Acute KD Research Articles

The Effectiveness of No or Low-Dose versus High-Dose Aspirin in Treating Acute Kawasaki Disease: A Systematic Review and Meta-Analysis.

This systematic review and meta-analysis assesses the effectiveness of no or low-dose versus high-dose aspirin on the incidence of coronary artery aneurysms (CAAs), intravenous immunoglobulin (IVIG) resistance, hospital stay length, and fever duration during the acute phase of Kawasaki disease. Our review adheres to the Preferred Reporting Items for Systematic Reviews guidelines. The PubMed and Google Scholar databases were comprehensively searched to identify relevant studies in the literature, including observational studies and randomized controlled trials (RCTs). The primary outcome was the incidence of CAAs. The secondary outcomes were the hospital stay length, fever duration, and IVIG resistance. The risk of bias was assessed using the Newcastle-Ottawa scale for cohort studies and Cochrane's Risk of Bias Tool for RCTs. The data were analyzed using the Review Manager software. Twelve studies with a total of 68,495 participants met the inclusion criteria. The incidences of CAAs (odds ratio [OR] = 0.93; 95% confidence interval [CI] = 0.64-1.34) and IVIG resistance (OR = 1.46; 95% CI = 1.00-2.12) did not differ significantly between no or low-dose versus high-dose aspirin in treating acute KD. Moreover, the fever durations (mean difference [MD] = 3.55 h; 95% CI = -7.99-15.10) and hospital stay lengths (MD = -0.54 days; 95% CI = -2.50-1.41) were similar in the no and low-dose aspirin group compared to the high-dose aspirin group. Our review indicates that there are no significant differences in the incidences of CAA and IVIG resistance, fever durations, and hospital stay lengths between no or low-dose versus high-dose aspirin in treating the acute phase of KD.

Identification of hub biomarkers and immune-related pathways participating in the progression of Kawasaki disease by integrated bioinformatics analysis

BackgroundKawasaki disease (KD) is a systemic vasculitis that commonly affects children and its etiology remains unknown. Growing evidence suggests that immune-mediated inflammation and immune cells in the peripheral blood play crucial roles in the pathophysiology of KD. The objective of this research was to find important biomarkers and immune-related mechanisms implicated in KD, along with their correlation with immune cells in the peripheral blood. Material/MethodsGene microarray data from the Gene Expression Omnibus (GEO) was utilized in this study. Three datasets, namely GSE63881 (341 samples), GSE73463 (233 samples), and GSE73461 (279 samples), were obtained. To find intersecting genes, we employed differentially expressed genes (DEGs) analysis and weighted gene co-expression network analysis (WGCNA). Subsequently, functional annotation, construction of protein–protein interaction (PPI) networks, and Least Absolute Shrinkage and Selection Operator (LASSO) regression were performed to identify hub genes. The accuracy of these hub genes in identifying KD was evaluated using the receiver operating characteristic curve (ROC). Furthermore, Gene Set Variation Analysis (GSVA) was employed to explore the composition of circulating immune cells within the assessed datasets and their relationship with the hub gene markers. ResultsWGCNA yielded eight co-expression modules, with one hub module (MEblue module) exhibiting the strongest association with acute KD. 425 distinct genes were identified. Integrating WGCNA and DEGs yielded a total of 277 intersecting genes. By conducting LASSO analysis, five hub genes (S100A12, MMP9, TLR2, NLRC4 and ARG1) were identified as potential biomarkers for KD. The diagnostic value of these five hub genes was demonstrated through ROC curve analysis, indicating their high accuracy in diagnosing KD. Analysis of the circulating immune cell composition within the assessed datasets revealed a significant association between KD and various immune cell types, including activated dendritic cells, neutrophils, immature dendritic cells, macrophages, and activated CD8 T cells. Importantly, all five hub genes exhibited strong correlations with immune cells. ConclusionActivated dendritic cells, neutrophils, and macrophages were closely associated with the pathogenesis of KD. Furthermore, the hub genes (S100A12, MMP9, TLR2, NLRC4, and ARG1) are likely to participate in the pathogenic mechanisms of KD through immune-related signaling pathways.

Diagnostic significance of noncoding RNAs in kawasaki disease: A systematic review and meta-analysis.

Kawasaki disease (KD) is a systemic vasculitis disease, and early effective intervention would reduce the occurrence of coronary artery lesions (CALs). Recently, many scholars have been committed to studying the relationship between noncoding RNAs and KD. This systematic review aimed to analyze the diagnostic value of noncoding RNAs(ncRNAs) in distinguishing different KD status. We searched for the literature about diagnostic values of ncRNAs in KD in CNKI, VIP, Wanfang, China Biomedical Literature Database as well as PubMed, Web of Science, Embase, and Cochrane Library up to April 15, 2022. All included studies were further analyzed using STATA 12.0, Meta-disc 1.4 and RevMan 5.4 software. A total of six studies investigating the diagnostic performance of ncRNAs in differentiating KD-CAL (n = 101) from KD-NCAL patients (n = 123) were included in this this meta-analysis. The calculated area under the curve(AUC) was 0.83 (0.80-0.86). Four studies on the diagnostic performance of ncRNAs in differentiating acute KD patients (n = 139) from convalescent KD patients (n = 109) were included. The calculated AUC was 0.87 (0.84-0.90). Four studies focused on the diagnostic performance of ncRNAs combined with other laboratory indexes in KD by assessing 137 KD patients and 152 febrile controls. The calculated AUC was 0.90 (0.87-0.92). Four studies assessed the diagnostic performance of ncRNAs in differentiating intravenous immunoglobulin (IVIG)-resistant KD patients from IVIG-responsive KD patients. The calculated AUC was 0.9135 ± 0.0307. These results indicated that ncRNAs have a good diagnostic efficacy in KD. This meta-analysis showed that ncRNAs have potential as a biomarker for distinguishing different KD status. However, since limited studies were included in this meta-analysis, larger and well-designed diagnostic studies should be conducted to validate these results. INPLASY.COM, identifier: doi: 10.37766/inplasy2022.10.0035.

A dual covariant biomarker approach to Kawasaki disease, using vascular endothelial growth factor A and B gene expression; implications for coronary pathogenesis

IntroductionKawasaki disease (KD) is the most common vasculitis in young children, with coronary artery lesions (CALs) and coronary aneurysms (CAAs) being responsible for most KD-related deaths. ObjectiveWe hypothesized that Vascular Endothelial Growth Factors (VEGFs) are pivotal in KD inflammation and coronary artery lesions. This study assessed VEGF-A and VEGF-B gene expression (GE) as potential biomarkers in KD inflammation. Study designWe analyzed NCBI-GEO datasets, categorizing gene expression patterns as "inflammatory" or "non-inflammatory". We focused on TNF-, NFKB1, VEGF-A, and VEGF-B GEs. Datasets were filtered based on differential changes in TNF and NFKB1 levels to isolate those with inflammatory shifts. ResultsInflammatory datasets (GSE63881, GSE73464, and GSE68004) displayed elevated TNF, NFKB1, and VEGF-A GE levels during acute KD. VEGF-B GE exhibited a distinctive trend: an initial drop and subsequent rise during recovery, a pattern that was missing in the non-inflammatory group. The treatment response was also studied, with intravenous immunoglobulin (IVIG) responders showing significant downregulation of NFKB1 GE after treatment: GSE16797 [IVIG ± methylprednisolone; p = 8.6443-03], GSE48498 [IVIG; p = 6.618e-02, infliximab; p = 3.240e-03], and GSE18606 [IVIG; p = 3.518e-02]. Considering the similar binding of VEGF-A and VEGF-B to the VEGFR1 receptor, a co-variate and inverse relationship is suggested. ConclusionTemporal VEGF-A, VEGF-B, and GE changes show promise as new post-inflammatory biomarkers for KD. Novelty results with the biomarker approach, with the potential for a dual temporal relationship between VEGF-A and VEGF-A. A comprehensive exploration of VEGF-A and VEGF-B genes and protein analysis in KD is warranted to understand the functional aspects of these changes and how best to utilize the pattern of changes for therapeutic benefit.

Platelet-derived microRNA-223 attenuates TNF-α induced monocytes adhesion to arterial endothelium by targeting ICAM-1 in Kawasaki disease.

BackgroundKawasaki disease (KD) is an acute vasculitis that may result in permanent coronary artery damage with unknown etiology. Endothelial cell (EC) dysfunction and platelet hyperactivity are the hallmarks of KD. Platelets are involved in the development of endothelial dysfunction. MiR-223 transferred by platelet microparticles (PMPs) has been found to involve in the functional regulation of endothelial cells in sepsis. However, the role of platelet-derived miR-223 in endothelial dysfunction has not yet been investigated in KD.ObjectivesWe seek to investigate the role of platelet-derived miR-223 in endothelial dysfunction of KD vasculopathy.Methods and resultsForty-five acute KD patients and 45 matched controls were randomly recruited in the study. When co-cultured with human coronary artery endothelial cells (HCAECs), KD platelets with higher levels of miR-223 were incorporated into HCAECs, resulting in the horizontal transfer of miR-223. Using KD platelets, PMPs, and platelet-releasate from the same amount of blood co-cultured with HCAECs, we found the increased expression of miR-223 in HCAECs was primarily derived from KD platelets, rather than PMPs or free miRNAs from platelet- releasate. KD platelet-derived miR-223 attenuated TNF-α induced intercellular cell adhesion molecule-1 (ICAM-1) expression in HCAECs. KD platelet-derived miR-223 also suppressed the monocyte adhesion to HCAECs. In vivo, platelet-specific miR-223 knockout (PF4-cre: miR-223flox/flox) C57BL/6 mice and miR-223flox/flox C57BL/6 mice were used. Using Lactobacillus casei cell wall extract (LCWE) to establish KD murine model, we showed that in LCWE-injected PF4-cre: miR-223flox/flox mice, deficiency of platelet-miR-223 exacerbates the medial thickening of the abdominal aorta, increased ICAM-1 expression with concomitant CD45+ inflammatory cells infiltration into the endothelium compared to LCWE-injected miR-223flox/flox mice.ConclusionsThe horizontal transfer of platelet-derived miR-223 suppresses the expression of ICAM-1 in HCAECs, which at least in part attenuates leukocyte adhesion, thereby reducing endothelial damage in KD vasculitis

Intravenous Immunoglobulin Nonresponse in Pediatric Patients Treated for Kawasaki Disease at a US Hospital.

Kawasaki disease (KD) is an acute febrile childhood vasculitis with a predilection for the coronary arteries treated with IVIG. In the United States, scoring systems to identify children at high-risk of persistent fever after initial IVIG treatment are lacking. Our study attempts to identify variables associated with IVIG non-response. Retrospective review of patients ages 0 to 18 admitted to an US academic children's hospital between August 1, 2010, and August 31, 2019, with the diagnosis of acute KD who received IVIG during hospitalization. A total of 64 patients were included, 73% male and 66% Caucasian with a mean age of 3.67 ± 3.35 years. Forty-eight patients (75%) received 1 dose of IVIG, and 16 (25%) received 2 doses of IVIG. The groups did not differ significantly at baseline. None had coronary artery aneurysms detected during hospitalization. Older age, female sex, Caucasian compared with African American race, leukocytosis, and hyponatremia were associated with a higher likelihood of IVIG non-response but none reached statistical significance. Patients who received ibuprofen (n = 26) were more likely to be IVIG non-responsive (p < 0.05). Aspirin dosing varied but was not predictive of IVIG non-response. In this study, risk factors to predict IVIG non-response in patients treated for KD were not identified. IVIG non-response was significantly more common in those receiving ibuprofen during the acute treatment phase. Larger studies are needed to validate the association of ibuprofen administration and IVIG non-response in patients with KD.

A phase I/IIa trial of atorvastatin in Japanese patients with acute Kawasaki disease with coronary artery aneurysm: Study protocol of a multicenter, single-arm, open-label trial

BackgroundKawasaki disease (KD) is a systemic vasculitis complicated with coronary artery abnormalities (CAAs). Intravenous immunoglobulin reduces the occurrence of CAAs, but significant number of KD patients with CAAs still exists. Thus, new approaches to prevent and attenuate CAAs are warranted. Atorvastatin has been shown to promote endothelial cell homeostasis and suppress vascular inflammation and has received enthusiasm as a potentially new candidate treatment for KD. In the United States, a phase I/IIa dose-escalation study of atorvastatin in KD patients with CAAs demonstrated the safety and pharmacokinetic data of atorvastatin. However, due to the uncertainty in the application of these results to other populations, we aim to examine the tolerability and generate pharmacokinetics data in Japanese KD patients. MethodsThis is a multicenter, single-arm, open-label, phase I/IIa study of atorvastatin in acute KD patients with CAAs in Japan. A minimum of 9 and a maximum of 18 KD patients (2 years–17 years old) will be recruited for a 3 + 3 dose-escalation study of a 6-week course of atorvastatin (0.125–0.5 mg/kg/day). The primary outcome will be safety of atorvastatin. The secondary outcomes will be pharmacokinetics of atorvastatin, activity of atorvastatin and echocardiographic assessment of CAAs. The activity of atorvastatin will include assessment of C-reactive protein or high sensitivity C-reactive protein and white blood cell levels. DiscussionThis study will provide evidence of the safety, tolerability, and pharmacokinetics of atorvastatin in Japanese KD patients and may lead new standard therapy for acute-phase KD associated with CAA complications. Trial registrationJapan Registry of Clinical Trials (JRCTs031180057). Registered December 19, 2018, https://jrct.niph.go.jp/en-latest-detail/jRCTs031180057.

Effects of immunoglobulin plus prednisolone in reducing coronary artery lesions in patients with Kawasaki disease: study protocol for a phase III multicenter, open-label, blinded-endpoints randomized controlled trial

BackgroundKawasaki disease (KD) is an acute systemic vasculitis of unclear etiology that mainly affects infants and young children. Strategies to reduce the incidence and severity of coronary artery lesions (CALs), the determinant factor in the long-term prognosis of KD, are currently a focus of studies on KD. Corticosteroids, preferred in the treatment of the majority of vasculitides, are controversial in the treatment of acute KD. In this trial, we will evaluate whether the addition of prednisolone to standard intravenous immunoglobulin (IVIG) plus aspirin therapy can reduce the occurrence of CAL in Chinese patients with KD.MethodsThis is a multicenter, prospective, open-label, randomized controlled trial, which is expected to be conducted in more than 20 hospitals in China and aims to assess the efficacy and safety of IVIG + prednisolone treatment versus standard treatment. Patients with KD who fulfill the inclusion and exclusion criteria will be recruited and randomized (1:1) to receive either a large dose of IVIG (2 g/kg over 12–24 h with a maximum dose of 60 g) + aspirin 30 mg/kg/d or IVIG (2 g/kg over 12–24 h) + aspirin 30 mg/kg/d + prednisolone (2 mg/kg/d with a maximum dose of 60 mg tapered over 15 days after normalization of C-reactive protein concentration). The primary outcome will be the occurrence of CAL at 1 month of illness. The follow-up duration for each participant will be set as 1 year. Patients and treating physicians will be unmasked to group allocation.DiscussionThis will be the first multicenter randomized controlled trial to evaluate the efficacy of IVIG + aspirin + prednisolone in Chinese pediatric patients with KD, which may provide high-level evidence for improving the initial treatment for acute KD.Trial registrationClinicalTrials.govNCT04078568. Registered on 16 August 2018.

Cognitive development of children with Kawasaki disease and the parenting stress of their caregivers in Taiwan: a case–control study

ObjectiveKawasaki disease (KD) is an acute form of febrile vasculitis that occurs in early childhood. The multisystemic vasculitis common in patients with KD may influence blood perfusion in the brain,...

Kawasaki disease presenting with hoarseness: A multinational study of the REKAMLATINA network.

Recently, hoarseness affecting the supraglottic structure has been reported in Kawasaki disease (KD). The objective of this study was to characterize the frequency of hoarseness in acute KD patients in Latin America. We used prospective data from the multinational Red de Enfermedad de Kawasaki en America Latina (REKAMLATINA) network. A total of 865 patients from 20 countries were enrolled during the 3 year study period. Data on hoarseness were available in 858 (99.2%) patients. The clinical and laboratory characteristics between hoarse and non-hoarse KD were compared. Hoarseness was documented in 100 (11.6%) patients. Hoarse patients were younger than those with KD without hoarseness (median age 18 vs 26months; P=0.002) and presented with lower hemoglobin (10.7g/dL vs 11.3g/dL; P=0.040) and hematocrit levels (32% vs 33%, P=0.048). Hoarseness was found to be prevalent as a presenting sign of acute KD in younger children. Anemia may indicate the presence of active inflammation.

Effects of changes on gut microbiota in children with acute Kawasaki disease.

BackgroundKawasaki disease (KD) is an acute febrile illness of early childhood. The exact etiology of the disease remains unknown. At present, research on KD is mostly limited to susceptibility genes, infections, and immunity. However, research on the correlation between gut microbiota and KD is rare.MethodsChildren with a diagnosis of acute KD and children undergoing physical examination during the same period were included. At the time of admission, the subjects’ peripheral venous blood and feces were collected. Faecal samples were analyzed for bacterial taxonomic content via high-throughput sequencing. The abundance, diversity, composition, and characteristic differences of the gut microbiota in KD and healthy children were compared by alpha diversity, beta diversity, linear discriminant analysis and LDA effect size analysis. Blood samples were used for routine blood examination, biochemical analysis, and immunoglobulin quantitative detection.ResultsCompared with the control group, the community richness and structure of gut microbiota in the KD group was significantly reduced (Chao1 richness estimator, mean 215.85 in KD vs. mean 725.76 in control, p < 0.01; Shannon diversity index, mean 3.32 in KD vs. mean 5.69 in control, p < 0.05). LEfSe analysis identified two strains of bacteria significantly associated with KD: Bacteroidetes and Dorea. Bacteroidetes were enriched in healthy children (mean 0.16 in KD vs. mean 0.34 in control, p < 0.05). Dorea was also enriched in healthy children but rarely existed in children with KD (mean 0.002 in KD vs. mean 0.016 in control, p < 0.05). Compared with the control, IgA and IgG in the KD group decreased (IgA, median 0.68 g/L in KD vs. median 1.06 g/L in control, p < 0.001; IgG, median 6.67 g/L in KD vs. median 9.71 g/L in control, p < 0.001), and IgE and IgM levels were not significantly changed.ConclusionsDysbiosis of gut microbiota occurs in children with acute KD and may be related to the etiology or pathogenesis of KD. It is worth noting that for the first time, we found that Dorea, a hydrogen-producing bacterium, was significantly reduced in children with acute KD. Overall, our results provide a theoretical basis for the prevention or diagnosis of KD based on intestinal microecology.

Elevated levels of pentraxin 3 correlate with neutrophil counts and coronary artery dilation during acute Kawasaki disease, as a predictor of coronary artery lesions

Abstract Kawasaki disease (KD) is the leading cause of acquired heart disease in the developed world. 25–30% of untreated patients will develop irreversible coronary artery lesions (CAL). Pentraxin-3 (PTX-3), has been well studied in inflammatory diseases, in particular vascular endothelial dysfunction associated with cardiovascular disease. We hypothesized that PTX-3 played an important role in the development of KD associated CAL, and investigated circulating levels of PTX-3 in KD patients serum. Children with acute KD were followed from diagnosis through normalization of the clinical markers of inflammation. Blood draws and echocardiograms were conducted at several timepoints, acute (prior to IVIG treatment), sub-acute (5–10 days after IVIG treatment), and convalescent (1–4 months after KD diagnosis) phases. In this study we enrolled 70 children with KD, which included 26 (37%) who presented with CAL. Patients included in this study came from a diverse ethnic background. We observed a significant increase in PTX-3 levels during acute KD phase, compared to sub-acute and convalescent phases. PTX-3 levels during acute KD were significantly higher among KD patients with CAL. Further, PTX-3 levels were significantly correlated to coronary artery diameter size during acute KD and neutrophil counts throughout KD progression regardless of coronary artery diameter. We observed PTX-3 levels highly correlated to neutrophil counts, a known source of PTX-3 in acute inflammation and an important player in the development of KD vasculitis. Therefore, we propose PTX-3 as a novel factor associated with acute KD and in the development of KD associated CAL, and neutrophil derived PTX-3 as an important player of KD vascular dysfunction.

Complement C3 deficiency exacerbates development of cardiovascular lesions in the LCWE-induced Kawasaki Disease Vasculitis

Abstract Background Kawasaki disease (KD) is an acute febrile systemic vasculitis of childhood, mostly affecting the coronary arteries. A central feature of acute KD is the activation of the innate and adaptive immune systems. Serum complement component C3 is significantly increased in children during the acute stages of KD, and a polymorphism of a C type lectin, mannose binding lectin (MBL), has been associated with KD vasculitis and coronary artery lesions. However, the role of the complement pathway in KD vasculitis remains unclear. Objective Determine if the complement system contributes to KD pathogenesis in the Lactobacillus casei cell wall extract (LCWE)-mediated murine model of KD vasculitis. Methods and Results Serum levels of various complement proteins were elevated in LCWE-injected mice. We observed that the LCWE extract, rich in rhamnose and a TLR2 ligand, was able to bind both C3 and MBL in-vitro. Genetic deletion of C3 exacerbated LCWE-induced aortitis, coronary arteritis and abdominal aorta dilation and aneurysm. Furthermore, in-vivo blockade of C5 with a C5aR1 antagonist (PMX205) also increased these cardiovascular lesions in LCWE mice. Conclusions Blocking key components of the complement system, such as C3 and C5aR, significantly enhanced cardiovascular lesions in the LCWE-induced KD vasculitis mouse model. Thus, C3 or downstream activation fragments may play a protective role in the development of KD cardiovascular lesions. Whether this protection is due to complement mediated increased clearance of immune complexes or common carbohydrate pathogen molecules will need to be determined and may help our current understanding of the immunopathologic mechanisms of KD vasculitis. (Supported by NIH grant AI072726 to MA)

IL-1 receptor antagonist, anakinra, prevents myocardial dysfunction in a mouse model of Kawasaki disease vasculitis and myocarditis.

SummaryKawasaki disease (KD) vasculitis is an acute febrile illness of childhood characterized by systemic vasculitis of unknown origin, and is the most common cause of acquired heart disease among children in the United States. While histological evidence of myocarditis can be found in all patients with acute KD, only a minority of patients are clinically symptomatic and a subset demonstrate echocardiographic evidence of impaired myocardial function, as well as increased left ventricular mass, presumed to be due to myocardial edema and inflammation. Up to a third of KD patients fail to respond to first‐line therapy with intravenous immunoglobulin (IVIG), and the use of interleukin (IL)‐1 receptor antagonist (IL‐1Ra, anakinra) is currently being investigated as an alternative therapeutic approach to treat IVIG‐resistant patients. In this study, we sought to investigate the effect of IL‐1Ra on myocardial dysfunction and its relation to myocarditis development during KD vasculitis. We used the Lactobacillus casei cell‐wall extract (LCWE)‐induced murine model of KD vasculitis and investigated the effect of IL‐1Ra pretreatment on myocardial dysfunction during KD vasculitis by performing histological, magnetic resonance imaging (MRI) and echocardiographic evaluations. IL‐1Ra pretreatment significantly reduced KD‐induced myocardial inflammation and N‐terminal pro B‐type natriuretic peptide (NT‐proBNP) release. Both MRI and echocardiographic studies on LCWE‐injected KD mice demonstrated that IL‐1Ra pretreatment results in an improved ejection fraction and a normalized left ventricular function. These findings further support the potential beneficial effects of IL‐1Ra therapy in preventing the cardiovascular complications in acute KD patients, including the myocarditis and myocardial dysfunction associated with acute KD.

Case report on kawasaki patient with immunoglobulin non-response successfully treated by infliximab

Background: Kawasaki disease KD is self-limiting acute multi-systemic vasculitis of unknown etiology.Current treatment recommendations for acute KD include IVIG and aspirin, but there are no evidence-basedguidelines for children who do not respond to IVIG treatment. Over the past few years, increasing knowledge ofthe pathophysiology of KD has resulted in the identification of key inflammation mediators and the use ofbiologic pathway targeting agents such as TNF and IL-1 inhibitors for children with IVIG-resistant disease.Case presentation: A 11-month-old girl was diagnosed with typical KD and treated with 3 IVIG and 1corticoid therapy, but the coronary arteries continued to dilate. After that, the patient was treated successfullywith Infliximab.

Risk factors and implications of progressive coronary dilatation in children with Kawasaki disease

BackgroundKawasaki disease (KD) is an acute systemic vasculitis that occurs in children and may lead to cardiovascular morbidity and mortality. Progressive coronary dilatation for at least 2 months is associated with worse late coronary outcomes in patients with KD having medium or giant aneurysms. However, the risk factors and occurrence of progressive coronary dilatation in patients with KD but without medium or giant aneurysms have been insufficiently explored.MethodsWe retrospectively enrolled 169 patients with KD from a tertiary medical center in Taiwan during 2009–2013. Medical records of all patients were reviewed. Echocardiography was performed during the acute KD phase and at 3–4 weeks, 6–8 weeks, 6 months, and 12 months after KD onset. Progressive coronary dilatation was defined as the progressive enlargement of coronary arteries on three consecutive echocardiograms. Logistic regression analysis was conducted to evaluate the potential risk factors for coronary aneurysms and progressive coronary dilatation.ResultsOf a total of 169 patients with KD, 31 (18.3%) had maximal coronary Z-scores of ≥ + 2.5 during the acute KD phase, 16 (9.5%; male/female: 9/7) had coronary aneurysms at 1 month after KD onset, and 5 (3.0%) satisfied the definition of progressive coronary dilatation. Multivariate logistic regression analysis revealed that an initial maximal coronary Z-score of ≥ + 2.5 [odds ratio (OR): 5.24, 95% confidence interval (CI): 1.31–21.3, P = 0.020] and hypoalbuminemia (OR: 4.83, 95% CI: 1.11–20.9, P = 0.035) were independent risk factors for coronary aneurysms and were significantly associated with progressive coronary dilatation. However, the association between intravenous immunoglobulin unresponsiveness and the development of coronary aneurysms at 1 month after KD onset didn’t reach the level of significance (P = 0.058).ConclusionsIn the present study, 3% (5/169) of patients with KD had progressive coronary dilatation, which was associated with persistent coronary aneurysms at 1 year after KD onset. Initial coronary dilatation and hypoalbuminemia were independently associated with the occurrence of progressive coronary dilatation. Therefore, such patients may require intensive cardiac monitoring and adjuvant therapies apart from immunoglobulin therapies.

A plasma mir-125a-5p as a novel biomarker for Kawasaki disease and induces apoptosis in HUVECs.

BackgroundKawasaki disease (KD) is a childhood systemic vasculitis that exhibits a specific preference for the coronary arteries. The aetiology remains unknown and there are no especially diagnostic tests. microRNAs (miRNAs) are 18 to 23 nucleotides non-coding RNAs that are negative regulator of gene expression and play a crucial role in the regulatory network of the genome. Recently, circulating miRNAs have been found presentation in human plasma and displayed some characteristics of the ideal biomarker. However, few researches explored differentially expressed miRNAs in the plasma of KD patients. Our study is to identify circulating miRNAs in KD plasma which can serve as potential biomarkers of KD diagnosis.Materials and methodsThe total of five pairs of acute KD and normal plasma samples were analyzed using ABI miRNAs TLDA Assay chip. Differentially expression of miR-125a-5p in plasma were confirmed by quantitative real-time PCR (qRT-PCR) in independent cohort (acute KD = 30, convalescent KD = 30 and healthy control = 32). After bioinformatics prediction, miR-125a-5p vector and inhibitor were transfected into HUVECs respectively, to observe MKK7 expression as a potential target gene. Flow cytometry was used to analyze apoptosis. The mRNA and protein levels of desired genes including MKK7, Caspase-3, Bax and Bcl2 were detected by qRT-PCR and western blotting.ResultsEighteen miRNAs were differentially expressed in acute KD’s plasma compared with healthy control. miR-125a-5p was significantly increased in plasma of KD patients (p = 0.000), but no variation between acute and convalescent KD (p = 0.357). Moreover, the results from the gain and loss functions of miR-125a-5p in HUVECs have shown that miR-125a-5p remarkably suppressed MKK7 expression, as a novel target gene. Importantly, miR-125a-5p also induced apoptosis in HUVECs through inhibition MKK7 levels to regulate Bax/Bcl2 pathway resulting to activate Caspase-3.ConclusionOur study indicated that the circulating miR-125a-5p levels in KD’s plasma have remarkably evaluated compared with healthy individuals. miR-125a-5p might play a role in the development of KD by regulating target gene MKK7 to induce apoptosis in vascular endothelial cells. Therefore, our findings have suggested that detected miR-125a-5p levels in plasma could be used as a potential biomarker in early KD diagnosis.

Measurements of Coronary Artery Aneurysms Due to Kawasaki Disease by Dual-Source Computed Tomography (DSCT).

Diameters of coronary artery aneurysms (CAAs) complicating acute phase KD can strongly predict the long-term prognosis of coronary artery lesions (CAL). Recently, computed tomographic angiography (CTA) has been used to detect CAL, and the purpose of this study was to determine whether coronary artery diameters measurements by CTA using dual-source computed tomography (DSCT) can be used instead of coronary angiogram (CAG) measurements. Twenty-five patients (22 males and three females) with CAL due to KD, who had undergone both CTA and CAG within one year, were retrospectively evaluated between 2007 and 2013. A prospective electrocardiogram-triggered CTA was performed on a DSCT (SOMATOM(®) Definition, Siemens Healthcare, Germany). Two pediatric cardiologists independently measured the diameters of CAAs twice in each maximum intensity projection (MIP), curved multiplaner reconstruction (MPR) and CAG. We measured 161 segments in total (segment 1-3, 5-7, 11, 13). Diagnostic accuracy was expressed as κ coefficient. A Bland-Altman analysis was also used to assess the intra-observer, inter-observer and inter-modality agreement. The diagnostic quality of CTA was excellent (κ = 0.93). Excellent inter-observer agreement for the diameters of CAAs was obtained for MIP, MPR and CAG and for the intra-observer agreement. The inter-modality agreement was also excellent in measurements of CAA (MPR-CAG: y = 0.9x + 0.40, r = 0.97, p < 0.0001 MIP-CAG: y = x + 0.1, r = 0.94, p < 0.0001). These values in normal coronary arteries were also obtained. We found a significant correlation between CTA and CAG in measuring the coronary arteries. We conclude that measuring coronary artery diameters by CTA is reliable and useful.

Abstract 102: Midkine, A New Functional Cytokine, Increased After IVIG May Protects From Vascular Injury In Acute Kawasaki Disease

Introduction: Midkine(MDK), found in 1988, a heparin-binding low-molecular protein, is a member of growth factors or cytokines, activated in the process of inflammatory disorders like RA, cancer development, angiogenesis and wound healing etc. It also acts as protecting cell apoptosis, stimulating fibrinolysis on endothelial cells, inhibiting or suppressing the development of regulatory T cells, improving myocardial dysfunction, and promoting the survival of ischemic myocardium. But, little is known in vasculitis syndrome. Methods: Serum samples, clinical data and laboratory values from17 patients with acute KD patients in before, 1-2 days, and 7-10 days after IVIG. Serum samples were analysed for MDK concentrations by EIA with anti-human MDK antibody. Results: MDK values were significantly elevated in serum from 1-2d after IVIG; 1-2d after IVIG; mean=1291.6 ± 1119.0, compared with Pre-IVIG; mean=530.9±468.6(p&lt;0.01), and rapidly decreased 7-10d after IVIG; mean=599.4±447.3 pg/ml(p&lt;0.05)(healthy human cut off level; &lt; 300pg/ml). CRP is significantly correlated with MDK (r=0.45, p&lt;0.05). MDK of 2 patient with CAA was higher than that of no CAA. Discussions and Conclusions: Knockout mice deficient in the MDK gene poorly develop neointima when the artery is damaged by ischemic shock. Clinically, MDK increases in human carcinoma and play a central role in inflammatory pathway. It promotes the migration of inflammatory leucocytes, namely macrophages and neutrophils. Recently, it is becoming a molecular target for the treatment or prevention of inflammatory diseases. Finally, this preliminary study suggests that MDK is a possible novel effector and may be relevant in the pathogenesis of arteritis in acute KD. IVIG may increase MDK levels and protect from inflammatory vascular damage.

Abstract O.17: Whole Genome Sequencing of a six-member African American family with two Kawasaki disease-affected siblings identifies novel susceptibility variants

We performed Whole Genome Sequencing (WGS) of a 6-member African American (AA) family with 2 of the 4 children affected with KD. We identified 6,712,158 unique variants across all 6 individuals. We first analyzed the quartet of mother/father/affected sibs and then compared variants in the 2 affected vs. 2 unaffected sibs. Assuming a recessive inheritance model, we identified 49,591 transmitted candidate homozygous variants exclusively present in both affected sibs compared to their parents. The affected vs. unaffected sibling analysis generated 64,187 candidate homozygous variants exclusive to the affected sibs. The intersection of the two analyses identified 20,943 variants of which 303 - in 117 genes - were predicted to be deleterious. We validated the findings in a cohort of 405 KD subjects and 6,252 normal controls using 4,060,864 imputed genotypes. Association analysis of the imputed GWAS dataset for KD susceptibility using the allelic and recessive tests in PLINK found 438,343 SNPs (nominal P-value &lt; 0.05). Of these, 17 variants in 10 genes were also among the 303 variants from the family analysis. These genes were ANGPT1, AS3MT, C10orf32, CMIP, CNNM2, LRIG2, MMP1, NT5C2, SLK, and TLR6. Of these, ANGPT1, MMP1, and TLR6 have been previously associated with KD. For SLK (serine-threonine protein kinase 2) rs10786779 located in the promoter, the homozygous A allele (risk) genotype showed significantly higher expression (p=0.04) in a cohort of 141 acute vs. convalescent mixed ethnic KD subjects on the Illumina HumanRef-12 V4 BeadChip. Conclusion: This is the first analysis of WGS in KD and the first to focus on genetic susceptibility in AA children who are second only to Asian children in susceptibility to KD. This exploratory analysis provides new validated variants that may likely contribute to KD susceptibility in AA children.