RESUMEN
Despite the lack of robust evidence of effectiveness, current treatment options for cancer-induced depression (CID) are limited to those developed for non-cancer related depression. Here, anhedonia-like and coping behaviours were assessed in female BALB/c mice inoculated with 4T1 mammary carcinoma cells. The behavioural effects of orally administered sulfasalazine (SSZ), a system xc- inhibitor, were compared with fluoxetine (FLX). FLX and SSZ prevented the development of anhedonia-like behaviour on the sucrose preference test (SPT) and passive coping behaviour on the forced swim test (FST). The SSZ metabolites 5-aminosalicylic acid (5-ASA) and sulfapyridine (SP) exerted an effect on the SPT but not on the FST. Although 5-ASA is a known anti-inflammatory agent, neither treatment with SSZ nor 5-ASA/SP prevented tumour-induced increases in serum levels of interleukin-1ß (IL-1ß) and IL-6, which are indicated in depressive disorders. Thus, the observed antidepressant-like effect of SSZ may primarily be attributable to the intact form of the drug, which inhibits system xc-. This study represents the first attempt at targeting cancer cells as a therapeutic strategy for CID, rather than targeting downstream effects of tumour burden on the central nervous system. In doing so, we have also begun to characterize the molecular pathways of CID.
Asunto(s)
Conducta Animal , Depresión/tratamiento farmacológico , Depresión/etiología , Ácido Glutámico/metabolismo , Neoplasias/complicaciones , Neoplasias/metabolismo , Sulfasalazina/uso terapéutico , Sistema de Transporte de Aminoácidos y+/metabolismo , Animales , Neoplasias Encefálicas/secundario , Línea Celular Tumoral , Cistina/metabolismo , Citocinas/biosíntesis , Depresión/sangre , Femenino , Ácido Glutámico/sangre , Ratones Endogámicos BALB C , Neoplasias/sangre , Análisis de Regresión , Sulfasalazina/farmacologíaRESUMEN
Depression is commonly comorbid in cancer patients and has detrimental effects on disease progression. Evidence suggests that biological mechanisms may induce the onset of cancer-induced depression (CID). The present investigation aims to establish a validated preclinical animal model of CID. Female BALB/c mice were allocated to four groups: control (n=12), chronic oral exposure to corticosterone (CORT) (n=12), CORT exposure followed by chronic low dose fluoxetine (FLX) treatment (n=12), and subcutaneous inoculation of 4T1 mammary carcinoma cells (n=13). Anhedonia was evaluated using the sucrose preference test (SPT), and behavioural despair was evaluated using the forced swim test (FST) and tail suspension test (TST). Sholl analyses were used to examine the dendritic morphology of Golgi-Cox impregnated neurons from the medial prefrontal cortex (mPFC). CORT exposure and tumor burden were both associated with decreased sucrose preference, increased FST immobility, and decreased basilar and apical dendritic branching of neurons in the mPFC. CORT-induced behavioural and dendritic morphological changes were reversible by FLX. No differences in TST immobility were observed between groups. On the secondary TST outcome measure, CORT exposure and tumor burden were associated with a trend towards decreased power of movement. CORT exposure induced a positive control model of a depressive-like state, with FLX treatment confirming the predictive validity of the model. This verified the sensitivity of behavioural and histological tests, which were used to assess the CID model. The induction of a depressive-like state in this model represents the first successfully validated animal model of CID.
Asunto(s)
Dendritas/patología , Trastorno Depresivo/patología , Trastorno Depresivo/fisiopatología , Neoplasias Mamarias Experimentales/patología , Neoplasias Mamarias Experimentales/fisiopatología , Corteza Prefrontal/patología , Anhedonia/efectos de los fármacos , Anhedonia/fisiología , Animales , Antidepresivos de Segunda Generación/farmacología , Línea Celular Tumoral , Corticosterona , Dendritas/fisiología , Trastorno Depresivo/tratamiento farmacológico , Sacarosa en la Dieta , Femenino , Fluoxetina/farmacología , Neoplasias Mamarias Experimentales/psicología , Ratones Endogámicos BALB C , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Trasplante de Neoplasias , Corteza Prefrontal/fisiopatología , Distribución AleatoriaRESUMEN
Cancer in bone is frequently a result of metastases from distant sites, particularly from the breast, lung, and prostate. Pain is a common and often severe pathological feature of cancers in bone, and is a significant impediment to the maintenance of quality of life of patients living with bone metastases. Cancer cell lines have been demonstrated to release significant amounts of the neurotransmitter and cell-signalling molecule l-glutamate via the system xC(-) cystine/glutamate antiporter. We have developed a novel mouse model of breast cancer bone metastases to investigate the impact of inhibiting cancer cell glutamate transporters on nociceptive behaviour. Immunodeficient mice were inoculated intrafemorally with the human breast adenocarcinoma cell line MDA-MB-231, then treated 14days later via mini-osmotic pumps inserted intraperitoneally with sulfasalazine, (S)-4-carboxyphenylglycine, or vehicle. Both sulfasalazine and (S)-4-carboxyphenylglycine attenuated in vitro cancer cell glutamate release in a dose-dependent manner via the system xC(-) transporter. Animals treated with sulfasalazine displayed reduced nociceptive behaviours and an extended time until the onset of behavioural evidence of pain. Animals treated with a lower dose of (S)-4-carboxyphenylglycine did not display this reduction in nociceptive behaviour. These results suggest that a reduction in glutamate secretion from cancers in bone with the system xC(-) inhibitor sulfasalazine may provide some benefit for treating the often severe and intractable pain associated with bone metastases.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Huesos/fisiopatología , Ácido Glutámico/metabolismo , Dolor/tratamiento farmacológico , Sulfasalazina/uso terapéutico , Adenocarcinoma/complicaciones , Adenocarcinoma/metabolismo , Animales , Antiinflamatorios no Esteroideos/farmacología , Benzoatos/uso terapéutico , Neoplasias de la Mama/complicaciones , Neoplasias de la Mama/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Antagonistas de Aminoácidos Excitadores/farmacología , Femenino , Glicina/análogos & derivados , Glicina/uso terapéutico , Humanos , Ratones , Ratones Desnudos , Dolor/etiología , Dolor/patología , Dimensión del Dolor , Sulfasalazina/farmacología , Análisis de Supervivencia , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Breast cancers are the most common source of metastases to bone, of which cancer-induced bone pain is a frequent pathological feature. Cancer-induced bone pain is a unique pain state with multiple determinants that remains to be well understood and managed. Current standard treatments are limited by dose-dependent side effects that can reduce the quality of life of patients. Glutamate is a neurotransmitter and bone cell-signalling molecule that is released via the system x(c)(-) cystine/glutamate antiporter from cancer cell types that frequently metastasize to bone, including breast cancers. In cancer cells, glutamate release is understood to be a side effect of the cellular response to oxidative stress that upregulates the expression and activity of system x(c)(-) to promote the increased import of cystine. Attenuation of glutamate release from cancer cells has been demonstrated to result in reductions in associated cancer-induced bone pain in animal models. This review examines the clinical implications of attenuating cystine uptake and glutamate release in the treatment of cancer-induced bone pain.
Asunto(s)
Neoplasias Óseas/metabolismo , Neoplasias Óseas/secundario , Neoplasias de la Mama/metabolismo , Dolor Nociceptivo/metabolismo , Estrés Oxidativo , Huesos/metabolismo , Neoplasias de la Mama/patología , Cisteína/metabolismo , Femenino , Ácido Glutámico/metabolismo , Humanos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Bone relies on multiple extracellular signaling systems to maintain homeostasis of its normal structure and functions. The amino acid glutamate is a fundamental extracellular messenger molecule in many tissues, and is used in bone for both neural and non-neural signaling. This review focuses on the non-neural interactions, and examines the evolutionarily ancient glutamate signaling system in the context of its application to normal bone functioning and discusses recent findings on the role of glutamate signaling as they pertain to maintaining healthy bone structure. The underlying mechanisms of glutamate signaling and the many roles glutamate plays in modulating bone physiology are featured, including those involved in osteoclast and osteoblast differentiation and mature cell functions. Moreover, the relevance of glutamate signaling systems in diseases that affect bone, such as cancer and rheumatoid arthritis, is discussed, and will highlight how the glutamate system may be exploited as a viable therapeutic target. We will identify novel areas of research where knowledge of glutamate communication mechanisms may aid in our understanding of the complex nature of bone homeostasis. By uncovering the contributions of glutamate in maintaining healthy bone, the reader will discover how this complex molecular signaling system may advance our capacity to treat bone pathologies.
RESUMEN
Glutamatergic intercellular communication is involved in many aspects of metabolic homeostasis in normal bone. In bone metastasis, the balance between bone formation and degradation is disrupted. Although the responsible mechanisms are not clear, we have previously identified that cancer cell lines used in bone tumour models secrete glutamate, suggesting that tumour-derived glutamate may disrupt sensitive signalling systems in bone. This study examines the role of glutamate in mature osteoclastic bone resorption, osteoblast differentiation, and bone nodule formation. Glutamate was found to have no effect on the survival or activity of mature osteoclasts, although glutamate transporter inhibition and receptor blockade increased the number of bone resorption pits. Furthermore, transporter inhibition increased the area of resorbed bone while significantly decreasing the number of osteoclasts. Alkaline phosphatase activity and extracellular matrix mineralization were used as measurements of osteoblast differentiation. Glutamate significantly increased osteoblast differentiation and mineralization, but transport inhibitors had no effect. These studies support earlier findings suggesting that glutamate may be more important for osteoclastogenesis than for osteoclast proliferation or functions. Since glutamate is capable of changing the differentiation and activities of both osteoclast and osteoblast cell types in bone, it is reasonable to postulate that tumour-derived glutamate may impact bone homeostasis in bone metastasis.
Asunto(s)
Comunicación Celular/fisiología , Ácido Glutámico/fisiología , Osteoblastos/fisiología , Osteoclastos/fisiología , Fosfatasa Alcalina/metabolismo , Sistema de Transporte de Aminoácidos y+/antagonistas & inhibidores , Animales , Resorción Ósea/inducido químicamente , Resorción Ósea/patología , Resorción Ósea/fisiopatología , Huesos/fisiopatología , Calcificación Fisiológica/efectos de los fármacos , Calcificación Fisiológica/fisiología , Bovinos , Comunicación Celular/efectos de los fármacos , Diferenciación Celular , Línea Celular , Línea Celular Transformada , Proliferación Celular/efectos de los fármacos , Supervivencia Celular , Humanos , Macrófagos/citología , Ratones , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/enzimología , Osteoclastos/citología , Osteoclastos/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidoresRESUMEN
Giant cell tumor of bone (GCT) is an aggressively osteolytic primary bone tumor that is characterized by the presence of abundant multinucleated osteoclast-like giant cells, hematopoietic monocytes, and a distinct mesenchymal stromal cell component. Previous work in our laboratory has shown that matrix metalloproteinase (MMP)-13 is the principal proteinase expressed by the stromal cells of GCT. The release of cytokines, particularly interleukin-1beta, by the giant cells of GCT acts on stromal cells to stimulate a surge in MMP-13 secretion. The purpose of this study was to determine the bone resorption capabilities of the cellular elements of GCT and the significance of the MMP-13 expression involved in GCT bone resorption. We present a 3-dimensional histomorphometric technique developed to analyze resorption pit depth and yield an accurate measurement of bone resorption with a direct physical view of lacunae on bone slices. In this study, we demonstrate that the mesenchymal stromal cells and the multinucleated giant cells of GCT are independently capable of bone resorption. However, coculture of these 2 cell fractions shows a synergistic increase in bone resorption. In addition, inhibition of MMP-13 reduces resorptive activity of the cells indicating that MMP-13 likely plays an important role in this tumor. This cell-cell cooperation involves giant cell-derived cytokine up-regulation of MMP-13 in the stromal cells, which in turn assists the giant cells in bone resorption. Future research will involve elucidation of the role of cell-cell/matrix communication pathways in bone resorption and tumorigenesis in GCT.
Asunto(s)
Neoplasias Óseas/enzimología , Tumor Óseo de Células Gigantes/enzimología , Metaloproteinasa 13 de la Matriz/metabolismo , Osteólisis/enzimología , Biomarcadores de Tumor , Neoplasias Óseas/patología , Neoplasias Óseas/cirugía , Comunicación Celular , Línea Celular Tumoral , Separación Celular , Tumor Óseo de Células Gigantes/patología , Tumor Óseo de Células Gigantes/cirugía , Células Gigantes/enzimología , Células Gigantes/patología , Humanos , Procesamiento de Imagen Asistido por Computador , Células Madre Mesenquimatosas/enzimología , Células Madre Mesenquimatosas/patología , Osteólisis/patología , Células del Estroma/enzimología , Células del Estroma/patología , Regulación hacia ArribaRESUMEN
Bone is a frequent site for metastasis of breast and prostate cancers, often resulting in pathologic changes in bone metabolism and severe pain. The mechanisms involved are not well understood, but tumour cells may release factors that interfere with bone homeostasis. Several observations have led us to hypothesize that the functional disruptions in bone metastasis are the result of a biological process common to many cell types. The high metabolic activity characteristic of cancer cells often upregulates oxidative stress protection mechanisms such as the antioxidant molecule glutathione. In maintaining redox balance, this normal metabolic response may result in unintended pathologic effects in certain sensitive organ sites. Malignant glioma cells kill surrounding neurons in the brain specifically by secreting the amino acid glutamate, an obligatory waste product of glutathione synthesis. We suggest that glutamate release is a plausible mechanism that may account for the pathologic changes in bone metastasis, since bone, like brain, is also highly sensitive to glutamatergic disruption. This report reviews the available evidence to draw a mechanistic connection between tumour cell oxidative stress and the pathology seen in patients with bone metastasis.
Asunto(s)
Neoplasias Óseas/metabolismo , Neoplasias Óseas/patología , Glutatión/biosíntesis , Osteoblastos/metabolismo , Animales , Neoplasias Óseas/secundario , Glutatión/metabolismo , Humanos , Osteoblastos/patología , Estrés Oxidativo/fisiología , Transducción de Señal/fisiología , Células Tumorales CultivadasRESUMEN
Although the amino acid glutamate is used as an intercellular signaling molecule for normal bone homeostasis, little is known regarding its possible role in the metabolic disruption characteristic of bone metastasis. We have previously shown in vitro that cancer cell lines relevant to bone metastasis release glutamate into the extracellular environment. This study demonstrates the expression of multiple glutamate transporters in cancer cell lines of non-central nervous system origin. Furthermore, we identify the molecular mechanism responsible for glutamate export and show that this system can be inhibited pharmacologically. By highlighting that glutamate secretion is a common biological feature of cancer cells, this study suggests that tumor-derived glutamate could interfere with glutamate-dependent intercellular signaling in normal bone. Pharmacological interference with cancer cell glutamate release may be a viable option for limiting host bone response to invading tumor cells in bone metastasis.
Asunto(s)
Sistema de Transporte de Aminoácidos y+/biosíntesis , Neoplasias Óseas/metabolismo , Transportador 1 de Aminoácidos Excitadores/biosíntesis , Transportador 2 de Aminoácidos Excitadores/biosíntesis , Ácido Glutámico/metabolismo , Proteína 1 de Transporte Vesicular de Glutamato/biosíntesis , Sistema de Transporte de Aminoácidos y+/genética , Animales , Neoplasias Óseas/secundario , Línea Celular Tumoral , Transportador 1 de Aminoácidos Excitadores/genética , Transportador 2 de Aminoácidos Excitadores/genética , Humanos , Ratones , Ratones Endogámicos , Metástasis de la Neoplasia , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Proteína 1 de Transporte Vesicular de Glutamato/genéticaRESUMEN
MDCK cells were engineered to express luciferase driven by cytomegalovirus (CMV) or hybrid ubiquitin B (UbB) promoter and encapsulated in alginate-poly-L-lysine-alginate microcapsules. In vitro experiments showed capsules could be monitored individually or in multi-layers quantitatively. When luciferase-expressing and non-luciferase expressing MDCK cells were mixed at different ratios and encapsulated, the signals increased linearly according to the number of capsules, in vitro and in vivo. For CMV-driven luciferase expression, the strongest signal was seen at 4 hours post-implantation, with a subsequent 50% decrease by 24 hours and then declined gradually to 10-20% until day 20. However, retrieved capsules showed good cell viability. When capsules contained plasmid driven by UbB promoter, there was no decline in signal. Our results indicate that luciferase could be used as a marker for microencapsulated cells to monitor the viability and gene expression of the implanted cells.
Asunto(s)
Cápsulas/química , Trasplante de Células/métodos , Terapia Genética/métodos , Luciferasas/administración & dosificación , Luciferasas/genética , Alginatos , Animales , Cápsulas/uso terapéutico , Línea Celular , Supervivencia Celular , Perros , Técnicas de Transferencia de Gen , Genes Reporteros , Vectores Genéticos , Implantes Experimentales , Luciferasas/biosíntesis , Ratones , Ratones Endogámicos BALB C , Farmacocinética , Polilisina/análogos & derivadosRESUMEN
Bone is one of the most frequent sites for metastasis of breast and prostate cancers. Bone metastases are associated with pathologic changes in bone turnover and severe pain. The mechanisms that trigger these effects are not well understood, but it is postulated that tumour cells release factors which interfere with signalling processes critical to bone homeostasis. We have identified that several cancer cell lines known to cause bone disruption in animal models of bone metastasis appear to secrete glutamate into their extracellular environment in vitro. Although these cells also express specific glutamate receptors, the implications of this potentially disruptive chemical signal are discussed in relation to normal glutamate-dependent communication processes in bone and a possible mechanistic connection is made between tumour cell glutamate release and the development of pathological changes in bone turnover.