Download Project-2

Pathogenesis of Inflammatory Bowel
Diseases: Genetic and Immune Aspects
Jae Hee Cheon MD, PhD1,2
1Department
of Internal Medicine and Institute of
Gastroenterology, Yonsei University College of Medicine, Seoul;
2Avison
Medical Research Center, Yonsei University, Seoul, Korea
Inflammatory Bowel Diseases (IBD)
Chronic inflammation of the bowel of unknown etiology
Ulcerative colitis (UC)
Crohn's disease (CD)
IBD Pathogenesis
Gaya et al. Lancet 2006;367:1271-84
3
Xavier RJ, et al Nature 2007
IBD Pathogenesis
• Genetic susceptibility (host)
• Immunological abnormalities (host)
• Environmental factors (host, environment)
–
–
–
–
Smoking
Breast feeding
Diet, drugs
Hygiene
• Microbes (environment, invironment)
Differential Responsiveness of Wild-type and
Mutant CARD15 to LPS
5
Ahmad et al. Gastroenterology 2004;126:1533-49
Ogura Y et al. Nature 2001;411:603-606
Types of Disease Genes
Gene frequency
Penetrance
AR/RR
Population AR
Study setting
Study type
Monogenic
(Mendelian)
Rare
Polygenic
(Complex)
Common (>1%)
High
Low
High
Low
Low
High
Family
Linkage
Population
Association
Genetic Studies in IBD
• Gene mutations from familial aggregation (linkage
analysis)
• Immunological approaches from animal models
• Hypothetical candidate gene approaches
• Non-biased array approaches (GWAS, Immunochip)
• Non-biased deep sequencing studies (NGS)
• Personal whole gene sequencing
Meta-analysis of GWAS in Caucasian
•
71 CD-, 47 UC-, and 28 both CD and UC-susceptibility loci/genes
Franke A. Nat Genet 2010; Anderson CA. Nat Genet 2011; Lees CW. Gut 2011
IBD Immunochip_163 loci associated to IBD
Cumulative IBD loci
150
100
GWAS metaanalyses
Immunochip
163 loci
NOD2
Single-center
GWAS
50
MHC in UC
0
2000 2002 2004 2006 2008 2010 2012
Jostins L et al. Nature 2012.
Year
Modified from Cheon JH. JGH 2013, Mathew CG. Nat Rev Genet 2008
Difficulties in studying genetics of IBD
• Polygenic
• Low penetrance rate
• Affected by environmental factors,
epigenetic changes, and ncRNAs…
• Microbiologic factors
• Ethnic differences
Cheon JH. J Gastroenterol Hepatol 2013
Genetic associations in Crohn’s disease
Gene association
Caucasian
Japanese
Korean
NOD-2
strong
no
no
ATG16L1
strong
no
no
IRGM
strong
no
yes
IL23R
strong
no
Yes
TNFSF15
weak
strong
strong
Yamazaki et al. J Hum Genet 2007
Yamazaki et al. J Hum Genet 2004
Inoue et al. Gastroenterol 2002
Yang et al. Inflamm Bowel Dis 2009
Yang et al. Am J Gastroenterol 2008
Croucher et al. Eur J Hum Genet 2003
Kim SW, Cheon JH, Kim WH et al. Gut 12
2011
Moon CM, Cheon JH, Kim WH et al IBD 2012
Purpose
IL23R Genetic polymorphisms in East Asians:
• Different IL23R genetic contributions between Korean and
Japan patients
• Recent Chinese and Korean data: the G149R SNP
• Different SNPs from the Western data
NOD2
IRGM
IL23R
China
No
Korea
No
Japan
No
?
Yes
Yes
Yes
No
No
Kim SW, Cheon JH et al. Gut 2011
Cheon JH. J Gastroenterol Hepatol132013
Yang SK et al. Gut 2013
GWAS of CD in Koreans: Asan Medical Center
• 4 previously reported loci: TNFSF15, IL23R, MHC region, and the
RNASET2-FGFR1OP-CCR6 region.
• 3 new susceptibility loci at genome-wide significance: ATG16L2 and/or
FCHSD2 as novel susceptibility genes for CD
Yang SK et al. Gut 2013
Results
Project-1: Clinical features, prognosis and genetic
markers for Korean CD patients
These findings offer new insights into the genetic architecture of CD and support the
complementary value of genetic studies in different populations.
Yang, et al. Inflamm Bowel Dis (in press)
Project-2
Distributions of genotype and allele frequencies of 4 miRNA
SNPs at Validation Stage
CD
No.(%)
Position
Genotype
UC
Additive
Codominant
Dominant
Recessive
P value
P value
P value
P value
OR
(95% CI)
0.31
OR
(95% CI)
OR
(95% CI)
0.47
1.32
(0.85-2.06)
1.17
(0.86-1.60)
1.33
(0.83-2.13)
1.29
(0.68-2.47)
0.18
0.25
0.33
0.80
(0.52-1.25)
0.77
(0.52-1.13)
0.87
(0.55-1.36)
0.37
(0.10-1.37)
0.6
0.84
0.73
0.93
(0.61-1.42)
0.92
(0.66-1.27)
0.96
(0.61-1.51)
0.80
(0.38-1.68)
0.92
0.95
0.8
0.94
(0.58-1.51)
0.98
(0.72-1.34)
0.91
0.92
(0.56-1.53)
0.98
(0.53-1.82)
0.91
1.05
(0.66-1.68)
1.02
(0.68-1.55)
1.08
(0.67-1.74)
0.82
(0.19-3.49)
(dbSNP ID)
mir146A
C/C
(rs2910164)
C/G
G/G
mir423
A/A
(rs6505162)
C/A
C/C
mir27A
C/C
(rs895819)
T/C
T/T
mir196A2
C/C
(rs11614913)
T/C
T/T
mir499
A/A
(rs3746444)
A/G
G/G
78
(40%)
88
(45%)
29
(15%)
10
(5%)
69
(35%)
117
(60%)
22
(11%)
85
(44%)
88
(45%)
41
(21%)
105
(53%)
53
(27%)
140
(71%)
53
(27%)
5
(3%)
OR
(95% CI)
0.22
OR
(95% CI)
0.75
113
(47%)
Additive
Codominant
Dominant
Recessive
P value
P value
P value
P value
OR
(95% CI)
0.013
OR
(95% CI)
OR
(95% CI)
0.03
OR
(95% CI)
0.0091
OR
(95% CI)
0.21
1.75
(1.14-2.67)
1.46
(0.81-2.63)
1.46
(1.08-1.97)
0.51
1.69
(1.08-2.65)
1.94
(1.02-3.67)
0.51
0.74
0.24
0.88
(0.61-1.28)
0.65
0.99
(0.64-1.53)
0.48
(0.13-1.78)
0.89
100 (41%)
1.10
(0.61-1.98)
0.13
0.38
(0.10-1.42)
0.57
0.99
(0.65-1.54)
0.82
No. (%)
Control
0.91
0.93
(0.61-1.43)
0.76
0.80
(0.19-3.40)
28
(12%)
10
(4%)
79
(33%)
153
(63%)
24
(10%)
108
(45%)
106
(45%)
52
(22%)
126
(52%)
62
(26%)
169
(7%)
69
(29%)
4
(2%)
0.48
(0.13-1.77)
0.93
(0.61-1.43)
0.64
0.82
0.91
(0.59-1.40)
0.88
(0.43-1.82)
0.90
(0.55-1.47)
0.90
(0.50-1.63)
0.91
(0.60-1.37)
0.92
(0.46-1.85)
0.95
(0.70-1.27)
0.91
1.02
(0.68-1.54)
0.93
(0.68-1.27)
0.72
0.66
0.66
0.89
0.90
(0.57-1.43)
0.97
(0.59-1.60)
0.96
0.96
0.79
1.00
(0.63-1.57)
1.23
(0.27-5.61)
1.01
(0.65-1.58)
1.23
(0.27-5.58)
No. (%)
50
(34%)
75
(50%)
24
(16%)
3
(2%)
49 (33%)
96
(65%)
14
(9%)
65
(44%)
70
(47%)
31
(21%)
75
(51%)
41
(28%)
103
(70%)
42 (28%)
3
(2%)
Project-2
Phenotype Analysis in UC: Infliximab-free survival rate
P < 0.01
Time to infliximab use
Project-2
UC phenotypes and miR-146a
miR-146A signaling pathway
Kim SW, Cheon JH, unpublished data
DNA methyltransferase inhibition accelerates the immunomodulation and migration
of human mesenchymal stem cells
Project-5
특허출원 : 10-2014-0001974 (출원 번호)
Lee et al. Scientific reports. (Aug., 2014 submitted)
Modern lifestyle linked with alterations of gut
microbial colonization
Manichanh C et al. Nat Rev Gastroneterol Hepatol 2012
Dysbiosis in iBD
Manichanh C et al. Nature Rev Gastroenterol Hepatol 2012
Area 3: Investigate the microbiome of Korean CD and
factors associated with change of microbiome
Phylum level
■ Firmicutes ■ Proteobacteria ■ Bacteroidetes
■ Actinobacteria ■ Fusobacteria
Class level
■ Bacilli ■ Clostridia ■ Negativicutes■ Gammaproteobacteria
■ Betaproteobacteria ■ Bacteroidia ■ Actinobacteria ■ Fusobacteria
 Global structure and individual bacterial abundance of stool and mucosal tissues
were different between healthy controls and Crohn’s disease patients
Area 3: Investigate the microbiome of Korean CD and
factors associated with change of microbiome
OTUs
Shannon
index
■ Firmicutes
■ Proteobacteria
■ Bacteroidetes
■ Actinobacteria
■ Fusobacteria
 The proportion of phylum Fusobacteria in stool and mucosal tissues was significantly
higher in active CD patients compared to HC and CD patients in remission.
Immunologic Dysregulation in IBD
• Dysfunction of epithelial barrier
• Innate immunity
– NOD2/CARD15
• Cellular response
– Autophagy
– ER stress
• Adaptive immunity
– T cells > B cells
Human Umbilical Cord Blood Mesenchymal Stem Cells Reduced Colitis in Mice via
Activation of NOD2 Signaling to COX2
Kim et al. Gastroenterology. 2013 Dec;145(6):1392-1403.
A p38 MAPK-Mediated Alteration of COX-2/PGE2 Regulates Immunomodulatory Properties in
Human Mesenchymal Stem Cell Aging.
Yu et al. PLoS One. 2014 Aug 4;9(8):e102426
ER Stress and IBD
Kaser A et al. Cell 2008
Hisamatsu T, et al Pharmacol Ther 2013
Model of the interplay between UPR-induced
autophagy, NF-kB signalling and inflammation
Adolph T et al. Nature 20123
Intestinal inflammation
1.
XBP-1 deficiency exaggerates NF-B
signaling in response to microbial TLR 2
ligands leading to inflammation in the gut
Gut
2.
Immune regulation in the gut with functional
impairment of CD4+ Treg to CCR7 deficiency
Commensal microbe
Wild-type
XBP-1
TLR2 ligand
TLR2
NF-B
Epithelial
XBP-1-/TLR2
NF-B
Wt
CCR7KO
5.05
CXCL1/KC
IL-10
CXCL1/KC
Gut homeostasis
Inflammation
Regulatory
plasmacytoid DC
IL-10 producing
CD8+ Treg
CD8
11.21
Research for protective agent against barrier dysfunction
Hirsutenone (M)
Project 2
C t-BH 1
5
10
20
ZO-1
-actin
In vitro study
In vivo study
Metabolism of Azathioprine/6-Mercaptopurine
6-methylthioITP
TPMT
6-thiouric acid
6-thioITP : allergic reaction
XO
ITPA
HGPRT
AZA
6-MP
IMPDH
6-thioIMP
6-Thioguanine
: Therapeutic effects
TPMT
6-Methyl MP
: hepatotoxicity
TPMT
6-methylthioIMP
myelotoxicity
Myelotoxicity of AZA in Korean Patients with IBD
Kim JH, Cheon JH, Kim WH. Korean J Gastroenterol. 2008
Ethnic Differences in Frequency of TPMT
Gene Mutation
No. of
subjects
2A
3A
3B
3C
Korean
812
0
0
0
2.1
Japanease
192
0
0
0
0.8
Chinese
192
0
0
0
2.3
American
Caucasian
282
0.2
3.2
0
0.2
British
199
0.5
4.5
0
0.25
French
304
0.7
3
0
0.4
German
1214
0.2
4.4
0
0.4
Italian
103
0.49
3.88
0
0.97
Swedish
800
0.06
3.7
0.13
0.44
Population
Cheon JH, Kim WH et al. Hepatogastroenterology 2009;56:421-3
Results
Project-1: Clinical features, prognosis and genetic
markers for Korean CD patients
Yang, et al. Nat Genet 2014
Results
Project-1: Clinical features, prognosis and genetic
markers for Korean CD patients
Yang, et al. Nat Genet 2014
IBD Pathogenesis
• A complex interplay between host genetic
susceptibility and environment/invironment
• Epithelial barrier abnormalities
• Innate or adaptive immune dysfunction
• Microbes (pathogens or commensals), diet,
parasites
• Heterogeneous immune disease