Intragenic deletions and a deep intronic mutation affecting pre-mRNA splicing in the dihydropyrimidine dehydrogenase gene as novel mechanisms causing 5-fluorouracil toxicity.
Dihydropyrimidine dehydrogenase (DPD) is the initial enzyme acting in the catabolism of the widely used antineoplastic agent 5-fluorouracil (5FU). DPD deficiency is known to cause a potentially lethal toxicity following administration of 5FU. Here, we report novel genetic mechanisms underlying DPD deficiency in patients presenting with grade III/IV 5FU-associated toxicity. In one patient a genomic DPYD deletion of exons 21-23 was observed. In five patients a deep intronic mutation c.1129-5923C>G was identified creating a cryptic splice donor site. As a consequence, a 44 bp fragment corresponding to nucleotides c.1129-5967 to c.1129-5924 of intron 10 was inserted in the mature DPD mRNA. The deleterious c.1129-5923C>G mutation proved to be in cis with three intronic polymorphisms (c.483 + 18G>A, c.959-51T>G, c.680 + 139G>A) and the synonymous mutation c.1236G>A of a previously identified haplotype. Retrospective analysis of 203 cancer patients showed that the c.1129-5923C>G mutation was significantly enriched in patients with severe 5FU-associated toxicity (9.1%) compared to patients without toxicity (2.2%). In addition, a high prevalence was observed for the c.1129-5923C>G mutation in the normal Dutch (2.6%) and German (3.3%) population. Our study demonstrates that a genomic deletion affecting DPYD and a deep intronic mutation affecting pre-mRNA splicing can cause severe 5FU-associated toxicity. We conclude that screening for DPD deficiency should include a search for genomic rearrangements and aberrant splicing.
van Kuilenburg
André B P
AB
Department of Clinical Chemistry, Academic Medical Center, Emma Children's Hospital, University of Amsterdam, Amsterdam, The Netherlands. a.b.vanKuilenburg@amc.uva.nl
Meijer
Judith
J
Mul
Adri N P M
AN
Meinsma
Rutger
R
Schmid
Veronika
V
Dobritzsch
Doreen
D
Hennekam
Raoul C M
RC
Mannens
Marcel M A M
MM
Kiechle
Marion
M
Etienne-Grimaldi
Marie-Christine
MC
Klümpen
Heinz-Josef
HJ
Maring
Jan Gerard
JG
Derleyn
Veerle A
VA
Maartense
Ed
E
Milano
Gérard
G
Vijzelaar
Raymon
R
Gross
Eva
E
D000328:Adult; D000368:Aged; D000964:Antimetabolites, Antineoplastic / Q000633:toxicity*; D055028:Comparative Genomic Hybridization; D042943:Dihydrouracil Dehydrogenase (NADP) / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005472:Fluorouracil / Q000633:toxicity*; D015321:Gene Rearrangement; D006801:Humans; D007438:Introns; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012326:RNA Splicing; D012333:RNA, Messenger; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion
D000964:Antimetabolites, Antineoplastic; D012333:RNA, Messenger; D042943:Dihydrouracil Dehydrogenase (NADP); D005472:Fluorouracil
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