A New Candidate Missense Mutation (Leu 1657 Ile) in an Apparently Asymptomatic Type 2A (Phenotype IIA) von Willebrand Disease Family

Summary

Type 2A von Willebrand disease (VWD) is mostly an autosomal dominantly inherited bleeding disorder characterised by a qualitative defect of von Willebrand factor (VWF). Mutation screening was used to screen the whole of VWF gene followed by direct sequencing to detect the mutation in a father and son diagnosed with type 2A (phenotype IIA) von Willebrand disease. A C5219 to A transversion was detected predicting Leucine to Isoleucine substitution in codon 1657. This novel missense mutation which was also identified by MboI restriction enzyme analysis, was found in both patient and his father but not in any other unaffected family member or 50 unrelated normal individuals. This substitution was reproduced by in vitro site directed mutagenesis of full-length VWF cDNA and transiently expressed in COS-7 cells. The corresponding recombinant VWF protein exhibited the full spectrum of VWF multimers, suggesting that the abnormal multimer seen in the patient results from increased proteolysis.

Key words

Type 2A VWD - mutation detection - chemical cleavage mismatch detection - recombinant VWF

• References

• 1 Rodeghiero F, Castaman G, Dini E. Epidemiological investigation of the prevalence of von Willebrand’s disease. Blood 1987; 69: 454-59.
• 2 Holmberg l, Nilsson IM. von Willebrand disease. Clin Haemat 1985; 14: 461-88.
• 3 Nichols WC, Ginsberg D. von Willebrand disease. Medicine 1997; 76: 1-20.
• 4 Sadler JE. A revised classification of von Willebrand disease. Thromb Haemost 1994; 71: 520-25.
• 5 Ginsberg D, Sadler JE. von Willebrand disease: A database of point mutation, insertions and deletions. Thromb Haemost 1993; 69: 177-84.
• 6 Beddall AC, Enayat SM, Williams CE, Short PE, Hill FGH. Simultaneous inheritance and expression of classical haemophilia A and type IIA von Willebrand’s disease. J Clin Path 1984; 37: 1035-39.
• 7 Naylor JA, Green PM, Montandon AJ, Rizza CR, Gianelli F. Detection of three novel mutations in two haemophilia A patients by rapid screening of whole essential region of factor VIII gene. Lancet 1991; 337: 635-39.
• 8 Jenkins V. Screening for candidate mutations causing von Willebrand’s disease. In: Hemostasis and Thrombosis. Perry DJ, Pasi KJ. eds. Totowa, New Jersey: Humana Press; 1999: 169-79.
• 9 Lyons SE, Burck ME, Bowie EJW, Ginsburg D. Impaired intracellular transportation produced by a subset of type IIA von Willebrand disease. J Biol Chem 1992; 267: 4424-30.
• 10 Inbal A, Selegsjohn U, Kornbrot N, Brenner B, Harison P, Randi A, Rabinowitz I, Sadler JE. Characterization of three mutations causing von Willebrand disease type IIA in five unrelated families. Thromb Haemost 1992; 67: 618-22.
• 11 Green PM, Montandon AJ, Bentley DR, Giannelli F. Genetics and molecular biology of haemophilia A and B. Blood Coag Fibrin 1991; 02: 539-65.
• 12 Burke RL, Pachl C, Quiroga M, Rosenberg S, Haigwood N, Nordfang O, Ezban M. The functional domains of coagulation factor VIII:C. J Biol Chem 1986; 261: 12574-78.
• 13 Wagner DD, Saffaripour S, Bonfanti R, Sadler JE, Cramer EM, Chapman B, Mayadas TN. Induction of specific storage organelles by von Willebrand factor propolypeptide. Cell 1991; 64: 403-13.
• 14 Deng WP, Nickoloff JA. Site-directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal Biochem 1992; 200: 81-8.
• 15 Holmes DS, Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Analyt Biochem 1981; 114: 193-95.
• 16 Kreigler M. Gene transfer and expression. A laboratory manual. New York, NY: Stockton Press; 1990
• 17 Graham FL, Van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 1973; 52: 456-67.
• 18 Dent JA, Burkowitz SD, Ware J, Kasper CK, Ruggeri ZM. Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor. Proc Natl Acad Sci USA 1990; 87: 6306-10.
• 19 Evan JSadler, Ginsberg D. A database of polymorphisms in the von Willebrand factor gene and pseudogene. Thromb Haemost 1993; 69: 185-91.
• 20 Theophilus BDM, Enayat MS, Higuchi M, Kazazian HH, Antonarakis SE, Hill FGH. Independent occurrence of the novel Arg2163His mutation in the factor VIII gene in three unrelated families with haemophilia A. Hum Mut. 1997 #126 On-line.A.
• 21 Furlan M, Robles R, Lämmle B. Purification and characterization of a protease from human plasma cleaving von Willebrand factor to fragments produced by in vivo proteolysis. Blood 1996; 87: 4223-34.
• 22 Ginsburg D. Molecular genetics of von Willebrand Disease. Thromb Haemost 1999; 82: 585-91.
• 23 Jenkins PV, Pasi KJ, Perkins SJ. Molecular modelling of ligand and mutation sites of the type 2A domains of human von Willebrand factor and their relevance to von Willebrand’s disease. Blood 1998; 91: 2032-44.
• 24 Tsai HM, Sussman II, Ginsburg D, Lankhof H, Sixma JJ, Nagel RL. Proteolytic cleavage of recombinant type 2A von Willebrand Factor mutants R834W and R834Q: Inhibition by doxycycline and by monoclonal antibody VP-1. Blood 1997; 89: 1954-62.