RESUMEN
PURPOSE: Since combination of Toll-like receptor (TLR) ligands could boost antitumor immunity, we evaluated the efficacy of dendritic cell (DC) vaccines upon dual activation of TLR9 and TLR7 in breast cancer models. METHODS: DCs were generated from mouse bone marrow or peripheral blood from healthy human donors and stimulated with CpG1826 (mouse TLR9 agonist), CpG2006 or IMT504 (human TLR9 agonists) and R848 (TLR7 agonist). Efficacy of antitumor vaccines was evaluated in BALB/c mice bearing metastatic mammary adenocarcinomas. RESULTS: CpG-DCs improved the survival of tumor-bearing mice, reduced the development of lung metastases and generated immunological memory. However, dual activation of TLRs impaired the efficacy of DC vaccines. In vitro, we found that R848 inhibited CpG-mediated maturation of murine DCs. A positive feedback loop in TLR9 mRNA expression was observed upon CpG stimulation that was inhibited in the presence of R848. Impaired activation of NF-κB was detected when TLR9 and TLR7 were simultaneously activated. Blockade of nitric oxide synthase (NOS) and indoleamine-pyrrole-2,3-dioxygenase (IDO) improved the activation of CpG-DCs. When we evaluated the effect of combined activation of TLR9 and TLR7 in human DCs, we found that R848 induced robust DC activation that was inhibited by TLR9 agonists. CONCLUSIONS: These observations provide insight in the biology of TLR9 and TLR7 crosstalk and suggest caution in the selection of agonists for multiple TLR stimulation. Blockade of NOS and IDO could improve the maturation of antitumor DC vaccines. R848 could prove a useful adjuvant for DC vaccines in human patients.
Asunto(s)
Adenocarcinoma/terapia , Neoplasias de la Mama/terapia , Vacunas contra el Cáncer/inmunología , Receptor Toll-Like 7/agonistas , Receptor Toll-Like 9/agonistas , Adyuvantes Inmunológicos/farmacología , Animales , Vacunas contra el Cáncer/farmacología , Células Dendríticas/inmunología , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BLRESUMEN
Myeloid-derived suppressor cells (MDSCs) are key regulatory cells that control inflammation and promote tumor-immune escape. To date, no specific immunomodulatory drug has proven efficacy in targeting the expansion and/or function of these cells in different pathophysiologic settings. In this study, we identified a context-dependent effect of the nonsteroidal anti-inflammatory drug indomethacin (IND) on MDSCs, depending on whether they were derived from tumor microenvironments (TME) or from tumor-free microenvironments (TFME). Treatment of mice bearing the LP07 lung adenocarcinoma with IND inhibited the suppressive activity of splenic MDSCs, which restrained tumor growth through mechanisms involving CD8(+) T cells. The same effect was observed when MDSCs were treated with IND and conditioned media from LP07 tumor cells in vitro. However, in the absence of a tumor context, IND enhanced the intrinsic suppressive function of MDSCs and amplified their protumoral activity. In a model of autoimmune neuroinflammation, IND-treated MDSCs differentiated in TFME attenuated inflammation, whereas IND-treated MDSCs differentiated in TME aggravated clinical symptoms and delayed resolution of the disease. Mechanistically, IND reduced arginase activity as well as NO and reactive oxygen species production in MDSCs differentiated in TME but not in TFME. Moreover, expression of the C/EBP-ß transcription factor isoforms correlated with the suppressive activity of IND-treated MDSCs. Our study unveils the dual and context-dependent action of IND, a drug that serves both as an anti-inflammatory and anticancer agent, which differentially affects MDSC activity whether these cells are derived from TME or TFME. These results have broad clinical implication in cancer, chronic inflammation and autoimmunity.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Microambiente Celular/efectos de los fármacos , Microambiente Celular/inmunología , Indometacina/farmacología , Células Mieloides/efectos de los fármacos , Células Mieloides/inmunología , Microambiente Tumoral/efectos de los fármacos , Microambiente Tumoral/inmunología , Animales , Autoinmunidad/efectos de los fármacos , Modelos Animales de Enfermedad , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/metabolismo , Femenino , Inmunofenotipificación , Ratones , Modelos Biológicos , Células Mieloides/metabolismo , Neoplasias/inmunología , Neoplasias/patología , Óxido Nítrico/metabolismo , Fenotipo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Carga Tumoral/efectos de los fármacos , Carga Tumoral/inmunologíaRESUMEN
Secretory leukocyte protease inhibitor (SLPI) is a serine protease inhibitor that was related to cancer development and metastasis dissemination on several types of tumors. However, it is not known the effect of SLPI on mammary and colon tumors. The aim of this study was to examine the effect of SLPI on mammary and colon tumor growth. The effect of SLPI was tested on in vitro cell apoptosis and in vivo tumor growth experiments. SLPI over-expressing human and murine mammary and colon tumor cells were generated by gene transfection. The administration of murine mammary tumor cells over-expressing high levels of SLPI did not develop tumors in mice. On the contrary, the administration of murine colon tumor cells over-expressing SLPI, developed faster tumors than control cells. Intratumoral, but not intraperitoneal administration of SLPI, delayed the growth of tumors and increased the survival of mammary but not colon tumor bearing mice. In vitro culture of mammary tumor cell lines treated with SLPI, and SLPI producer clones were more prone to apoptosis than control cells, mainly under serum deprivation culture conditions. Herein we demonstrated that SLPI induces the apoptosis of mammary tumor cells in vitro and decreases the mammary but not colon tumor growth in vivo. Therefore, SLPI may be a new potential therapeutic tool for certain tumors, such as mammary tumors.
Asunto(s)
Neoplasias de la Mama/metabolismo , Neoplasias del Colon/metabolismo , Neoplasias Mamarias Animales/metabolismo , Inhibidor Secretorio de Peptidasas Leucocitarias/metabolismo , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Línea Celular Tumoral , Neoplasias Colorrectales/metabolismo , Femenino , Silenciador del Gen , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Mamarias Animales/tratamiento farmacológico , Ratones , Ratones Endogámicos BALB C , Inhibidor Secretorio de Peptidasas Leucocitarias/farmacología , Transfección , Neoplasias del Cuello Uterino/metabolismoRESUMEN
Currently, an in vivo spontaneous model of estrogen dependent, tamoxifen sensitive breast cancer does not exist. We show here the characterization of the M05 mammary tumor that appeared spontaneously in a 1-year-old virgin female BALB/c mouse in our animal facility. The M05 tumor is a semi-differentiated adenocarcinoma that expresses estrogen and progesterone receptors. When it was transplanted to either male or ovariectomized female mice it did not grow. Moreover, ovariectomy or treatment with tamoxifen of tumor bearing mice led to a halt in tumor growth. Treatment of ovariectomized mice that had been inoculated with the M05 tumor showed that only estradiol, but not progesterone, promoted the re-growth of the tumor. Finally, after passage nine, tumor growth was achieved in male and ovariectomized female mice suggesting that the tumor had progressed to hormone independence. However, like often found in the clinic, expression of estrogen and progesterone receptors was maintained. This model mimics the biology of estrogen receptor positive breast cancer in humans and presents itself as an invaluable tool for the study of endocrine resistance in a physiologically relevant setting.
Asunto(s)
Neoplasias Mamarias Experimentales/tratamiento farmacológico , Tamoxifeno/uso terapéutico , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/inmunología , Adenocarcinoma/patología , Animales , División Celular , Femenino , Inmunocompetencia , Masculino , Neoplasias Mamarias Experimentales/inmunología , Neoplasias Mamarias Experimentales/patología , Ratones , Ratones Endogámicos BALB C , Trasplante de Neoplasias/métodos , Trasplante de Neoplasias/patología , Ovariectomía , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/metabolismoRESUMEN
Few studies have explored the mechanistic basis for the apparent paradoxical effects of nitric oxide and its interrelated redox species (NO(X)) in cancer biology. Our aim was to determine the differential effects of the redox state and kinetics of nitrosative species on the key cancer processes of apoptosis. Therefore, a murine lung adenocarcinoma cell line was exposed to various NO(X) donor compounds differing in redox state and delivery kinetics. DNA strand breaks (DSBs) were measured by the alkaline single-cell gel electrophoresis assay (the COMET assay) and correlated with cell viability by the MTT and soft agar colony assays, while caspase enzymatic activity was measured using an in vitro fluorogenic caspase assay. Finally, cDNA microarrays defined apoptosis-related gene expression alterations resultant from these NO(X) donors. Exogenous NO(X) differentially influences DSBs, and apoptosis-related cell death and expression based on the redox state and kinetics of NO(X) delivery. In our murine lung adenocarcinoma model we have demonstrated differential effects of NO(X) based on the mode of delivery and redox state. These data suggest that the development of NO(X)-based cancer chemotherapy must consider the redox state and kinetics of delivery into their logical design.
Asunto(s)
Adenocarcinoma/metabolismo , Neoplasias Pulmonares/metabolismo , Óxido Nítrico/metabolismo , Adenocarcinoma/genética , Animales , Apoptosis/genética , Biomarcadores de Tumor/metabolismo , Caspasas/metabolismo , Ensayo de Unidades Formadoras de Colonias , Ensayo Cometa , Daño del ADN/efectos de los fármacos , ADN de Neoplasias/análisis , Cinética , Neoplasias Pulmonares/genética , Ratones , Ratones Endogámicos BALB C , Donantes de Óxido Nítrico/farmacología , Nitritos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Oxidación-Reducción , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
Mice bearing LP07 lung adenocarcinoma present some characteristics similar to those shown in patients with several malignant diseases. LP07 tumor bearers develop paraneoplastic syndromes such as cachexia, leukocytosis, and hypercalcemia, partly due to a systemic inflammatory response. We analyzed some of the mechanisms involved in the effectiveness of the association of the appetite-stimulant medroxiprogesterone acetate (MPA) and the nonselective cyclooxigenase (COX) inhibitor indomethacin (INDO) in LP07 tumor bearing mice. INDO and INDO plus MPA treatments significantly inhibited tumor growth, which was not inhibited by MPA. The number of lung metastatic nodules was decreased with all treatments, being most effective INDO alone and INDO plus MPA. A significant decrease of plasmatic levels of the matrix metalloproteinases MMP-9 and MMP-2 correlated with these results. Paraneoplastic syndromes, leukocytosis, and cachexia were abolished by all treatments. We determined effects of the treatments on circulating cytokines shown to regulate cachexia and inflammation. Both treatments alone, and INDO plus MPA, reduced circulating IL-6 throughout tumor evolution. A pronounced increase in serum IL-1ss levels was detected in untreated tumor bearers. These levels decreased and were closer to normal serum values when LP07 mice were treated with INDO plus MPA. The combination of a nonsteroidal antiinflammatory drug as INDO and MPA showed to be effective in inhibiting tumor and metastatic growth and diminishing paraneoplastic symptoms and SIR. A variety of specific molecules are implicated as playing a role in cancer-induced cachexia and hematological alterations.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica , Neoplasias Pulmonares/tratamiento farmacológico , Síndromes Paraneoplásicos/prevención & control , Adenocarcinoma/complicaciones , Adenocarcinoma/patología , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antineoplásicos Hormonales/administración & dosificación , Estimulantes del Apetito/administración & dosificación , Caquexia/etiología , Caquexia/prevención & control , Ensayo de Inmunoadsorción Enzimática , Femenino , Hipercalcemia/etiología , Hipercalcemia/prevención & control , Indometacina/administración & dosificación , Interleucina-1/sangre , Interleucina-6/sangre , Neoplasias Pulmonares/complicaciones , Neoplasias Pulmonares/patología , Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 2 de la Matriz/efectos de los fármacos , Metaloproteinasa 9 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/efectos de los fármacos , Acetato de Medroxiprogesterona/administración & dosificación , Ratones , Ratones Endogámicos BALB CRESUMEN
Mice bearing LP07 lung adenocarcinoma show some characteristics that are similar to those present in patients with NSCLC. LP07 tumor-bearing mice develop the paraneoplastic syndromes of cachexia, leukocytosis and hypercalcemia. These symptoms may be partly due to a systemic inflammatory response. Our aim was to determine if treatment with NSAIDs would lower tumor and metastasis growth and their accompanying syndromes. The nonselective COX inhibitor indomethacin and the selective COX-2 inhibitor celecoxib reduced tumor growth and metastasis outcome in s.c. LP07 tumor-bearing mice. Both drugs also inhibited the development of leukocytosis and the weight loss associated with LP07 progression. Serum levels of the inflammatory cytokines IL-1beta and IL-6, mediators of cachexia, were modulated by NSAIDs. Inhibition of in vitro migration and invasion and reduction in angiogenesis were attained when cells were treated with either indomethacin or celecoxib. MMP-9 activity was also reduced in conditioned media from LP07 cells treated with celecoxib. These data suggest that several processes implicated in tumor progression can be modulated with NSAID treatment. Improvement in performance status through modulation of cachexia may offer a possibility for combining anti-inflammatory treatments with more aggressive therapies.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Antiinflamatorios no Esteroideos/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Síndromes Paraneoplásicos/prevención & control , Adenocarcinoma/patología , Animales , División Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Quimiotaxis/efectos de los fármacos , Inhibidores de la Ciclooxigenasa/uso terapéutico , Citocinas/análisis , Progresión de la Enfermedad , Femenino , Indometacina/uso terapéutico , Lactonas/uso terapéutico , Neoplasias Pulmonares/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Invasividad Neoplásica/prevención & control , SulfonasRESUMEN
BACKGROUND: The normal duct and lobular system of the mammary gland is lined with luminal and myoepithelial cell types. Although evidence suggests that myoepithelial cells might suppress tumor growth, invasion and angiogenesis, their role remains a major enigma in breast cancer biology and few models are currently available for exploring their influence. Several years ago a spontaneous transplantable mammary adenocarcinoma (M38) arose in our BALB/c colony; it contains a malignant myoepithelial cell component and is able to metastasize to draining lymph nodes and lung. METHODS: To characterize this tumor further, primary M38 cultures were established. The low-passage LM38-LP subline contained two main cell components up to the 30th subculture, whereas the higher passage LM38-HP subline was mainly composed of small spindle-shaped cells. In addition, a large spindle cell clone (LM38-D2) was established by dilutional cloning of the low-passage MM38-LP cells. These cell lines were studied by immunocytochemistry, electron microscopy and ploidy, and syngeneic mice were inoculated subcutaneously and intravenously with the different cell lines, either singly or combined to establish their tumorigenic and metastatic capacity. RESULTS: The two subpopulations of LM38-LP cultures were characterized as luminal and myoepithelium-like cells, whereas LM38-HP was mainly composed of small, spindle-shaped epithelial cells and LM38-D2 contained only large myoepithelial cells. All of them were tumorigenic when inoculated into syngeneic mice, but only LM38-LP cultures containing both conserved luminal and myoepithelial malignant cells developed aggressive papillary adenocarcinomas that spread to lung and regional lymph nodes. CONCLUSION: The differentiated histopathology and metastatic ability of the spontaneous transplantable M38 murine mammary tumor is associated with the presence and/or interaction of both luminal and myoepithelial tumor cell types.
Asunto(s)
Adenocarcinoma/genética , Neoplasias Mamarias Animales/genética , Mioepitelioma/genética , Adenocarcinoma/enzimología , Adenocarcinoma/secundario , Adenocarcinoma Papilar/enzimología , Adenocarcinoma Papilar/genética , Adenocarcinoma Papilar/patología , Adenocarcinoma Papilar/ultraestructura , Animales , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Proliferación Celular , ADN de Neoplasias/genética , Modelos Animales de Enfermedad , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/secundario , Ganglios Linfáticos/patología , Metástasis Linfática/genética , Metástasis Linfática/patología , Neoplasias Mamarias Animales/enzimología , Neoplasias Mamarias Animales/patología , Neoplasias Mamarias Experimentales/enzimología , Neoplasias Mamarias Experimentales/genética , Neoplasias Mamarias Experimentales/patología , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica de Transmisión/métodos , Trasplante de Neoplasias/métodos , Péptido Hidrolasas/biosíntesis , Ploidias , Esferoides Celulares/química , Esferoides Celulares/metabolismo , Esferoides Celulares/ultraestructura , Células Tumorales CultivadasRESUMEN
Systemic syndromes characterized by a persistent activity of circulating mediators (cytokines) are frequently present with advanced cancer. We grouped under the general heading of "Systemic Immune-Metabolic Syndrome (SIMS)" a particular variety of distressing systemic syndrome characterized by dysregulation of the psycho-neuro-immune-endocrine homeostasis, with overlapping clinical manifestations. SIMS may include cachexia, anorexia, nausea, early satiety, fatigue, tumor fever, cognitive changes and superinfection. The aim of this study was to ameliorate some of the SIMS symptoms in a homogeneous group of lung adenocarcinoma patients using a multitargeted therapy. Fifteen patients with evidence of SIMS were studied. SIMS was defined as the presence of weight loss, anorexia, fatigue performance status>/=2 and acute-phase protein response. Patients received medroxyprogesterone (MPA) (500 mg twice daily), celecoxib (200 mg twice daily), plus oral food supplementation for 6 weeks. After treatment, 13 patients either had stable weight (+/- 1%) or had gained weight. There were significant differences in improvement of body-weight-change rate, nausea, early satiety, fatigue, appetite and performance status. Patients who had any kind of lung infection showed higher levels of IL-10 compared to non-infected patients (P=0.039). Our results suggest that patients with advanced lung adenocarcinoma, treated with MPA, celecoxib and dietary intervention, might have considerable improvement in certain SIMS outcomes. This multitargeted symptomatic approach deserves further study.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Antineoplásicos/uso terapéutico , Caquexia/terapia , Fatiga/terapia , Medroxiprogesterona/uso terapéutico , Sulfonamidas/uso terapéutico , Sobreinfección/terapia , Adenocarcinoma/complicaciones , Adulto , Anciano , Caquexia/dietoterapia , Caquexia/etiología , Celecoxib , Fatiga/dietoterapia , Fatiga/etiología , Femenino , Humanos , Neoplasias Pulmonares/complicaciones , Masculino , Persona de Mediana Edad , Proyectos Piloto , Pirazoles , Sobreinfección/dietoterapia , Sobreinfección/etiología , SíndromeRESUMEN
Durante el crecimiento de los tumores el papel del sistema inmune es controvertido, y a pesar de las grandes expectativas en poder estimularlo para que fuera eficiente en el rechazo de los mismos, este objetivo aún no se ha cumplido. En este revisión se resumen y se analizan los estudios inmunológicos realizados con modelos de tumores murinos en nuestro laboratorio. Nosotros hemos trabajado con tumores de mama murinos, y en nuestros primeros estudios hemos determinado que el sistema inmune de los portadores de tumor durante las primeras etapas de su evolución reconoce en forma específica los antígenos tumorales, y que sus linfocitos están activados para respuestas de hipersensibilidad retardada in vivo e in vitro y angiogénesis linfocitaria. Sin embargo, esta funcionalidad no se correlaciona con los mecanismos efectores de rechazo tumoral. El reconocimiento y activación linfocitarias desaparecem a medida que tumor crece. Las células tumorales y linfocitos del portador de tumor secretan factores solubles que exacerban el crecimiento tumoral y metastásico. Las poblaciones de neutrófilos y mastocitos también se encuentran alteradas durante el crecimiento del tumor. Postulamos que las células tumorales secretan factores que inducen poblaciones celulares a producir inmunosupresores que favorecem el desarrollo del mismo tumor.
Asunto(s)
Animales , Ratones , Neoplasias Mamarias Animales/inmunología , Antígenos de Neoplasias , Neoplasias Mamarias Animales/metabolismo , Neoplasias Mamarias Animales/patologíaRESUMEN
Durante el crecimiento de los tumores el papel del sistema inmune es controvertido, y a pesar de las grandes expectativas en poder estimularlo para que fuera eficiente en el rechazo de los mismos, este objetivo aún no se ha cumplido. En este revisión se resumen y se analizan los estudios inmunológicos realizados con modelos de tumores murinos en nuestro laboratorio. Nosotros hemos trabajado con tumores de mama murinos, y en nuestros primeros estudios hemos determinado que el sistema inmune de los portadores de tumor durante las primeras etapas de su evolución reconoce en forma específica los antígenos tumorales, y que sus linfocitos están activados para respuestas de hipersensibilidad retardada in vivo e in vitro y angiogénesis linfocitaria. Sin embargo, esta funcionalidad no se correlaciona con los mecanismos efectores de rechazo tumoral. El reconocimiento y activación linfocitarias desaparecem a medida que tumor crece. Las células tumorales y linfocitos del portador de tumor secretan factores solubles que exacerban el crecimiento tumoral y metastásico. Las poblaciones de neutrófilos y mastocitos también se encuentran alteradas durante el crecimiento del tumor. Postulamos que las células tumorales secretan factores que inducen poblaciones celulares a producir inmunosupresores que favorecem el desarrollo del mismo tumor. (AU)