RESUMEN
In cancer patients, the prevalence of myeloid-derived suppressor cells (MDSCs) is correlated with the degree of malignancy. In the present study, we investigated the role of circulating M-MDSCs in premetastatic niche formation using a mouse syngeneic tumor model and found that there was an increased frequency of M-MDSCs in the peripheral blood of tumor-bearing mice. M-MDSCs tracking and lung tissue histological analyses revealed that the malignant conditions promote the residence of circulating M-MDSCs and increased tumor cell arrest in the lungs. We further found that MMP-9 expression was increased in the circulating M-MDSCs and the administration of an MMP-9 inhibitor suppressed M-MDSCs transplantation-induced tumor cell arrest in the lung. Therefore, our findings suggest that the expansion of circulating M-MDSCs during tumor progression contributes to premetastatic niche formation by increasing MMP-9 expression.
Asunto(s)
Neoplasias Pulmonares/enzimología , Neoplasias Pulmonares/patología , Pulmón/patología , Metaloproteinasa 9 de la Matriz/metabolismo , Monocitos/patología , Células Supresoras de Origen Mieloide/patología , Secuencia de Aminoácidos , Animales , Progresión de la Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica , Terapia de Inmunosupresión , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/genética , Masculino , Metaloproteinasa 9 de la Matriz/química , Melanoma Experimental/patología , Ratones Endogámicos C57BL , Metástasis de la Neoplasia , Péptidos/químicaRESUMEN
The tumor premetastatic niche initiated by primary tumors is constructed by multiple molecular factors and cellular components and provides permissive condition that allows circulating tumor cells to successfully metastasize. Myeloid-derived suppressor cells (MDSCs), a population of immature cells in pathological conditions, play a critical role in the formation of the premetastatic niche. However, few researches are focused on the function of monocytic MDSCs (mo-MDSCs), a subtype of MDSCs, in the construction of the niche. Here, we show that the number of mo-MDSCs is significantly increased in the premetastatic lungs of tumor-bearing mice, thus promoting tumor cell arrest and metastasis. Before the arrival of tumor cells, the lung-recruited mo-MDSCs produced IL-1ß, thereby increasing E-selectin expression and promoting tumor cell arrest on endothelial cells. Depletion of mo-MDSCs in the premetastatic lungs decreased IL-1ß production, resulting in reduced E-selectin expression. In addition, compared with alveolar macrophages and interstitial macrophages, mo-MDSCs were the major source of IL-1ß expression in the premetastatic lungs. Cytokine array analyses and transwell experiments revealed that CCL12 recruits mo-MDSCs to premetastatic lungs. CCL12 knockdown in tumor-bearing mice significantly decreased mo-MDSC infiltration into the premetastatic lungs, leading to reduced E-selectin expression. Overall, the permissive conditions produced by the infiltrated mo-MDSCs correlated with increased tumor cell arrest and metastasis. These results reveal a novel role of mo-MDSCs in constructing the premetastatic niche. Thus, inhibition of mo-MDSCs infiltration may change the premetastatic niche to normal condition and attenuate tumor metastasis.
Asunto(s)
Selectina E/biosíntesis , Interleucina-1beta/fisiología , Melanoma Experimental/secundario , Monocitos/fisiología , Células Supresoras de Origen Mieloide/fisiología , Proteínas de Neoplasias/biosíntesis , Células Neoplásicas Circulantes , Nicho de Células Madre , Microambiente Tumoral , Animales , Adhesión Celular , Movimiento Celular , Técnicas de Cocultivo , Selectina E/genética , Endotelio Vascular/patología , Regulación Neoplásica de la Expresión Génica/inmunología , Técnicas de Silenciamiento del Gen , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/secundario , Macrófagos/metabolismo , Melanoma Experimental/inmunología , Melanoma Experimental/patología , Ratones , Ratones Endogámicos C57BL , Proteínas Quimioatrayentes de Monocitos/genética , Proteínas Quimioatrayentes de Monocitos/fisiología , Células Supresoras de Origen Mieloide/clasificación , Proteínas de Neoplasias/genética , Especificidad de Órganos , Células Tumorales CultivadasRESUMEN
During malignant progression, primary tumors rebuild leukocyte profile and suppress the host anti-tumor immune response. Tumor-associated neutrophils (TAN) increased in the cancer patients and emerged as an important participant and regulator of immune responses. The aim of this study is to investigate the role of circulating TAN (cTAN) in the metastatic process of advanced malignancy. We tested circulating neutrophils from patients (n = 180) with various types of cancer using flow cytometry analyses. We also used B16F10 cell-implanted C57BL/6 tumor-bearing mice model to simulate the advanced malignancy. Peripheral neutrophils were isolated by ficoll density gradient centrifugation, and in vitro tumor-leukocyte co-culture model was used to test tumor cell survival under leukocyte challenge condition. Here, we showed that neutrophils increased in the peripheral blood under the pathological condition of advanced malignancy both in cancer patients and in tumor-bearing mice. In mouse model, the malignantly increased neutrophils were identified as TAN according to the gene transcriptional analyses. We also showed that cTAN enhance tumor metastasis and cTAN could inhibit the activation of the peripheral leukocytes and rescue tumor cells from leukocyte challenge. In conclusion, our finding suggests that the abundance of cTAN in advanced cancer patients contributes to the circulating tumor cell survival by suppressing peripheral leukocyte activation.
Asunto(s)
Leucocitos/metabolismo , Neoplasias/sangre , Células Neoplásicas Circulantes/metabolismo , Neutrófilos/metabolismo , Anciano , Animales , Supervivencia Celular/genética , Femenino , Voluntarios Sanos , Humanos , Leucocitos/patología , Masculino , Melanoma Experimental/sangre , Melanoma Experimental/patología , Ratones , Persona de Mediana Edad , Metástasis de la Neoplasia , Neoplasias/genética , Neoplasias/patología , Células Neoplásicas Circulantes/patología , Neutrófilos/patologíaRESUMEN
Nanoparticles have been investigated as drug carriers and promising agents for cancer therapy. However, the tumor microenvironment (TME), which is formed by the tumor, is considered a barrier for nanocarriers to enter the internal tumor tissue. Therefore, the evaluation of the biological distribution of nanocarriers in TME can provide useful information on their role in tumor-targeted drug delivery. Although the tumor-bearing mouse model is commonly used to investigate the distribution of nanocarriers in the TME, there is currently a lack of a testing system to predict the distribution of nanocarriers in tumor tissues, especially in patients. This study revealed that the macrophages and dendritic cells (DCs) were more distributed in the peripheral part than the central part of the tumor, which might be an obstacle to the uniform distribution of nanoparticles in the tumor. In addition, the cellular uptake of gold nanoparticles (AuNR and AuNS) in macrophages and DCs cell lines (RAW264.7 and DC1.2) was markedly different from that in the TME. Hence, the study model of the interaction between nanoparticles and macrophages and DCs has an important impact on the accuracy of the results. The vibratome sections of tumor tissues preserved the spatial distribution of immune cells and tumor cells, and had very little effects on their morphologies and activities. More importantly, we found that the distribution of nanocarriers in vibratome sections was similar to that in tumors in vivo. In all, ex vivo analysis using vibratome sections of tumor tissues provides a more convenient and stable method for elucidating the influences of TME on the distribution of nanocarriers.
RESUMEN
Integrin regulation in neutrophil adhesion is essential for innate immune response. c-Abl kinase is a nonreceptor tyrosine kinase and is critical for signaling transduction from various receptors in leukocytes. Using neutrophils and dHL-60 (neutrophil-like differentiation of HL-60) cells, we show that c-Abl kinase is activated by beta(2) integrin engagement and is required for beta(2) integrin-dependent neutrophil sustained adhesion and spreading. The expression of beta(2) integrin on neutrophils induced by TNF-alpha is not affected by c-Abl kinase inhibitor STI571, suggesting that c-Abl kinase is not involved in TNF-alpha-induced integrin activation. The recruitment of c-Abl kinase to beta(2) integrin is dependent on talin head domain, which constitutively interacts with beta(2) integrin cytoplasmic domain. After activated, c-Abl kinase increases the tyrosine phosphorylation of Vav. The SH3 domain of c-Abl kinase is involved in its interaction with talin and Vav. Thus, c-Abl kinase plays an essential role in the activation of Vav induced by beta(2) integrin ligation and in regulating neutrophil-sustained adhesion and spreading.
Asunto(s)
Antígenos CD18/fisiología , Neutrófilos/citología , Neutrófilos/inmunología , Proteínas Proto-Oncogénicas c-abl/fisiología , Antígenos CD18/metabolismo , Adhesión Celular/inmunología , Citoplasma/enzimología , Citoplasma/inmunología , Citoplasma/metabolismo , Células HL-60 , Humanos , Activación Neutrófila/inmunología , Neutrófilos/enzimología , Estructura Terciaria de Proteína , Proteínas Proto-Oncogénicas c-abl/metabolismo , Transducción de Señal/inmunología , Talina/fisiología , Factor de Necrosis Tumoral alfa/fisiologíaRESUMEN
Thymic involution is an important factor leading to the aging of the immune system. Most of what we know regarding thymic aging comes from mouse models, and the nature of the thymic aging process in humans remains largely unexplored due to the lack of a model system that permits longitudinal studies of human thymic involution. In this study, we sought to explore the potential to examine human thymic involution in humanized mice, constructed by transplantation of fetal human thymus and CD34+ hematopoietic stem/progenitor cells into immunodeficient mice. In these humanized mice, the human thymic graft first underwent acute recoverable involution caused presumably by transplantation stress, followed by an age-related chronic form of involution. Although both the early recoverable and later age-related thymic involution were associated with a decrease in thymic epithelial cells and recent thymic emigrants, only the latter was associated with an increase in adipose tissue mass in the thymus. Furthermore, human thymic grafts showed a dramatic reduction in FOXN1 and AIRE expression by 10 weeks post-transplantation. This study indicates that human thymus retains its intrinsic mechanisms of aging and susceptibility to stress-induced involution when transplanted into immunodeficient mice, offering a potentially useful in vivo model to study human thymic involution and to test therapeutic interventions.
Asunto(s)
Envejecimiento/fisiología , Timo/inmunología , Timo/metabolismo , Animales , Biomarcadores , Movimiento Celular , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Técnica del Anticuerpo Fluorescente , Expresión Génica , Humanos , Inmunofenotipificación , Ratones , Ratones Transgénicos , Modelos Animales , Reacción en Cadena en Tiempo Real de la Polimerasa , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Timo/citologíaRESUMEN
Although the involvement of protein arginine methyltransferase 1 (PRMT1) in tumorigenesis has been reported, its roles in breast cancer progression and metastasis has not been elucidated. Here we identified PRMT1 as a key regulator of the epithelial-mesenchymal transition (EMT) in breast cancer. We showed that the EMT program induced by PRMT1 endowed the human mammary epithelial cells with cancer stem cell properties. Moreover, PRMT1 promoted the migratory and invasive behaviors in breast cancer cells. We also demonstrated that abrogation of PRMT1 expression in breast cancer cells abated metastasis in vivo in mouse model. In addition, knockdown of PRMT1 arrested cell growth in G1 tetraploidy and induced cellular senescence. Mechanistically, PRMT1 impacted EMT process and cellular senescence by mediating the asymmetric dimethylation of arginine 3 of histone H4 (H4R3me2as) at the ZEB1 promoter to activate its transcription, indicating the essential roles of this epigenetic control both in EMT and in senescence. Thus, we unraveled a dual function of PRMT1 in modulation of both EMT and senescence via regulating ZEB1. This finding points to the potent value of PRMT1 as a dual therapeutic target for preventing metastasis and for inhibiting cancer cell growth in malignant breast cancer patients.