Jiahong Xia

The expression levels of plasma micoRNAs in atrial fibrillation patients.

Background MicroRNA (miRNA) has been found in human blood. It has been increasingly suggested that miRNAs may serve as biomarkers for diseases. We examined the potential of circulating miRNA to serve as predictors of atrial fibrillation (AF). Methodology/Principal Findings During the discovery stage of this project, we used massively parallel signature sequencing (MPSS) to carry out an in-depth analysis of the miRNA expression profile (miRNome) in 5 healthy controls, 5 patients with paroxysmal atrial fibrillation (PAF) alone, and 5 patients with persistent atrial fibrillation (PersAF) alone. Twenty-two specific miRNAs were found to be dysregulated in each PAF group, PersAF group, or control group. Four candidate microRNAs (miRNA-146a, miRNA-150, miRNA-19a, and miRNA-375) met our selection criteria and were evaluated in an independent cohort of 90 plasma samples using TaqMan miRNA quantitative reverse transcriptase–polymerase chain reaction (qRT-PCR). We found miRNA-150 levels to be reduced by a factor of approximately 17 in PAF relative to controls and a factor of approximately 20 in PersAF relative to controls (P<.0001). Logistic regression analyses were carried out to evaluate the reduced miRNA-150 expression levels (odds ratio [OR] 1.96, 95% confidence interval [CI] 1.5 to 3.57, P<0.001), age (OR 1.1, 95% CI 1.36 to 2.73, P<0.001), and Left atrial diameter (LAD) (OR 1.5, 95% CI 1.36 to 1.8, P<0.001). Each was independently associated with AF. Much of the identified target genes related to AF were part of the inflammatory response system. We found that plasma levels of CRP were negatively correlated with the plasma levels of miRNA-150. Conclusions/Significance In summary, we firstly found that plasma miRNA-150 levels in from AF patients were substantially lower than that from healthy people. Circulating reduced miRNA-150 was significantly associated with AF.

MiRNAs as biomarkers of myocardial infarction: a meta-analysis.

Background Recent studies have demonstrated that acute myocardial infarction induces a distinctive miRNA signature, suggesting that miRNAs may serve as diagnostic markers. Although many studies have investigated the use of miRNAs in the detection of cardiac injury, some had small sample sizes (<100 patients) or reported different results for the same miRNA. Here, the role of circulating miRNAs for use as biomarkers of myocardial infarction is summarized and analyzed. Methods and Results Medline, SCI, Embase, and Cochrane databases were searched up to January 2013 for studies that evaluated associations between miRNAs and myocardial infarction. Relevant publications were identified by searching for combinations of “myocardial infarction,” “miRNAs,” and their synonyms. Methodological quality was scored using a standardized list of criteria, and diagnostic performance was assessed using estimates of test sensitivity and specificity. These values were summarized using summary receiver-operating characteristic curves. Nineteen studies met the inclusion criteria: 15 studies reported sensitivity, specificity, and AUC, but 4 studies did not. Total miRNAs: sensitivity: 0.78 (95%CI: 0.77–0.80; P = 0.0000); specificity: 0.82 (95%CI: 0.80–0.83; P = 0.0000). miR-499: sensitivity: 0.88 (95%CI:0.86–0.90; P = 0.0000); specificity: 0.87 (95%CI:0.84–0.90; P = 0.0000). miR-1: sensitivity: 0.63 (95%CI:0.59–0.66; P = 0.0000); specificity: 0.76 (95%CI:0.71–0.80; P = 0.0000). miR-133a: sensitivity: 0.89 (95%CI:0.83–0.94; P = 0.0047); specificity: 0.87 (95%CI:0.79–0.92; P = 0.0262). miR-208b: sensitivity: 0.78 (95%CI:0.76–0.81; P = 0.0581); specificity: 0.88 (95%CI:0.84–0.91; P = 0.0000). The correlation between miRNAs and other diagnostic biomarkers of myocardial infarction was obvious. Conclusion MiRNAs, especially miR-499 and miR-133a, may be suitable for use as diagnostic biomarkers of myocardial infarction.

Foxp3+ T regulatory cells (Tregs) are increased in nasal polyps (NP) after treatment with intranasal steroid

The pathogenesis of chronic rhinosinusitis (CRS) with nasal polyps(NP) is still poorly understood. To evaluate the role of Foxp3+ T regulatory cells (Tregs) in the pathogenesis and management of NP, we investigated the location and expression of Foxp3 in NP before and after treatment with intranasal steroid. NP specimens were obtained from 14 patients with NP before and after intranasal administration of mometasone (50 microg/day for 4 weeks). Foxp3 was detected by double immunofluorescence stain, quantitative reverse transcriptase polymerase chain reaction (RT-PCR), flow cytometry and western blot. The concentration of interleukin(IL)-10 in supernatants of homogenized tissue was measured by enzyme-linked immunosorbent assay (ELISA). We found that Foxp3 and IL-10 were downregulated in NP compared to the control mucosa (P<0.05). Foxp3 and IL-10 expression were increased significantly after intranasal steroid treatment (P<0.05). And Foxp3 was tightly correlated with IL-10 in NP (P<0.05) after treatment. These data suggest that Foxp3 is downregulated in NP and intranasal steroid attenuates the chronic inflammatory response by enhancing the expression and function of Foxp3 in NP.

GM-CSF contributes to aortic aneurysms resulting from SMAD3 deficiency

Heterozygous loss-of-function SMAD3 (Mothers against decapentaplegic homolog 3) mutations lead to aneurysm-osteoarthritis syndrome (AOS). In the present study, we found that mice lacking Smad3 had a vascular phenotype similar to AOS, marked by the progressive development of aneurysms. These aneurysms were associated with various pathological changes in transmural inflammatory cell infiltration. Bone marrow transplants from Smad3-/- mice induced aortitis and aortic root dilation in irradiated WT recipient mice. Transplantation of CD4+ T cells from Smad3-/- mice also induced aortitis in Smad3+/+ recipient mice, while depletion of CD4+ T cells in Smad3-/- mice reduced the infiltration of inflammatory cells in the aortic root. Furthermore, IFN-γ deficiency increased, while IL-17 deficiency decreased, disease severity in Smad3+/- mice. Cytokine secretion was measured using a cytokine quantibody array, and Smad3-/- CD4+ T cells secreted more GM-CSF than Smad3+/+ CD4+ T cells. GM-CSF induced CD11b+Gr-1+Ly-6Chi inflammatory monocyte accumulation in the aortic root, but administration of anti-GM-CSF mAb to Smad3-/- mice resulted in significantly less inflammation and dilation in the aortic root. We also identified a missense mutation (c.985A>G) in a family of thoracic aortic aneurysms. Intense inflammatory infiltration and GM-CSF expression was observed in aortas specimens of these patients, suggesting that GM-CSF is potentially involved in the development of AOS.

Treatment With Interleukin-12/23p40 Antibody Attenuates Acute Cardiac Allograft Rejection

Background. Interleukin (IL)-12 and -23 share the p40 subunit and are crucial for the development of T helper (Th) 1- and Th17-cell responses in acute graft rejection. However, little is known about the impact of treatment with antagonistic anti-p40 antibody in inhibiting rejection of cardiac allografts. Methods. C57BL/6 mice were transplanted with syngeneic or allogeneic (BALB/c) hearts and treated with 100 or 200 &mgr;g or 400 &mgr;g anti-P40 monoclonal antibody on postoperative days 1 and 3, respectively. The survival of grafts was monitored daily by abdominal palpation until the complete cessation of cardiac contractility (endpoint). The severity of acute rejection was evaluated by histology and immunohistochemistry. The expression of transcription factors within the grafts were measured by quantitative real-time polymerase chain reaction. Systemically, the lymphocytes were characterized by flow cytometry, and the serum levels of cytokines were determined by ELISA. Results. In comparison with mice treated with isotype IgG or saline, treatment with anti-p40 significantly alleviated acute phase allograft rejection and resulted in prolonged survival of cardiac allografts (P<0.05). These changes were associated with reduced infiltration of inflammatory cells and down-regulation of Th1- and Th17-specific transcription factors and cytokines. Furthermore, treatment with anti-p40 significantly reduced the percentages of splenic Th1 and Th17 cells, but not Th2 and regulatory T cells (P<0.05), with concomitant reduction of serum interferon-&ggr; and IL-17 levels (P<0.05). Conclusion. Our data indicated that treatment with anti-p40 inhibited Th1- and Th17-cell responses and prolonged the survival of cardiac allografts in mice.

The Treg/Th17 Imbalance in Patients with Idiopathic Dilated Cardiomyopathy

To assess whether Treg/Th17 balance was broken in patients with idiopathic dilated cardiomyopathy (DCM). We studied 25 patients who were diagnosed as idiopathic DCM (18 men and seven women, mean age 35.6 ± 5.2) and 25 normal persons (18 men and seven women, mean age 33.8 ± 4.9). Then, we detected Treg/Th17 functions on different levels including cell frequencies, related cytokine secretion and key transcription factors in patients with idiopathic DCM and controls. The results demonstrated that patients with idiopathic DCM revealed significant increase in peripheral Th17 number, Th17‐related cytokines (IL‐17, IL‐6, IL‐23) and transcription factor (RORγt) levels and obvious decrease in Treg number, Treg‐related cytokines (TGF‐β1 and IL‐10) and transcription factor (Foxp3) levels when compared to normal persons. Results indicated that Treg/Th17 functional imbalance existed in patients with idiopathic DCM, suggesting a potential role for Treg/Th17 imbalance in the development of idiopathic DCM.

MicroRNA-150 Protects the Heart From Injury by Inhibiting Monocyte Accumulation in a Mouse Model of Acute Myocardial Infarction

Background—MicroRNAs (miRs) and inflammatory monocytes participate in many cardiac pathophysiological processes including acute myocardial infarction (AMI). Recently, we observed that miR-150 is downregulated in injured mouse plasma after AMI as well as in human infarcted monocytes. However, the precise functional role of miR-150 in response to AMI remains unknown. Methods and Results—In a mouse model of AMI and in human subjects with AMI, we found that miR-150 expression was reduced in monocytes. In vitro studies showed that ectopic expression of miR-150 markedly reduced monocyte migration and proinflammatory cytokine production, whereas blockade of miR-150 had opposing effects. In vivo studies showed that overexpression of miR-150 in mice resulted in improved cardiac function, reduced myocardial infarction size, inhibition of apoptosis, and reduced inflammatory Ly-6Chigh monocyte invasion levels after AMI. Wild-type mice transplanted with miR-150 null (−/−) bone marrow cells could reverse this protective effect. Mechanistic studies demonstrated that miR-150 inhibited the expression of chemokine receptor 4 (CXCR4), thereby promoting monocyte migration. Conclusions—Our findings indicate that miR-150 acts as a critical regulator of monocyte cell migration and production of proinflammatory cytokines, leading to cardioprotective effects against AMI-induced injury. Thus, miR-150 may be a suitable target for therapeutic intervention in the setting of ischemic heart disease.

CCR5 blockade in combination with cyclosporine increased cardiac graft survival and generated alternatively activated macrophages in primates.

Maraviroc (MVC), a specific antagonist of CCR5 expressed on macrophages and activated T cells, may modulate inflammation and may be useful in patients with HIV infection. In this study we used nonhuman primates to examine the effect and mechanism of MVC alone or in combination with cyclosporine (CsA) to prolong cardiac allograft survivals. In an established rhesus monkey cardiac allograft model, recipients treated with MVC plus CsA showed significantly prolonged survival of heart allografts (>240 d, p < 0.001). These in vivo results in the MVC/CsA group correlated with delayed alloantibody response and markedly decreased graft infiltration by CCR5+, CD4+, CD8+, and CD68+ cells (p < 0.05), as compared with other groups. Furthermore, grafts from the MVC/CsA group had elevated numbers of alternatively activated macrophages (AAMs) and the expression of peroxisome proliferator-activated receptor γ (PPARγ). Blockade of PPARγ abrogated the prolonged allograft survival (median survival time, 45 d) and the upregulated AAMs in MVC/CsA-treated recipients. In conclusion, MVC/CsA protects cardiac allograft in primates and this effect is associated with generating AAMs through activation of the PPARγ nuclear receptor.

Dynamic changes in Th1, Th17, and FoxP3+ T cells in patients with acute cellular rejection after cardiac transplantation

Wang S, Li J, Xie A, Wang G, Xia N, Ye P, Rui L, Xia J. Dynamic changes in Th1, Th17, and FoxP3+ T cells in patients with acute cellular rejection after cardiac transplantation. 
Clin Transplant 2011: 25: E177–E186.

Retinoic acid attenuates acute heart rejection by increasing regulatory T cell and repressing differentiation of Th17 cell in the presence of TGF‐β

Retinoic acid (RA), in a transforming growth factor beta (TGF‐β)‐dependent manner, promotes differentiation of regulatory T cells (Tregs) but inhibits the differentiation of Th17 cells in vitro from naive CD4+T cells. In addition, transfer of induced Tregs (iTregs) reduces rejection. We therefore examined whether RA could attenuate acute cardiac transplant rejection in vivo in a mouse model by regulating the reciprocal differentiation of Tregs and Th17 cells. The iTregs and naive T cells were respectively transferred into congenic mice. Two weeks later, the percentages of transferred cells and Forkhead box P3 (FoxP3)+ Tregs were measured in spleen. Mice with cardiac transplants were treated with TGF‐β alone, RA alone, both or none. The percentage of Tregs or Th17 cells in CD4+T cells, the level of FoxP3 protein or serous interleukin (IL)‐17A, or suppressive function of Tregs from recipient mice were assessed. The percentage of Th17 cells and level of serum IL‐17A both increased significantly during acute rejection. RA favored differentiation to Tregs over Th17 cells. Unlike naive T cells, only a few transferred iTregs remained after transfer. Treatment with RA plus TGF‐β prolonged graft survival, increased the percentage of Tregs, and decreased the percentage of Th17 cells in peripheral T cells. Tregs from all recipients had normal suppressive function. In conclusion, treatment with RA plus TGF‐β attenuates acute rejection by promoting the differentiation of Tregs and inhibiting the differentiation of Th17 cells.