in the in vitro growth rate and c-myc gene expression between lymphoma+ and lymphoma− cells. An in vitro cytotoxicity assay showed that the lymphoma+ cell line was sensitive to MHC Class I—restricted CTL-mediated lysis, whereas the lymphoma− line was refractory to it. Both were susceptible to NK cell—mediated lysis, comparable to the level shown by YAC-1 cells. Flow cytometry revealed that lymphoma+ reacted positively for cell-surface MHC Class I molecules, whereas lymphoma− had no reaction. Four- to 72-hour REAs, performed using either cell line, disclosed no clearance of radiolabeled tumor cells from the brain in independent groups of untreated and T cell—depleted mice; this contrasted with eradication of radioactivity from the lungs. In NK cell—depleted mice, however, there was no elimination of radiolabeled tumor cells from the brain or lungs. The MHC Class I expression on lymphoma+ cells was enhanced after intracerebral inoculation, rendering them less sensitive to NK cells. By contrast, lymphoma− cells remained negative for cell-surface MHC expression, being sensitive to NK cells and refractory to CTLs after intracerebral inoculation. These results indicate the absence of NK cell—mediated lytic activity in the brain. This allows even NK cell—sensitive tumor cells to escape intracerebral immunosurveillance. Conclusions. These experiments have refined the information that the brain may lack NK cell—mediated defense immunity against intracerebrally growing tumors, representing a characteristic aspect of this immunologically privileged organ.

Combined cimetidine and temozolomide, compared with temozolomide alone: significant increases in survival in nude mice bearing U373 human glioblastoma multiforme orthotopic xenografts

2005 ◽  
Vol 102 (4) ◽  
pp. 706-714 ◽  
Author(s):  
Florence Lefranc ◽  
Syril James ◽  
Isabelle Camby ◽  
Jean-François Gaussin ◽  
Francis Darro ◽  
...  
Keyword(s):  
Glioblastoma Multiforme ◽  
Tumor Cells ◽  
Nude Mice ◽  
Malignant Gliomas ◽  
Glioma Cells ◽  
Computer Assisted ◽  
Human Glioblastoma ◽  
Content Type ◽  
Human Glioblastoma Multiforme ◽  
Fine Print

Object. Malignant gliomas consist of both heterogeneous proliferating and migrating cell subpopulations, with migrating glioma cells exhibiting less sensitivity to antiproliferative or proapoptotic drugs than proliferative cells. Therefore, the authors combined cimetidine, an antiinflammatory agent already proven to act against migrating epithelial cancer cells, with temozolomide to determine whether the combination induces antitumor activities in experimental orthotopic human gliomas compared with the effects of temozolomide alone. Methods. Cimetidine added to temozolomide compared with temozolomide alone induced survival benefits in nude mice with U373 human glioblastoma multiforme (GBM) cells orthotopically xenografted in the brain. Computer-assisted phase-contrast microscopy analyses of 9L rat and U373 human GBM cells showed that cimetidine significantly decreased the migration levels of these tumor cells in vitro at concentrations at which tumor growth levels were not modified (as revealed on monotetrazolium colorimetric assay). Computer-assisted microscope analyses of neoglycoconjugate-based glycohistochemical staining profiles of 9L gliosarcomas grown in vivo revealed that cimetidine significantly decreased expression levels of endogenous receptors for fucose and, to a lesser extent, for N-acetyl-lactosamine moieties. Endogenous receptors of this specificity are known to play important roles in adhesion and migration processes of brain tumor cells. Conclusions. Cimetidine, acting as an antiadhesive and therefore an antimigratory agent for glioma cells, could be added in complement to the cytotoxic temozolomide compound to combat both migrating and proliferating cells in GBM.

A comparative study on the pharmacokinetics and biodistribution of boronated porphyrin (BOPP) and sulfhydryl boron hydride (BSH) in the RG2 rat glioma model

1995 ◽  
Vol 83 (1) ◽  
pp. 86-92 ◽  
Author(s):  
Crister P. Ceberg ◽  
Arne Brun ◽  
Stephen B. Kahl ◽  
Myoung Seo Koo ◽  
Bertil R. R. Persson ◽  
...  
Keyword(s):  
Body Weight ◽  
Tumor Cells ◽  
Brain Tissue ◽  
Malignant Gliomas ◽  
Normal Brain ◽  
Boron Hydride ◽  
Content Type ◽  
Rat Glioma ◽  
Glioma Model ◽  
Fine Print

✓ Boron neutron capture therapy is a treatment modality for cancer that depends on the specific uptake of boron by the tumor cells. The infiltrative growth of malignant gliomas requires that boron reach and accumulate in migrating cells outside the margin of the tumor; thus, it is important that the biodistribution of new boron compounds is also studied in the surrounding healthy brain tissue. This study is undertaken in the present work, in which the biodistribution and pharmacokinetics of sulfhydryl boron hydride (BSH) and boronated porphyrin (BOPP) in the RG2 rat glioma model are investigated. This model mimics the characteristics of human glioma with cells migrating into the surrounding brain. The animals were infused intravenously with either BSH (25 µg or 175 µg of boron per gram of body weight) or BOPP (12 µg of boron per gram body weight). For the low dose of BSH, the maximum tumor—boron content was 8 ppm at approximately 9 hours after the infusion with a tumor-to-blood ratio of 0.6. At the higher dose, the corresponding figures were 15 ppm after 12 hours with a tumor-to-blood ratio of 0.5. For BOPP, a tumor—boron concentration of 81 ppm was achieved 24 hours after the infusion and sustained in that range for at least 72 hours. The tumor-to-blood ratio at 24 hours was slightly above 6, but continued to increase as the blood was cleared. These results indicate that both compounds are spread into the normal brain tissue following the same pathways as the migrating tumor cells and in this way can be taken up even in distant tumor cell foci.

Presence and significance of NK cells in glioblastomas

1989 ◽  
Vol 70 (5) ◽  
pp. 728-731 ◽  
Author(s):  
Jesús Vaquero ◽  
Santiago Coca ◽  
Santiago Oya ◽  
Roberto Martínez ◽  
Josefa Ramiro ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Nk Cells ◽  
Survival Time ◽  
Tumor Cells ◽  
Lymphocytic Infiltration ◽  
Surface Marker ◽  
Content Type ◽  
Hematoxylin And Eosin ◽  
Hematoxylin And Eosin Staining ◽  
Fine Print

✓ A monoclonal antibody against the surface marker IOT-10 of natural killer (NK) cells was used to investigate the presence of these cells in a series of 25 glioblastomas. In 40% of the tumors, IOT-10-positive NK cells were found in small numbers scattered among the tumor cells. The presence of IOT-10-positive NK cells was not related to the degree of lymphocytic infiltration in the tumor as demonstrated by hematoxylin and eosin staining, nor did it appear to influence the survival time of the patients studied.

Thymidine kinase-mediated killing of rat brain tumors

1993 ◽  
Vol 79 (5) ◽  
pp. 729-735 ◽  
Author(s):  
David Barba ◽  
Joseph Hardin ◽  
Jasodhara Ray ◽  
Fred H. Gage
Keyword(s):  
Gene Therapy ◽  
Brain Tumors ◽  
Tumor Cells ◽  
Wild Type ◽  
Cns Disorders ◽  
Content Type ◽  
Potential Applications ◽  
Ganciclovir Treatment ◽  
The Brain ◽  
Fine Print

✓ Gene therapy has many potential applications in central nervous system (CNS) disorders, including the selective killing of tumor cells in the brain. A rat brain tumor model was used to test the herpes simplex virus (HSV)-thymidine kinase (TK) gene for its ability to selectively kill C6 and 9L tumor cells in the brain following systemic administration of the nucleoside analog ganciclovir. The HSV-TK gene was introduced in vitro into tumor cells (C6-TK and 9L-TK), then these modified tumor cells were evaluated for their sensitivity to cell killing by ganciclovir. In a dose-response assay, both C6-TK and 9L-TK cells were 100 times more sensitive to killing by ganciclovir (median lethal dose: C6-TK, 0.1 µg ganciclovir/ml; C6, 5.0 µg ganciclovir/ml) than unmodified wild-type tumor cells or cultured fibroblasts. In vivo studies confirmed the ability of intraperitoneal ganciclovir administration to kill established brain tumors in rats as quantified by both stereological assessment of brain tumor volumes and studies of animal survival over 90 days. Rats with brain tumors established by intracerebral injection of wild-type or HSV-TK modified tumor cells or by a combination of wild-type and HSV-TK-modified cells were studied with and without ganciclovir treatments. Stereological methods determined that ganciclovir treatment eliminated tumors composed of HSV-TK-modified cells while control tumors grew as expected (p < 0.001). In survival studies, all 10 rats with 9L-TK tumors treated with ganciclovir survived 90 days while all untreated rats died within 25 days. Curiously, tumors composed of combinations of 9L and 9L-TK cells could be eliminated by ganciclovir treatments even when only one-half of the tumor cells carried the HSV-TK gene. While not completely understood, this additional tumor cell killing appears to be both tumor selective and local in nature. It is concluded that HSV-TK gene therapy with ganciclovir treatment does selectively kill tumor cells in the brain and has many potential applications in CNS disorders, including the treatment of cancer.

The use of tissue culture in differentiation between angioblastic meningioma and hemangiopericytoma

1971 ◽  
Vol 34 (3) ◽  
pp. 341-348 ◽  
Author(s):  
Jans Muller ◽  
John Mealey
Keyword(s):  
Tissue Culture ◽  
Tumor Cells ◽  
Cerebellopontine Angle ◽  
Content Type ◽  
Angioblastic Meningioma ◽  
Fine Print

✓ A solid, extrinsic hemangiopericytoma of the cerebellopontine angle was studied histologically and by means of tissue culture. The explanted tumor cells formed classic meningiomatous whorls indicative of the meningeal derivation of this neoplasm. Whorls were entirely absent in the histological preparations, however. The cases reported under the diagnosis of intracranial hemangiopericytoma and angioblastic meningioma have been reviewed; no valid histological distinction between these two types could be made.

The deposition of Hg203-chlormerodrin in experimental brain tumors

1971 ◽  
Vol 35 (3) ◽  
pp. 303-308 ◽  
Author(s):  
Tatsuya Kobayashi ◽  
Louis Bakay ◽  
Joseph C. Lee
Keyword(s):  
Brain Tumors ◽  
Tumor Cells ◽  
Normal Brain ◽  
Intracranial Tumors ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography ◽  
Content Type ◽  
Meningeal Tumors ◽  
Vacuolar System ◽  
Fine Print

✓ The deposition of Hg203-chlormerodrin was studied in intracranial tumors in mice induced by implantation of 20-methyl cholanthrene by tissue assay, as well as light microscopic and electron microscopic autoradiography. The investigations were carried out in astrocytomas, glioblastomas, and meningeal tumors. The chlormerodrin content of the tumors exceeded that of normal brain with a significant tumor/brain ratio ranging from 5.8 to 22.5. It was found that the chlormerodrin molecule becomes rapidly incorporated in the tumor cells, with a preference for that portion of the cytoplasm associated with the vacuolar system.

Administration of interleukin-12 and -18 enhancing the antitumor immunity of genetically modified dendritic cells that had been pulsed with Semliki Forest virus—mediated tumor complementary DNA

2002 ◽  
Vol 97 (5) ◽  
pp. 1184-1190 ◽  
Author(s):  
Ryuya Yamanaka ◽  
Naoki Yajima ◽  
Naoto Tsuchiya ◽  
Junpei Honma ◽  
Ryuichi Tanaka ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Antitumor Immunity ◽  
Semliki Forest Virus ◽  
Malignant Gliomas ◽  
Glioma Cells ◽  
Antitumor Effects ◽  
Content Type ◽  
Immunogene Therapy ◽  
Fine Print

Object. Immunogene therapy for malignant gliomas was further investigated in this study to improve its therapeutic efficacy. Methods. Dendritic cells (DCs) were isolated from bone marrow and pulsed with phosphate-buffered saline or Semliki Forest virus (SFV)—mediated 203 glioma complementary (c)DNA with or without systemic administration of interleukin (IL)-12 and IL-18 to treat mice bearing the 203 glioma. To study the immune mechanisms involved in tumor regression, the authors investigated tumor growth of an implanted 203 glioma model in T cell subset—depleted mice and in interferon (IFN) γ—neutralized mice. To examine the protective immunity produced by tumor inoculation, a repeated challenge of 203 glioma cells was given by injecting the cells into the left thighs of surviving mice and the growth of these cells was monitored. The authors demonstrated that the combined administration of SFV-cDNA, IL-12, and IL-18 produced significant antitumor effects against the growth of murine glioma cells in vivo and also can induce specific antitumor immunity. The synergic effects of the combination of SFV-cDNA, IL-12, and IL-18 in vivo were also observed to coincide with markedly augmented IFNγ production. The antitumor effects of this combined therapy are mediated by CD4+ and CD8+ T cells and by NK cells. These results indicate that the use of IL-18 and IL-12 in DC-based immunotherapy for malignant glioma is beneficial. Conclusions. Immunogene therapy combined with DC therapy, IL-12, and IL-18 may be an excellent candidate in the development of a new treatment protocol. The self-replicating SFV system may therefore provide a novel approach for the treatment of malignant gliomas.

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