Transcript Diapositiva 1 - PneumoTrieste 2015

BPCO e carenza ereditaria di
AAT
Ilaria Ferrarotti
Centro per la Diagnosi del Deficit di Alfa1-Antitripsina
Fondazione IRCCS Policlinico S. Matteo
Dipartimento di Medicina Molecolare
Università di Pavia
Alpha-1 antitrypsin
Deficiency
Common
genetic disease among Caucasians (1/5000)
Autosomal
co-dominant heredity
Reduced
or complete lack of alpha-1-antitrypsin secretion from
hepatocytes into the bloodstream
Severe
AAT deficiency is most often associated with homozygous Zmutation (PiZZ); over 120 genetic variants of the Pi gene are currently
known
Lung,
liver, and skin (very rare) can be affected
Often
remains unrecognized or is wrongly diagnosed as COPD
CASE-FINDING
Severe AATD is clearly associated with disease risk accounting for 1-2%
of all COPD cases.
Reference
N° subjects
Severe AATD
Intermediate AATD
screened
detection rate
detection rate
US
965
0.01554
0.15855
Spain
2137
0.00514
0.02386
Lithuania
1167
0.00943
0.06855
Denden, COPD 2010
Tunisia
100
0
0.04
Molina, Ther Adv Resp Dis 2011
Spain
596
0.00503
0.16611
US
3457
0.00578
0.10095
Lieberman, Chest 1986
De la Roza, ERJ 2005
Sitkauskiene, Resp Med 2008
Rahaghi, COPD 2012
site
Nevertheless...
.... large genome-wide association studies (GWAS) on these pulmonary
outcomes have so far failed to point to this chromosomal region.
Missing heritability
Manolio TA et al, Nature 2009
• The first GWAS on AAT serum
concentration in a subset of the
SAPALDIA (Swiss Cohort Study of Air
Pollution and Lung Disease in Adults)
general population sample
Detection of associated
chromosomal loci
•Fine mapping of the SERPINA1 locus
(genotyping of additional SNPs in the
whole study sample - exon
sequencing in subjects with
unexplained low levels of circulating
AAT ).
Addictionally asssessment of
contribution of this locus to the
trait’s variability
Conclusions:
• GWAS on AAT blood levels, not yet present in the literature, identified a
region that includes the gene SERPINA1 as the only associated locus in
the genome.
• Strong signals of common variants completely disappeared after
conditioning on two low frequent causal variants
•Rare SERPINA1 alleles much rather than common alleles contribute to
the further variability of this blood marker.
Manhattan-Plot of genome-wide p-values for association with blood AAT levels
Manhattan-Plot conditioning
of genome-wide
for association with
on SP-values
and Z alleles.
blood AAT levels
p<5*10-8
14q32.1
Inference:
• this locus is suggestive to contribute to the missing heritability of
pulmonary outcomes.
The absence of this locus among the statistically significantly associated
signals in corresponding GWAS may reflect
1) small effect sizes of the common variants in combination with
insufficient sample size and stringent correction for multiple testing
leading to a high risk of type II errors
2) poor tagging of the underlying causal variants by nearby SNPs in the
chip arrays would further reduce the chance to find this locus among
the significantly associated results.
LOSS OF FUNCTION
Normal lung
degradation
AATD lung
degradation
HNE
HNE
AAT
AAT
Luisetti, Breathe 2007
Alpha1-antitrypsin serum level,
SERPINA1 genotype , and risk for COPD
Dahl & Nordestgaard, Int J COPD 2008;4:157-167
GAIN OF FUNCTION
AATD as conformational disease
Gooptu B, Lomas D, Ann Rev Biochem, 2009
Polymers in the lung
McElvaney, Lung 2007
Tan et al,
Annual Congress, European Respiratory Society, Vienna, 1-5 Settembre, 2012
Effect of augmentation therapy on immune function of patients
with severe Alpha1-antitrypsin deficiency
Ferrarotti I1, Carroll TP2, Inghilleri S1, Ottaviani S1, Luisetti M1, McElvaney NG2
1Center for Diagnosis
of Inherited Alpha1-antitrypsin Deficiency, Dept of Molecular Medicine, Section of Pneumology, IRCCS San Matteo Hospital Foundation, University of Pavia, Italy;
Medicine, Royal College of Surgeons in Ireland, Education and Research Centre, Beaumont Hospital, Dublin, Ireland.
2Respiratory Research Division, Department of
‘We acknowledge the support of the European Respiratory Society (ERS) and the National Society (AIMAR), joint
ERS/AIMAR Fellowship STRTF 87-2010’
A
B
C
D
Augmentation therapy appeared to
attenuate IL-6 and IL-8 production
from ZZ and Q0/Q0 monocytes.
(***p<0.001, **p<0.01)
Relative IL-8 and IL-6 levels
measured by ELISA in monocyte
supernatants collected after 24h
incubation in the presence (+) or
absence of LPS.
A-B. Comparison among MM, ZZ
and ZZ treated with aumentation
therapy (ZZa.t.).
C-D. Comparison among MM, M/Q0
and
Q0/Q0
treated
with
aumentation therapy (Q0/Q0 a.t.)
Reduced chemotactic activity of AATD individuals (PI*ZZ) receiving augmentation
therapy in comparison to untreated PI*ZZ patients and controls
.
Chemotaxis MM vs ZZ (no Prolastin)
400
P< 0.005
p=0.005
n° cells
300
p=0.009
200
100
0
MM
MM fMLP
ZZ
Patient PI*ZZ:
male, 49y, 20pack/y
FEV1%29 - FVC%68
no aug. therapy
ZZ fMLP
Patient PI*ZZ:
male, 42y, 20pack/y
FEV1%31-FVC%60,
aug.therapy 2 years
MM controls: healthy, matched for age and gender
Reduced superoxide production from AATD individuals (PI*ZZ and PI*Q0/Q0) on
augmentation therapy in comparison to untreated PI*ZZ and controls.
N
25
24
150
53
9
272
1.168
54
688
291
8
1,2
15,8
6,7
0.2
Netherlands
1.4
New
PI*RR
30
0.7
Zealand
PI*NullNull
24
0.6
Poland
Spain
PI*ZNull
15
0.4
Sweden
§ R denotes non-Z and non-S deficiency variants
349
8,0
287
3
447
675
6,6
0,1
10,3
15,5
Switzerland
86
2,0
AATD subjects enrolled 5,511
(Update 20th August 2013)
Genotype
n
PI*ZZ
3,667
PI*SZ
494
PI*ZR §
Argentina
Australia
Austria
Belgium
Brazil
Canada
Denmark
Finland
%
Germany
Italy
85.4
Israel
11.4
62
%
0.6
0,5
3,4
1,2
0,2
6,2
DISTRIBUTION OF DIFFERENT
CLINICAL RESPIRATORY PHENOTYPES
ACCORDING TO GENOTYPES
100%
90%
80%
70%
60%
bronchiectasis
50%
asthma
40%
emphysema
30%
chronic bronchitis
20%
other lung diseases
10%
0%
* R denotes non-Z and non-S deficiency variants
DISTRIBUTION OF EARLY-ONSET
EMPHYSEMA ACCORDING TO
GENOTYPES
50
% of early-onset emphysema
50
45
40
35
30
20
25
14.98
20
15
10
6.67
8.06
3.04
5
0
SZ
RR
ZR
ZZ
Early onset emphysema
* R denotes non-Z and non-S deficiency variants
Z/Null
Null/Null
Severe AATD Italian Registry
413 patients (december 2013)
Genotype
SZ
17%
SS
1%
ZZ
62%
Null/Null
11%
RR
16%
RR
20%
ZR
61%
SR
12%
IRCCS San Matteo Hospital Foundation,
University of Pavia:
Pulmonology Unit
Head of Department: Maurizio Luisetti
Laboratory of Biochemistry and Genetics of Lung Disease
Stefania Ottaviani
Marina Gorrini
Michele Zorzetto
Ilaria Campo
Simona Inghilleri
Simona Ferrari
Unit of Pulmonology
Francesca Mariani
Zamir Kadija
Elena Paracchini