322.

DISORDERS OF COAGULATION OR FIBRINOLYSIS: POSTER II | NOVEMBER 29, 2018

The Highly Prevalent Deletions in F8 Intron 13 Found

in French Mild Haemophilia a Patients Result of
Both Founder Effect and Recurrent De Novo Events
*,1,2 *,1 *,3
Yohann Jourdy, PhD PharmD, Mathilde Fretigny, PharmD, Fanny Lassalle, PharmD,
4 5 *,6 *,6
David Lillicrap, MD, Claude Negrier, Patrice Bouvagnet, MD, Christine Vinciguerra, PharmD
1
Hospices Civils de Lyon, BRON, France
2
EA 4609 Hémostase et cancer, Université Claude Bernard Lyon 1, LYON, France
3
Centre Hospitalier Régional Universitaire de Lille, Lille, France
4
Department of Pathology and Molecular Medicine, Queen's University, Kingston, Canada
5
Haematology Department, Hôpital Edouard Herriot, Lyon, France
6
Hospices Civils de Lyon, Bron, France

Blood (2018) 132 (Supplement 1) : 2489.

http://doi.org/10.1182/blood-2018-99-111791

Abstract
Background: Genetic variations that are found at a relatively high frequency can be the consequence
of a founder effect or repeated de novo events. Recently, our group has identified an intronic deletion,
c.2113+461_2113+473del [DEL13T], in the F8 intron 13, in two mild haemophilia A patients. This
deletion removed a part of the poly(T)-tail from the right arm of antisens AluY element and led to AluY
exonisation. Then, deletions in the poly(T)-tail of AluY in F8 intron 13, from 10 to 14 pb in size, were found
in approximately 6% of all cases of mild haemophilia in France.

Aim: In this study, we determined whether these highly prevalent deletions are the result of recurrent
molecular mechanism or of a founder effect.

Methods: Haplotype reconstruction was performed after analysis of F8 extragenic and intragenic
polymorphic markers in 71 unrelated French mild haemophilia A patients carrying a deletion
in the poly(T)-tail of AluY in F8 intron 13 (c.2113+460_2113+473del [DEL14T] n=1; DEL13T,
n=62; c.2113+462_2113+473del [DEL12T], n=2; c.2113+463_2113+473del [DEL11T], n=5 and
c.2113+464_2113+473del, n=1 [DEL10T]) and in 50 non-haemophilia A subjects. The ESTIAGE tool was
used to estimate the age of the DEL13T. Nineteen genetically unresolved mild Haemophilia A patients
from Queen's University, Canada, were also included in the study. All patients and controls gave informed
consent for genetic studies.

Results: Two intragenic (STR13 and STR22) and 3 extragenic (DXS8061, ST14 and DF2) microsatellites
were investigated in the DNA of the 71 mild haemophilia A patients. This set of polymorphisms markers
covered a genomic region of 2,779,113 nucleotides (4.17 cM).

Among the 62 patients carrying the DEL13T, 60 patients had the same allele for the marker directly
flanking the deletion on the centromeric side (STR13, genetic distance 0.0062 cM). Two of them differed
from the others at the next centromeric maker STR22 (genetic distance 0.2047 cM) and 20 patients
differed from the others at ST14 (genetic distance 3.4237 cM). On the telomeric side, only 8 patients
differed from the others at DF2 (genetic distance 0.0292 cM). None of control individuals shared such
haplotypes with these 60 patients. These results provided evidence that the founder effect hypothesis
was very plausible for the variant DEL13T. The ESTIAGE tool estimated that the DEL13T occurred about
61 generations ago (95% CI : 51-74 generations). Assuming that a generation spanned 25 years, the
French common ancestor carrying the c.2113+461_2113+473del was estimated to have lived between
th th
the 2 and the 8 century.

However, two patients carrying the DEL13T and 9 patients carrying the other deletions (DEL10T,
DEL11T, DEL12T and DEL14T) had a different haplotype suggesting that these deletions arose
independently. In order to support the hypothesis of a recurrent molecular mechanism, we have
investigated the presence of these deletions in other geographies. F8 intron 13 deletions were found
in 3/19 Canadian patients included in this study (DEL13T, n=2 and DEL10T, n=1). Haplotype analysis
performed in these three patients suggested a de novo mechanism for two of them.

Conclusion: This study supports both a founder effect for the c.2113+461_2113+473del in the French
mild haemophilia A patients and a recurrent molecular mechanism leading to deletion in the poly(T)-
tail of AluY in F8 intron 13. We recommended that these deletions be specifically investigated in all mild
haemophilia A patients in whom no genetic abnormality has been detected by standard genetic analysis.
Finally, these results suggest that large poly(T)-tail of inverted Alu elements may be a mutational hot spot
and such deletions leading to alu-exonization, could occur in other genes.

Disclosures

Negrier: Sobi/Bioverativ: Honoraria, Membership on an entity's Board of Directors or advisory

committees, Research Funding; Octapharma: Honoraria, Research Funding; CSL Behring: Consultancy,
Honoraria, Membership on an entity's Board of Directors or advisory committees; Novo Nordisk:
Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding;
Alnylam: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research
Funding; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees;
LFB: Honoraria, Membership on an entity's Board of Directors or advisory committees; Baxalta/Shire:
Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding;
Bayer: Honoraria, Research Funding; Roche: Honoraria, Membership on an entity's Board of Directors or
advisory committees, Research Funding.

Author notes
* Asterisk with author names denotes non-ASH members.

© 2018 by the American Society of Hematology