RESUMEN
BACKGROUND: The Fos-related antigen 1 (FRA-1) transcription factor promotes tumor cell growth, invasion and metastasis. Phosphorylation of FRA-1 increases protein stability and function. We identify a novel signaling axis that leads to increased phosphorylation of FRA-1, increased extracellular matrix (ECM)-induced breast cancer cell invasion and is prognostic of poor outcome in patients with breast cancer. METHODS: While characterizing five breast cancer cell lines derived from primary human breast tumors, we identified BRC-31 as a novel basal-like cell model that expresses elevated FRA-1 levels. We interrogated the functional contribution of FRA-1 and an upstream signaling axis in breast cancer cell invasion. We extended this analysis to determine the prognostic significance of this signaling axis in samples derived from patients with breast cancer. RESULTS: BRC-31 cells display elevated focal adhesion kinase (FAK), SRC and extracellular signal-regulated (ERK2) phosphorylation relative to luminal breast cancer models. Inhibition of this signaling axis, with pharmacological inhibitors, reduces the phosphorylation and stabilization of FRA-1. Elevated integrin αVß3 and uPAR expression in these cells suggested that integrin receptors might activate this FAK-SRC-ERK2 signaling. Transient knockdown of urokinase/plasminogen activator urokinase receptor (uPAR) in basal-like breast cancer cells grown on vitronectin reduces FRA-1 phosphorylation and stabilization; and uPAR and FRA-1 are required for vitronectin-induced cell invasion. In clinical samples, a molecular component signature consisting of vitronectin-uPAR-uPA-FRA-1 predicts poor overall survival in patients with breast cancer and correlates with an FRA-1 transcriptional signature. CONCLUSIONS: We have identified a novel signaling axis that leads to phosphorylation and enhanced activity of FRA-1, a transcription factor that is emerging as an important modulator of breast cancer progression and metastasis.
Asunto(s)
Neoplasias de la Mama/genética , Proteínas Proto-Oncogénicas c-fos/genética , Receptores del Activador de Plasminógeno Tipo Uroquinasa/genética , Activador de Plasminógeno de Tipo Uroquinasa/genética , Neoplasias de la Mama/patología , Matriz Extracelular/genética , Femenino , Humanos , Integrina alfaVbeta3/administración & dosificación , Integrina alfaVbeta3/genética , Células MCF-7 , Invasividad Neoplásica/genética , Invasividad Neoplásica/patología , Fosforilación , Transducción de Señal/efectos de los fármacos , Vitronectina/administración & dosificaciónRESUMEN
Glioblastoma (GBM) is the most common primary malignant astrocytoma characterized by extensive invasion, angiogenesis, hypoxia, and micrometastasis. Despite the relatively leaky nature of GBM blood vessels, effective delivery of antitumor therapeutics has been a major challenge due to the complications caused by the blood-brain barrier (BBB) and the highly torturous nature of newly formed tumor vasculature (blood tumor barrier-BTB). External beam radiotherapy was previously shown to be an effective means of permeabilizing central nervous system (CNS) barriers. By using targeted short-ranged radionuclides, we show for the first time that our targeted actinium-225-labeled αvß3-specific liposomes (225Ac-IA-TLs) caused catastrophic double stranded DNA breaks and significantly enhanced the permeability of BBB and BTB in mice bearing orthotopic GBMs. Histologic studies revealed characteristic α-particle induced double strand breaks within tumors but was not significantly present in normal brain regions away from the tumor where BBB permeability was observed. These findings indicate that the enhanced vascular permeability in these distal regions did not result from direct α-particle-induced DNA damage. On the basis of these results, in addition to their direct antitumor effects, 225Ac-IA-TLs can potentially be used to enhance the permeability of BBB and BTB for effective delivery of systemically administered antitumor therapeutics. Mol Cancer Ther; 16(10); 2191-200. ©2017 AACR.
Asunto(s)
Sistemas de Liberación de Medicamentos , Glioblastoma/tratamiento farmacológico , Glioblastoma/radioterapia , Neovascularización Patológica/tratamiento farmacológico , Neovascularización Patológica/radioterapia , Actinio , Partículas alfa/uso terapéutico , Animales , Transporte Biológico/genética , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/efectos de la radiación , Permeabilidad Capilar/efectos de los fármacos , Permeabilidad Capilar/efectos de la radiación , Línea Celular Tumoral , Daño del ADN/efectos de los fármacos , Daño del ADN/efectos de la radiación , Glioblastoma/genética , Glioblastoma/patología , Humanos , Integrina alfaVbeta3/administración & dosificación , Liposomas/administración & dosificación , Liposomas/química , Ratones , Neovascularización Patológica/genética , Neovascularización Patológica/patologíaRESUMEN
Multifunctional near-infrared (NIR) nanoparticles demonstrate great potential in tumor theranostic applications. To achieve the sensitive detection and effective phototherapy in the early stage of tumor genesis, it is highly desirable to improve the targeting of NIR theranostic agents to biomarkers and to enhance their accumulation in tumor. Here we report a novel targeted multifunctional theranostic nanoparticle, internalized RGD (iRGD)-modified indocyanine green (ICG) liposomes (iRGD-ICG-LPs), for molecular imaging-guided photothermal therapy (PTT) and photodynamic therapy (PDT) therapy against breast tumor. The iRGD peptides with high affinity to αvß3 integrin and effective tumor-internalized property were firstly used to synthesize iRGD-PEG2000-DSPE lipopeptides, which were further utilized to fabricate the targeted ICG liposomes. The results indicated that iRGD-ICG-LPs exhibited excellent stability and could provide an accurate and sensitive detection of breast tumor through NIR fluorescence molecular imaging. We further employed this nanoparticle for tumor theranostic application, demonstrating significantly higher tumor accumulation and tumor inhibition efficacy through PTT/PDT effects. Histological analysis further revealed much more apoptotic cells, confirming the advantageous anti-tumor effect of iRGD-ICG-LPs over non-targeted ICG-LPs. Notably, the targeting therapy mediated by iRGD provides almost equivalent anti-tumor efficacy at a 12.5-fold lower drug dose than that by monoclonal antibody, and no tumor recurrence and obvious treatment-induced toxicity were observed in our study. Our study provides a promising strategy to realize the sensitive detection and effective treatment of tumors by integrating molecular imaging into PTT/PDT therapy.
Asunto(s)
Colorantes/uso terapéutico , Verde de Indocianina/uso terapéutico , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Imagen Molecular/métodos , Oligopéptidos/uso terapéutico , Fotoquimioterapia/métodos , Animales , Apoptosis/efectos de los fármacos , Línea Celular , Supervivencia Celular , Colorantes/farmacocinética , Sistemas de Liberación de Medicamentos/métodos , Femenino , Verde de Indocianina/farmacocinética , Integrina alfaVbeta3/administración & dosificación , Integrina alfaVbeta3/uso terapéutico , Liposomas , Ratones , Ratones Endogámicos BALB C , Nanopartículas , Polietilenglicoles/química , Espectroscopía Infrarroja Corta , Nanomedicina Teranóstica , Distribución TisularRESUMEN
Chronic obstructive pulmonary disease (COPD) is an intractable pulmonary disease, causes widespread and irreversible alveoli collapse. In search of a treatment target molecule, which is able to regenerate collapsed alveoli, we sought to identify a factor that induces differentiation in human alveolar epithelial stem cells using all-trans retinoic acid (ATRA), whose alveolar repair capacity has been reported in animal experiments. When human alveolar epithelial stem cells were exposed to ATRA at a concentration of 10µM for over seven days, approximately 20% of the cells differentiated into each of the type-I and type-II alveolar epithelial cells that constitute the alveoli. In a microarray analysis, integrin-α1 and integrin-ß3 showed the largest variation in the ATRA-treated group compared with the controls. Furthermore, the effect of the induction of differentiation in human alveolar epithelial stem cells using ATRA was suppressed by approximately one-fourth by siRNA treatments with integrin α1 and integrin ß3. These results suggested that integrin α1 and ß3 are factors responsible for the induction of differentiation in human alveolar epithelial stem cells. We accordingly investigated whether integrin nanoparticles also had a regenerative effect in vivo. Elastase-induced COPD model mouse was produced, and the alveolar repair effect of pulmonary administration using nanoparticles of integrin protein was evaluated by X-ray CT scanning. Improvement in the CT value in comparison with an untreated group indicated that there was an alveolar repair effect. In this study, it was shown that the differentiation-inducing effect on human alveolar epithelial stem cells by ATRA was induced by increased expression of integrin, and that the induced integrin enhanced phosphorylation signaling of AKT, resulting in inducing differentiations. Furthermore, the study demonstrated that lung administration of nanoparticles with increased solubility and stability of integrin repaired the alveolus of an Elastase-induced COPD model mouse. Those results show that those integrin nanoparticles are effective as novel COPD treatment target compounds.
Asunto(s)
Integrina alfa1beta1/administración & dosificación , Integrina alfaVbeta3/administración & dosificación , Nanopartículas/administración & dosificación , Alveolos Pulmonares/fisiología , Células Madre/efectos de los fármacos , Tretinoina/administración & dosificación , Administración por Inhalación , Animales , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Glucósidos/química , Humanos , Integrina alfa1beta1/genética , Integrina alfaVbeta3/genética , Masculino , Ratones Endogámicos ICR , Nanopartículas/química , Elastasa Pancreática , Alveolos Pulmonares/citología , Alveolos Pulmonares/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/inducido químicamente , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Regeneración/efectos de los fármacos , Células Madre/citologíaRESUMEN
A new cyclic peptide containing the isoDGR motif that, after coupling to albumin, selectively binds αvß3, an integrin overexpressed in the tumor vasculature. IsoDGR-tagged albumin binds tumor vessels and can be exploited as a carrier for the preparation of tumor vasculature-selective nanomedicines, such as gold nanoparticles (Au) carrying tumor necrosis factor α (TNF), a potent vascular damaging agent.
Asunto(s)
Integrina alfaVbeta3/química , Neoplasias/metabolismo , Oro/química , Humanos , Integrina alfaVbeta3/administración & dosificación , Nanopartículas del Metal/química , Factor de Necrosis Tumoral alfa/químicaRESUMEN
This work was to report preparation and quality control of targeted microbubbles (MB(t)) via biotin-avidin bridge, specifically verification of the "biotin-avidin-biotin" sandwich structure. (1) Lipid microbubbles filled with sulphur hexafluoride were produced by sonication and compared with commercially available microbubbles. (2) MB(t) were produced via biotin-avidin bridge. Specifically, the "biotin-avidin-biotin" sandwich structure on the surface of MB(t) was verified in vitro using fluorescence. (3) Adhesion of alpha(v)beta(3)-integrin targeted MB(t) (MB(αvß3)) to human umbilical vein endothelial cells (HUVECs) was tested using the parallel plate flow chamber (PPFC). "Biotin-avidin" is a reliable method to attach molecular probe onto the surface of microbubbles.
Asunto(s)
Avidina/química , Biotina/química , Células Endoteliales/metabolismo , Integrina alfaVbeta3/administración & dosificación , Microburbujas , Células Cultivadas , Medios de Contraste , Humanos , Aumento de la Imagen , Integrina alfaVbeta3/química , Neoplasias/irrigación sanguínea , Neovascularización Patológica/diagnósticoRESUMEN
Blood vessel loss and inflammation cause secondary degeneration following spinal cord injury. Angiopoietin-1 through the Tie2 receptor, and other ligands through alphavbeta3 integrin, promote endothelial cell survival during developmental or tumour angiogenesis. Here, daily intravenous injections with an alphavbeta3-binding peptide named C16 or an angiopoietin-1 mimetic following a spinal cord contusion at thoracic level 9 in mice rescued epicentre blood vessels, white matter and locomotor function, and reduced detrimental inflammation. Preserved vascularity and reduced inflammation correlated with improved outcomes. C16 and angiopoietin-1 reduced leukocyte transmigration in vitro. Growth factor receptors and integrins facilitate each others' function. Therefore, angiopoietin-1 and C16 were combined and the effects were additive, resulting in almost complete functional recovery. The treatment had lasting effects when started 4 h following injury and terminated after one week. These results identify alphavbeta3 integrin and the endothelial-selective angiopoietin-1 as vascular and inflammatory regulators that can be targeted in a clinically relevant manner for neuroprotection after central nervous system trauma.
Asunto(s)
Angiopoyetina 1/administración & dosificación , Integrina alfaVbeta3/administración & dosificación , Fármacos Neuroprotectores/administración & dosificación , Fragmentos de Péptidos/administración & dosificación , Traumatismos de la Médula Espinal/prevención & control , Médula Espinal/irrigación sanguínea , Médula Espinal/efectos de los fármacos , Secuencia de Aminoácidos , Animales , Movimiento Celular/efectos de los fármacos , Movimiento Celular/fisiología , Quimioterapia Combinada , Femenino , Humanos , Inyecciones Intravenosas , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Recuperación de la Función/efectos de los fármacos , Recuperación de la Función/fisiología , Traumatismos de la Médula Espinal/fisiopatología , Vértebras Torácicas , Factores de TiempoRESUMEN
Our objectives were 1) to characterize angiogenesis in the MDA-MB-435 xenograft mouse model with three-dimensional (3D) MR molecular imaging using alpha(5)beta(1)(RGD)- or irrelevant RGS-targeted paramagnetic nanoparticles and 2) to use MR molecular imaging to assess the antiangiogenic effectiveness of alpha(5)beta(1)(alpha(nu)beta(3))- vs. alpha(nu)beta(3)-targeted fumagillin (50 mug/kg) nanoparticles. Tumor-bearing mice were imaged with MR before and after administration of either alpha(5)beta(1)(RGD) or irrelevant RGS-paramagnetic nanoparticles. In experiment 2, mice received saline or alpha(5)beta(1)(alpha(nu)beta(3))- or alpha(nu)beta(3)-targeted fumagillin nanoparticles on days 7, 11, 15, and 19 posttumor implant. On day 22, MRI was performed using alpha(5)beta(1)(alpha(nu)beta(3))-targeted paramagnetic nanoparticles to monitor the antiangiogenic response. 3D reconstructions of alpha(5)beta(1)(RGD)-signal enhancement revealed a sparse, asymmetrical pattern of angiogenesis along the tumor periphery, which occupied <2.0% tumor surface area. alpha(5)beta(1)-targeted rhodamine nanoparticles colocalized with FITC-lectin corroborated the peripheral neovascular signal. alpha(5)beta(1)(alpha(nu)beta(3))-fumagillin nanoparticles decreased neovasculature to negligible levels relative to control; alpha(nu)beta(3)-targeted fumagillin nanoparticles were less effective (P>0.05). Reduction of angiogenesis in MDA-MB-435 tumors from low to negligible levels did not decrease tumor volume. MR molecular imaging may be useful for characterizing tumors with sparse neovasculature that are unlikely to have a reduced growth response to targeted antiangiogenic therapy.
Asunto(s)
Neoplasias de la Mama/patología , Integrina alfaVbeta3/administración & dosificación , Angiografía por Resonancia Magnética , Neovascularización Patológica/patología , Inhibidores de la Angiogénesis/administración & dosificación , Animales , Neoplasias de la Mama/tratamiento farmacológico , Adhesión Celular , Medios de Contraste , Ciclohexanos/uso terapéutico , Ácidos Grasos Insaturados/uso terapéutico , Fibronectinas/metabolismo , Citometría de Flujo , Ratones , Microscopía Confocal , Nanopartículas , Trasplante de Neoplasias , Neovascularización Patológica/diagnóstico , Oligopéptidos , Tamaño de la Partícula , Receptores de Vitronectina/antagonistas & inhibidores , Receptores de Vitronectina/biosíntesis , Sesquiterpenos/uso terapéuticoRESUMEN
BACKGROUND: Drug eluting stents prevent vascular restenosis but can delay endothelial healing. A rabbit femoral artery model of stenosis formation after vascular injury was used to study the effect of intramural delivery of alpha(v)beta(3)-integrin-targeted rapamycin nanoparticles on vascular stenosis and endothelial healing responses. METHODS AND RESULTS: Femoral arteries of 48 atherosclerotic rabbits underwent balloon stretch injury and were locally treated with either (1) alpha(v)beta(3)-targeted rapamycin nanoparticles, (2) alpha(v)beta(3)-targeted nanoparticles without rapamycin, (3) nontargeted rapamycin nanoparticles, or (4) saline. Intramural binding of integrin-targeted paramagnetic nanoparticles was confirmed with MR molecular imaging (1.5 T). MR angiograms were indistinguishable between targeted and control arteries at baseline, but 2 weeks later they showed qualitatively less luminal plaque in the targeted rapamycin treated segments compared with contralateral control vessels. In a first cohort of 19 animals (38 vessel segments), microscopic morphometric analysis of the rapamycin-treated segments revealed a 52% decrease in the neointima/media ratio (P<0.05) compared to control. No differences (P>0.05) were observed among balloon injured vessel segments treated with alpha(v)beta(3)-targeted nanoparticles without rapamycin, nontargeted nanoparticles with rapamycin, or saline. In a second cohort of 29 animals, endothelial healing followed a parallel pattern over 4 weeks in the vessels treated with alpha(v)beta(3)-targeted rapamycin nanoparticles and the 3 control groups. CONCLUSIONS: Local intramural delivery of alpha(v)beta(3)-targeted rapamycin nanoparticles inhibited stenosis without delaying endothelial healing after balloon injury.
Asunto(s)
Angioplastia de Balón/efectos adversos , Antibacterianos/administración & dosificación , Constricción Patológica/prevención & control , Integrina alfaVbeta3/administración & dosificación , Nanopartículas/uso terapéutico , Sirolimus/administración & dosificación , Animales , Antibacterianos/farmacología , Constricción Patológica/tratamiento farmacológico , Constricción Patológica/patología , Stents Liberadores de Fármacos , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/patología , Arteria Femoral/efectos de los fármacos , Arteria Femoral/patología , Imagen por Resonancia Magnética , Masculino , Nanomedicina/métodos , Nanopartículas/administración & dosificación , Conejos , Sirolimus/farmacologíaRESUMEN
The association of the alpha(v)beta(3) integrin with tumor metastasis and tumor related angiogenesis has been suggested. Therefore, by imaging the alpha(v)beta(3) receptor with PET, information concerning the tumor status could be obtained. Cyclic peptides including the RGD sequence, were radiolabeled by direct electrophilic fluorination with [(18)F]AcOF. In tumor-bearing mice, the labeled peptides accumulated at the tumor with a high tumor to blood ratio. These findings suggest that an assessment of tumor characteristics may be obtained by using these (18)F-labeled peptides.