genetic studies on hlh in koreaplan.medone.co.kr/70_icksh2019/data/ss08-3_jongjin_seo.pdf · 2019....
Post on 20-Aug-2020
0 Views
Preview:
TRANSCRIPT
Genetic Studies on HLH in Korea
Jong Jin Seo, MD, PhD
Asan Medical CenterSeoul, Korea
2019. 3. 15ICKSH & KSH Annual Meeting
I have no personal or financial interests to declare:
I have no financial support from an industry source at the current presentation.
대한혈액학회 Korean Society of Hematology
COI disclosureName of author ： Jong Jin Seo
Brief introduction on genetic aspects of familialHLH (FHL)
Previous Korean studies on the genetics of HLH Search for unidentified gene mutations in Korean
patients with HLH (preliminary result) Genotype-phenotype study of Korean FHL
patients (preliminary result)
Contents of presentation
Primary HLH is caused by a gene mutation, either at the FHL loci or in a gene responsible for one of several immunodeficiency syndromes
The incidence of 5 FHL types varies in different ethnic groups, some with founder effect
Patients with HLH gene mutation tend to present at a younger age than those without mutation
Previous studies showed that deleterious non-sense mutations are associated with earlier onset and a more severe clinical course suggesting possible genotype/ phenotype correlation
Genetic overview of FHL (1)
FHL is usually inherited in an autosomal recessive manner (ie, inheritance of a mutation at both alleles of a gene is required to manifest the disease)
Occasionally, patients may have compound heterozygotes (ie, a different mutation in each allele of the same gene), or they may show digenic inheritance(ie, separate mutations in 2 different genes)
The presence of homozygous, compound heterozygous, or digenic mutations is sufficient for diagnosis of FHL
Genetic overview of FHL (2)
Diagnostic criteria (HLH-2004)
Expanding spectrum of genes associated with primary HLH
J Pediatr 2013 163(5):1253-9
Locus Gene % Worldwide* % in Japanese‡ % in Korean
FHL1 N/A 4 consanguineous Pakistani families Not found
FHL2 PRF1 20%-30% worldwide(>50% in AA) 59 ?
FHL3 UNC13D ~20%-30% worldwide 34
FHL4 STX11 ~20% of Turkish/Kurdish familieslow in other ethnic groups Not found
FHL5 STXBP216% in Central Europeans, Turks, and Saudis~20% in North American
7
The incidence of FHL subtypes varies among different ethnic groups
* GeneReviews ‡ Plos One 2010;5(11):e14173
Genetic cornerstones of HLH
2005FHL4/STX11
2009FHL5/
STXBP2
1999FHL2/PRF1
1999of FHL1
2003FHL3/
UNC13D
2004 HLHdiagnostic
criteria
2010 Hematologica40 HLH pt.
8 FHL3 & 1 FHL2
2013 Ann Hematol72 HLH pt.
17 FHL3 & 3 FHL2Identification of
deep intronic mutation of UNC13D with
founder effect
2016 Clin GenetThe 1st FHL5
2016 Ann Lab MedThe 1st FHL4
Incorporation ofGenetic tests
1994 HLHdiagnostic
criteria
(International) (Korean)
2018 (Unpublished) Search for unidentified
genetic mutation Geno-phenotype
study
Genetic studies for HLH in Korea
1st
(2010, Haematologica)
• 40 pt with HLH
• FHL2 to FHL4 (PRF1,
UNC13D, STX11)
2nd
(2013, Ann Hematol)
• 32 new pt with HLH
• Pt without FHL
mutation / and
single mutation in
UNC13D (FHL3)
from the 1st set
• UNC13D deep
intronic mutation
3rd
(2016, Clin Genet)
• Pt without FHL
mutation from the
1st & 2nd set
• STXBP2 (FHL5)
(from the Korean Registry for Histiocytosis)
Genetic studies for HLH in Korea
1st
(2010, Haematologica)
• 40 pt with HLH
• FHL2 to FHL4 (PRF1,
UNC13D, STX11)
2nd
(2013, Ann Hematol)
• 32 new pt with HLH
• Pt without FHL
mutation / and
single mutation in
UNC13D (FHL3)
from the 1st set
• UNC13D deep
intronic mutation
3rd
(2016, Clin Genet)
• Pt without FHL
mutation from the
1st & 2nd set
• STXBP2 (FHL5)
(from the Korean Registry for Histiocytosis)
1st Set(n=40)
PRF1(n=1)
UNC13D(n=8)
No mutation(n=31)
Biallelic mutations (n=4)
Monoallelic mutation only(n=4)
STX11(n=0)
Haematologica 2010
► UNC13D is the predominant causative gene in the Korean population
Genetic subtypes of FHL in Korea (1st set)
Meeths M et al. Blood 2011;118:5783-5793
Genetic studies for HLH in Korea
1st
(2010, Haematologica)
• 40 pt with HLH
• FHL2 to FHL4 (PRF1,
UNC13D, STX11)
2nd
(2013, Ann Hematol)
• 32 new pt with HLH
• Pt without FHL
mutation / and
single mutation in
UNC13D (FHL3)
from the 1st set
• UNC13D deep
intronic mutation
3rd
(2016, Clin Genet)
• Pt without FHL
mutation from the
1st & 2nd set
• STXBP2 (FHL5)
(from the Korean Registry for Histiocytosis)
2nd Set(n=32)
1st Set(n=40)
PRF1(n=1)
UNC13D(n=8)
No mutation(n=31)
2 mutations (n=4)
1 mutation only(n=4)
Investigation for
c.118-308C>T
UNC13D STX11PRF1STX11(n=0)
1st set(n=40)
UNC13D mutation
(n=8)
No mutation
(n=31)
PRF1 mutation (n=1)
2 mutations
(n=4)
1 mutation only
(n=4)
c.118-308C>T in 3/4 (75%)
c.118-308C>T in 2 in homozygous status
23%(9/40)
28%(11/40)
Mutation detection rate
Predominance of FHL3 in Korea due to 2 recurrent mutations of UNC13D
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32
c.247C>TR83*
(3%; 1/34)
c.754-1G>C(26%; 9/34)
c.1993-2A>G(3%; 1/34)
c.1693delGE565Sfs*7(6%; 2/34)
c.2954+5G>A(6%; 2/34)
c.2367+1G>A(3%; 1/34)
c.118-308C>T(41%; 14/34)
*c.1055+1G>A(9%; 3/34)
c.292C>TQ98*
(3%; 1/34)
Haplotype analyses revealed significantly shared haplotypes in both mutations, indicating the
presence of “founder effect”
This explains the unusual predominance of FHL3 in Korea
Genetic studies for HLH in Korea
1st
(2010, Haematologica)
• 40 pt with HLH
• FHL2 to FHL4 (PRF1,
UNC13D, STX11)
2nd
(2013, Ann Hematol)
• 32 new pt with HLH
• Pt without FHL
mutation / and
single mutation in
UNC13D (FHL3)
from the 1st set
• UNC13D deep
intronic mutation
3rd
(2016, Clin Genet)
• Pt without FHL
mutation from the
1st & 2nd set
• STXBP2 (FHL5)
(from the Korean Registry for Histiocytosis)
Mutation (+)(n=20)
72 patients (1st & 2nd set)
UNC13D(n=17)
PRF1(n=3)
STX11(n=0)
Mutation (-)(n=52)
STXBP2 mutation analysis(n=50)
No DNA samples for 2 patients
1 patient with 2 causative mutations in STXBP2 Identified mutations
- c.184A>G (p.Asn62Asp), heterozygous- c.577A>C, heterozygous
Frequency of FHL5 in Korean patients with HLH- 5% (1/21) of patients with FHL
FHL5 is rare in Korea
Summary of this Study
Search for unidentified genetic mutation in Korean pediatric patients
with HLH using gene panel test
- Preliminary results -
Analysis using gene panel test in Korean patients with no definitive diagnosis of FHL
Genes included in 14-Gene Panel Test
Variants identified by HLH-associated gene panel test in 72 patients
P122
P154
P157
P158
P161
P162
P163
P167
P168
P169
P18
P19
P21
P22
P24
P25
P26
P27
P28
P30
P31
P32
P33
P34
P38
P39
P40
P46
P47
P48
P50
P52
P55
P58
P59
P62
P64
P67
P68
P74
P78
P83
P87
P91
P92
P93
P94
P96
P97
P99
P100
P102
P103
P106
P107
P110
P112
P118
P120
P121
P3
P7
P9
P10
P13
P13
P14
P15
P16
P17
P170
P8
AP3B1
BLOC1S6
CD27
ITK
LYST
MAGT1
PRF1
RAB27A
SH2D1A
SLC7A7
STX11
STXBP2
UNC13D
XIAP
HLH-associated mutation Variant of unknown significance
Distribution of the genetic defects in Korean patients with HLH (n=101)
Targeted gene panel test further identified 10 Korean
patients with primary HLH who were previously not
diagnosed with conventional FHL2-5 genetic tests
- 2 male patients with XLP type 1 and type 2
- 8 patients with 2 HLH-associated variants
(6 of them with digenic defects)
17% of the registered patients were found to have
monoallelic mutation
Summary of this study
Functional analysis of identified variants Clinical correlation according to the types of
mutation Identification of unknown genetic
background in patients not enrolled to Korean HLH Registry using gene panel test
More extended gene panel test (>14 genes) for genetically undiagnosed patients
Further workups
Identification of genetic mutation with gene panel test : The registry study of
Korean pediatric patients with hemophagocytic lymphohistiocytosis
1Hyery Kim, 2Ja Young Seo, 1Kyung Nam Koh, 1Ho Joon Im, 3Ji Won Lee, 3Kun Hee Yoo, 3Ki Woong Sung, 3Hong Hoe Koo, 4Jae Min Lee, 5Hyoung Jin
Kang, 5Hee Young Shin,1Jong Jin Seo
1Department of Pediatrics, University of Ulsan College of Medicine, Asan Medical Center, Republic of Korea2Department of Laboratory Medicine, Gachon University Gil medical center, Republic of Korea3Department of Pediatrics, Sungkyunkwan University School of Medicine, Republic of Korea4Department of Pediatrics, Yeungnam University College of Medicine, Republic of Korea5Department of Pediatrics, Seoul National University College of Medicine, Republic of Korea
(Abstract OP09-6, 3/15 )
- Preliminary results -
Nationwide retrospective multicenter study
by reviewing medical records
July, 2005~March, 2017
Inclusion criteria
- Patients diagnosed with HLH according to
the HLH-2004 diagnostic criteria, and
- Patients with identified gene mutation
Total 48 patients from 10 institutions
Median age at Dx : 3.2 months (7 days-13 yrs) 77.0% <1 yr (n=37), 89.6% <3 yr (n=43)
0
2
4
6
8
10
12
1 mon 2 mon 3 mon 4 mon 6 mon 7 mon 8 mon 9 mon 1 yr 2 yr 6 yr 7 yr 9 yr 13 yr
3
11
10
5
3
2 2
1
3 3
1
2
1 1
OS 68.0%
PRF1 (N=7)
UNC13D (N=37)
STX11 (N=1)
STXBP2 (N=2)
SH2D1A (N=2) P=0.870
71.4%
50.0%
Overall Survival (months)
63.1%
48.8%
PRF1 (N=7)UNC13D (N=36)STX11 (N=1)STXBP2 (N=2)SH2D1A (N=2)
P=0.228
58.3% 50.5%
Sex Age at Dx(mon) Mutation Mutation
DescriptionFamily
Hx HSCT Reactivation Survival OS(mon)
F 71.9 c.[1693delG(p.Asp565Serfs*7)];[2954+5G>A] Frameshift/Splicing No Yes No Dead in CR 5.3M 5.1 c[1693delG(p.Asp565Serfs*7)];[2954+5G>A] Frameshift/Splicing Yes Yes Before HSCT Alive 135.6F 85.5 c.[1693delG(p.Asp565Serfs*7)];[2954+5G>A] Frameshift/Splicing Yes Yes No Alive 94.9F 108.8 c.[1693delG];[2954+5G>A] Frameshift/Splicing Yes Yes No Alive 51.6F 1.2 c.[118-308C>T];[2447+1G>A] Regulatory/? No No Before HSCT DOD 3.9F 1.9 c.[118-308C>T];[766C>T] Regulatory/Nonsense No Yes No Alive 52M 1.9 c.[118-308C>T];[2695C>T(p.Arg899*)] Regulatory/Nonsense No Yes After HSCT Dead in CR 8.6M 0.3 c.[292C>T(p.Gln98*)];[118-308C>T] Regulatory/Nonsense No Yes Before HSCT Alive 116.8F 26.1 c.[118-308C>T];[2695C>T(p.Arg899*)] Regulatory/Nonsense Yes Yes Before HSCT Alive 33.1F 6.9 c.[118-308C>T];[247C>T(p.Arg83*)] Regulatory/Nonsense No Yes Before HSCT Alive 77.7F 1.7 c.[118-308C>T];[118-308C>T] Regulatory/Regulatory No Yes No Alive 73.8M 2.1 c.[118-308C>T];[118-308C>T] Regulatory/Regulatory No Yes No Alive 47.5F 1.3 c.[118-308C>T];[754-1G>C] Regulatory/Splicing No Yes Before HSCT Alive 102.8F 1.6 c.[118-308C>T];[754-1G>C] Regulatory/Splicing No Yes No Alive 50.4M 3.2 c.[118-308C>T];[754-1G>C] Regulatory/Splicing No Yes No Alive 47.9F 3.8 c.[118-308C>T];[754-1G>C] Regulatory/Splicing No Yes No Alive 99.8F 2.1 c.[118-308C>T];[754-1G>C] Regulatory/Splicing No Yes After HSCT DOD 39.7F 11.6 c.[118-308C>T];[754-1G>C] Regulatory/Splicing Yes Yes No Dead in CR 9.8M 7.4 c.[118-308C>T];[1055+1G>A] Regulatory/Splicing No Yes No Dead in CR 8.2M 12.7 c.[118-3C>A];[602A>G(p.His201Arg)] Splicing/Missense No Yes No Alive 41.5M 2.5 c.[754-1G>C];[1055+1G>A] Splicing/Splicing No Yes Before HSCT Alive 113.4F 31.3 c.[754-1G>C];[754-1G>C] Splicing/Splicing No No Before HSCT DOD 9.5M 2.9 c.[754-1G>C];[2367+1G>A] Splicing/Splicing No No Before HSCT DOD 11.5F 3.9 c.[754-1G>C];[1993-2A>G] Splicing/Splicing No No No DOD 1.6M 5.2 c.[754-1G>C];[754-1G>C] Splicing/Splicing No Yes No DOD 3.7
Overall Survival (months) Reactivation free survival (months)
P=0.101
Biallelic splicing mutations(N=5)
58.7% Others (N=21)
Biallelic splicing mutations (N=5)
71.1% Others (N=21)
P=0.03920.0%
51.2% Total (N=26)
61.3% Total (N=26)
The prevalence of UNC13D was 75.5% of Korean FHL patients, and it was followed by PRF1 mutation (14.3%)
There was no statistical difference in the phenotype at diagnosis between genotype groups in Korean patients with FHL
Approximately 35% of patients showed reactivation, and there was no difference in reactivation rates by genotype
There was no difference in the post-transplant OS rates by genotype
Patients with biallelic splicing mutations showed significantly lower OS and lower reactivation free survival rates compared to other patients
This Korean study needs further analysis in larger cohort
Locus Gene % Worldwide* % in Japanese‡ % in Korean
FHL1 N/A4 consanguineous Pakistani families Not found Not found
FHL2 PRF1 20%-30% worldwide(>50% in AA) 59 ~15
FHL3 UNC13D ~20%-30% worldwide 34 75~80
FHL4 STX11 ~20% of Turkish/Kurdish familieslow in other ethnic groups Not found ~2 (1 case)
FHL5 STXBP216% in Central Europeans, Turks, and Saudis~20% in North American
7 ~5 (2 cases)
Frequencies of FHL Gene Mutations
* GeneReviews ‡ Plos One 2010;5(11):e14173
Genetic heterogeneity and expanding spectrum of HLH necessitates advanced molecular diagnostic technique
Multi-gene panel test may become more readily available in expanded spectrum of genes
Genotype-phenotype correlation of FHL patients needs to be clarified with further studies
Comprehensive molecular testing can provide detailed genetic information, and thereby may help to reveal unanswered molecular pathophysiology which may improve the treatment outcome of HLH
Prospects
Acknowledgement
AMC PHO Ho Joon ImKyung Nam KohHyery KimAMC LabChan-Jeoung ParkSeongsoo JangYoung-Uk Cho
SNUH PHOHyoung Jin Kang
Kyung Hee Univ. MC PHOHoi Soo Yoon
Young Nam Univ. MC PHOJae Min Lee
SMC PHO/LabKeon Hee Yoo Hee-Jin Kim
Gil MC LabJa-Young Seo
CMC PHONack Gyun Chung
Severance Hospital PHOChuhl Joo Lyu
Chonnam NUH PHOHoon KookHee Jo Baek
Thanks to all the members of Korea Histiocytosis WP
Thank you for your attention!
top related