Recently， microRNAs have been shown to be involved in hematopoietic cell development， but their role in eosinophilopoiesis has not yet been described. In this article， we show that miR-223 is upregulated during eosinophil differentiation in an ex vivo bone marrow-derived eosinophil culture system. Targeted ablation of miR-223 leads to an increased proliferation of eosinophil progenitors. We found upregulation of a miR-223 target gene， IGF1R， in the eosinophil progenitor cultures derived from miR-223(-/-) mice compared with miR-223(+/+) littermate controls. The increased proliferation of miR-223(-/-) eosinophil progenitors was reversed by treatment with an IGF1R inhibitor (picropodophyllin). Whole-genome microarray analysis of differentially regulated genes between miR-223(+/+) and miR-223(-/-) eosinophil progenitor cultures identified a specific enrichment in genes that regulate hematologic cell development. Indeed， miR-223(-/-) eosinophil progenitors had a delay in differentiation. Our results demonstrate that microRNAs regulate the development of eosinophils by influencing eosinophil progenitor growth and differentiation and identify a contributory role for miR-223 in this process.
A novel regulator of macrophage activation: miR-223 in obesity-associated adipose tissue inflammation.
Next Section Abstract Background— Macrophage activation plays a crucial role in regulating adipose tissue inflammation and is a major contributor to the pathogenesis of obesity-associated cardiovascular diseases. On various types of stimuli， macrophages respond with either classic (M1) or alternative (M2) activation. M1- and M2-mediated signaling pathways and corresponding cytokine production profiles are not completely understood. The discovery of microRNAs provides a new opportunity to understand this complicated but crucial network for macrophage activation and adipose tissue function. Methods and Results— We have examined the activity of microRNA-223 (miR-223) and its role in controlling macrophage functions in adipose tissue inflammation and systemic insulin resistance. miR-223 −/− mice on a high-fat diet exhibited an increased severity of systemic insulin resistance compared with wild-type mice that was accompanied by a marked increase in adipose tissue inflammation. The specific regulatory effects of miR-223 in myeloid cell–mediated regulation of adipose tissue inflammation and insulin resistance were then confirmed by transplantation analysis. Moreover， using bone marrow–derived macrophages， we demonstrated that miR-223 is a novel regulator of macrophage polarization， which suppresses classic proinflammatory pathways and enhances the alternative antiinflammatory responses. In addition， we identified Pknox1 as a genuine miR-223 target gene and an essential regulator for macrophage polarization. Conclusion— For the first time， this study demonstrates that miR-223 acts to inhibit Pknox1， suppressing proinflammatory activation of macrophages; thus， it is a crucial regulator of macrophage polarization and protects against diet-induced adipose tissue inflammatory response and systemic insulin resistance.
MicroRNA-223 antagonizes angiogenesis by targeting β1 integrin and preventing growth factor signaling in endothelial cells.
Endothelial cells in situ are largely quiescent， and their isolation and culture are associated with the switch to a proliferative phenotype.To identify antiangiogenic microRNAs expressed by native endothelial cells that are altered after isolation and culture， as well as the protein targets that regulate responses to growth factors.Profiling studies revealed that miR-223 was highly expressed in freshly isolated human， murine， and porcine endothelial cells， but those levels decreased in culture. In primary cultures of endothelial cells， vascular endothelial cell growth factor and basic fibroblast growth factor further decreased miR-223 expression. The overexpression of precursor-miR-223 did not affect basal endothelial cell proliferation but abrogated vascular endothelial cell growth factor-induced and basic fibroblast growth factor-induced proliferation， as well as migration and sprouting. Inhibition of miR-223 in vivo using specific antagomirs potentiated postnatal retinal angiogenesis in wild-type mice， whereas recovery of perfusion after femoral artery ligation and endothelial sprouting from aortic rings from adult miR-223(-/y) animals were enhanced. MiR-223 overexpression had no effect on the growth factor-induced activation of ERK1/2 but inhibited the vascular endothelial cell growth factor-induced and basic fibroblast growth factor-induced phosphorylation of their receptors and activation of Akt. β1 integrin was identified as a target of miR-223 and its downregulation reproduced the defects in growth factor receptor phosphorylation and Akt signaling seen after miR-223 overexpression. Reintroduction of β1 integrin into miR-223-ovexpressing cells was sufficient to rescue growth factor signaling and angiogenesis.These results indicate that miR-223 is an antiangiogenic microRNA that prevents endothelial cell proliferation at least partly by targeting β1 integrin.
MicroRNAs-223 Antagonises Angiogenesis by Targeting β1 Integrin and Preventing Growth Factor Signaling in Endothelial Cells.
ABSTRACT Rationale： Endothelial cells in situ are largely quiescent and their isolation and culture is associated with the switch to a proliferative phenotype. Objective： To identify anti-angiogenic microRNAs (miRNAs) expressed by native endothelial cells that are altered after isolation and culture， as well as the protein targets that regulate responses to growth factors. Methods and Results： Profiling studies revealed that miR-223 was highly expressed in freshly isolated human， murine and porcine endothelial cells but that levels decreased in culture. In primary cultures of endothelial cells， vascular endothelial cell growth factor (VEGF) and basic fibroblast growth factor (bFGF) further decreased miR-223 expression. The overexpression of pre-miR-223 did not affect basal endothelial cell proliferation but abrogated VEGF- and bFGF-induced proliferation as well as migration and sprouting. Inhibition of miR-223 in vivo using specific antagomirs potentiated postnatal retinal angiogenesis in wild-type mice while recovery of perfusion after hindlimb ligation and endothelial sprouting from aortic rings from adult miR-223(-/y) animals were enhanced. MiR-223 overexpression had no effect on the growth factor-induced activation of ERK1/2 but inhibited the VEGF- and bFGF-induced phosphorylation of their receptors and activation of Akt. β1 integrin was identified as a target of miR-223 and its downregulation reproduced the defects in growth factor receptor phosphorylation and Akt signaling seen after miR-223 overexpression. Reintroduction of β1 integrin into miR-223 ovexpressing cells was sufficient to rescue growth factor signaling and angiogenesis. Conclusions： These results indicate that miR-223 is an anti-angiogenic miRNA that prevents endothelial cell proliferation at least partly by targeting β1 integrin.
Abstract Inflammasomes are multiprotein signaling platforms that form upon sensing microbe- or damage-associated molecular patterns. Upon their formation， caspase-1 is activated， leading to the processing of certain proinflammatory cytokines and the initiation of a special type of cell death， known as pyroptosis. Among known inflammasomes， NLRP3 takes on special importance because it appears to be a general sensor of cell stress. Moreover， unlike other inflammasome sensors， NLRP3 inflammasome activity is under additional transcriptional regulation. In this study， we identify the myeloid-specific microRNA miR-223 as another critical regulator of NLRP3 inflammasome activity. miR-223 suppresses NLRP3 expression through a conserved binding site within the 3' untranslated region of NLRP3， translating to reduced NLRP3 inflammasome activity. Although miR-223 itself is not regulated by proinflammatory signals， its expression varies among different myeloid cell types. Therefore， given the tight transcriptional control of NLRP3 message itself， miR-223 functions as an important rheostat controlling NLRP3 inflammasome activity.
MicroRNA-223 (miR-223) has been shown to be a potential diagnostic and prognostic marker for several cancers. In addition， miR-223 has been reported to suppress osteosarcoma cell proliferationin vitro. However， the clinical value of miR-223 is still unknown. We detected the expression of miR-223 expression in the serum of osteosarcoma patients and in osteosarcoma cancer cells using RT-PCR. We compared the serum expression of miR-223 with the clinicopathological characteristics and survival of osteosarcoma patients. Finally， we explored the role of miR-223 on the invasion of osteosarcoma cancer cells using cell migration and invasion assays. We observed that the expression of miR-223 was significantly decreased in the serum of osteosarcoma patients and osteosarcoma cancer cells compared to healthy controls (P<0.01). Moreover， a receiver operating characteristic (ROC) curve analysis indicated that serum miR-223 is a potential diagnostic marker of osteosarcoma with an area under the ROC curve (AUC) of 0.956. Importantly， the patients with a lower expression of miR-223 tended to have distant metastasis (P<0.001) and a more advanced clinical stage (P<0.001). In addition， the survival time of patients with low miR-223 expression was significantly shorter compared to patients with high miR-223 expression (P<0.001). Furthermore， we found that miR-223 could inhibit the migration and invasion of osteosarcoma cells. miR-223 might be related to the metastasis of osteosarcoma and could be used as a potential diagnostic and prognostic biomarker in osteosarcoma.
Patient-reported quality of life (QOL) analysis of radium-223 dichloride (Ra-223) evaluating pain relief from the phase 3 ALSYMPCA study.
Ra-223， a first-in-class α-emitter， significantly improved overall survival vs placebo (pbo) and was well tolerated (Parker. 2013; ALSYMPCA). Ra-223 had a positive impact on pain， significantly delaying time to external beam radiotherapy and reducing pain and opioid use (Nilsson. ASCO GU 2013). This post hoc analysis assessed Ra-223 treatment (tx) effects on QOL Functional Assessment of Cancer Therapy-Prostate (FACT-P) pain-related score (PRS)， with subgroup assessments by prior docetaxel use (pD) and baseline number of bone mets. Pain was assessed at baseline and wk 16 and 24 with FACT-P; PRS was derived from 4 pain-related questions. In responders， baseline PRS increased 2 points (improved pain). Nonresponders had neutral response (baseline PRS change -1， 0， +1) and failure (2-point baseline PRS decrease [worse pain]， including pts starting opioids during tx). Chi-square tests calculated values of responder rates between tx groups. Logistic regression determined odds ratios (ORs) (Ra-223 vs pbo) adjusted for stratification factors (pD， baseline total ALP， concurrent bisphosphonate) and baseline PRS as covariates. Of 921 pts (Ra-223， n = 614; pbo， n = 307)， analysis included pts with recorded PRS at each assessment visit (wk 16; Ra-223， n = 405; pbo， n = 175， wk 24， Ra-223， n = 305; pbo， n = 118， wk 16 and 24， Ra-223， n= 283; pbo， n = 109). Significantly more Ra-223 vs pbo pts were responders; pain improved at each visit (wk 16， = 0.028; wk 24， = 0.007) and across visits (improved for wk 16 and 24， = 0.013). Ra-223 tx significantly increased odds of improved pain vs pbo at wk 16 (OR， 1.70; 95% CI， 1.08-2.70; = 0.023) and wk 24 (OR， 2.18; 95% CI， 1.17-4.06; = 0.014); Table shows increased odds for wk 16 and 24. For all assessed subgroups and time points， a nominally higher % of Ra-223 pts had improved pain vs pbo pts. Ra-223 was associated with greater odds of pain relief vs pbo based on FACT-P PRS.
On May 15， 2013， the U.S. Food and Drug Administration () approved Ra 223 dichloride (Ra-223; Xofigo injection; Bayer HealthCare Pharmaceuticals Inc.) for the treatment of patients with castration-resistant (CRPC)， symptomatic ， and no known visceral . The review was based on clinical trial -06， which randomly allocated patients (2：1) to either Ra-223 plus best standard of care (BSoC) or placebo plus BSoC. The primary endpoint was overall survival (OS) with a key secondary endpoint of time to first symptomatic skeletal event (SSE). A statistically significant improvement in OS was demonstrated [HR， 0.70; 95% confidence interval， 0.55-0.88， P = 0.0019]. At the prespecified interim analysis， the median OS durations were 14.0 and 11.2 months in the Ra-223 and placebo arms， respectively. The improvement in OS was supported by a delay in time to first SSE favoring the Ra-223 arm. The most common (>10%) adverse reactions in patients receiving Ra-223 were nausea， diarrhea， vomiting， and peripheral edema. The most common (>10%) hematologic laboratory abnormalities were ， ， ， ， and . Ra-223 is the first α-emitting radiotherapeutic and the first radiopharmaceutical to demonstrate an OS advantage in .
BACKGROUND Radium-223 prolongs overall survival in patients with castration-resistant prostate cancer (CRPC) and symptomatic bone metastases， regardless of prior docetaxel. Whether or not chemotherapy can be safely administered following radium-223 treatment is of clinical importance. An exploratory analysis of prospectively collected data， from the ALSYMPCA (ALpharadin in SYMptomatic Prostate CAncer) patient subgroup who received chemotherapy after radium-223 or placebo treatment， was conducted to evaluate the safety and efficacy of chemotherapy following radium-223. METHODS In ALSYMPCA， CRPC patients with symptomatic bone metastases and no visceral metastases were randomized 2：1 to receive six injections of radium-223 (50 kBq/kg IV) or placebo plus best standard of care， stratified by prior docetaxel， baseline alkaline phosphatase， and current bisphosphonate use. In this exploratory analysis， chemotherapy agents administered following study treatment were identified; timing and duration were calculated. Hematologic safety was reviewed， and overall survival analyzed. RESULTS Overall， 142 radium-223 and 64 placebo patients received subsequent chemotherapy; most common were docetaxel (70% radium-223， 72% placebo) and mitoxantrone (16% radium-223， 20% placebo). The majority of patients (61% radium-223， 58% placebo) had received prior docetaxel. Radium-223 patients started subsequent chemotherapy later than placebo patients; chemotherapy duration was similar between groups. In radium-223 and placebo patients receiving subsequent chemotherapy， median hematologic values (hemoglobin， neutrophils， and platelets) remained nearly constant up to 18 months following start of chemotherapy， regardless of prior docetaxel treatment. A low percentage of patients in both groups had grades 3–4 hematologic values (<10%). Platelet count decline， from last measurement before chemotherapy， was numerically greater in radium-223 versus placebo patients. Median overall survivals from start of chemotherapy were 16.0 and 15.8 months following radium-223 and placebo， respectively. CONCLUSIONS Chemotherapy following radium-223， regardless of prior docetaxel， is feasible and appears to be well tolerated in patients with CRPC and symptomatic bone metastases. Prostate . © 2016 Wiley Periodicals， Inc.
Glycogen synthase kinase (GSK)-3， a negative regulator of cardiac hypertrophy， is inactivated in failing hearts. To examine the histopathological and functional consequence of the persistent inhibition of GSK-3223; in the heart in vivo， we generated transgenic mice with cardiac-specific overexpression of dominant negative GSK-3223; (Tg-GSK-3223;-DN) and tetracycline-regulatable wild-type GSK-3223;. GSK-3223;-DN significantly reduced the kinase activity of endogenous GSK-3223;， inhibited phosphorylation of eukaryotic translation initiation factor 2B， and induced accumulation of 223;-catenin and myeloid cell leukemia-1， confirming that GSK-3223;-DN acts as a dominant negative in vivo. Tg-GSK-3223;-DN exhibited concentric hypertrophy at baseline， accompanied by upregulation of the -myosin heavy chain gene and increases in cardiac function， as evidenced by a significantly greater Emax after dobutamine infusion and percentage of contraction in isolated cardiac myocytes， indicating that inhibition of GSK-3223; induces well-compensated hypertrophy. Although transverse aortic constriction induced a similar increase in hypertrophy in both Tg-GSK-3223;-DN and nontransgenic mice， Tg-GSK-3223;-DN exhibited better left ventricular function and less fibrosis and apoptosis than nontransgenic mice. Induction of the GSK-3223; transgene in tetracycline-regulatable wild-type GSK-3223; mice induced left ventricular dysfunction and premature death， accompanied by increases in apoptosis and fibrosis. Overexpression of GSK-3223;-DN in cardiac myocytes inhibited tumor necrosis factor-–induced apoptosis， and the antiapoptotic effect of GSK-3223;-DN was abrogated in the absence of myeloid cell leukemia-1. These results suggest that persistent inhibition of GSK-3223; induces compensatory hypertrophy， inhibits apoptosis and fibrosis， and increases cardiac contractility and that the antiapoptotic effect of GSK-3223; inhibition is mediated by myeloid cell leukemia-1. Thus， downregulation of GSK-3223; during heart failure could be compensatory.