PREIMPLANTATION DIAGNOSIS OF INCONTINENTIA

PREIMPLANTATION DIAGNOSIS OF INCONTINENTIA PIGMENTI
Raskova D.1, Eliasova I.1, Putzova M. 1, Feldmar P.1, Mika J.1, Stejskal D.1, Hejtmankova M.1, Malikova M.2
Centre for Medical Genetics and Reproductive Medicine GENNET, Kostelní 9, Prague 7, Czech Republic
2
Institute of Biology and Medical Genetics, 2nd Medical Faculty, Charles University, Prague 5, Czech Republic
1
Introduction
In our centre we have been providing preimplantation genetic diagnosis
(PGD) since 2007. We have accomplished 117 IVF cycles followed by
PGD in 43 monogenic diagnoses of all inheritance types up to now.
On average 5-6 embryos have been biopsied per IVF cycle. The success
rate of PGD procedures (the pregnancy confirmed by the fetal heart beat)
is approximately 43%. PGD list of diagnoses is being continually updated.
THE NAME OF DISEASE
DISEASE ABBREVIATION
GENE
CHROMOZOME
FIRST INVESTIGATED
CMT 1A
PMP22
17p12
8/2007
CF
CFTR
7q31.2
6/2007
FBN1
15q21.1
3/2008
Charcot - Marie - Tooth1A
Cystic fibrosis
Marfan syndrome
Huntington disease
HCH
IT15
4p16.3
1/2008
RhD aloimunisation
RhD
RHD
chr. 1
7/2008
Myotonic dystrofy
MD1
DMPK
19q13.32
5/2008
Fragile X
FRAXA
FMR1
Xq27.3
4/2008
XL SCID
SCID
IL2RG
Xq13.1
5/2008
Factor VIII
Xq28
1/2008
PKHD1
6p12.2
5/2008
GJB2 (Connexin26)
13q12.11
9/2008
SPTLC1
9q22.31
1/2009
FGFR2
10q26.13
11/2008
BRCA1
BRCA1
17q21.31
4/2009
Duchenne muscular dystrophy
DMD
dystrofin
Xp21.1
4/2009
Neurofibromatosis 1
NF1
NF1
17q11.2
4/2009
Spinal muscular atrophy
SMA
SMN1
5q12.2-13.3
7/2009
HLXB9
7q36.3
7/2009
Hemophylia A
Polycystic kidney disease – autosomal recesive
ARPKD
Deafness autosomal recesive
Hereditay sensory neuropathy1
HSN1
Crouzon syndrome
BRCA1
The Case Report
The case report shows PGD in a 34 year old woman with incontinentia
pigmenti (IP2), verified by the mutation in the NEMO gene, which maps
to Xq28 (MIM ID 308300). IP is genodermatosis that segregates as
an X-linked dominant disorder and is usually lethal prenatally in males.
In affected females it causes highly variable abnormalities of the skin,
hair, nails, teeth, eyes and central nervous system. The risk of recurrence
is 50% for daughters and it is lethal for 50% of sons.
Currarino syndrome
Treacher Collins syndrome
TCS
TCOF1
5q33.1
9/2009
Protoporphyria erythropoietic
EPP
FECH
18q21.3
9/2009
IPEX syndrome
IPEX
FOXP3
Xp11.23
9/2009
Propionic acidemia
PA
PCCB
3q21-3q22.3
11/2009
Alagille syndrome
AS
JAG-1
20p12
11/2009
Epidermolysis bullosa
EB
keratin 14 (KRT14)
17q21.2
12/2009
HNPCC
MLH1
3p22.2
1/2010
PMD
PLP1
Xq22.2
1/2010
CMTX1
GJB1,Cx32
Xq13.1
2/2010
Leiden mutation
FV
Factor V
1q24.2
4/2010
Fabry disease
FD
GLA
Xq22.1
4/2010
ADPKD
PKD1
16p13.3
5/2010
Factor IX
Xq27.1
7/2010
JEB
LAMB3
1q32.2
7/2010
BRCA2
BRCA2
13q13.1
7/2010
LCHAD
HADHA
2p23.3
8/2010
COL4A5
Xq22.3
8/2010
ADSL
22q13.1-q13.2
8/2010
VHL
3p25-p26
8/2010
ADA
20q13.12
9/2010
Lynch syndrome
Pelizaeus - Merzbacher syndrome
Charcot - Marie - Tooth – XL
Polycystic kidney disease -autosomal dominant
Hemophylia B
Our patient developed erythema, blisters and hyperpigmentation in
infancy, but all of these symptoms later completely disappeared. She
had 3 miscarriages and gave birth to one healthy daughter. In 1 of the
miscarriages a karyotype 45, X with intragene deletion in the NEMO gene
was confirmed.
Methods
Genetic pre-case haplotyping (PGH) is an initial and important constituent
for determination of disease-associated haplotype by comparison with the
haplotype of other members of the family.
During the IVF cycle, we use a genetic haplotyping technique by multiplex
PCR on products of MDA (multiple displacement amplification) from
1 blastomere biopsied from the cleavage-stage embryo.
In case of ambiguous results from the blastomere, analysis of the
trofoectoderm can be repeated and the embryo can still be transferred
on day 5.
Results
PGH was concluded in terms of healthy haplotype mother-daughter
because we did not keep at disposition DNA of other family member with
disease - associated haplotype.
Epidermolysis bullosa junctionalis
BRCA2
Long chain 3 - hydroxyacyl CoA dehydrogenace deficiency (LCHAD)
Alport syndome
Adenylosukcinatlyasis deficienty
Von Hippel - Lindau syndrome
VHL
Adenosindeaminasis deficiency
Mukopolysacharidosis I
MPS1
IDUA
4p16.3
9/2010
Incontinentia pigmenti
IP
NEMO
Xq28
11/2010
MYH9
22q12.3
01/2011
MYH9 related disorders (e.g. Epstain syndrome)
Facioscapulohumeral muscular dystrophy
FSHD
FRG1
4q35
02/2011
Adrenal hyperplasia congenital
CAH
CYP21
6p2.1-3
4/2011
Pre-case haplotyping
Inconentia pigmenti
(GD)
mother
DXS8086
DXS8069
DXS7423
DXS8011
DXS8103
DXS1684
DXS8061
DXS8087
NEMO(IKBKG)
DXS1073
KI I I R
20xAT
26xAC
21xAT
22xGT
DXS1108
24xGT
23xGT
DXYS154
AMXY
SRY
marker
DXS8086
DXS8069
DXS7423
DXS8011
DXS8103
DXS1684
DXS8061
DXS8087
NEMO(IKBKG)
DXS1073
KI I I R
20xAT
26xAC
21xAT
22xGT
DXS1108
24xGT
23xGT
DXYS154
AMXY
SRY
i nf. a l e l a
273
277
331
241
323
192
243
296
109
111
293
270
158
268
154
XX-OK
273
209
331
182
241
294
198
275
271
209
327
182
247
294
198
283
224
144
109
239
171
293
270
158
253
186
108
/
236
147
133
237
171
286
259
156
253
180
108
/
277
209
323
192
243
296
198
275
273
209
331
182
241
294
198
275
224
144
111
239
171
286
268
154
253
186
108
/
224
144
109
239
171
293
270
158
253
186
108
/
XY-OK
273
209
331
182
241
294
198
275
224
144
109
239
171
293
270
158
253
186
108
/
father
271
209
327
182
247
294
198
283
236
147
133
237
171
286
259
156
253
180
108
/
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
192
114
108
mater.patol
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
192
114
108
XX affected embryo
277
209
323
192
243
296
198
275
271
209
327
182
247
294
198
283
224
144
111
239
171
286
268
154
253
186
108
/
236
147
133
237
171
286
259
156
253
180
108
/
ma te r. OK
pa te r. OK
XY affected embryo
277
209
323
192
243
296
198
275
224
144
111
239
171
286
268
154
253
186
108
/
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
192
114
108
Figure II: Methodical overview
At the first IVF cycle, we analyzed nine embryos of which four were
affected. Monosomy of chromosome X was determined in one embryo.
In the next embryo, the outcome could not be unambiguously established
due to proof of the crossing-over in the NEMO gene area. Selection of
healthy embryos for transfer was recommended in two cases. One male
embryo was transferred and the resulting pregnancy was confirmed by
an ultrasound investigation.
Conclusion
Preimplantation haplotyping (PGH) as a clinical method within PGD
represents a successful alternative in the reproductive choices of families
at risk of transmitting genetic disorders.
References
1.Renwick P. J. et al.: Proof of principle and first cases using preimplantation genetic
haplotyping–a paradigm shift for embryo diagnosis. RBMOnlineVol.13No 1.2006
2.PGDIS: Guidelines for good practise in PGD: programme requirements and laboratory
quality assurance, RBMOnline-Vol16. No 1. 2008 134-147
3.Hellani A, Coskun S, Benkhalifa M et al.: Multiple displacement amplification on single
cell and possible PGD applications. Molecular Human Reproduction 10, 2004 847–852.
4.Bodak, N., Hadj-Rabia, S., Hamel-Teillac, D., de Prost, Y., Bodemer, C. Late recurrence of
inflammatory first-stage lesions in incontinentia pigmenti: an unusual phenomenon and a
fascinating pathologic mechanism. Arch. Derm. 139: 201-204, 2003. [PubMed:12588226]
5.Landy, S. J., Donnai, D. Incontinentia pigmenti (Bloch-Sulzberger syndrome). J. Med. Genet.
30: 53-59, 1993. [PubMed:8423608]
6.Smahi, A., Hyden-Granskog, C., Peterlin, B., Vabres, P., Heuertz, S., Fulchignoni-Lataud,
M. C., Dahl, N., Labrune, P., Le Marec, B., Piussan, C., Taieb, A., von Koskull, H., HorsCayla, M. C. The gene for the familial form of incontinentia pigmenti (IP2) maps to the
distal part of Xq28. Hum. Molec. Genet. 3: 273-278, 1994. [PubMed:8004094]
7.The International Incontinentia Pigmenti Consortium Genomic rearrangement in NEMO
impairs NF-kappa-B activation and is a cause of incontinentia pigmenti. Nature 405: 466472, 2000. [PubMed:10839543]

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