Study Of Metabolomics And Clinical Heterogenous Mechanisms Of Clinical Heterogeneity In The Different HRCT-Classified Phenotypes Of COPD With Treatment Of Tiotropium Bromide

Part ? 1H-NMR-based metabolic profiling of healthy individuals and high-resolution CT-classified phenotypes of COPD with treatment of tiotropium bromideObjective:Heterogeneity of COPD results in different therapeutic effects for different patients receiving the same treatment.COPD patients need to be individually treated according to their own characteristics.The purpose of this study was to explore the differences in different CT phenotypic COPD by molecular metabolites through the use of metabolomics.Methods:According to the characteristics of CT imaging,67 COPD patients were grouped into phenotype E(n=35)or phenotype M(n=32),Each COPD patient received tiotropium bromide powder for inhalation for a therapeutic period of 3 months.All subjects were assigned into phenotype E in pre-therapy(EB,n=35),phenotype E in post-therapy(EA,n=35),phenotype M in pre-therapy(MB,n=32),phenotype M in post-therapy(MA,n=32),or normal control(N,n=34).The method of metabolomics based on 1H nuclear magnetic resonance(1H-NMR)was used to compare the changes in serum metabolites between COPD patients and normal controls and between different phenotypes of COPD patients in pre-and post-therapy.Results:Patients with COPD phenotype E responded better to tiotropium bromide than patients with COPD phenotype M in terms of pulmonary function and COPD assessment test scores.There were differences in metabolites in COPD patients vs normal control people.Differences were also observed between different COPD phenotypic patients receiving the treatment in comparison with those who did not receive treatment.The changes of metabolites involved lactate,phenylalanine,fructose,glycine,asparagine,citric acid,pyruvic acid,proline,acetone,ornithine,lipid,pyridoxine,maltose,betaine,lipoprotein,and so on.These identified metabolites covered the metabolic pathways of amino acids,carbohydrates,lipids,genetic materials,and vitamin.Conclusion:The efficacy of tiotropium bromide on COPD phenotype E is better than that of phenotype M.Metabolites detected by 1H-NMR metabolomics have potentialities of differentiation of COPD and healthy people,discrimination of different COPD phenotypes,and giving insight into the individualized treatment of COPD.Part ? Clinical study of CHRM2 and CHRM3 gene polymorphisms in high-resolution CT-classified phenotypes of COPD with treatment of tiotropium bromideObjective:Chronic obstructive pulmonary disease is a heterogeneous disease which results from self-genetic factors and environmental factors.The genetic polymorphisms of muscarinic-cholinergic receptor 2(CHRM2)and 3(CHRM3)in HRCT classified COPD patients are analyzed to correlate them with the severity of the disease and the efficacy of treatment with tiotropium bromide.Thus,to discuss whether the different therapeutic efficacy of tiotropium on different phenotypes of COPD is associated with the genetic factors of the genes of receptor.Methods:Four hundred and fifty-five patients with COPD were screened and divided into phenotype E(n=238)and phenotype M(n=217)according to the HRCT classification criteria.The changes in related clinical characteristics and evaluative indicators of efficacy of treatment with tiotropium bromide between the two phenotypes were compared.At the same time,the CHRM2 and CHRM3 genetic polymorphisms was detected in COPD patients,and the genotypic distribution frequencies of the two genes were also compared between the different COPD phenotypes respectively.Meanwhile,the related clinical characteristics and the evaluative indicators of the therapeutic efficacy of tiotropium bromide were compared within genotypes of CHRM2 and CHRM3 respectively to find out whether the different therapeutic efficacy of tiotropium bromide on different phenotypes of COPD was associated with certain genotype.Results:The phenotype E was higher on the changes of FEV1,FEV1/FVC,FEV1/predicted%and CAT scores than the phenotype M.Genotypic distribution frequencies of CHRM2 rs1824024 and CHRM3 rs481036 had no significant differences between phenotype E and phenotype M.The three genotypes of CHRM2 rs1824024 showed significant differences in smoking history,numbers of acute exacerbations and changes from FEV1,predicted FEV1%and CAT scores.But among three genotypes of CHRM3 rs481036,there were no significant differences in gender,age,BMI,existing smokers,smoking history,numbers of acute exacerbations,AFEV1,AFEV1/FVC,AFEV1/predicted%,ACAT and AmMRC.Conclusion:The efficacy of tiotropium bromide on COPD phenotype E is better than that of phenotype M.CHRM2 rs1824024 and CHRM3 rs481036 are not associated with HRCT-classified phenotypes of COPD patients.Presence of severe airway wall thickening in COPD patients has no correlation with the genetic polymorphisms of CHRM2 and CHRM3.The single nucleotide polymorphism mutation homozygous genotype A/A in CHRM2 rs1824024 of COPD patients is less effective in treating with tiotropium bromide than the wild homozygous genotype C/C and the mutant heterozygous genotype C/A.The single nucleotide polymorphism genotypes in CHRM3 rs481036 of COPD patients have no different therapeutic responses to tiotropium bromide,thus the CHRM3 genetic polymorphism may not relate to the therapeutic efficacy of tiotropium bromide in COPD patients.Part ?:Non-neuronal cholinergic system in HRCT-classified phenotypes of COPD patients with treatment of tiotropium bromideObjective:Different HRCT-classified phenotypes of COPD patients have different therapeutic responses to the treatment of tiotropium bromide.Aiming at identifying whether there are differences in the main components(VAChT,OCTs,CHT1 and ChAT)of non-neuronal cholinergic system before and after treatment with tiotropium bromide in patients with different HRCT phenotypes to explore whether the difference in treatment efficacy of tiotropium bromide is due to the involvement in non-neuronal cholinergic system.Methods:Two hundred and nineteen patients with COPD were screened and divided into phenotype E(n=116)and phenotype M(n=103)according to HRCT classification criteria.The changes of mRNA expression and protein expression in VAChT,OCT1,OCT2 and CHT1 were compared between COPD patients with different phenotypes before and after treatment with tiotropium bromide.Meanwhile,the changes of enzymatic activity of ChAT between different phenotypic COPD patients before and after treatment with tiotropium bromide was also compared.Results:Expression of mRNA,the mRNA expression levels of VAChT,OCT1 and OCT2 between phenotype E and phenotype M were significantly different before and after treatment with tiotropium bromide.The protein expression of CHT1 between phenotype E and phenotype M had no significant differences before and after treatment with tiotropium bromide.Expression of protein,the protein expression levels of VAChT,OCT1 and OCT2 between phenotype E and phenotype M were significantly different before and after treatment with tiotropium bromide.But the protein expression of CHT1 between phenotype E and phenotype M did not change before and after treatment with tiotropium bromide.Activity of enzyme,the enzymatic activity of ChAT between phenotype E and phenotype M had significant differences before and after treatment with tiotropium bromide.Conclusion:The non-neuronal cholinergic system is involved in the heterogeneity of treatment with tiotropium bromide in COPD patients with different HRCT phenotypes.Differences exist on VAChT,OCT1,and OCT2 of non-neuronal cholinergic systems in COPD patients with different HRCT phenotypes.ChAT activity of non-neuronal cholinergic system in phenotype E could be reduced by treatment with tiotropium bromide so that indirectly mediate the heterogeneity in different phenotypes.The difference in therapeutic efficacy of tiotropium bromide in COPD patients with different HRCT phenotypes may be caused by the anti-inflammatory and anti-airway remodeling effects of tiotropium bromide.