RESUMEN
Fulvestrant ('Faslodex') is a new oestrogen receptor (ER) antagonist with no agonist effects. This report describes the experience of a single centre including 126 postmenopausal women with advanced breast cancer (ABC) in a fulvestrant Compassionate Use Programme. All patients had previously received endocrine treatment for early or ABC. Patients received fulvestrant as first- (n=7), second- (n=51), third- (n=50) or fourth-line endocrine therapy (n=18) for ABC (median duration of treatment: 4 months [range 3-27(+) months], follow-up: 13 months [range 1-38(+) months]). Twelve patients had partial responses (PR) and 43 patients experienced stable disease (SD) > or = 6 months (objective response rate: 9.5%; clinical benefit [CB] rate: 43.6%). Ten of 12 patients with a PR had HER2-negative tumours, and 9/12 had ER-positive and progesterone receptor (PgR)-positive disease (two patients had unknown HER2 status and one had unknown ER and PgR status). Nine of the 18 patients with HER2-positive tumours experienced CB with fulvestrant. Although CB rates were similar when fulvestrant was given as first- to fourth-line endocrine treatment, the proportion of those experiencing CB who had a PR appeared to decrease when fulvestrant was used later in the sequence. Fulvestrant was well tolerated; six patients experienced adverse events (all grade I/II). These data demonstrate that fulvestrant is an effective and well-tolerated therapy for patients with ABC progressing on prior therapies.
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Antineoplásicos Hormonales/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Estradiol/análogos & derivados , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos Hormonales/efectos adversos , Progresión de la Enfermedad , Estradiol/efectos adversos , Estradiol/uso terapéutico , Femenino , Fulvestrant , Genes erbB-2 , Humanos , Persona de Mediana Edad , Posmenopausia , Receptores de Estrógenos/metabolismo , Resultado del TratamientoRESUMEN
Gemcitabine is a chemotherapeutic agent with proven antitumor effects in pancreatic and non-small cell lung cancer; however, studies establishing the definite significance in other solid tumors are still in progress. We herein present three female patients with advanced breast cancer who received gemcitabine as salvage chemotherapy. Gemcitabine at a dose of 1250 mg/m2 was scheduled for days 1, 8 and 15 with a subsequent rest for 1 week. However, within 1 week after the very first administration of gemcitabine myelotoxicity WHO grade IV occurred in all patients, leading to discontinuation of therapy. In two patients this gemcitabine-induced hematotoxicity could be overcome by means of vigorous supportive care, but one patient died after cerebral bleeding due to severe thrombocytopenia. We conclude that gemcitabine in heavily pretreated breast cancer patients should only be used with extreme caution with special focus on platelet counts until solid data from clinical studies for doses and schedules are available.
Asunto(s)
Antimetabolitos Antineoplásicos/efectos adversos , Neoplasias de la Mama/tratamiento farmacológico , Desoxicitidina/efectos adversos , Células Mieloides/efectos de los fármacos , Adulto , Neoplasias de la Mama/patología , Neoplasias de la Mama/secundario , Desoxicitidina/análogos & derivados , Femenino , Pruebas Hematológicas , Humanos , Masculino , Persona de Mediana Edad , GemcitabinaRESUMEN
PURPOSE: Isolated limb or liver perfusion with tumor necrosis factor (TNF) and melphalan results in regression of advanced cancers in the majority of treated patients. However, the contribution of TNF to the efficacy of isolation perfusion with melphalan has not been demonstrated conclusively in random assignment trials. Furthermore, TNF is an inflammatory cytokine and may be associated with significant systemic and regional toxicity. This study was conducted to characterize the toxicity and secondary cytokine production attributable to TNF by comparing these parameters in patients undergoing isolated hepatic perfusion (IHP) using melphalan with or without TNF. EXPERIMENTAL DESIGN: Thirty-two patients with unresectable colorectal cancer confined to the liver underwent a 60-min hyperthermic IHP using 1.5 mg/kg melphalan alone (n = 17) or with 1.0 mg of TNF (n = 15) with inflow via the gastroduodenal artery and outflow via an isolated segment of inferior vena cava. Complete vascular isolation was confirmed using the I-131 radiolabeled albumin-monitoring technique. Post-IHP parameters of hepatic and systemic toxicity and cytokine levels [TNF, interleukin (IL)-6 and IL-8] in perfusate and serum were measured. RESULTS: Levels of IL-6 and IL-8 in perfusate at the end of the 60-min IHP were significantly higher in TNF-treated patients (P < or = 0.001). Peak systemic IL-6 and IL-8 levels post-IHP were also significantly higher in TNF-treated compared with non-TNF-treated patients (P < 0.0001) by 28- and 268-fold, respectively. The peak levels of these cytokines were associated with significantly lower systolic blood pressure and higher heart rate and mean pulmonary artery blood pressure in TNF-treated patients during the first 48 h post-IHP (P < or = 0.03). Serum bilirubin levels were significantly higher (P = 0.017) and platelets lower (P = 0.03) in TNF-treated compared with non-TNF-treated patients. However, elevations in aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were not significantly different between groups and returned toward baseline within 1 week after IHP. CONCLUSIONS: Addition of TNF to melphalan during IHP results in significant differences in post-IHP production of IL-6 and IL-8 with associated changes in mean arterial blood pressure and greater regional toxicity, as reflected in higher levels of serum bilirubin. However, these measurable differences were transient and did not appear to be of major clinical consequence. Prior to its routine use, the benefit of TNF in isolation perfusion should be demonstrated in random assignment trials.
Asunto(s)
Neoplasias Colorrectales/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Hígado/efectos de los fármacos , Factor de Necrosis Tumoral alfa/toxicidad , Adulto , Anciano , Análisis de Varianza , Antineoplásicos Alquilantes/uso terapéutico , Neoplasias Colorrectales/tratamiento farmacológico , Femenino , Humanos , Hígado/metabolismo , Masculino , Melfalán/uso terapéutico , Persona de Mediana Edad , PerfusiónRESUMEN
Endothelial monocyte activating polypeptide-II (EMAP-II) is an inflammatory cytokine known to have a role in neutrophil and macrophage chemotaxis and in apoptosis. It is a tumour-derived cytokine that sensitizes tumour vasculature to the effects of systemic TNF. In order to gain insight into the mechanism by which EMAP-II sensitizes vessels to TNF, we focused on its effects on TNF receptor expression. In human umbilical vein endothelial cells (HUVEC), TNF-R1 mRNA is increased four-fold following incubation with recombinant EMAP-II. Conditioned media from cell lines known to produce high levels of EMAP-II upregulated TNF-R1 but not TNF-R2 by up to twenty-fold compared to media controls and low expressing cell lines; this effect was blocked by anti-EMAP-II antibody. Recombinant EMAP-II upregulated TNF-R1 expression by approximately six-fold. Analysis of HUVEC lysates by ELISA showed increased expression of TNF-R1 within 2 h; TNF-R2 expression was unaffected by recombinant EMAP-II. Finally, immunohistochemistry of human melanomas in vivo showed that TNF-R1 staining is increased on the vessels of tumours known to express high levels of EMAP-II compared to low EMAP-II expressing tumours. These results suggest that EMAP-II upregulates TNF-R1 expression by endothelial cells both in vitro and in vivo. This induction of TNF-R1 expression may be the mechanism by which EMAP-II sensitizes tumour endothelium to the effects of TNF leading to haemorrhagic necrosis.
Asunto(s)
Antígenos CD/genética , Citocinas , Proteínas de Neoplasias/metabolismo , Proteínas de Unión al ARN/metabolismo , Receptores del Factor de Necrosis Tumoral/genética , Regulación hacia Arriba , Animales , Antígenos CD/biosíntesis , Endotelio Vascular/citología , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Citometría de Flujo/métodos , Fluorescencia , Humanos , Líquido Intracelular , Ratones , Ratones Endogámicos C3H , Ratones Desnudos , Proteínas de Neoplasias/genética , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/genética , Receptores del Factor de Necrosis Tumoral/biosíntesis , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis Tumoral , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Células Tumorales CultivadasRESUMEN
BACKGROUND: Endostatin is a potent angiogenesis inhibitor, which is currently being used in Phase I trials as an antitumor agent. The purpose of this study was to determine whether endostatin has an effect on wound healing in a murine model. MATERIALS AND METHODS: The function of endostatin was confirmed using a human microvascular endothelial cell (HMVEC) proliferation assay in which cells are treated for 4 days with growth media plus or minus endostatin. Full-thickness incisions were made on the dorsum of athymic nude mice and closed primarily with skin staples. PVA sponges were implanted in some wounds to determine vascular ingrowth. Subsequently, mice were treated with recombinant human endostatin at 20 mg/kg/day or 50 mg/kg/dose BID versus control for a total of 14 days. On Days 2, 4, 8, 12, and 16, three mice per group had serum samples drawn and were sacrificed. Perpendicular breaking strength (N) was determined using an Instron 5540 tensometer. Wound strength was determined by dividing breaking strength by wound area (N/cm(2)). Vascular density in sponges was determined using CD31 immunohistochemistry. Serum endostatin concentrations were determined using a commercially available ELISA kit. RESULTS: Endostatin caused a significant reduction of endothelial cell proliferation after 4 days compared to media alone (72%, P = 0.031). At all time points tested, there was no statistical difference in the wound-breaking strength between endostatin and control-treated mice at either the low or high dose. Serum endostatin levels were consistently 10-fold higher in endostatin-treated mice than in controls. No differences in vascular density were seen in endostatin versus control-treated mice as determined by CD31 immunohistochemistry of PVA sponges. CONCLUSION: Therapy with human endostatin does not induce a significant decrease in breaking strength of cutaneous wounds in mice.
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Antineoplásicos/farmacología , Colágeno/farmacología , Neovascularización Fisiológica/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Piel/lesiones , Cicatrización de Heridas/efectos de los fármacos , Animales , División Celular/efectos de los fármacos , Endostatinas , Endotelio Vascular/citología , Femenino , Humanos , Ratones , Ratones Desnudos , Proteínas Recombinantes/farmacología , Piel/citología , Suturas , Resistencia a la TracciónRESUMEN
The application of hyperthermia (HT) and tumour necrosis factor alpha (TNF) in isolation perfusion of the limb or liver results in regression of advanced cancers confined to these regions of the body in most patients and are thought to exert anti-tumour effects primarily on tumour neovasculature. However, the individual contribution of either treatment factor on endothelial cells (EC) are not known. In this study, we investigated the in vitro effects of moderate and severe HT on human umbilical vein EC (HUVEC) with and without TNF in clinically relevant doses. HUVEC were exposed to normothermia (37 degrees C) or moderate (39 degrees C) and severe (41 degrees C) HT for 90 or 180 min with or without TNF (1 microg/ml). Cell viability, cytokine secretion (IL-6, IL-8, VEGF, ICAM-1, VCAM-1, RANTES, E-selectin, P-selectin, L-selectin, and PECAM-1), and induction of procoagulant activity as reflected in tissue factor (TF) production were assessed at the end of the treatment period and at several time points thereafter. Neither HT nor TNF exerted significant cytotoxic effects on EC at the doses and temperatures used. HT resulted in increased production of PECAM-1 with little or no additional effect when combined with TNF. TNF caused increased secretion of IL-6, IL-8, ICAM-1, and VCAM-1 with little or no additional effect from HT. Increased E-selectin and RANTES levels were observed with TNF and HT only at 24 h after treatment. HT and TNF had mainly antagonistic effects on VEGF secretion with HT causing primarily decreased production and TNF causing increased VEGF secretion under all temperatures. Most notably, there was a rapid, prolonged and synergistic peak increase in procoagulant activity when TNF and HT were used in combination compared to TNF or HT treatment alone. These results indicate that TNF and HT exert primarily independent effects on inflammatory cytokine production in EC but synergistically increase procoagulant activity as reflected in TF production. These data provide a possible mechanism for the thrombotic effects in tumour neovasculature seen following isolation perfusion with these agents and provide a rationale for their combined use in this treatment setting.
Asunto(s)
Coagulantes/metabolismo , Endotelio Vascular/metabolismo , Hipertermia Inducida , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Tromboplastina/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Quimiocina CCL5/metabolismo , Selectina E/metabolismo , Factores de Crecimiento Endotelial/metabolismo , Endotelio Vascular/citología , Endotelio Vascular/inmunología , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Selectina L/metabolismo , Linfocinas/metabolismo , Melfalán/farmacología , Selectina-P/metabolismo , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Molécula 1 de Adhesión Celular Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
OBJECTIVE: Tumor infiltrating lymphocytes (TILs) stimulated with interleukin-2 (IL-2) ex vivo have been successfully used therapeutically in some cancer patients, but their potency in eliciting an effective anti-tumor response is variable. We have tried to augment killing activity of tumor infiltrating lymphocytes derived from hepato-cellular carcinoma (HCC) using autologous monocytes derived dendritic cells. METHODS: Tumor infiltrating lymphocytes (TILs) from 6 patient with hepatocellular carcinoma were isolated and the phenotype were further characterized. From the same patients, autologous dendritic cells were generated from CD14+ monocytes that were cultured for 6 days in the presence of granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4). Those professional antigen presenting cells were pulsed with whole autologous hepatoma tumor lysates (pDC). TILs were cocultured with pDC or unpulsed DC. To assess the cytotoxic potency of TILs, the ability to lyse the tumor cell targets K652, Daudi and an allogeneic HCC celline was determined in a standard cytotoxic assay. RESULTS: Tumor cells targets in vitro are poorly lysed by tumor infiltrating lymphocytes indicating T-cell hyporesponsiveness. In contrast, the killing activity of HCC derived TILs against Daudi (9.15% +/- 7.5) and allogeneic HCC tumor target (18.2% +/- 9.2) could be significantly augmented when stimulated with pDC (Daudi: 38% +/- 6.8 and allogeneic HCC: 55% +/- 10). The killing activity of TILs against K562 was unaffected by pDC. CONCLUSION: The low cytotoxic activity profile of HCC derived TILs in vitro can be increased by tumor lysate pulsed dendritic cells and may therefore be more effective in vivo when used for adoptive immunotherapy.
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Antígenos de Neoplasias/inmunología , Carcinoma Hepatocelular/inmunología , Células Dendríticas/inmunología , Neoplasias Hepáticas/inmunología , Activación de Linfocitos/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Linfocitos T Citotóxicos/inmunología , Humanos , Inmunofenotipificación/métodos , Vacunación/métodosRESUMEN
BACKGROUND: Hyperthermic isolated hepatic perfusion (IHP) has been shown to cause significant regression of advanced unresectable liver metastases in patients. Although there are different agents and treatment modalities used in IHP, the contribution of perfusion hyperthermia is unknown. PURPOSE: A large animal model of unresectable liver metastases and a technical standard for IHP in this model were established. This model was used to assess the effects of hyperthermia on vascular permeability of tumors and normal liver tissue during IHP. METHODS: Sixty-five New Zealand White rabbits were used in a series of experiments. Disseminated liver tumors were established by direct injection of 1 x 10(6) VX-2 cells into the portal vein by laparotomy in anesthetized animals. Several surgical perfusion techniques were explored to determine a reliable and reproducible IHP model. Vascular permeability in tumor versus liver was then assessed with Evan's Blue labeled bovine albumin under normothermic (tissue temperature 36.5 degrees C +/- 0.5 degree C), moderate hyperthermic (39 degrees C +/- 0.5 degree C), or severe hyperthermic (41 degrees C +/- 0.5 degree C) conditions. RESULTS: Tumor model and perfusion techniques were successfully established with inflow through the portal vein and outflow through an isolated segment of the inferior vena cava. A gravity driven perfusion circuit with stable perfusion parameters and complete vascular isolation was used. Vascular permeability was higher in tumor than in normal tissues (P = .03) at all time points during IHP. Hyperthermia resulted in a significant (up to 5-fold) increase in permeability of neovasculature; when severe hyperthermia was used, tumor vascular permeability was increased even more than normal liver permeability (P = .01). CONCLUSIONS: The VX-2/New Zealand White rabbit system can be used as a reproducible large-animal model for IHP of unresectable liver metastases. It can be used to characterize the contribution and mechanism of action of different treatment parameters used in IHP. Hyperthermia preferentially increases vascular permeability in tumors compared with liver tissue in a dose-dependent fashion, thus providing a mechanism for its presumed benefit during isolated organ perfusion.
Asunto(s)
Permeabilidad Capilar , Hipertermia Inducida , Circulación Hepática , Neoplasias Hepáticas Experimentales/irrigación sanguínea , Neoplasias Hepáticas Experimentales/secundario , Neovascularización Patológica/metabolismo , Animales , Vasos Sanguíneos/metabolismo , Bovinos/sangre , Femenino , Hipertermia Inducida/métodos , Perfusión , Conejos , Valores de ReferenciaRESUMEN
BACKGROUND: Several approaches to gene therapy for cancer have yielded promising results in rodent models. The translation of these results to the clinical realm has been delayed by the lack of tumor models in large animals. We investigated the pattern of transgene (i. e., foreign or introduced gene) expression and virus vector elimination after systemic gene delivery using a thymidine kinase-negative vaccinia virus in a rabbit model of disseminated liver metastases. METHODS: VX-2 rabbit carcinoma cells were maintained by serial transplantation in the thigh muscles of New Zealand White rabbits, and disseminated liver metastases were established by direct injection of tumor cells into the portal vein of the animals. Different doses of a recombinant thymidine kinase-negative vaccinia virus vector encoding the firefly luciferase reporter gene (i.e., transgene) were injected into tumor-bearing rabbits. Transgene activity in tumors and other organs was measured at multiple time points thereafter. The pattern of development of antibodies against the vaccinia virus vector was also examined. Two-tailed Student's paired t test was used for comparisons of transgene activity. RESULTS: Transgene expression was increased in tumors by at least 16-fold in comparison with expression in other tissues by day 4 after vector injection (all P<. 001) and was maintained for approximately 1 week, providing evidence of tumor-specific gene delivery in this model. Rapid elimination of the circulating vector by the host immune system was observed. Anti-vector antibodies were detectable in serum as early as day 6 and were maintained for more than 3 months. CONCLUSIONS: Tumor-specific gene delivery is possible after systemic injection of a thymidine kinase-negative vaccinia virus vector in a model of rabbit liver metastases. Although the period of transgene expression appears limited because of a rapid immune response, the therapeutic window might be sufficient for an enzyme/prodrug gene therapy approach in clinical application.
Asunto(s)
Expresión Génica , Terapia Genética/métodos , Vectores Genéticos , Neoplasias Hepáticas Experimentales/genética , Neoplasias Hepáticas Experimentales/terapia , Transgenes/genética , Virus Vaccinia/genética , Animales , Anticuerpos Antivirales/sangre , Línea Celular , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Genes de Insecto/genética , Genes Reporteros/genética , Genes Virales/genética , Vectores Genéticos/uso terapéutico , Haplorrinos , Células HeLa , Humanos , Neoplasias Hepáticas Experimentales/secundario , Luciferasas/genética , Plásmidos , Conejos , Proteínas Recombinantes/genética , Timidina Quinasa/deficiencia , Timidina Quinasa/genética , Factores de Tiempo , Virus Vaccinia/enzimología , Virus Vaccinia/inmunologíaRESUMEN
OBJECTIVE: Specific and efficient tumor-targeted gene delivery is the major goal for successful cancer gene therapy. SUMMARY BACKGROUND DATA: A recombinant thymidine kinase-deleted vaccinia virus (vv) encoding the firefly luciferase (luc) reporter gene or the prodrug converter gene cytosine deaminase (CD) was constructed. The authors compared the extent, duration, and pattern of transgene (luc) expression in vivo after portal venous, intraperitoneal, or intravenous virus administration and survival after treatment with the vv containing CD followed by the prodrug 5-fluorocytosine (5-FC) in a murine model of disseminated liver metastases from colon cancer. METHODS: Recombinant vv containing the luc transgene within the thymidine kinase locus was administered to mice with isolated liver metastases from an MC38 adenocarcinoma. Transgene expression was determined in tumor and organs at various time points. Tumor-bearing mice were treated with recombinant vv containing CD and 5-FC or with appropriate controls and followed for survival. RESULTS: Tumor-specific gene delivery was achieved irrespective of administration route, with gene expression in tumors increased by up to 100,000-fold compared with normal tissues. There was significantly increased transgene expression in tumor after portal venous or intraperitoneal virus administration (p = 0.001 vs. systemic). Treatment using a CD-expressing vv and systemic 5-FC resulted in a significant survival benefit in all treatment groups compared with controls (p < 0.007); there was no additional benefit for portal venous or intraperitoneal virus administration. CONCLUSIONS: Suicide gene therapy using vv with the CD/5-FC system leads to tumor-specific gene expression and improved survival and can result in cure of established liver metastases.
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Adenocarcinoma/secundario , Adenocarcinoma/terapia , Terapia Genética , Neoplasias Hepáticas/secundario , Neoplasias Hepáticas/terapia , Virus Vaccinia/genética , Adenocarcinoma/mortalidad , Animales , Citosina Desaminasa , Escherichia coli/genética , Expresión Génica/genética , Técnicas de Transferencia de Gen , Neoplasias Hepáticas/mortalidad , Luciferasas/genética , Ratones , Nucleósido Desaminasas/genética , Tasa de Supervivencia , TransfecciónRESUMEN
Tumor necrosis factor alpha (TNF-alpha) is a proinflammatory cytokine with potent experimental antitumor activity. Its clinical use in cancer treatment is severely limited by its considerable toxicity after systemic administration, and it is currently confined to isolated limb and organ perfusion settings. In this report, we introduce a novel concept of TNF-alpha-based gene therapy using the TNF-sensitizing properties of endothelial cell monocyte-activating polypeptide II (EMAP-II). We hypothesized that transfer of the EMAP-II gene into established TNF-resistant human melanomas would render these tumors sensitive to subsequent systemic TNF-alpha treatment. To achieve tumor selective gene delivery, we constructed a recombinant vaccinia virus encoding the human EMAP-II gene (vvEMAP). In vitro transfection of human melanoma cells led to the production of EMAP-II by these cells. Supernatants of vvEMAP-transfected tumor cells mediated the induction of tissue factor in endothelial cells. We characterized the pattern of gene expression after systemic administration of a recombinant vaccinia virus encoding a reporter gene in a murine in vivo model of s.c. human melanoma. Gene expression in tumor tissue was increased 100-fold as compared with normal tissue, providing evidence for tumor-selective gene delivery. Finally, human melanomas in nude mice were sensitized in vivo by transferring the EMAP-II gene using vvEMAP. Subsequent systemic administration of TNF-alpha led to tumor regression and growth inhibition of these previously TNF-resistant tumors (P < 0.05). This approach using gene therapy to sensitize primarily unresponsive tumors toward TNF-alpha may enhance the usefulness of TNF-alpha in clinical treatment strategies by increasing the window for the therapeutic application of the cytokine, thus reducing the dose necessary for antitumor responses and subsequently reduce toxicity.
Asunto(s)
Citocinas , Resistencia a Antineoplásicos , Terapia Genética , Melanoma/terapia , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/fisiología , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/fisiología , Neoplasias Cutáneas/terapia , Factor de Necrosis Tumoral alfa/toxicidad , Animales , Línea Celular , Células Cultivadas , Medios de Cultivo Condicionados , Endotelio Vascular/fisiología , Femenino , Genes Reporteros , Inhibidores de Crecimiento/genética , Inhibidores de Crecimiento/fisiología , Humanos , Luciferasas/genética , Melanoma/patología , Ratones , Ratones Desnudos , Proteínas Recombinantes/metabolismo , Tromboplastina/genética , Transfección , Células Tumorales Cultivadas , Virus VacciniaRESUMEN
Suicide gene therapy using the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) has shown promising results for the treatment of colon carcinoma cells in vitro. Efficient viral infection and tumor-specific gene delivery is crucial for clinically measurable treatment effects. After proving efficient gene transfer in vitro, we demonstrate here that genes can be delivered to metastatic liver tumors in vivo in a highly selective manner using systemic delivery of a thymidine kinase-deleted (TK-) recombinant vaccinia virus (Western Reserve strain). When the vector was administered systemically in C57BL/6 mice or nude/athymic mice with established disseminated MC38 liver metastases, transgene expression in tumors was usually 1,000 to 10,000-fold higher compared with other organs (n = 160; P < 0.0001). This tumor-specific gene transfer leads to significant tumor responses and subsequent survival benefits after the transfer of the CD gene to liver metastases and subsequent systemic treatment with the prodrug 5-FC (P < 0.0001). We describe reporter gene and survival experiments both in immunocompetent and athymic nude mice, establishing a gene expression pattern over time and characterizing the treatment effects of the virus delivery/prodrug system. Cure rates of up to 30% in animals with established liver metastases show that suicide gene therapy using TK- vaccinia virus as a vector may be a promising system for the clinical application of tumor-directed gene therapy.
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Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/secundario , Antimetabolitos Antineoplásicos/uso terapéutico , Neoplasias del Colon/patología , Flucitosina/uso terapéutico , Regulación Neoplásica de la Expresión Génica , Terapia Genética , Vectores Genéticos/genética , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/secundario , Nucleósido Desaminasas/genética , Profármacos/uso terapéutico , Virus Vaccinia/genética , Adenocarcinoma/genética , Adenocarcinoma/patología , Animales , Antimetabolitos Antineoplásicos/farmacocinética , Citosina Desaminasa , Femenino , Flucitosina/farmacocinética , Genes Reporteros , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/prevención & control , Ratones , Ratones Endogámicos C57BL , Ratones Desnudos , Trasplante de Neoplasias , Nucleósido Desaminasas/biosíntesis , Profármacos/farmacocinética , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genéticaRESUMEN
Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) is widely used in the treatment or prevention of neutropenia induced by cytostatic regimens. Recent studies with this cytokine have shown several local and/or systemic side effects. We herein report on four patients with different tumor entities receiving GM-CSF as a part of their intensified cytostatic regimen. All four patients developed immune phenomena (sicca syndrome, seropositive arthralgia, hyperthyroidism, and pneumonitis, respectively) during or after subcutaneous treatment with GM-CSF. Pathologic alterations in immunologic serum parameters as well as histopathologic findings accompanied the clinical symptoms. These observations suggest that the therapeutic application of GM-CSF might be involved in the clinical emergence of autoimmune diseases.
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Enfermedades Autoinmunes/inducido químicamente , Factor Estimulante de Colonias de Granulocitos y Macrófagos/efectos adversos , Neoplasias/tratamiento farmacológico , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Autoanticuerpos/análisis , Enfermedades Autoinmunes/patología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/uso terapéutico , Humanos , Masculino , Persona de Mediana Edad , Neoplasias/patología , Neutropenia/inducido químicamente , Neutropenia/tratamiento farmacológico , Proteínas Recombinantes , Sarcoma/tratamiento farmacológico , Sarcoma/patologíaRESUMEN
p53 and c-erbB-2 expression, and their correlation with cell proliferation and steroid hormone receptors, were investigated in 121 carcinomas, 23 lobular in situ carcinomas (LCIS), 74 intraductal carcinomas (DCIS) and 24 minimal invasive carcinomas. DCIS were classified according to the EORTC classification. All markers were measured immunohistochemically on paraffin sections. None of the LCIS, 9 DCIS and 9 minimal invasive cancers showed nuclear positivity for p53. A strong association between histological type and p53 expression was found. Proliferation rates correlated with p53 expression. c-erbB-2 positivity was found in 1 LCIS, 27 DCIS and 12 minimal invasive cancers. There was a significant correlation between p53 expression and c-erbB-2. Both parameters were associated with high proliferation rate and negativity for steroid hormone receptor status. Nuclear pleomorphism could become a comparable prognostic marker in DCIS as it is for infiltrating carcinomas.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/metabolismo , Carcinoma in Situ/metabolismo , Carcinoma Ductal de Mama/metabolismo , Carcinoma Lobular/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Neoplasias de la Mama/patología , Carcinoma in Situ/patología , Carcinoma Ductal de Mama/patología , Carcinoma Lobular/patología , División Celular , Femenino , Humanos , Proteínas de Neoplasias/metabolismo , Pronóstico , Receptor ErbB-2/metabolismo , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/metabolismoRESUMEN
Clinical trials utilising interleukin (IL)-2 with tumor-infiltrating lymphocytes (TIL) have demonstrated efficacy in the treatment of metastatic renal cell carcinoma (RCC). Several cytokines, as well as growth factors have demonstrated modulatory effects upon the biological properties of TIL from RCC, suggesting a potentially important role for cytokines other than IL-2 in the development of active and tumor-specific TIL. IL-12 was recently characterized as a natural-killer-cell-stimulatory factor or cytotoxic-T-cell-maturation factor. These properties of IL-12 prompted us to investigate the impact of this cytokine upon the activation of TIL from human RCC. In an attempt to enhance the in vitro growth and activity of renal TIL, we have grown eight renal TIL cultures in varying concentrations of IL-2 (8, 40, 80, 400 U/ml) and IL-12 (200 U/ml). In addition, IL-12 (200 U/ml) was added to TIL cultures pre-activated with IL-2 (400 U/ml). Growth, cell expansion, and the ability of TIL to release certain cytokines upon tumor stimulation were determined. Proliferation assays, phenotypic analysis, and cytotoxicity assays were performed at an early and a late culture stage. IL-12, alone and when added to suboptimal concentrations of IL-2, failed to induce TIL growth. While the addition of IL-12 to optimal doses of IL-2 suppressed TIL culture expansion, sequential culture exposure first to IL-2 and then to IL-2+IL-12 increased the number of cells expressing CD3+/CD56+ and these cultures demonstrated enhanced in vitro lysis of autologous tumor. IL-12 clearly demonstrated a sequence-dependent impact of the biological behaviour of TIL from RCC. The optimal use of IL-12 in the in vitro expansion of renal TIL may result in cells with an enhanced specific anti-tumor effect.
Asunto(s)
Carcinoma de Células Renales/inmunología , Interleucina-12/farmacología , Neoplasias Renales/inmunología , Linfocitos Infiltrantes de Tumor/efectos de los fármacos , Complejo CD3/análisis , Antígeno CD56/análisis , Carcinoma de Células Renales/patología , Citotoxicidad Inmunológica , Humanos , Interleucina-2/farmacología , Neoplasias Renales/patología , Linfocitos Infiltrantes de Tumor/inmunología , Linfocitos Infiltrantes de Tumor/patología , Células Tumorales CultivadasRESUMEN
The application of recombinant colony stimulating factors for chemotherapy induced granulocytopenia is becoming common in clinical oncology. Here we report on localized cutaneous side effects after subcutaneous administration of recombinant human granulocyte-macrophage colony-stimulating factor (rh GM-CSF) in 11 patients with breast cancer receiving cytostatic treatment. Seven patients suffering from inflammatory breast cancer received cytostatic chemotherapy with mitoxantrone/cyclophosphamide, whereas four patients suffering from noninflammatory breast cancer received high-dose epirubicin/cyclophosphamide, respectively. rh GM-CSF was applicated subcutaneously in a dose of 5 micrograms/kg/d for at least ten days. In all patients, sharply demarked, maculous itching and burning erythemas restricted to the injection sites occurred after three to four injections of rh GM-CSF. These eruptions cleared within 2 to 3 weeks, but reappeared after reexposure to rh GM-CSF. In contrast to previous sporadic reports, no generalized erythemas were observed. Because of this unexpected and subjectively intolerable side effect, rh GM-CSF administration had to be interrupted in all patients. Histopathological findings revealed skin infiltration with lymphocytes, monocytes/macrophages, neutrophils, and occasionally eosinophils, respectively. Since GM-CSF is known to alter immune functions, it seems likely that the eruptions were at least in part due to local immune reactions.
Asunto(s)
Adenocarcinoma/inmunología , Agranulocitosis/tratamiento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Neoplasias de la Mama/inmunología , Eritema/inducido químicamente , Eritema/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/efectos adversos , Adenocarcinoma/tratamiento farmacológico , Adulto , Agranulocitosis/inducido químicamente , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Eritema/patología , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/uso terapéutico , Humanos , Persona de Mediana Edad , Estudios Prospectivos , Proteínas Recombinantes/efectos adversos , Proteínas Recombinantes/uso terapéutico , Teratoma/tratamiento farmacológico , Teratoma/inmunologíaAsunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Mama/terapia , Paclitaxel/uso terapéutico , Premedicación , Ciclofosfamida/administración & dosificación , Resistencia a Múltiples Medicamentos , Epirrubicina/administración & dosificación , Femenino , Fluorouracilo/administración & dosificación , Humanos , Mastectomía SegmentariaRESUMEN
1. The pharmacokinetics of [R]-leucovorin ([R]-LV), [S]-leucovorin ([S]-LV) and the circulating metabolite [S]-5-methyltetrahydrofolate ([S]-5-MTHF) were studied after administration of racemic LV and [S]-LV in 21 subjects. 2. After intravenous infusion of 600 mg m-2 rac-LV (group 1, n = 7) or 300 mg m-2 [S]-LV (group 3, n = 7), the decay of [S]-LV in plasma was biexponential with a distribution half-life of 0.8 to 1 h and an elimination half-life of 11 to 23 h. When rac-LV was administered as a 2 h i.v. infusion (400 mg m-2) following a loading dose of 200 mg m-2 (group 2, n = 7), the plasma concentrations of [R]-LV and [S]-5-MTHF decayed monoexponentially with mean (+/- s.d.) half-lives of 10 +/- 3 h and 7 +/- 2 h, respectively. 3. The AUC of [S]-5-MTHF was significantly higher after infusion of 300 mg m-2 [S]-LV than after infusion of 600 mg m-2 rac-LV (83 +/- 22 micrograms ml-1 h vs 53 +/- 22 micrograms ml-1 h; P = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Neoplasias Gastrointestinales/metabolismo , Leucovorina/farmacocinética , Tetrahidrofolatos/sangre , Adulto , Anciano , Femenino , Semivida , Arteria Hepática , Humanos , Infusiones Intraarteriales , Infusiones Intravenosas , Inyecciones Intravenosas , Leucovorina/administración & dosificación , Leucovorina/sangre , Masculino , Persona de Mediana Edad , EstereoisomerismoRESUMEN
In an attempt to evaluate the feasibility of 5-fluorouracil (FU) treatment modulated by (R,S)-leucovorin (LV) and interferon alpha (IFN alpha) in patients with advanced colorectal cancer, we conducted a phase I trial with increasing doses of subcutaneous IFN alpha (3 x 1 x 10(6) U, 3 x 3 x 10(6) U, 3 x 3 x 10(6) U, 3 x 5 x 10(6) U and 3 x 10 x 10(6) U/week) and 500 mg/m2 LV i.v. as a 2-h infusion with 600 mg/m2 FU i.v. as a midpoint injection. Unacceptable side-effects occurred in all 3 patients at the highest dose level of IFN alpha, while toxicity was tolerable at 3 x 5 x 5 x 10(6) U IFN alpha/week. Thus, this dose was defined as the maximal tolerable dose for IFN alpha in combination with FU and LV. In a subsequent phase II study a total of 83 treatment courses (median: 2.8, range: 2-10) were administered to 30 evaluable patients. Side-effects were acceptable with no WHO grade IV toxicities. Grade III toxicities consisted in thrombopenia (2/30), stomatitis (2/30), diarrhoea (3/30) and nausea/vomiting (4/30). After a median observation time of 17 months (range: 8-22 months), no complete remission was observed and 9 patients experienced a partial response lasting for a median of 6.6 months (range: 3-13+ months), for an overall response rate of 30% (95% confidence interval: 15%-49%). These results show that the described regiment of FU doubly modulated by LV and IFN alpha is active in colorectal cancer and can be safely administered in an out patient setting with acceptable toxicity. Thus, this regimen is suitable to be used for further evaluation in clinical trials.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias Colorrectales/tratamiento farmacológico , Fluorouracilo/administración & dosificación , Interferón-alfa/administración & dosificación , Leucovorina/administración & dosificación , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Femenino , Fluorouracilo/efectos adversos , Humanos , Interferón alfa-2 , Interferón-alfa/efectos adversos , Leucovorina/efectos adversos , Masculino , Persona de Mediana Edad , Proteínas RecombinantesRESUMEN
In situ determination of proliferative activity was performed on 184 consecutive primary invasive breast cancers. Methods used were monoclonal antibody Ki-67 in immunohistochemistry and thymidine labeling index. Tumor proliferation correlated between both methods (p = 0.0001). For thymidine labeling index and Ki-67, respectively, significant correlations existed with histologic tumour grade and steroid hormone receptors (Tumor grade: TLI p = 0.0001; Ki-67 p = 0.0001. ER-ICA: TLI = 0.0001; Ki-67 p = 0.014. PgR-ICA: TLI p = 0.0001; Ki-67 p = 0.0008). For thymidine labeling index a significant correlation was demonstrated for overall survival (p = 0.001) and recurrence free survival (p = 0.01). No statistical significance was observed for clinical outcome and Ki-67 (overall survival p = 0.18; recurrence free survival p = 0.1). None of the factors, TLI or Ki-67, was an independent prognostic factor as demonstrated by multivariate analysis.