A population structure analysis using two chronic obstructive pulmonary disease related gene polymorphisms

Chronic obstructive pulmonary disease (COPD) is a multifactorial disease characterized by airflow obstruction that is usually progressive and associated with an abnormal inflammatory response of the lungs to noxious particles and gases. Cigarette smoke (CS) is the main risk factor, but only a small proportion of smokers (15-20%) develop symptomatic disease, this suggests that there are individual susceptibility factors involved in disease onset and progression. Considering the impact of environmental and genetic risk factors in COPD, this dissertation sought to uncover genetic susceptibility biomarkers in a population affected by COPD and explore tissue metabolic alterations induced by chronic CS exposure in a mouse model. A case-control study was carried on in a COPD population, aiming to investigate whether polymorphisms of microsomal epoxide hydrolase (EPHX1) gene, and related phenotypes, had any bearing on individual susceptibility to COPD onset and severity. DNA of COPD patients and controls was genotyped for functional polymorphisms of EPHX1 gene (exon 3 Tyr113His, exon 4 His139Arg), and haplotype analysis was performed using PCR-RFLP and PCR-RT techniques. The statistical analysis did not show any significant result about the potential relationship between analyzed SNPs, related phenotypes, and COPD risk and severity. Stable isotope-resolved metabolomics approach was used to study glycolytic pathway alterations induced by chronic CS exposure in lung and liver tissue of an emphysema mouse model. C57BL/6J mice, after CS exposure, were injected via IP injection with glucose tracer containing carbon stable isotope - 13C6-glucose – then, tissues were collected and the incorporation of 13C into metabolites was monitored by mass spectrometry and nuclear magnetic resonance spettroscopy. Lung tissue analyses did not reveal any significant alteration in lung tissue glycolysis of mice exposed to CS. On the other hand, CS may contribute to dysregulated glycolysis, PPP, glycogen synthesis and utilization, in emphysema mouse model liver tissue.

Objective In this study,smokers in the nicotine receptor 3 (CHRNA3) gene promoter region polymorphism and smoking-related chronic obstructive pulmonary disease (COPD) susceptibility.Methods A case-control study of smoking were collected 60 cases of patients with COPD and 60 healthy smokers,using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to detect single gene CHRNA3 nucleotide polymorphism rs6495309 (T＞C) genotypes,with SPSS 13.0 software for non-conditional logistic regression calibration confounding factors,analysis of genetic variation and incidence of COPD in smokers.Results The genotype frequencies were in Hardy-Weinberg equilibrium ( P ＞0.05).CHRNA3 rs6495309 (T＞C) genotype with smoking COPD patients:three genotypes in smoking patients with COPD and healthy smoking group in the distribution of a statistically significant difference,P =0.043.CHRNA3 rs6495309 CC genotype associated with susceptibility to COPD in smokers ( P =0.014),and smoking in the male population,rs6495309 CC genotype associated with COPD,more obvious,the smoking men carrying the CC genotype individuals suffering from COPD is 4.22 times than the TT genotype (OR =4.222,95％ CI for the 1.369-13.020,P =0.012),and TC genotype not associated with COPD.And with the smoking index increase rs6495309 CC genotype was higher than it in the proportion of patients with COPD accounted for.However,the incidence of COPD severity according to further analysis available,rs6495309 CC genotype in Ⅲ,Ⅳ-class people with COPD Ⅰ,Ⅱ grade COPD population difference was not statistically significant,the rs6495309 CC genotype is not associated with severity of COPD.Conclusions CHRNA3 gene promoter polymorphism has susceptibility to COPD male smokers,and the rs6495309 CC genotype has significant correlated with the susceptibility of COPD in male smokers.The change of CHRNA3 gene promoter polymorphism was positively correlated with the severity of smoking,but is not relevant with COPD severe degree. Key words: CHRNA3; Chronic obstructive pulmonary disease; Smokers; Genetic polymorphism

Etiology of chronic obstructive pulmonary disease remains unknown but despite some inconsistencies in reports on inflammatory cells,mediators and proteases involved in the pathogenesis of chronic obstructive pulmonary disease,genetic risk factors were proposed as a cause of susceptibility to the disease.Results of many studies suggested polygenic inheritance,with the genetic component consisting of several genes of a small effect each,rather than of single major gene.We are going to review the clinical importance of alpha-1 antitrypsin,matrix metalloproteinase,Tissue inhibitor of metalloproteinases-2,Vitamin-D-binding protein,glutathione S-transferase,microsomal epoxide hydrolase,heine oxygenase-1 genes,Surfacant protein and IL-8,TNF-α polymorphisms associated with susceptibility and progression of the chronic obstructive pulmonary disease.It was reported that there are combination with different genes or different locus in the same gene.Animal model genetics may help in clarifying some aspects of pathogenesis.If this type of study were followed by quantitative trait locus analysis it may help to identify candidate genes for further study in humans.Genome-wide association analysis may nOW be performed looking for up to 500000 SNPs at any one time to identify regions in linkage disequilibrium(LD)with features of COPD.This approach does,however,have limitations.It means there are millions of data to analysis,but Software available from industry and academia can help to deal with that.The number of areas being investigated will raise the potential for false positive results,SO confirmation of any positive results in multiple independent populations should be sought.As more genes are identified we may be able to characterize patients with COPD more accurately and target therapies to those subgroups most likely to benefit. Key words: Chronic obstructive pulmonary disease; Oxidative/anti-oxidative; Proteinase/antiproteinase; Inflammation; Gene polymorphism

Understanding COPD Patients' Barriers to Behavioural and Lifestyle Changes Using a Resilience, Readiness to Change, and Self-Management Model

Chronic obstructive pulmonary disease(COPD) is a complex human disease, its etiology remains unknow. Results of many studies suggests multiple genes and environmental effect it. Hereditary risk factors are considered as a risk factor of this disease. Investigatin on the relationship between gene polymorphism and COPD will help people understand its etilolgy and mechanism of COPD and also can provide help for treatment and prevention COPD. Key words: Chronic obstructive pulmonary disease; Susceptibility gene; Genetic polymorphisrn

Objective To study the relationship between tumor necrosis factor-α+489G/A gene polymorphisms and susceptibility to development of chronic obstructive pulmonary disease(COPD).Methods Polymerase chain reaction-restriction fragment length polymorphism method was used to detect the polymorphism of TNF-α+489G/A gene in 5 5 cases of COPD,47 cases of sex,age and smoke history matched controls and 55 cases of body-check healthy subjects and their susceptibility to development of COPD was analyzed.Results The frequencies of GA/AA genotype in COPD group,control group and body-check healthy group were 23.6%,14.3%and 9.1%,respectively.There were not statistic difference between the groups(P＞0.05).Statistical analysis showed that COPD group had a significantly higher allele A frequency than that of body-check healthy group(P＜0.05).The prevalence of chronic pulmonary heart disease in the GA/AA genotype was higher than the GG genotype in COPD group.Conclusions The peoples who carry allele A in TNF-α+489 may increase the risk of COPD and poor progress. Key words: Tumor necrosis factor-α gene; Polymorphisms; Chronic obstructive pulmonary disease

Objective To detect the correlations between polymorphisms of vitamin D receptor (VDR) genes and chronic obstructive pulmonary disease (COPD) in smokers. Methods Fifty smoking COPD patients and fifty smoking volunteers were enrolled in the study.Polymorphisms of VDR genes including Apa I, Bsm I and Taq I were detected by using polymerase chain reaction-restriction fragments length polymorphism (PCR-RFLP). Correlations between VDR gene polymorphisms and the susceptibility to COPD in smokers were analyzed. Results There was a statistically significant difference in the frequency distributions of aa, Aa and AA genotypes between the smoking COPD group and the smoking control group (P<0.05). Using the Aa+ AA genotype as a reference, individuals carrying the aa genotype increased the risk of COPD in smokers (OR=2.786, 95%CI: 1.215-6.389). There was a statistically significant difference in the frequency distributions of a and A alleles between the two groups (P<0.05). Compared with carrying allele A, individuals carrying allele a increased the risk of COPD in smokers (OR=1.897, 95% CI: 1.042-3.457). There was no statistically significant difference in genotype frequency distribution of Bsm I and Taq I between the two groups. Conclusions Apa I gene polymorphism may increase the susceptibility of COPD in smokers, while Bsm I and Taq I gene polymorphisms may not be associated with the susceptibility of COPD in smokers. Key words: Pulmonary disease, chronic obstructive; Vitamin D receptor; Polymorphism; Smokers

Pulmonary hypertension is a major complication of chronic obstructive pulmonary disease (COPD), affecting the prognosis of patients with COPD. Chronic hypoxia is the main pathogenesis. In recent years, through the research to certain gene's polymorphism with pulmonary hypertension,have found that certain genetic polymorphism is a risk factor about COPD patients with pulmonary hypertension. This article is to introduce pulmonary hypertension in COPD gene polymorphism makes a summary. Key words: Gene polymorphism; Chronic obstructive pulmonary disease; Pulmonary hypertension

Objective To investigate whether polymorphisms and haplotypes of S1,S2 locus alleles in ADAM33 gene has any bearingon chronic obstructive pulmonary disease (COPD) and lung function in Han population of northern China.Methods The genotypes of S1,S2 locus alleles in 90 COPD patients and 90 healthy controls were tested with PCR-direct sequeneing analysis. Haplotypes and their frequencies were established and analyzed by SHEsis online software.Results ①There was no significant difference between the COPD group and the control group in the frequencies of S1 locus genotypes and alleles ( P ＞0.05).There was a significant difference between the COPD group and the control group in the frequencies of S2 locus genotypes and alleles ( P ＜0.05).②Logistic regression analysis showed:there was not statistically significant difference in the relative risk of COPD in S1 locus genotype ( P ＞0.05).There was a statistically significant difference in the relative risk of COPD in S2 locus genotype ( P ＜0.05),and the odds ratio ( OR ) of G/G+C/G genotype was 2.364 (95％ CI 1.251-4.466).③In COPD group,the relationship between ADAM33 gene S2 locus genotype and clinical parameters of lung function display:the difference of FEV1 ％ in genotypes was no statistically significant,but the difference of FEV1/FVC in genotypes was statistically significant,and G/G genotype with FEV1/FVC decline was more significantly than C/C,C/G genotype.④SHEsis online software analysis for S1,S2 loci haplotype showed that haplotype C/G in COPD and control groups was statistically significant difference ( P ＜ 0.05).Conclusions In Han population of northern China,the polymorphism in ADAM33 gene have association with the susceptibility to COPD,and have no association with the severity of COPD. Key words: Chronic obstructive pulmonary disease; A disintegrin and metalloprotease; ADAM33 gene ; Single nucleotide polymorphism; Haplotype

Objective To investigate correlation of tumor necrosis factor-α-308 (TNF-α-308) loci gene polymorphism and chronic obstructive pulmonary disease (COPD). Methods One hundred and forty-three COPD patients treated in our hospital from February 2014 to March 2017 were selected, and 150 healthy volunteers were selected as control group.The two groups of TNF-α-308 loci gene polymorphism were detected by sequencing. Results TNF-α-308 loci genotype were three: GG type, GA type and AA type.The frequencies of genotype AA/GA and allele A in the observation group were 29.37% and 17.13%, which were significantly higher than 12.00% and 6.67% in the control group.The genotype frequencies of TNF-α-308 loci gene GA/AA were not significantly related to the sex, age, smoking and severity of COPD patients (χ2=0.001, 0.009, 0.035, 0.001, all P>0.05). The bronchial wall thickness, ratio of bronchial wall thickness and pulmonary artery wall in GA/AA genotype patients were (0.30±0.11) cm and 0.40±0.12, significantly higher than those in type GG patients (t=11.231, 6.667, both P 0.05). Conclusions TNF-α-308 loci gene may be associated with the susceptibility to COPD, the A allele may be a susceptible gene, the bronchial wall thickness is increased significantly, however, the polymorphism of this locus is not related to the sex, age and severity of COPD patients. Key words: Tumor necrosis factor-α; 308 loci; Gene polymorphism; Chronic obstructive pulmonary disease

Objective To investigate the relationship between the variants of the class A scavenger receptor (SR-A) and chronic obstructive pulmonary disease (COPD) with or without lung cancer in China. Methods From January 1, 2016, to June 20, 2016, 100 patients with COPD with lung cancer, 100 patients with COPD without lung cancer, and 100 healthy smokers were enrolled into Shanghai Ruijin Hospital for genotyping of eight single nucleotide polymorphisms (SNPs: ex3P36A_C>G, ex3S41Y_C>A, ex4V113A_T>C, ex4P174Y_G>T, ex6P275A_C>G, ex6R293X_C>T, ex10G369S_G>A, and ex11H441R_A>G) by gene sequencing. Results The genotype frequencies of the aforementioned eight SNPs between patients with COPD with and without lung cancer and healthy controls were not significantly different.However, during DNA sequencing, a new promising SNP, rs13306550 (IVS4+ 3A>G), was found in the splice donor site, which was significantly associated with an increased risk of COPD. Conclusions This study demonstrated that the variant rs13306550 was a risk factor for COPD susceptibility, but had little influence on the pathogenesis of lung cancer in patients with COPD. Key words: Chronic obstructive pulmonary disease; Lung cancer; Class A scavenger receptor; Single-nucleotide polymorphism

Objective To investigate the correlation between matrix metalloproteinase-9 (MMP-9)gene polymorphism and the susceptibility of chronic obstructive pulmonary disease(COPD).Methods Examine the frequency of the genetype of MMP-9 in 69 patients and 61 control subjects by reslriction fragment length polymorphism.Results ①The frequency of MMP-9 QQ,R/Q genotype were 31.8％,14.6％ in COPD group,respectively,and 37.8％,26.2％ in control group,respectively.There were no significant differences between the two groups ( P ＞0.05).The distribution frequencies of RR genotypes were significantly different between two groups ( P ＜0.05).The differences in allele frequencies were also significant between the two groups ( P ＜0.05).②Smokers,there were no significant differences on QQ,R/ Q genotypes in COPD group and in controls.There were significantly different on RR genotypes in COPD group and in controls ( P ＜0.05).In non-smokers,there were no significant differences on QQ,R/Q,RR genotypes in two group.Smokers,there were significantly different on the frequencies allele of R and Q between two groups.In non-smokers,there were no significant differences on the frequencies allele of R and Q between two groups.Conclusious ①MMP-9 R279Q polymorphisms maybe a genetic risk factor for COPD.②The allele gene of R maybe a susceptibility factor for COPD in smokers. Key words: Matrix metalloproteinases; Chronic obstructive pulmonary disease; Polymorphism; Smoking

Objective To investigate the correlation between the mierosatellite polymorphism of heme oxygenase-1(HOX-1)gene promoter and susceptibility to chronic obstructive pulmonary disease(COPD)in Han nationality of Southwest China and CT imaging.Method The alleles frequencies with varying number of(GT)n repeatings in the Hox-1 gene in 180 smokers with COPD and in 150 healthy smokers were analyzed.Results Polymorphisms of the(GT)n repeats were grouped into three classes:S(≤25 repeats),M(26-31 repeats),L(≥32repeats).The proportion of genotypic frequencies in the group with class L alleles(L/S,L/M,I/L)was significantly higher in the smokers with COPD than in healthy smokers(29.4%vs18.7%,P=0.023,OR=1.8,95%CI 1.1-3.1).However,there was no relationship between the polymorphism of HOX-1 gene and subtypes of COPD by CT imaging in COPD patients(P＞0.05).Conclusion Genetic polymorphism in Hox-1 is associated with development of COPD in Han nationality of Southwest China.But the polymorphism of HOX-1 gene is not associated with subtypes of COPD by CT imaging. Key words: Chronic obstructive pulmonary disease; Heme Oxygenase-1; Polymorphism; CT

Chronic obstructive pulmonary disease (COPD) is a major cause of chronic morbidity and mortality. It is influenced by both environmental and genetic factors. Aquaporin5 plays a critical role in the maintenance of normal lung water homeostasis. We investigated whether polymorphisms in the gene aquaporin5 had any bearing on individual susceptibility to the development of COPD. 332 COPD patients and 373 unrelated, age-matched healthy people were recruited for the study. All participants were Chinese Han people. We designed denaturing high performance liquid chromatography (DHPLC) to detect SNP in exons1, 2, 3 of AQP5 and DNA sequencing to confirm it. The allele +2254A>G (rs3736309) in intron3 and +3088A>G (Thr-Thr, rs41308104) in exon4 from NCBI dbSNP were genotyped by PCR-based restriction fragment length polymorphism. We found no SNPs in exons1-4 of the AQP5 gene in our study population. However, the genotype frequency of +2254A>G SNP in intron3 was significantly different in cases and controls. The frequencies of AA AG GG in cases were 45.2%, 40.7%, and 14.1%, while in controls they were 34.5%, 51.6%, and 13.9% (chi2 = 9.899, P = 0.007), respectively. Higher OR for COPD was seen for persons with +2254AA genotype against +2254AG genotype (OR = 2.73; 95%CI = 1.88-3.97). Carriers of the variant allele +2254G had a lower risk of COPD than homozygous wild type carriers (OR = 0.44; 95%CI = 0.307-0.631). The +2254A to G variant in intron 3 of AQP5 was associated with a decreased risk of COPD in a Chinese population.

Men are different, women too. A concept which has long been neglected in biomedical research. Except for reproductive differences women were regarded as smaller men, clinical guidelines did not differentiate between men and women. There are, however, further differences. Every cell has a sex, which might influence regulation of gene expression, disease phenotype and drug toxicity. In drug research it is important to understand the pathomechanism and clinical presentation of a disease as well as the mechanism of action of a drug. Information on drug safety and efficacy are collected in preclinical and clinical studies. Natural history of disease studies provide valuable information on the clinical presentation of a disease. It is important to provide this information gender stratified to be able to offer best care to future patients. Chronic obstructive pulmonary disease (COPD) has traditionally been regarded as a disease of white men but today almost as many women are affected by the disease as men. The burden of COPD is still projected to increase, particularly in women. Despite this there are few studies comparing the clinical manifestation and clinical course in men and women with COPD. It was the aim of this thesis to contribute new data to the natural history of COPD with a special focus on the effect of gender. The studies of this thesis were conducted with data from the General Practice Research Database (GPRD), a large population-based database in the United Kingdom. The GPRD provides anonymized medical information on a 5% representative sample of the UK population. This thesis presents six studies focussing on a population of 35,772 COPD patients, aged 40-79 years, who received their incident COPD diagnosis between 1995 and 2005 and the same number of randomly matched COPD-free patients for comparison. In a case-control analysis we compared the prevalence of co-morbidities and respiratory drug utilization in men and women with COPD. In nested case control analyses COPD and COPD-free patients were compared with respect to their risk to develop cardiovascular or gastrointestinal outcomes, depression or cancer. The first study described the COPD population with respect to co-morbidities, drug use and survival. Patients with COPD had more co-morbidities and a lower survival than COPD-free patients. In COPD patients the prevalence of diabetes, myocardial infarction, stroke / transient ischemic attack (TIA), arrhythmia and peptic ulcer were higher in men than in women while depression and osteoporosis were more prevalent in women. We observed small but significant gender differences in drug utilization and survival. The second study analysed in more detail the association between COPD and the prevalence of diabetes. The prevalence of diabetes was lower in the COPD group than in the COPD-free comparison group. This association was significant in men but not women and mainly seen in users of sulfonylurea. Studies 3-6 were follow-up and nested case control analyses evaluating the risk of the cardiovascular outcomes (arrhythmia, pulmonary embolism, deep vein thrombosis, myocardial infarction and stroke / TIA), gastro-oesophageal reflux disease, peptic ulcer, depression and cancer risk in the population of COPD patients and compared it to a COPD-free population. The incidence of most cardiovascular diseases was higher in patients with COPD. COPD had a stronger impact on the risk of MI and stroke / TIA in women than in men. Relative risks of PE, DVT and arrhythmia were similar in men and women. Severe COPD materially increased the risk of MI and PE in both men and women. The incidence rates of GORD were slightly higher in men than in women while peptic ulcer incidence rates were higher in men. COPD did not materially alter the risk of GORD or peptic ulcer. Current use of long-acting beta agonists was associated with a decreased risk of peptic ulcer. Patients with COPD had a higher risk of cancer than COPD-free patients. The increased risk was mainly driven by a high lung cancer risk among COPD patients, which was higher in women than in men. This effect was seen independent of smoking status. Many patients with COPD developed depression during follow-up, particularly patients with severe COPD. The risk of depression was higher in women than in men but COPD seemed to have a greater impact in men than in women. The studies of this thesis provide further evidence that patients with COPD are at an increased risk of depression, most cardiovascular diseases and lung cancer. They also demonstrate that gender-stratified analyses are important to adequately address the risk for a disease.