Novel asthma therapeutics: insights from whole-genome studies

Asthma is characterized by recurrent and reversible airflow obstruction as well as bronchial hyper-responsiveness. Airway inflammation is central to asthma pathogenesis. Bronchodilators are advocated as rescue treatments for acute asthma symptoms, whereas anti-inflammatory drugs such as Inhaled Corticosteroids (ICS) are common controller therapies for chronic asthma. Leukotriene modifiers are widely prescribed alternatives to ICS. During the past several years, a number of pharmacogenomic studies adopting whole-genome arrays have identified novel gene targets that contribute to the heterogeneity in responses to these anti-asthma drugs. These gene chips contain dense probes that capture either genotypes of single-nucleotide polymorphisms or the expression of genes across the entire human genome. Through these approaches, CLCA1, periostin, serpinB2, FKBP51, NFKB, GLCCI1 and T gene were reported to modulate ICS response in asthma patients whereas ARG1, CRHR2, SPATS2L and COL22A1were novel genes for bronchodilator responses. Some of these therapeutic targets were replicated in independent populations and/or supported by downstream in vitro and in vivo experiments on their functionality. Adequate bioinformatics support was essential in pharmacogenomics research in view of the enormous amount of whole-genome data involved. These whole-genome findings will ultimately facilitate personalized asthma pharmacotherapy that allows us to choose among treatment options that will likely be effective to any particular patient. However, more resources and collaborative efforts are required to advance the pharmacogenomics research

Airway inflammatory and spirometric measurements in obese children

Objectives: To investigate the association between obesity and airway inflammation and spirometric parameters in local children. Design: Cross-sectional and observational study. Setting: Paediatric clinics of a university-affiliated teaching hospital in Hong Kong. Patients: Chinese subjects aged 6 to 18 years were recruited from the paediatric clinics. Obesity was defined as being 120% or more of the median weight-for-height. Main outcome measures: Airway inflammation assessed by exhaled nitric oxide concentration; lung function evaluated by measuring forced expiratory flow in 1-second and forced vital capacity using spirometry; and peak expiratory flow rate measured by using a mini-Wright peak flow meter. Results: Fifty-five subjects were recruited into four groups as follows: 13 non-obese controls, 16 obese non-asthmatics, 15 non-obese asthmatics, and 11 obese asthmatics. The median (interquartile range) exhaled nitric oxide concentrations of these groups were 17.6 (14.4-20.9), 33.3 (26.1-75.4), 65.7 (32.0-110.0) and 49.2 (41.1-82.6) parts per billion, respectively (P=0.001 for trend). Post-hoc analysis revealed higher exhaled nitric oxide concentration in the latter three groups (obese and/or asthmatic subjects) than controls (P\u3c or =0.002). Exhaled nitric oxide concentration did not differ among obese non-asthmatics, non-obese asthmatics, and obese asthmatics (P\u3e0.1 for all). In non-asthmatics, exhaled nitric oxide concentration correlated positively with age (P=0.048), weight-for-height z-score (P=0.001), and forced vital capacity (P=0.009). Weight-for-height z-score correlated positively with forced vital capacity (P=0.041), but inversely with the forced expiratory flow in 1-second/forced vital capacity ratio (P=0.049). Such correlations were not observed in asthmatic children. Conclusion: Increased airway inflammation as revealed by exhaled nitric oxide concentration was found in obese non-asthmatic children. Weight-for-height z-score as an indicator of childhood obesity correlated with exhaled nitric oxide concentration and spirometric parameters in children without asthma. Nonetheless, concomitant obesity does not influence exhaled nitric oxide concentration in asthmatic children. Further studies are needed to identify the pathophysiologic mechanisms for such associations

Differences in asthma genetics between Chinese and other populations

Asthma is caused by complex gene-gene and gene-environment interactions. Most asthma genes are not replicable across populations, which is possibly because of differences in the epidemiology of these genes. Our case-control association and next-generation sequencing studies revealed substantial discrepancies in the frequencies of single nucleotide polymorphisms (SNPs) and haplotype blocks for asthma genes between Chinese and other populations. The minor allele frequencies for nearly half of our studied SNPs differed by 0.2 or greater between southern Chinese subjects in Hong Kong and European white populations, African populations, or both. Because genome-wide association studies for asthma have not been performed in Chinese subjects, we cannot tell whether the genomic findings of recent consortium-based genome-wide association studies are applicable to our population. In addition, our group performed Roche 454 pyrosequencing on a 100-kb area spanning each of 10 asthma loci in 24 healthy Hong Kong children. For the 17q21 locus, there was substantial variation in the haplotype structures that were constructed from 224 common SNPs among Hong Kong subjects and 6 ethnic groups under the 1000 Genomes Project. Sixteen mostly small haplotype blocks were formed in Hong Kong, whereas 6 haplotype blocks were identified in Han Chinese in Beijing and central European subjects and 11 and 19 blocks were identified in Puerto Rican and Yoruba African subjects. In conclusion, differences in allele frequencies of asthma genes and haplotype structures of asthma loci are found between Chinese subjects and other ethnic groups. These sequence variations must be considered during the selection of tagging SNPs for replicating genetic associations between populations

Eczema phenotypes are associated with multiple vitamin D pathway genes in Chinese children

Background: Vitamin D is increasingly recognized to play crucial roles in cutaneous immunity, and vitamin D treatment improved eczema control in small clinical trials. Several vitamin D-related genes were associated with asthma, but there are no data for eczema. Methods: Twenty-three single-nucleotide polymorphisms (SNPs) of five vitamin D-related genes (CYP27A1, CYP2R1, CYP27B1, GC and VDR) were genotyped in 1442 Chinese children with eczema and 1231 non-allergic controls. SNPs that followed Hardy–Weinberg equilibrium and yielded ≥95% genotyping call-rate were included. Haplotypic associations and SNP–SNP interactions for eczema diagnosis and subphenotypes were analysed. Results: Atopic eczema was associated with rs4674343 of CYP27A1 (odds ratio 0.66, 95% confidence interval 0.53–0.83, P = 0.0004). Increased eosinophil percentage was associated with CYP2R1 rs2060793A (P = 0.001) and rs1933064A (P = 0.001). Two CYP2R1 haplotypes increased eczema risk whereas one VDR haplotype lowered eczema risk. GC rs7041 and CYP2R1 rs7935792 interacted to modulate total IgE (cross-validation consistency 10/10, P = 0.047). Specifically, high-risk eczema patients had higher log-transformed total IgE than low-risk patients (2.76 ± 0.76 vs 2.60 ± 0.80, P = 0.002). Conclusion: A vitamin D-related SNP rs4674343 on CYP27A1 was found to be protective against atopic eczema. CYP2R1 and VDR haplotypes altered eczema susceptibility and eosinophil percentage, and GC and CYP2R1 interacted to determine total IgE among eczema patients

Asthma and atopy are associated with chromosome 17q21 markers in Chinese children

Background: Single‐nucleotide polymorphism (SNP)‐based genome‐wide association study revealed that markers on chromosome 17q21 were linked to childhood asthma but not atopy in Caucasians, with the strongest signal being detected for the SNP rs7216389 in the gene. Such association was unknown in Chinese. This study delineated the allele and genotype frequencies of 10 SNPs at chromosome 17q21, and investigated the relationship between these SNPs and asthma and plasma IgE in southern Chinese children. Methods: Asthmatic children and non‐allergic controls were recruited from pediatric clinics. Their plasma total and aeroallergen‐specific IgE concentrations were measured by immunoassay. Ten SNPs on 17q21 region were genotyped by multiplex SNaPshot™, and their genotype associations with asthma traits analyzed using multivariate regression. Results: 315 patients and 192 controls were enrolled. The allele frequency for C allele of rs7216389 varied significantly from 0.232 in our controls, 0.389 in Han Chinese to 0.536 in Caucasians. Asthma diagnosis was associated with rs11650680 and five other SNPs including rs7216389 (=0.019–0.034), whereas atopy was associated only with rs11650680 (=0.0004). Linear regression revealed the covariates for plasma total IgE to be significant for rs11650680 (=0.008–0.0002). Haplotypic associations were found with atopy and increased plasma total IgE, with the respective odds ratios and 95% confidence intervals for TTTCCGTT haplotype to be 0.21 and 0.09–0.52 (=0.0002) and 0.41 and 0.18–0.90 (=0.025). Conclusion: Childhood asthma and atopy are associated with chromosome 17q21 in Chinese, but such association may involve genes other than in this region

Determinants of, and reference equation for, exhaled nitric oxide in the Chinese population

Measurement of fractional exhaled nitric oxide concentration (FeNO) has been proposed as a useful biomarker for monitoring and management of airway diseases. Limited information is available regarding reference levels of FeNO levels in Chinese adults. This study aimed to investigate the reference equation and determinants of FeNO in Chinese adults. 1093 (577 males) healthy nonsmoking subjects aged 18-90 years were recruited. FeNO was measured online using a chemiluminescence analyser. Other assessments included spirometry, skin prick tests, total serum IgE levels and eosinophil count in peripheral blood. The geometric mean FeNO was 32.6 (95% reference interval (RI) 31.4-33.7) ppb for all subjects. FeNO values were higher in males than females (geometric mean (95% RI) 38.3 (36.5-40.2) ppb versus 27.1 (25.8-28.5) ppb, p\u3c0.0001), and in atopic than nonatopic subjects (34.6 (33.0-36.3) ppb versus 29.8 (28.3-31.4) ppb, p\u3c0.0001). FeNO correlated with age (r(2) = 0.23), height (r(2) = 0.20), IgE level (r(2) = 0.18) and percentage eosinophil count (r(2) = 0.18) (all p\u3c0.0001), but not with spirometric parameters. Based on multiple regression modelling, the reference equation of FeNO value was: log(FeNO) = 0.781 + 0.104(sex) + 0.004(age) + 0.084(atopy) + 0.003(height in cm), where for sex 1 = male and 0 = female, age is measured in years, for atopy 1 = atopic and 0 = nonatopic, and height is measured in cm. The FeNO of Chinese adults is higher than that of the Caucasian population, and is affected by age, sex, height and atopic status. This study provides useful references for the interpretation of FeNO

Childhood asthma and spirometric indices are associated with polymorphic markers of two vitamin D 25-hydroxylase genes

Background: Polymorphic markers of vitamin D pathway genes have been associated with asthma traits in different White populations. This study investigated the relationship between asthma phenotypes and single nucleotide polymorphisms (SNPs) of vitamin D receptor (VDR), vitamin D binding protein (GC), two 25-hydroxylases (CYP2R1 and CYP27A1), and 1α-hydroxylase (CYP27B1) in Hong Kong Chinese children. Methods: 23 SNPs of the five vitamin D pathway genes were successfully genotyped in 914 asthmatic children and 1231 non-allergic controls. Genotypic and haplotypic associations with asthma phenotypes (diagnosis, spirometric indices, total IgE, and eosinophil percentage) were analyzed by multivariate regression. Generalized multifactor dimensionality reduction was used to detect epistatic interactions between SNPs for asthma phenotypes. Results: Several SNPs of CYP27A1, CYP27B1, GC, and CYP2R1 were associated with asthma or spirometric indices, although only the association between FEV1 and CYP2R1 rs7935792 passed Bonferroni correction (p = 2.73 × 10(-4) ). Patients with CC genotype of rs7935792 had higher FEV1 than those with the other two genotypes. Asthma was also associated with TT haplotype of CYP27A1 and AGGATA haplotype of CYP2R1 (p = 0.021 and 0.024, respectively). Besides, strong association was found between FEV1 and GATAG of CYP2R1 (β = 13.37, p = 4.83 × 10(-4) ). GMDR failed to identify any 2-locus to 4-locus interaction that modulated asthma or spirometric indices. Conclusions: Several SNPs and haplotypes of CYP2R1 are associated with asthma diagnosis and FEV1 in children. Asthma is also modestly associated with a CYP27A1 haplotype. These two 25-hydroxylase genes may be genetic determinants for asthma phenotypes in children

Identifying uncontrolled asthma in young children: clinical scores or objective variables?

Objective: Several international asthma guidelines emphasize the importance of assessing asthma control. However, there is limited data on the usefulness of available assessment tools in indicating disease control in young asthmatics. This study investigated the ability of Chinese version of Childhood Asthma Control Test (C-ACT) and other disease-related factors in identifying uncontrolled asthma (UA) in young children. Methods: During the same clinic visit, asthma patients 4 to 11 years of age completed C-ACT and underwent exhaled nitric oxide and spirometric measurements. Blinded to these results, the same investigator assigned Disease Severity Score (DSS) and rated asthma control according to Global Initiative for Asthma. Results: The mean (SD) age of 113 recruited patients was 9.1 (2.0) years, and 35% of them had UA. C-ACT, DSS and forced expiratory volume in 1 second (FEV1) differed among patients with different control status (p \u3c 0.001 for C-ACT and DSS; p = 0.014 for FEV1). Logistic regression confirmed that UA was associated with DSS (p \u3c 0.001), PEF (p = 0.002), C-ACT (p = 0.011), and FEV1 (p = 0.012). By ROC analysis, C-ACT and DSS were the best predictors for UA (p \u3c 0.001), followed by PEF (p = 0.006) and FEV1 (p = 0.007). When analyzed by the Classification and Regression Tree (CART) approach, the sequential use of DSS and C-ACT had 77% sensitivity and 84% specificity in identifying UA. Conclusions: C-ACT is better than objective parameters in identifying young Chinese children with UA

Domestic exposure to aeroallergens in Hong Kong families with asthmatic children

Indoor aeroallergen exposures increased asthma symptoms in Caucasians, but their determinants and relationship to asthma and allergy in Asians are unclear. This study investigated exposures to cat, cockroach, and Blomia tropicalis allergens in 115 Hong Kong families with asthmatic children. Patients underwent exhaled nitric oxide and spirometric measurements. Home visits were made within 2 weeks during which parents completed a standardized questionnaire. Fel d 1, Bla g 2, and Blo t 5 in dust samples collected from patients\u27 mattresses, bedroom floors, and living room floors were measured by immunoassays. These aeroallergens were only detectable in some homes (38-55% for Fel d 1; 9-21% for Bla g 2, and 7-14% for Blo t 5). The presence of cat and/or dog was a strong determinant for Fel d 1 in all indoor sites. The timing and frequency of bedding change was associated with Bla g 2 levels, whereas the timing of bedroom floor cleaning was a consistent factor for Blo t 5 levels. Asthmatic children in families with high allergen exposure were more likely to have ≥4 wheezing attacks in preceding 12 months and exercise-induced wheezing than those with normal allergen exposure (P = 0.051 and 0.030, respectively). Mattress levels of all three allergens were also associated with severity of several allergy symptoms (P = 0.025-0.005). None of these aeroallergens correlated with exhaled nitric oxide and spirometric parameters. This study identifies determinants for cat, cockroach, and B. tropicalis levels in Hong Kong families with asthmatic children. These exposures are associated with severity of allergy symptoms