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Hereditary Periodic Fever Syndromes A group of inherited disorders characterized by recurrent, self-limited episodes of fever and synovial, serosal, and/or oculocutaneous inflammation. 169 Claudius Galen 1790 Heberden 1895 Osler 1908 Janeway & Mosenthal 1940 Kile & Rusk 1945 Siegal 1949 Reimann 1958 Heller 1962 Muckle & Wells 1981 Prieur, Griscelli 1982 Williamson et al 1984 van der Meer et al
Missense Mutations in the TNFRSF1A Extracellular Domains
Mutation Exon Amino Acid Ethnic Background
175 TC 2 Cys30Arg Irish
185 GA 2 Cys33Tyr Irish/Scottish
236 CT 3 Thr50Met Irish
French-Canadian
242 GT 3 Cys52Phe Irish/English/German
349 TC 4 Cys88Arg Scottish (Australian)
350 GA 4 Cys88Tyr Finnish
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2) constitutive activation, formation of intermolecular disulfide bonds between unpaired cysteines in mutant receptors (Santoro, 1995) -Il-6 induction - crude test - normal
3) resistance of mutant TNFRSF1A to the normal homeostatic effects of activation-induced cleavage
.
- FACS analysis - reduced/delayed TNFRSF1A shedding in affected members of family with C52F mutation
1) increased affinity of mutant TNFRSF1A for ligand (i.e TNF) -binding and affinity studies - normal
.
Possible Disease Mechanisms in TRAPS
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Periodic fever family with amyloidosis of kidney
affected
unaffected
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TNF Receptors on Leukocytes from Patients with the C52F MutationTNF Receptors on Leukocytes from Patients with the C52F Mutation
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Leukocytes Bearing the C52F Mutation Have Defective Clearance of TNFRSF1A (CD120a)Leukocytes Bearing the C52F Mutation Have Defective Clearance of TNFRSF1A (CD120a)
SCREENING METHODS for TRAPSSCREENING METHODS for TRAPS Genotyping: microsatellites within (TNFRp55)
and flanking (D12S99 and D12S77)TNFRSF1A locus used to genotype family members.
DHPLC: we screened 20 ADRF families and 184 sporadic cases forTNFRSF1A mutations using the Transgenomic WAVE® System.
Sequencing: ABI 373A sequencer.
ELISA assays: used to determine serum levels of TNFRSF1A and TNFRSF1B (sTNFRSF1A and sTNFRSF1B).
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DHPLC elution profileDHPLC elution profile
0.0
1.5
3.0
. 1. 2. 3. 4. 5. 6. 7. 8. 9.
Time (min)
0.0
2.5
. 1. 2. 3. 4. 5. 6. 7. 8. 9.
Time (min)
Wt C70R
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a b05 0903 0708 13
MARKERSD12S99TNFRp55D12S77
a d05 0403 0708 12
a d05 0403 0708 12
c d04 0407 0708 12
a c05 0403 0708 08
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3
831
875 326
Israeli Arab family A
612
1114
Periodic fever
C70R M 1 2 3 4 5 C
500
800
Not genotyped
4 5
De-novo C70R mutation (Israeli Arab family)
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II
IV
III
I
c d04 04A A06 0606 11
e f04 08G G02 0211 12
a e04 04G G02 0205 11
a e/f04 04G G02 0205 12*
d g04 06-- --06 0111 10
d b04 04A A06 0711 11
c j04 01A A06 0706 11
c j04 01A A06 0706 11
826 731
g h 04 04 G A 06 01 06 03
a c04 04G A02 0605 06
Order of markers D12S99 +36 A/G TNFRp55 D12S77
Irish Family
727
781 11401450
1450
650
650
Soluble TNFRSF1A levels n= 746-1966pg/ml
Periodic fever in an Irish Family - unlinked to all known HPF loci
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7 8 9 10 11 12 13 14 S2
sTN
FR
SF
1A
(p
g/m
l)
s TNFRSF1A levels in affected family members of
(A) TRAPS (B) families without mutation s TNFRSF1A levels in affected family members of
(A) TRAPS (B) families without mutation
(A)
(B)
746
1966
746
1966
unaffected
Periodic fever
Probably affected
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A total of 8 novel TNFRSF1A mutations (T37I, D42, G46E, T50K, F60L,C70R, C70S and R92P), and 5 known (c.193-14 G>A, P46L, T50M, C88R,
and R92Q) mutations in a study of 20 families. The C70S mutation found in a Japanese patient
Affected members in 4 of the 7 families without TNFRSF1A mutations have low sTNFRSF1A
These findings indicate the presence of genetic heterogeneity in ADRFs.
Summary of TRAPS and ADRF families
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Etanercept (a recombinant p75 TNFR:Fc fusion protein), in TRAPS: the Promise of Molecular MedicineEtanercept (a recombinant p75 TNFR:Fc fusion protein), in TRAPS: the Promise of Molecular Medicine
Patient 3 (14yo, M, H22Y)
0
10
20
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 3 (14yo, M, H22Y)
0
10
20
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 4 (32yo, M, T50M)
0
20
40
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 4 (32yo, M, T50M)
0
20
40
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 2 (15yo, F, T50M)
0
30
60
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 2 (15yo, F, T50M)
0
30
60
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 1 (50yo, M, T50M)
0
25
50
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
Patient 1 (50yo, M, T50M)
0
25
50
Q1 Q2 Q3 Q4
Study Phase
Att
ack
Sco
re
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Apoptosis -
A type of programmed cell death in which the cell dies (commits suicide) through a series of cellular events including enzymatic fragmentation of DNA.
# necrosis - cells bursts and discharges contents. e.g. infarction
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Pyrin/marenostin is mainly expressed in leucocytesPyrin/marenostin is mainly expressed in leucocytes
Cell 90; 797, 1997
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Pyrin Interaction with ASC proteinPyrin Interaction with ASC protein
Apoptosis-Associated speck-like protein with a caspase recruitment domain(CARD)
Interacts with pyrintwo-hybrid, IP, andfluorescence microscopy
Aggregates during Apoptosis in HL-60 cells
Proapoptotic
Interacts with pyrin by yeasttwo-hybrid, IP, andfluorescence microscopy
Proapoptotic
N-terminal pyrin domain, C- terminal CARD
GFP-pyrin + ASC-myc(Cy3 label) merged
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Monocytes from Pyrin Truncation MiceManifest a Defect in ApoptosisMonocytes from Pyrin Truncation MiceManifest a Defect in Apoptosis
24 hr 48 hr 72 hr
+/+ -/- +/+ -/- +/+ -/-
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Connecting Pathways with Patients: Mutations in a Pyrin-Binding Protein Cause PAPA Syndrome
*
E250Q
Pyrin
B CC B30.2Pyrin domain
1 781
CC SH31 415
PSTPIP1/CD2P1
*A230T
PAPA Syndrome: Pyogenic Arthritis, Pyoderma Gangrenosum, and Acne
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Autosomal dominant disorders of unknown cause, variably associated with arthralgias, rashes and conjunctivitis