F8 genetic analysis strategies when standard approaches fail

Summary

Haemophilia A is a common X-linked recessive disorder caused by mutations in F8 leading to deficiency or dysfunction of coagulant factor VIII (FVIII). Despite tremendous improvements in mutation screening methods, in a small group of patients with FVIII deficiency suffering from haemophilia A, no DNA change can be found. In these patients, analysis reveals no causal mutations even after sequencing the whole coding region of F8 including the flanking splice sites, as well as the promoter and the 3’ untranslated region (UTR). After excluding the mutations mimicking the haemophilia A phenotype in interacting partners of the FVIII protein affecting the half life and transport of the protein, mutations or rearrangements in non-coding regions of F8 have to be considered responsible for the haemophilia A phenotype.

In this review, we present the experiences with molecular diagnosis of such cases and approaches to be applied for mutation negative patients.

Zusammenfassung

Hämophilie A, eine häufige, X-chromosomal rezessiv vererbte Erkrankung, verursacht einen Mangel oder eine Dysfunktion des Blutgerinnungsfaktors VIII (FVIII). Trotz großer Fortschritte in Mutationsscreeningverfahren kann bei einer kleinen Gruppe von Patienten, die unter einem Hämophilie-A-ähnlichen Phänotyp leiden, keine genetische Veränderung im F8 nachgewiesen werden. Nach Sequenzierung der gesamten kodierenden F8-Region einschließlich flankierender Spleißstellen, Promotorbereich und 3’-UTR Region kann in diesen Fällen keine ursächliche Mutation nachgewiesen werden. Nach Ausschluss von Mutationen in Genen der Interaktionspartner des FVIII-Proteins, die Halbwertszeit und Transport beeinflussen, sollte nach der Ursache für den Hämophilie-A-ähnlichen Phänotyp in nicht-kodierenden F8-Bereichen untersucht werden.

In dieser Übersicht fassen wir Erfahrungen in der Diagnostik solcher Fälle zusammen und stellen neben molekulargenetischen Mechanismen, die zur Ausprägung eines solchen Phänotyps führen, eine gestaffelte genetischen Untersuchung im Rahmen der Diagnostik vor.

Keywords

Haemophilia A - mutation negative patients - extended diagnostic flowchart

Schlüsselwörter

Hämophilie A - Mutation-negativen Patienten - erweiterte Diagnostik - Flussdiagramm

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