Identification of rheumatoid arthritis-associated
PADI4, SLC22A4/A5, FCRL3 & PTPN22
International Symposium on Inflammatory Diseases of Barrier Organs 3-4 June 2005, Kiel, Germany
Ryo Yamada Center for Genomic Medicine Kyoto University Kyoto, Japan
Genetics and Genetic Analysis of Rheumatoid Arthritis
• Twin and family studies
– Relative risk to monozygotic twin ( λMZ )
• 12~62 • 2~17
– Relative risk to siblings (λsib) – HLA locus explains 1/3-1/2 of total genetic components. – There are multiple non-HLA genes.
• Multiple linkage studies • Many candidate-approach studies
Major findings from SNP-based studies
• PADI4 : RA
– Post-translational enzyme to produce targets of the most RA-specific autoantibodies.
• SLC22A4 /A5 : RA & Crohn
– Ergothioneine or carnitine transporter expressed in hematologic lineages.
• PTPN22 : T1DM, SLE, RA & AITD
– Lymphoid-specific intracellular phophatase
• FCRL3 : RA, SLR & AITD
– Fc receptor homolog on B-cell membrane
ＰＡＤＩ４ Ca2+-dependent post-translational modification
Arginine Citrulline
NH2 C=O NH CH2 NH2 C=NH2 + NH CH2
CH2 CH2 HCNH3+ COO-
PADIs
CH2 CH2 HCNH3+ COO-
Loss of ionic NH2+ of Arg residue
Effects on intra- and intermolecular interactions
ＰＡＤＩ４ Ca2+-dependent post-translational modification
K Arita M Sato et al. Nat Str & Mol Biol 2004
PADI4 Citrullination Antigenicity
この部分だけ！
Human and Mouse PADI Gene cluster
Association Plots in the PADI Cluster
PADI4
-log10(P)＝５
Enzyme substrate
Hypothetical mechanism of RA-susceptible variant
Following studies
• Case-control association studies
– UK (Barton, A. J., Worthington et al. 2004 Arthritis & Rheum):
• Similar allele frequency • No statistical significance
– Japanese (Kuwahara, M., Kamatani, N et al. 2004 ACR Poster)
• Allelic effect on expression
– Higher concentration of PADI4 in PBMC from individuals with more copies of “RA-susceptible haplotype”(Harney, S.M.,
Brown, M.A. et al. 2004 ACR Poster)
• Allelic effect on autoantibody-production
– Combinatorial effect on anti-CCP Ab production between HLA and PADI4 polymorphisms”(Constantin, A., Cantagrel, A., et al. 2004
ACR Poster)
SLC22A4(/A5)
* Nucleotide binding motif * C
* *
+
+ +
+
1
2
3
4
5
6
7
8
9
10
11
+
N Sugar transporter signature sequence motif
The SNPs associated with RA or Crohn disease
10kb
Genes Exons
RIL
SLC22A4
SLC22A5
IRF1
RA & Crohn
SNPs
Crohn
Psoriatic Arthritis
Heat shock elementRUNX1 binding site L503F binding site Tokuhiro, S. et al. Peltekova, V. D. et al. Barton, A. et al.
Japanese
Caucasian
Ethnic difference in allele frequency of disease-related SNPs
10kb
Genes Exons SNPs
RIL
SLC22A4
SLC22A5
IRF1
intron1 intron2 Leu→Phe promoter (slc2F2 ) (slc2F1)
J vs. C J
SNP1 SNP2 0.29 0.29
J vs. A
P value FST
C vs. A
P value 0.14 0.14 FST 0.012 0.012
C
0.09 0.09
A
0.04 0.04
P value
FST
<0.00001§ 0.064* <0.00001§ 0.12* <0.00001§ 0.066* <0.00001§ 0.12*
SNP3
SNP4
0.001 0.39
0.001 0.52
0.04
0.58
<0.00001§ 0.24*
<0.00001§ 0.35*
<0.00001§ 0.018 <0.00001§ 0.19*
<0.00001§ 0.40* <0.00001§ 0.0038
§P<0.01, ＊FST> 0.05
Mori, et al. J Hum Genet 2005 May 10
Assumption of common responsible variant
Japanese
Caucasian
0W0 1M0
Segregation
0W0 1M0 0M1 1M1
FCRL3
Fc Receptor-like 3
Transmembranous protein （734aa) FcγR homolog ６ Ig-like domains outside Multiple tyrosin motifs (ITAM, ITIM, hemiITAM) Expressed by B cells in germinal centers
FCRL3 and others in 1q21-23
マウス
CIA（Mcia2) EAE,TMEVD (Eae3, Tmevd2) NOD (Idd10, Idd17)
1q21 mCh3
ヒト
FcγRI
Psoriasis FCRL1~5 Multiple sclerosis
CD1
SLE
Lupus models (sle1, swrl1)
1q23 mCh1
FcγRII/III
CD3Z
RA
NFkB regulates FCRL3 expression
Binds C > T
Lymphoid organs and B cell- specific expression
Expression of FCRL3 in lymphocyte aggregates in RA synovial tissue
T-cell
B-cell CD20 100x
CD3
100x
FCRL3 Antisense 100x
400x
RAにおけるFCRL3の働き(仮説）
PTPN22
• 1p13
• 110kDa intracellular tyrosine phosphatase • Functional SNP （1858 C->T, R620W) • Positive association – IDDM (Caucasian, Italian) Bottini N et al. – RA (Caucasian) Begovich AB et al. – SLE (Caucasian) Kyogoku, C. et al.
• No Tryptophan (W) allele in Japanese
PTPN22
10kb Genes Exons SNPs
PTPN22 Chromosome 1p13 Arg→Trp
J vs. C
J vs. A
P value FST
C vs. A
P value FST
J
SNP1 0.00
C
A
P value FST
0.08 0.02 <0.00001§ 0.040
0.00032§ 0.011
0.16
0.016
§P<0.01 ＊FST >0.05
Mori, et al. J Hum Genet 2005 May 10
Connection of multiple autoimmune diseases by susceptible genes/loci
Entry of Antigen
Environmental
Production of Antigen
PADI4
Body
Genetic
Response to Antigen Presentation of Antigen Recognition of Presented Antigen Immune System Transient Autoreaction Manifestation of phenotype Maintained Autoreaction
PTPN22
RUNX1 SLC22A4/A5
From the Atlas of Renal Pathology American Journal of Kidney Diseases, Volume 32, Issue 1 Medical Encyclopedia
MΦ
Anti-oxydant transporter
対象とする自己免疫疾患関連遺伝子
locus gene PADI4
1
SNP数・機能
4SNPs シトルリン化酵素 PDCD1 Crohn
SLC22A4 3SNPs
2 有機カチオントランスポーター 代表基質：Ergothioneine
SLC22A4/A5
SLE T1D RA
SLC22A5 1SNP
有機カチオントランスポーター 代表基質：carnitine
3 PTPN22
PADI4
PDCD1 PTPN22
1SNP 免疫グロブリンスーパーファミリー 分子 1SNP TCRシグナル抑制性制御酵素
4
Crohn：クローン病 SLE：全身性エリテマトーデス T1D：Ⅰ型糖尿病
疾患感受性SNP アレル頻度 人種による比較
J C
A
人種差 疾患 なし
既報での関連
あり なし
PADI4
RA RA Crohn
J・C J
C C
SLC22A4/5
あり
SLC22A4/5
あり
Crohn
C
－
(J:多型なし)
PDCD1
あり
RA, SLE C T1D
－
PTPN22
あり
RA, SLE T1D
C
－
Caucasian
PDCD1 MIF1 CTLA4 SLC22A4/A5
PTPN22
TNFRSF1
PADI4
Preliminary calculation with random-effect model Susceptible allele
Japanese
• Lab. Rheumatic Diseases
– – – – – – – – – – – Dr. K. Yamamoto Dr. A. Suzuki Dr. X. Chang Dr. Y. Kochi Dr. S. Tokuhiro Ms. R. Kawaida Ms. M. Mori Ms. K. Kobayashi Ms. M. Ohtake Ms. E. Kanno Ms. K. Komakine
• SRC, RIKEN
– – – – – Dr. A. Sekine Dr. T. Tsunoda Dr. Y. Nakamura Mr. H. Kawakami Mr. T. Kawaguchi
• Clinical Institutes
– Dr. T. Sawada
• And many other collaborators
Thank you all !