Background
Methods
Study design
Ethics approval and consent to participate
Adult cohorts and participants
Children and adolescent cohort and participants
Lung function
DNA methylation measurement
Methylation signals considered in SERPINA gene cluster
Covariate information
Statistical analyses
Cohort-specific analyses on lung function and its decline in adults
Meta-analysis of adult cohort results
Cohort-specific analysis on lung function and its growth in children and adolescents
Sensitivity analyses
Results
Characteristics of adult cohort participants
SAPALDIA2 T1, 2001 | SAPALDIA3 T2, 2010 | ECRHS 2 T1, 1998 | ECRHS 3 T2, 2008 | NFBC 1966 T1, 1997 | NFBC 1966 T2, 2012 | ||
---|---|---|---|---|---|---|---|
N | 561 | 561 | 267 | 267 | 248 | 248 | |
Female, % | 48.3 | 48.3 | 52.1 | 52.1 | 49.8 | 48.2 | |
Age (years), mean (SD) | 50.4 (10.8) | 58.7 (10.7) | 44.0 (6.0) | 55.0 (6.0) | 31.0 (0.3) | 46.3 (0.4) | |
Height (cm), mean (SD) | 170.2 (9.2) | 169.5 (9.3) | 170.0 (9.0) | 170.0 (9.0) | 172.0 (8.7) | 172.0 (8.6) | |
Weight (kg), mean (SD) | 73.9 (14.8) | 75.6 (15.6) | 74 (15.0) | 77 (16.0) | 72.1 (12.8) | 80.8 (15.3) | |
Body mass index (kg/m2), mean (SD) | 25.4 (4.2) | 26.2 (4.6) | 25.2 (4.2) | 26.8 (4.5) | 24.3 (3.6) | 27.2 (4.5) | |
Smoking status, % | |||||||
Ex-smoker | 52.0 | 63.0 | 54.7 | 69.3 | 46.8 | 69.2 | |
Current smoker | 48.0 | 37.0 | 45.3 | 30.7 | 53.2 | 30.8 | |
Pack-years, mean (SD)a | 20.4 (20.2) | 22.6 (22.1) | 17 (17.0) | 20 (21.0) | 7.7 (5.9) | 11.0 (9.6) | |
Education (%) | |||||||
SAPALDIA | ECRHS & NFBC | ||||||
Lowb | ≤16 yearsc | 5.0 | 4.0 | 10.9 | 10.9 | 0.4 | 0.4 |
Intermediate | 17–19 years | 66.0 | 67.0 | 30.5 | 30.5 | 61.9 | 61.9 |
High | 20+ years | 29.0 | 29.0 | 58.6 | 58.6 | 37.7 | 37.7 |
FVC (L), mean (SD) | 4.4 (1.0) | 4.1 (1.0) | 4.4 (1.0) | 4.0 (1.0) | 4.9 (1.0) | 4.6 (0.9) | |
FVC % Predictedd, mean (SD) | 106.2 (13.4) | 106.0 (13.4) | 100.7 (13.0) | 99.2 (13.5) | 101.8 (10.8) | 104.1 (11.2) | |
FEV1 (L), mean (SD) | 3.3 (0.8) | 3.0 (0.8) | 3.4 (0.8) | 3.0 (0.7) | 4.1 (0.8) | 3.5(0.7) | |
FEV1% Predictedd, mean (SD) | 99.3 (14.2) | 97.9 (16.0) | 96.5 (13.7) | 93.8 (14.2) | 102.1 (11.5) | 100.1 (11.8) | |
FEV1/FVC, mean (SD) | 0.8 (0.08) | 0.7 (0.08) | 0.8 (0.06) | 0.8 (0.1) | 0.8 (0.05) | 0.8 (0.06) | |
FEV1/FVC % Predictedd, mean (SD) | 93.2 (9.4) | 92.0 (9.9) | 95.4 (7.7) | 94.2 (7.7) | 99.9 (6.8) | 95.8 (7.0) | |
CRP (mg/L), mean (SD) | 1.92 (2.8) | – | – | – | – | – | |
AAT (g/L), mean (SD) | 1.27 (0.2) | – | – | – | – | – | |
COPD, defined by FEV1/FVC < LLNe | 15.3 | 17.1 | 10.5 | 12.7 | 2.2 | 11.4 | |
Doctor-diagnosed asthma, % | 15.7 | 19.1 | 14.6 | 17.2 | 10.5 | 17.1 | |
Respiratory medication (last 12 months), % | 22.5 | 25.8 | 12.4 | 14.3 | NA | NA | |
Spirometer brand and model, % | |||||||
SensorMedics 2200 | 100.0 | – | – | – | – | – | |
EasyOne NDD Medical Technologies | – | 100.0 | – | 100.0 | – | – | |
SensorMedics 2400 | – | – | 36.0 | – | – | – | |
Jaeger Pneumotach | – | – | 16.1 | – | – | – | |
Biomedin Spirometer | – | – | 47.9 | – | – | – | |
Vitalograph P Model | – | – | – | – | 100.0 | – | |
MasterScreen Pneumo | – | – | – | – | – | 100.0 |
Association of methylation at 119 CpGs in the SERPINA1 cluster with lung function in adult ever smokers
SERPINA1
CpG | CpG Position | Gene Group | SAPALDIA | Meta- Analysis of Lung Function Association in SAPALDIA, NFBC and ECRHS | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Circulating AAT | Cross-Sectional T1 | Cross-Sectional T2 | Repeat Cross-Sectional | Change | ||||||||||||
Coef. | P.valuea | Coef. | P.valuea | Directionb | Coef. | P.valuea | Directionb | Coef. | P.valuea | Directionb | Coef. | P.valuea | Directionb | |||
cg10761141 | 94,844,776 | 3UTR | −0.30 | 0.25 | 0.75 | 0.13 | +++ | 0.63 | 0.21 | + − + | −0.04 | 0.84 | + − – | 0.02 | 0.48 | +++ |
cg25042671 | 94,849,061 | Body |
−0.59
|
0.04
| −0.72 | 0.25 | − + − | − 0.80 | 0.21 | – − − | −0.22 | 0.52 | ++− | −0.03 | 0.44 | − + − |
cg20267408 | 94,849,703 | 5UTR | −0.44 | 0.21 | −0.15 | 0.82 | ++− | −0.05 | 0.94 | ++− | 0.10 | 0.79 | ++− | 0.00 | 0.97 | + − – |
cg26938334 | 94,849,747 | 5UTR | 1.16 | 0.42 | − 0.01 | 0.99 | − + − | − 0.35 | 0.74 | − + − | − 0.33 | 0.34 | – − + | − 0.01 | 0.71 | – – – |
cg16110645 | 94,850,433 | 5UTR | − 0.64 | 0.34 | 0.39 | 0.67 | −++ | 0.55 | 0.59 | + − + | 0.08 | 0.85 | + − + | −0.02 | 0.75 | – − + |
cg10832639 | 94,850,965 | 5UTR | −0.25 | 0.72 | 1.24 | 0.16 | +++ | 0.01 | 0.99 | −++ | 0.18 | 0.61 | −++ | 0.00 | 0.95 | ++− |
cg02126235 | 94,855,034 | 5UTR | −0.30 | 0.58 | 0.76 | 0.45 | +++ | 0.67 | 0.55 | ++− | − 0.26 | 0.55 | – – – | 0.02 | 0.69 | ++− |
cg22828990 | 94,855,099 | 5UTR | −0.09 | 0.82 | 0.19 | 0.79 | – − + | 0.30 | 0.68 | – − + | 0.69 | 0.03 | + − + |
−0.09
|
0.04
| – – – |
cg09968361 | 94,855,344 | 5UTR | −0.33 | 0.44 |
1.43
|
0.04
| +++ | 0.65 | 0.36 | −++ |
0.72
|
0.02
| +++ |
−0.10
|
0.01
| – |
cg13826459 | 94,855,505 | TSS1500 | −0.08 | 0.76 | 0.16 | 0.76 | −++ |
0.98
|
0.05
| +++ | −0.05 | 0.83 | − + − | 0.04 | 0.15 | +++ |
cg25968219 | 94,855,590 | TSS1500 |
−1.81
|
0.05
|
3.29
|
0.003
| +++ | −1.29 | 0.37 | – − + | 0.37 | 0.59 | ++− | − 0.08 | 0.15 | – − + |
cg01606800 | 94,855,990 | TSS1500 |
−1.08
|
0.001
| 0.47 | 0.47 | −++ | −0.13 | 0.85 | – − + | − 0.11 | 0.73 | – − + | − 0.01 | 0.75 | – − + |
cg02181506 | 94,856,984 | 1stExon | −0.78 | 0.08 | 0.11 | 0.89 | – − + | 0.65 | 0.40 | + − + | 0.28 | 0.47 | + − + | 0.02 | 0.71 | + − – |
cg10070185 | 94,857,151 | TSS200 | −0.05 | 0.91 | 0.37 | 0.59 | – − + | 1.08 | 0.16 | +++ | 0.24 | 0.39 | −++ | 0.04 | 0.29 | + − + |
cg24621042 | 94,857,275 | TSS1500 |
−0.68
|
0.04
| 0.87 | 0.14 | +++ | 1.16 | 0.06 | +++ | −0.15 | 0.60 | − + − | − 0.01 | 0.87 | + − + |
cg00802237 | 94,858,066 | TSS1500 | −0.07 | 0.81 | −0.01 | 0.99 | + − – | − 0.48 | 0.40 | – − + | 0.12 | 0.67 | + − + | −0.03 | 0.35 | – − + |
cg04179148 | 94,858,339 | TSS1500 | −0.70 | 0.06 |
1.80
|
0.01
| +++ | 0.29 | 0.67 | +++ | 0.36 | 0.25 | +++ | −0.03 | 0.47 | – – – |
CpG | Position | Gene Group | SAPALDIA | Meta- Analysis of Lung Function Association in SAPALDIA, NFBC and ECRHS | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Circulating AAT | Cross-Sectional T1 | Cross-Sectional T2 | Repeat Cross-Sectional | Change | ||||||||||||
Coef. | P.valuea | Coef. | P.valuea | Directionb | Coef. | P.valuea | Directionb | Coef. | P.valuea | Directionb | Coef. | P.valuea | Directionb | |||
cg10761141 | 94,844,776 | 3UTR | −0.30 | 0.25 | 0.10 | 0.14 | −++ | −0.02 | 0.79 | + − + | −0.01 | 0.79 | – − + | 0.00 | 0.82 | + − – |
cg25042671 | 94,849,061 | Body |
−0.59
|
0.04
| −0.09 | 0.29 | − + − | − 0.12 | 0.22 | – – – |
−0.12
|
0.03
| + − – | 0.00 | 0.62 | + − + |
cg20267408 | 94,849,703 | 5UTR | −0.44 | 0.21 | −0.05 | 0.57 | – – – | 0.05 | 0.59 | −++ | 0.01 | 0.88 | + − + | 0.00 | 0.78 | −++ |
cg26938334 | 94,849,747 | 5UTR | 1.16 | 0.42 | 0.10 | 0.24 | − + − | −0.10 | 0.47 | −++ | 0.01 | 0.84 | ++− | −0.01 | 0.33 | – − + |
cg10832639 | 94,850,965 | 5UTR | −0.25 | 0.72 | 0.16 | 0.14 | + − + | 0.02 | 0.86 | − + − | 0.08 | 0.18 | +++ | 0.01 | 0.32 | +++ |
cg02126235 | 94,855,034 | 5UTR | −0.30 | 0.58 | 0.26 | 0.06 | +++ | 0.06 | 0.71 | − + − | 0.06 | 0.41 | ++− | 0.00 | 0.90 | – − + |
cg09968361 | 94,855,344 | 5UTR | −0.33 | 0.44 | 0.11 | 0.23 | +++ | 0.00 | 0.99 | ++− | 0.05 | 0.25 | ++− | −0.01 | 0.37 | – − + |
cg13826459 | 94,855,505 | TSS1500 | −0.08 | 0.76 | 0.07 | 0.34 | + − + |
0.15
|
0.05
| + − + | 0.04 | 0.32 | +++ | 0.01 | 0.24 | −++ |
cg25968219 | 94,855,590 | TSS1500 |
−1.81
|
0.05
|
0.31
|
0.02
| +++ | −0.16 | 0.43 | – − + | − 0.01 | 0.90 | −++ | 0.00 | 0.59 | + − + |
cg01606800 | 94,855,990 | TSS1500 |
−1.08
|
0.001
|
0.17
|
0.05
| + − + | 0.01 | 0.96 | – − + | 0.00 | 0.97 | – − + | 0.00 | 0.73 | ++− |
cg02181506 | 94,856,984 | 1stExon | −0.78 | 0.08 | −0.01 | 0.89 | + − – | 0.05 | 0.64 | + − – | −0.03 | 0.58 | + − – | 0.00 | 0.80 | ++− |
cg10070185 | 94,857,151 | TSS200 | −0.05 | 0.91 | 0.07 | 0.45 | +++ | 0.19 | 0.09 | −++ | −0.01 | 0.81 | – − + | 0.00 | 0.63 | – − + |
cg24621042 | 94,857,275 | TSS1500 |
−0.68
|
0.04
|
0.18
|
0.02
| +++ | 0.16 | 0.08 | +++ | 0.07 | 0.09 | +++ | −0.01 | 0.09 | + − – |
cg00802237 | 94,858,066 | TSS1500 | −0.07 | 0.81 | 0.03 | 0.71 | + − + | −0.14 | 0.09 | – – – | 0.00 | 0.96 | −++ | 0.00 | 0.50 | + − – |
cg04179148 | 94,858,339 | TSS1500 | −0.70 | 0.06 |
0.20
|
0.03
| +++ | 0.04 | 0.73 | ++− |
0.12
|
0.02
| +++ | 0.00 | 0.57 | – – – |
SERPINA gene cluster
Association of methylation at 119 CpGs with circulating alpha-1 antitrypsin in adult ever smokers in SAPALDIA
Association of methylation at 119 CpGs with lung function in tobacco-smoke exposed children and adolescents
Children Cross-Sectional T1 (2000) | Adolescents Cross-Sectional T2 (2007) | |
---|---|---|
N | 259 | 259 |
Females (%) | 53.7 | 53.7 |
Age (years), mean (SDb) | 8.6 (0.2) | 15.4 (0.3) |
Height (cm), mean (SD) | 133.3 (6.0) | 170.2 (8.0) |
Weight (kg), mean (SD) | 31.3 (6.3) | 62.8 (11.0) |
BMI | 17.4 (2.5) | 21.7 (3.3) |
Maternal Education (%) | ||
Ordinary Level or Lower | 56.4 | 56.4 |
Advanced Level | 27.8 | 27.8 |
University | 15.8 | 15.8 |
Asthmatics (%) | 21.6 | 28.9 |
Smoking Exposure (%) | ||
In-utero smoking (smoking mother) | 25.1 | 25.1 |
Second-hand exposure at home | 56.0 | 56.0 |
Self-exposure (smoked > 2 times in a life-time) | 52.1 | 52.1 |
COPD [FEV1/FVC < LLNc] (%) | 5.0 | 4.6 |
Asthma medications in the last 12 months (%) | 14.0 | 14.0 |
FVC(L), mean (SD) | 2.0 (0.3) | 3.9 (0.9) |
FEV1(L), mean (SD) | 1.7 (0.3) | 3.4 (0.7) |
FEV1/FVC(L), mean (SD) | 0.9 (0.06) | 0.9 (0.07) |
SERPINA1
SERPINA gene cluster
Replication of previously reported COPD signals
Circulating AATa | cg02181506 | cg24621042 | ||
---|---|---|---|---|
Coef. |
P
value
| Coef. |
P
value
| |
− 0.78 | 0.08 | − 0.68 | 0.04 | |
Adults (SAPALDIA, NFBC66, ECRHS) | ||||
FEV1, repeat cross-sectional | 0.28 | 0.47 | −0.15 | 0.60 |
FEV1, change | 0.02 | 0.71 | −0.01 | 0.87 |
Children and Adolescents (ALSPAC) | ||||
FEV1, repeat cross-sectional | 1.42 | 0.35 | 0.31 | 0.29 |
FEV1, change | −0.54 | 0.26 | − 0.17 | 0.06 |
Adults (SAPALDIA, NFBC66, ECRHS) | ||||
FEV1/FVC, repeat cross-sectional | −0.03 | 0.58 | 0.07 | 0.09 |
FEV1/FVC, change | 0.00 | 0.80 | −0.01 | 0.09 |
Children and Adolescents (ALSPAC) | ||||
FEV1/FVC, repeat cross-sectional | 0.01 | 0.98 | 0.08 | 0.07 |
FEV1/FVC, change | −0.12 | 0.06 | 0.00 | 0.71 |
Sensitivity analyses in the SAPALDIA cohort
cg02181506 | cg24621042 | cg08257009 | ||||
---|---|---|---|---|---|---|
Coef. |
P
value
| Coef. |
P
value
| Coef. |
P
value
| |
FEV1/FVC repeat cross-sectional | ||||||
Main modela | −0.01 | 0.93 | 0.05 | 0.54 | 0.07 | 0.12 |
Adjustment for PiS and PiZ genotype | −0.01 | 0.94 | 0.05 | 0.54 | 0.07 | 0.12 |
Adjustment for neutrophils at T1 and T2 | −0.00 | 0.97 | 0.05 | 0.52 | 0.07 | 0.11 |
Men | 0.03 | 0.86 | 0.07 | 0.47 | 0.10 | 0.11 |
Women | −0.07 | 0.65 | 0.06 | 0.62 | 0.05 | 0.49 |
Not obese (BMI < 30 kg/m2) | −0.06 | 0.58 | 0.04 | 0.60 |
0.12
|
0.01
|
Obese (BMI ≥ 30 kg/m2) | 0.19 | 0.25 | 0.06 | 0.68 | −0.12 | 0.14 |
No self-report of ever asthma at T1 and T2 | −0.03 | 0.78 | 0.07 | 0.35 | 0.08 | 0.09 |
Asthma ever self-report at T1 or T2 | 0.14 | 0.44 | −0.05 | 0.72 | −0.01 | 0.88 |
FEV1/FVC changeb | ||||||
Main modelc | −0.01 | 0.60 | −0.01 | 0.49 |
0.02
|
0.005
|
Adjustment for circulating hs-crp and AATd | −0.008 | 0.56 | −0.007 | 0.01 |
0.02
|
0.006
|
Adjustment for PiS and PiZ genotype | −0.01 | 0.57 | −0.01 | 0.48 |
0.02
|
0.005
|
Adjustment for neutrophils at T1 and T2 | −0.01 | 0.44 | −0.01 | 0.36 |
0.02
|
0.01
|
Men | 0.01 | 0.77 | −0.00 | 0.74 |
0.02
|
0.01
|
Women | −0.03 | 0.14 | −0.01 | 0.56 | 0.02 | 0.08 |
Not obese (BMI < 30 kg/m2) at T1 | −0.00 | 0.84 | −0.00 | 0.87 |
0.02
|
0.01
|
Obese (BMI ≥ 30 kg/m2) at T1 | −0.04 | 0.13 | −0.04 | 0.07 | 0.02 | 0.17 |
No self-report of ever asthma at T1 and T2 | −0.01 | 0.57 | −0.01 | 0.40 |
0.01
|
0.04
|
Asthma ever self-report at T1 or T2 | −0.03 | 0.28 | 0.01 | 0.65 |
0.03
|
0.04
|