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Background: Five clinical clusters (early-onset increasing severity clusters 1, 2, and 4, late-onset severe cluster 3, and late-onset severe obstructed cluster 5) have been identified in the SARP cross-sectional cohort (n=726; Moore 2010). This approach demonstrated asthma heterogeneity and showed differential responses to biologic therapy (Bleecker 2019 ERS). Objective: Investigate longitudinal phenotypes of clinical clusters in the SARP longitudinal cohort. Methods: Subjects in longitudinal cohort (n=594) with 3-year (n=461) and 5-year (n=272) were assigned to clinical clusters using an 11-variable discriminant function. Phenotypes were compared using Kruskal-Wallis test. Results: Baseline clinical characteristics of five clusters in the longitudinal cohort were similar to the cross-sectional cohort. From baseline to Year 5, 2/3 of subjects remained in the same cluster and 1/6 each changed to more severe or milder clusters. Cluster 5 had consistently higher numbers of asthma exacerbations than clusters 1-4. Responses to a corticosteroid evoked phenotype (before and after 40 mg intramuscular triamcinolone): severe asthma clusters 3-5 had greater % increase in FEV1 than non-severe clusters (p Conclusions: SARP clinical clusters are reproducible in the longitudinal cohort. Severe clusters have better responses to corticosteroid. Cluster 5 has consistently greater asthma exacerbations over time. Longitudinal changes in FEV1 primarily determine cluster progression.

Background: Severe asthma is heterogeneous, with different phenotypes and endotypes. Aims and Objectives: We identified subsets of benralizumab-treated patients (pts) with severe asthma by assignment to Severe Asthma Research Program (SARP) clinical clusters. Methods: Pts (N=2,281) from SIROCCO (Lancet 2016:2115; n=1,082) and CALIMA (Lancet 2016:2128; n=1,199) were assigned to clusters via a discriminant function based on 11 predictors. Drug responses were compared with the Kruskal-Wallis test (nominal p-values). Results: Pts met criteria for 4 of 5 SARP clusters. Cluster 2 pts (n=393) were 81% atopic with early onset moderate asthma (mean age 15 yrs). Cluster 4 pts (n=386) were 82% atopic with early onset severe asthma (mean age 11 yrs). Cluster 3 pts (n=641) were 50% atopic with late-onset severe asthma (mean age 47 yrs). Cluster 5 pts (n=861) were 55% atopic with late-onset asthma (mean age 33 yrs) and persistent airflow obstruction. All clusters had annual exacerbation rate reductions for combined benralizumab arms vs. placebo, which were greater in late-onset asthma clusters (Cluster 3, −48%; 5, −50%; p Conclusions: Benralizumab improved lung function and reduced exacerbations in all clusters, with greater effects in late-onset asthma Clusters 3 and 5 vs. early onset Clusters 2 and 4. This supports the importance of understanding asthma heterogeneity and responsiveness to targeted therapies.

Combination of IL-6 (non-Type 2 asthma) and FeNO or blood eosinophil count (Type 2 asthma) identified asthma endotypes related to asthma severity, exacerbations, and responsiveness to corticosteroids and potential for response to anti-Type 2 and anti-IL-6 treatment.

Idiopathic pulmonary fibrosis (IPF) is a progressive disease, with a median survival of 3-5 years following diagnosis. Lung remodeling by invasive fibroblasts is a hallmark of IPF. In this study, we demonstrate that inhibition of vimentin intermediate filaments (VimIFs) decreases the invasiveness of IPF fibroblasts and confers protection against fibrosis in a murine model of experimental lung injury. Increased expression and organization of VimIFs contribute to the invasive property of IPF fibroblasts in connection with deficient cellular autophagy. Blocking VimIF assembly by pharmacologic and genetic means also increases autophagic clearance of collagen type I. Furthermore, inhibition of expression of collagen type I by siRNA decreased invasiveness of fibroblasts. In a bleomycin injury model, enhancing autophagy in fibroblasts by an inhibitor of VimIF assembly, withaferin A (WFA), protected from fibrotic lung injury. Additionally, in 3D lung organoids, or pulmospheres, from patients with IPF, WFA reduced the invasiveness of lung fibroblasts in the majority of subjects tested. These studies provide insights into the functional role of vimentin, which regulates autophagy and restricts the invasiveness of lung fibroblasts.

Background: CC16 is a protein produced by bronchial epithelial cells (BEC) that participates in host defense. Objectives: Our purpose is to investigate CC16 as a biomarker for COPD using systems biology in the SPIROMICS cohort. Methods: Genome-wide pQTL analyses of CC16 serum protein levels were performed in non-Hispanic Whites (n=1,874) and African Americans (n=403). Spearman correlation analyses between CC16 mRNA levels and other protein-coding genes (n=14,746) were performed using RNAseq data from BEC (n=131). Association analyses between CC16 SNPs/mRNA/protein and COPD-related phenotypes were performed using a generalized linear model. Results: The A allele of rs3741240 was associated with lower CC16 protein levels in non-Hispanic Whites (p=9x10-6) and African Americans (p=0.04). CC16 mRNA levels were positively correlated with CC16 protein levels (p=0.02). CC16 mRNA levels were positively correlated with MUC5B, IL12A, C3, TLR5, IL6, DPP4, and SFTPD, and negatively correlated with MUC5AC, IL18R1, and TLR6 at genome-wide significant level (p 1 vs. 0, OR=0.8, p=0.02). Conclusions: rs3741240 is a pQTL SNP for CC16 protein levels. CC16 mRNA levels are positively correlated with inflammation genes and negatively correlated with atopic genes. These findings are consistent with eQTL and co-expression analyses from SARP asthma cohort (n=107) (Li, 2018, ATS abstract). CC16 protein levels were associated with reduced future COPD exacerbations and will be further investigated longitudinally in SPIROMICS.

BACKGROUND The release of various inflammatory mediators into the bronchial lumen is thought to reflect both the type and degree of airway inflammation, eosinophilic Th2, and Th9, or neutrophilic Th1, and Th17, in patients with asthma. AIMS We investigated whether cytokines and chemokines differed in sputum from subjects with more severe compared with milder asthma and whether unbiased factor analysis of cytokine and chemokine groupings indicates specific inflammatory pathways. METHODS Cell-free supernatants from induced sputum were obtained from subjects with a broad range of asthma severity (n = 158) and assessed using Milliplex® Cytokines/Chemokine kits I, II and III, measuring 75 individual proteins. Each cytokine, chemokine or growth factor concentration was examined for differences between asthma severity groups, for association with leucocyte counts, and by factor analysis. RESULTS Severe asthma subjects had 9 increased and 4 decreased proteins compared to mild asthma subjects and fewer differences compared to moderate asthma. Twenty-six mediators were significantly associated with an increasing single leucocyte type: 16 with neutrophils (3 interleukins [IL], 3 CC chemokines, 4 CXC chemokines, 4 growth factors, TNF-α and CX3CL1/Fractalkine); 5 with lymphocytes (IL-7, IL-16, IL-23, IFN-α2 and CCL4/MIP1β); IL-15 and CCL15/MIP1δ with macrophages; IL-5 with eosinophils; and IL-4 and TNFSF10/TRAIL with airway epithelial cells. Factor analysis grouped 43 cytokines, chemokines and growth factors which had no missing data onto the first 10 factors, containing mixes of Th1, Th2, Th9 and Th17 inflammatory and anti-inflammatory proteins. CONCLUSIONS Sputum cytokines, chemokines and growth factors were increased in severe asthma, primarily with increased neutrophils. Factor analysis identified complex inflammatory protein interactions, suggesting airway inflammation in asthma is characterized by overlapping immune pathways. Thus, focus on a single specific inflammatory mediator or pathway may limit understanding the complexity of inflammation underlying airway changes in asthma and selection of appropriate therapy.

Preventing asthma exacerbations is a goal of National Asthma Guidelines, but identification of at-risk individuals is difficult, especially in older asthmatics. We aimed to characterize exacerbation-prone asthma in older adults to identify characteristics and predictive biomarkers associated with high-risk phenotypes to facilitate precision medicine approaches. 70 non-smoking subjects ≥60 years old in the Severe Asthma Research Program underwent characterization (comprehensive questionnaires, lung function with bronchodilator reversibility, atopy assessment, biomarker collection). Using patient-reported exacerbations two phenotypes were defined; Exacerbation Prone Asthma (EPA,≥1/past year) and Very Frequent Exacerbations (VFE,≥3/past year). Risk and biomarker associations were assessed with odds ratios (OR) using logistic regression. 44% of older asthmatics had EPA. Healthcare utilization and treatment with higher corticosteroid doses were more frequent in EPA compared to non-exacerbating subjects (p

Idiopathic pulmonary fibrosis (IPF) is a fatal progressive fibrotic lung disease characterized by the presence of invasive myofibroblasts in the lung. Currently, there are only two FDA-approved drugs (pirfenidone and nintedanib) for the treatment of IPF. There are no defined criteria to guide specific drug therapy. New methodologies are needed not only to predict personalized drug therapy, but also to screen novel molecules that are on the horizon for treatment of IPF. We have developed a model system that exploits the invasive phenotype of IPF lung tissue. This ex vivo 3D model uses lung tissue from patients to develop pulmospheres. Pulmospheres are 3D spheroids composed of cells derived exclusively from primary lung biopsies and inclusive of lung cell types reflective of those in situ, in the patient. We tested the pulmospheres of 20 subjects with IPF and 9 control subjects to evaluate the responsiveness of individual patients to antifibrotic drugs. Clinical parameters and outcomes were also followed in the same patients. Our results suggest that pulmospheres simulate the microenvironment in the lung and serve as a personalized and predictive model for assessing responsiveness to antifibrotic drugs in patients with IPF.

Autoimmunity has been implicated in the pathogenesis of idiopathic pulmonary fibrosis (IPF); however, the repertoire of autoantigens involved in this disease and the clinical relevance of these autoimmune responses are still being explored. Our initial discovery assays demonstrated that circulating and intrapulmonary vimentin levels are increased in IPF patients. Subsequent studies showed native vimentin induced HLA-DR–dependent in vitro proliferation of CD4 T cells from IPF patients and enhanced the production of IL-4, IL-17, and TGF-β1 by these lymphocytes in contrast to normal control specimens. Vimentin supplementation of IPF PBMC cultures also resulted in HLA-DR–dependent production of IgG with anti-vimentin specificities. Circulating anti-vimentin IgG autoantibody levels were much greater in IPF subjects from the University of Alabama at Birmingham (n = 102) and the University of Pittsburgh (U. Pitt., n = 70) than in normal controls. Anti-vimentin autoantibody levels in IPF patients were HLA biased and inversely correlated with physiological measurements of lung function (i.e., forced expiratory volumes and diffusing capacities). Despite considerable intergroup differences in transplant-free survival between these two independent IPF cohorts, serious adverse outcomes were most frequent among the patients within each population that had the highest anti-vimentin autoantibody levels (University of Alabama at Birmingham: hazard ratio 2.5, 95% confidence interval 1.2–5.3, p = 0.012; University of Pittsburgh: hazard ratio 2.7, 95% confidence interval 1.3–5.5, p = 0.006). These data show that anti-vimentin autoreactivity is prevalent in IPF patients and is strongly associated with disease manifestations. These findings have implications with regard to the pathogenesis of this enigmatic disease and raise the possibility that therapies specifically directed at these autoimmune processes could have therapeutic efficacy.

Background: Smokers has been clustered into COPD and non-COPD clusters (Li, ATS 2016). Smokers with preserved lung function may have COPD symptoms, exacerbations, and usage of respiratory medicine (Woodruff, NEJM 2016, 374:1811-21). Objectives: We aim to compare clinical characteristics of four clusters mainly composed of smokers with preserved lung function. Methods: Clinical characteristics were compared using the Kruskal-Wallis test and chi-square test. Results: C1 consists of resistant smokers with normal lung function, early emphysema, lower HCU and inhaled steroid (ICS) use, and lower symptom scores. C2 consists of heavy smokers with normal lung function, early emphysema, higher HCU and ICS use, and higher blood neutrophils. C3 has normal lung function, no emphysema, and lower HCU and ICS use. C4 has lower lung function, no emphysema, higher HCU and ICS use, higher “label” of asthma, higher inflammation, and higher symptom scores. Conclusions: Four distinct endophenotypic clusters mainly composed of smokers with preserved lung function have been identified. This study supports heterogeneity of smokers and indicates the distinction between smokers with preserved lung function and COPD is not as simple as measuring spirometry alone .

Background: The SARP1/2 cluster analysis identified 5 clinical asthma phenotypes that reflect inflammatory mechanisms: a spectrum of early onset allergic asthma (Clusters 1,2,4), late onset severe asthma (Cluster 3) and severe asthma with COPD characteristics (Cluster 5). Aims: Evaluate reproducibility and stability of SARP1/2 clinical cluster phenotypes in SARP3, a 3-year longitudinal cohort enriched for severe asthma. Methods: SARP3 subjects > 12 years old (n= 594) were assigned to SARP1/2 clusters using 11 variables and compared to the SARP1/2 cohort (n=1176). Subjects in both SARP1/2 and SARP3 (n=68) were assigned to clusters twice using unique SARP1/2 and 3 data. Results: The SARP3 cohort is skewed toward severe asthma clusters 3(16%), 4(15%) and 5(19%). SARP3 subjects are older with longer disease duration (p Conclusions: All five SARP1/2 clinical cluster phenotypes are reproducible in SARP3 with a shift toward more severe clusters (3,4,5). In a subset of SARP subjects with longitudinal data there was heterogeneity in the progression of disease severity that was associated with changes in baseline lung function. These results suggest that a clinical approach guided by monitoring of lung function could identify patients at risk for disease progression.

Objective: Genome-wide association studies (GWASs) have identified genes associated with asthma, however expression of these genes in asthma-relevant tissues has not been studied. This study tested expression and correlation between GWAS-identified asthma genes and asthma or asthma severity. Methods: Correlation analyses of expression levels of GWAS-identified asthma genes and asthma-related biomarkers were performed in cells from human bronchial epithelial biopsy (BEC, n = 107) and bronchial alveolar lavage (BAL, n = 94). Results: Expression levels of asthma genes between BEC and BAL and with asthma or asthma severity were weakly correlated. The expression levels of IL18R1 were consistently higher in asthma than controls or in severe asthma than mild/moderate asthma in BEC and BAL (p < 0.05). In RAD50-IL13 region, the expression levels of RAD50, not IL4, IL5, or IL13, were positively correlated between BEC and BAL (ρ = 0.53, P = 4.5 × 10−6). The expression levels of IL13 were positively correlated with IL5 in BEC (ρ = 0.35, P = 1.9 × 10−4) and IL4 in BAL (ρ = 0.42, P = 2.5 × 10−5), respectively. rs3798134 in RAD50, a GWAS-identified SNP, was correlated with IL13 expression and the expression levels of IL13 were correlated with asthma (P = 0.03). rs17772583 in RAD50 was significantly correlated with RAD50 expression in BAL and BEC (P = 7.4 × 10−7 and 0.04) but was not associated with asthma. Conclusions: This is the first report studying the expression of GWAS-identified asthma genes in BEC and BAL. IL13, rather than RAD50, IL4, or IL5, is more likely to be the asthma susceptibility gene. Our study illustrates tissue-specific expression of asthma-related genes. Therefore, whenever possible, disease-relevant tissues should be used for transcription analysis.

The National Heart, Lung, and Blood Institute has sponsored several asthma clinical networks, but the Severe Asthma Research Program (SARP) is unique, because it is not a clinical trials network, and it includes both adults and children. Investigators in SARP have comprehensively characterized 1,644 patients with asthma over the past 10 years, including 583 individuals with severe asthma and 300 children below the age of 18 years. The diversity in clinical characteristics, physiologic measures, and biomarkers in a large number of subjects across the ages provides an ideal cohort in which to investigate asthma heterogeneity. Using both biased and unbiased approaches, multiple asthma phenotypes have been described in SARP. These phenotypic analyses have improved our understanding of heterogeneity in asthma, and may provide a starting point to transform clinical practice through the evidence-based classification of disease severity. Although these new phenotypes strive to make order out of a heterogeneous group of patients, they are limited by that heterogeneity. There may be large groups of patients, especially those with milder asthma, that can be grouped into a clinical phenotype to guide therapy, but there will always be patients on the “edge” of a phenotype who will not fit into these groupings. In the SARP cluster analysis, subjects on the “edge” of a phenotype frequently had lung function that was better or worse than other subjects in the same cluster, despite similar clinical characteristics. This suggests that different pathophysiologic mechanisms may be responsible for decrements in lung function in some subjects. This is extremely important for subjects with severe asthma who may be on the “edge” of two phenotypes that may be driven by different pathobiologic mechanisms that warrant different therapeutic approaches.

Background Clinical cluster analysis from the Severe Asthma Research Program (SARP) identified 5 asthma subphenotypes that represent the severity spectrum of early-onset allergic asthma, late-onset severe asthma, and severe asthma with chronic obstructive pulmonary disease characteristics. Analysis of induced sputum from a subset of SARP subjects showed 4 sputum inflammatory cellular patterns. Subjects with concurrent increases in eosinophil (≥2%) and neutrophil (≥40%) percentages had characteristics of very severe asthma. Objective To better understand interactions between inflammation and clinical subphenotypes, we integrated inflammatory cellular measures and clinical variables in a new cluster analysis. Methods Participants in SARP who underwent sputum induction at 3 clinical sites were included in this analysis (n = 423). Fifteen variables, including clinical characteristics and blood and sputum inflammatory cell assessments, were selected using factor analysis for unsupervised cluster analysis. Results Four phenotypic clusters were identified. Cluster A (n = 132) and B (n = 127) subjects had mild-to-moderate early-onset allergic asthma with paucigranulocytic or eosinophilic sputum inflammatory cell patterns. In contrast, these inflammatory patterns were present in only 7% of cluster C (n = 117) and D (n = 47) subjects who had moderate-to-severe asthma with frequent health care use despite treatment with high doses of inhaled or oral corticosteroids and, in cluster D, reduced lung function. The majority of these subjects (>83%) had sputum neutrophilia either alone or with concurrent sputum eosinophilia. Baseline lung function and sputum neutrophil percentages were the most important variables determining cluster assignment. Conclusion This multivariate approach identified 4 asthma subphenotypes representing the severity spectrum from mild-to-moderate allergic asthma with minimal or eosinophil-predominant sputum inflammation to moderate-to-severe asthma with neutrophil-predominant or mixed granulocytic inflammation.

Background Sputum eosinophil percentages are a strong predictor of airway inflammation and exacerbations and aid asthma management, whereas sputum neutrophil percentages indicate a different severe asthma phenotype that is potentially less responsive to T H 2-targeted therapy. Variables, such as blood eosinophil counts, total IgE levels, fraction of exhaled nitric oxide (Feno) levels, or FEV 1 percent predicted, might predict airway eosinophil percentages, whereas age, FEV 1 percent predicted, or blood neutrophil counts might predict sputum neutrophil percentages. Availability and ease of measurement are useful characteristics, but accuracy in predicting airway eosinophil and neutrophil percentages either individually or combined is not established. Objectives We sought to determine whether blood eosinophil counts, Feno levels, and IgE levels accurately predict sputum eosinophil percentages and whether age, FEV 1 percent predicted, and blood neutrophil counts accurately predict sputum neutrophil percentages. Methods Subjects in the Wake Forest Severe Asthma Research Program (n = 328) were characterized by blood and sputum cell counts, health care use, lung function, Feno levels, and IgE levels. Multiple analytic techniques were used. Results Despite significant association with sputum eosinophil percentages, blood eosinophil counts, Feno levels, and total IgE levels did not accurately predict sputum eosinophil percentages, and combinations of these variables did not improve prediction. Age, FEV 1 percent predicted, and blood neutrophil counts were similarly unsatisfactory for the prediction of sputum neutrophil percentages. Factor analysis and stepwise selection found Feno levels, IgE levels, and FEV 1 percent predicted, but not blood eosinophil counts, correctly predicted 69% of sputum eosinophil percentages of less than 2% or 2% and greater. Likewise, age, asthma duration, and blood neutrophil counts correctly predicted 64% of sputum neutrophil percentages of less than 40% or 40% and greater. A model to predict both sputum eosinophil and neutrophil percentages accurately assigned only 41% of samples. Conclusion Despite statistically significant associations, Feno levels, IgE levels, blood eosinophil and neutrophil counts, FEV 1 percent predicted, and age are poor surrogates, both separately and combined, for accurately predicting sputum eosinophil and neutrophil percentages.

Background The IL6R single nucleotide polymorphism (SNP) rs4129267 has recently been identified as an asthma susceptibility locus in subjects of European ancestry but has not been characterized with respect to asthma severity. The SNP rs4129267 is in linkage disequilibrium ( r 2 = 1) with the IL6R coding SNP rs2228145 (Asp 358 Ala). This IL6R coding change increases IL-6 receptor (IL-6R) shedding and promotes IL-6 transsignaling. Objectives We sought to evaluate the IL6R SNP rs2228145 with respect to asthma severity phenotypes. Methods The IL6R SNP rs2228145 was evaluated in subjects of European ancestry with asthma from the Severe Asthma Research Program (SARP). Lung function associations were replicated in the Collaborative Study on the Genetics of Asthma (CSGA) cohort. Serum soluble IL-6R levels were measured in subjects from SARP. Immunohistochemistry was used to qualitatively evaluate IL-6R protein expression in bronchoalveolar lavage cells and endobronchial biopsies. Results The minor C allele of IL6R SNP rs2228145 was associated with a lower percent predicted FEV 1 in the SARP cohort ( P = .005), the CSGA cohort ( P = .008), and in a combined cohort analysis ( P = .003). Additional associations with percent predicted forced vital capacity (FVC), FEV 1 /FVC ratio, and PC 20 were observed. The rs2228145 C allele (Ala 358 ) was more frequent in severe asthma phenotypic clusters. Elevated serum soluble IL-6R levels were associated with lower percent predicted FEV 1 ( P = .02) and lower percent predicted FVC ( P = .008) (n = 146). IL-6R protein expression was observed in bronchoalveolar lavage macrophages, airway epithelium, vascular endothelium, and airway smooth muscle. Conclusions The IL6R coding SNP rs2228145 (Asp 358 Ala) is a potential modifier of lung function in subjects with asthma and might identify subjects at risk for more severe asthma. IL-6 transsignaling might have a pathogenic role in the lung.

The Severe Asthma Research Program cohort includes subjects with persistent asthma who have undergone detailed phenotypic characterization. Previous univariate methods compared features of mild, moderate, and severe asthma.To identify novel asthma phenotypes using an unsupervised hierarchical cluster analysis.Reduction of the initial 628 variables to 34 core variables was achieved by elimination of redundant data and transformation of categorical variables into ranked ordinal composite variables. Cluster analysis was performed on 726 subjects.Five groups were identified. Subjects in Cluster 1 (n = 110) have early onset atopic asthma with normal lung function treated with two or fewer controller medications (82%) and minimal health care utilization. Cluster 2 (n = 321) consists of subjects with early-onset atopic asthma and preserved lung function but increased medication requirements (29% on three or more medications) and health care utilization. Cluster 3 (n = 59) is a unique group of mostly older obese women with late-onset nonatopic asthma, moderate reductions in FEV(1), and frequent oral corticosteroid use to manage exacerbations. Subjects in Clusters 4 (n = 120) and 5 (n = 116) have severe airflow obstruction with bronchodilator responsiveness but differ in to their ability to attain normal lung function, age of asthma onset, atopic status, and use of oral corticosteroids.Five distinct clinical phenotypes of asthma have been identified using unsupervised hierarchical cluster analysis. All clusters contain subjects who meet the American Thoracic Society definition of severe asthma, which supports clinical heterogeneity in asthma and the need for new approaches for the classification of disease severity in asthma.

Because three-axle heavy-vehicle with front-wheel steering has big radius at low speed and bad stability at high speed, in order to improve heavy vehicle steering performance at different speed, the multi-axle dynamic steering technology is put forward. Selecting zero side-slip angle of mass center and proportional control strategy to control vehicle, Using MATLAB, the steering performance of the three-axle vehicle with different steering modes are simulated. The result shows that multi-axle dynamic steering can decrease the steering radius at low speed and improve vehicle stability at high speed.

Background Genome-wide association studies (GWAS) of asthma and asthma-related traits, including our previous TENOR study1, have consistently identified ORMDL3-GSDMB, IL33, IL1RL1-IL18R1, RAD50-IL13, TSLP-WDR36, and HLA-DR/DQ regions.2 Methods In this study, GWAS of asthma was performed in non-Hispanic white population from STAMPEED study (813 cases and 1564 controls). Our GWAS results were compared with the published GWAS of asthma and autoimmune diseases (AD). Results Multiple SNPs in TNFAIP3 interacting protein 1 (TNIP1) on chromosome 5q32-q33.1 were associated with asthma in STAMPEED: rs1422673 (P = 3.44 × 10−7) and rs10036748 (P = 1.41 × 10−6). rs1422673 was weakly associated with asthma in the published GABRIEL study (P = 0.018 for meta-analysis)2 but not in the TENOR study (P = 0.18 but same trend).1 TNIP1 may interact with TNFAIP3 and inhibit TNFα-induced NFκB inflammation pathway. Joint analyses were performed on 6 SNPs in GSDMB (rs2872507), IL33 (rs3939286), IL1RL1 (rs13431828), IL13 (rs20541), TSLP (rs1837253), and HLA-DRA (rs2395185) in STAMPEED and TENOR populations, but only limited variance can be explained (percentage of deviance = 1.5–1.9%; the area under the receiver operating characteristic curve (AUC) = 0.58–0.59). Minor allele T of rs20541 in IL13 is the risk allele for asthma but the protective allele for psoriasis. Minor allele A of rs2872507 in GSDMB is the protective allele for asthma but the risk allele for rheumatoid arthritis, Crohn's disease and ulcerative colitis. T allele of rs10036748 in TNIP1 is the minor protective allele for asthma, but the minor or major risk allele for systemic lupus erythematosus in non-Hispanic white or Chinese population, respectively. Conclusions Our study provides genetic evidence that asthma and AD have opposite immunopathogenesis directions.

Background The T helper 2 (Th2) inflammatory pathway, including the Th2-activating cytokine interleukin 33 and its receptor interleukin 1 receptor-like 1 have been strongly implicated in asthma susceptibility (Moffatt MF, et al NEJM 2010). However, the role of Th2 pathway genetic variation in asthma progression and severity is not well understood. Our research group recently developed a clustering algorithm based on comprehensive phenotype information to assign subjects with asthma in the Severe Asthma Research Program (SARP) to 5 primary clusters; 3 of which represent increasing severe allergic asthma (Moore WC, et al AJRCCM, 2010). We hypothesized that common and potentially deleterious rare variation in this pathway would be associated with severe asthma based on SARP cluster designation. Methods To evaluate common variants (minor allele frequency or MAF >5%), 419 SARP non-Hispanic white participants with a cluster assignment were genotyped for 182 single nucleotide polymorphisms (SNPs) in Th2 pathway genes using whole-genome SNP data. Individual SNPs and a cumulative model of significant SNPs were evaluated using contingency tables with a chi-square test for trend and ordinal regression models adjusted for age, sex, and principal components. Rare (MAF

Background Recent meta-analyses of genome-wide association studies in general populations of European descent have identified 28 loci for lung function. Objective We sought to identify novel lung function loci specifically for asthma and to confirm lung function loci identified in general populations. Methods Genome-wide association studies of lung function (percent predicted FEV 1 [ppFEV 1 ], percent predicted forced vital capacity, and FEV 1 /forced vital capacity ratio) were performed in 4 white populations of European descent (n = 1544), followed by meta-analyses. Results Seven of 28 previously identified lung function loci ( HHIP , FAM13A , THSD4 , GSTCD , NOTCH4-AGER , RARB , and ZNF323 ) identified in general populations were confirmed at single nucleotide polymorphism (SNP) levels ( P IL12A , IL12RB1 , STAT4 , and IRF2 ) associated with ppFEV 1 ( P −4 ) belong to the T H 1 or IL-12 cytokine family pathway. By using a linear additive model, these 4 T H 1 pathway SNPs cumulatively explained 2.9% to 7.8% of the variance in ppFEV 1 values in 4 populations ( P = 3 × 10 −11 ). Genetic scores of these 4 SNPs were associated with ppFEV 1 values ( P = 2 × 10 −7 ) and the American Thoracic Society severe asthma classification ( P = .005) in the Severe Asthma Research Program population. T H 2 pathway genes ( IL13 , TSLP , IL33 , and IL1RL1 ) conferring asthma susceptibility were not associated with lung function. Conclusion Genes involved in airway structure/remodeling are associated with lung function in both general populations and asthmatic subjects. T H 1 pathway genes involved in anti-virus/bacterial infection and inflammation modify lung function in asthmatic subjects. Genes associated with lung function that might affect asthma severity are distinct from those genes associated with asthma susceptibility.

Background Genome-wide association studies (GWASs) of asthma have consistently implicated the ORM1-like 3 and gasdermin B (ORMDL3-GSDMB) , IL33 , IL-1 receptor–like 1 and IL-18 receptor 1 (IL1RL1-IL18R1) , RAD50-IL13 , thymic stromal lymphopoietin and WD repeat domain 36 region (TSLP-WDR36) , and HLA-DR/DQ regions. Objective A GWAS of asthma was performed in a non-Hispanic white population. Methods A GWAS was performed in 813 Severe Asthma Research Program/Collaborative Studies on the Genetics of Asthma/Chicago Asthma Genetics Study cases and 1564 control subjects. The GWAS results were compared with those of the published GWASs of autoimmune diseases. Results Multiple single nucleotide polymorphisms in the TNFAIP3 interacting protein 1 (TNIP1) gene, which interacts with TNFAIP3 and inhibits the TNF-α–induced nuclear factor κB inflammation pathway, were associated with asthma: rs1422673 ( P = 3.44 × 10 −7 ) and rs10036748 ( P = 1.41 × 10 −6 , r 2 = 0.67). rs1422673 was also associated with asthma in the published GABRIEL ( P = .018) and EVE ( P = 1.31 × 10 −5 ) studies. The minor allele T of rs20541 in IL13 is the risk allele for asthma but the protective allele for psoriasis. The minor allele T of rs2395185 in HLA-DRA is the risk allele for asthma but the protective allele for ulcerative colitis. The minor allele A of rs2872507 in GSDMB is the protective allele for asthma but the risk allele for rheumatoid arthritis, Crohn disease, and ulcerative colitis. The T allele of rs10036748 in the TNIP1 gene is the minor protective allele for asthma but the minor or major risk allele for systemic lupus erythematosus and systemic sclerosis in non-Hispanic white or Chinese subjects, respectively. Conclusions Our study suggests that single nucleotide polymorphisms associated with both asthma and autoimmune diseases might have opposite effects on immunopathogenesis.

Background Asthma in children is a heterogeneous disorder with many phenotypes. Although unsupervised cluster analysis is a useful tool for identifying phenotypes, it has not been applied to school-age children with persistent asthma across a wide range of severities. Objectives This study determined how children with severe asthma are distributed across a cluster analysis and how well these clusters conform to current definitions of asthma severity. Methods Cluster analysis was applied to 12 continuous and composite variables from 161 children at 5 centers enrolled in the Severe Asthma Research Program. Results Four clusters of asthma were identified. Children in cluster 1 (n = 48) had relatively normal lung function and less atopy. Children in cluster 2 (n = 52) had slightly lower lung function, more atopy, and increased symptoms and medication use. Cluster 3 (n = 32) had greater comorbidity, increased bronchial responsiveness, and lower lung function. Cluster 4 (n = 29) had the lowest lung function and the greatest symptoms and medication use. Predictors of cluster assignment were asthma duration, the number of asthma controller medications, and baseline lung function. Children with severe asthma were present in all clusters, and no cluster corresponded to definitions of asthma severity provided in asthma treatment guidelines. Conclusion Severe asthma in children is highly heterogeneous. Unique phenotypic clusters previously identified in adults can also be identified in children, but with important differences. Larger validation and longitudinal studies are needed to determine the baseline and predictive validity of these phenotypic clusters in the larger clinical setting.

Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants.This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations.Stratified by sputum granulocytes,2% oror = 2% eosinophils and40% oror = 40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased2-fold in sputa withor = 40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1beta, and macrophage inflammatory protein 3alpha/CCL20 for those withor = 40% neutrophils; these mediators demonstrated positive associations with neutrophil counts.Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils.