Lethal factor VII deficiency due to novel mutations in the F7 promoter: Functional analysis reveals disruption of HNF4 binding site

Summary

Hereditary factor VII (FVII) deficiency is a rare autosomal recessive disorder. Deleterious mutations that prevent the synthesis of any amount of functional FVII have been associated with life-threatening haemorrhage in neonates. Here we report two infants, of Maghrebian origin, who suffered a fatal spontaneous cerebral haemorrhage. Investigation of the molecular basis for their severe FVII deficiency revealed novel mutations in a homozygous state within the F7 gene promoter: a single nucleotide substitution (c.-65G>C) and a 2bp deletion (c.-60_-59delTT). To determine whether these promoter variants were responsible for the FVII deficiency, computer–assisted sequence analyses were performed. The data predicted a disrupted binding of both HNF4 and COUP-TF transcription factors with each variant. Concordantly, experimental results revealed an altered HNF4-induced transactivation in the promoter mutated variants. The execution of functional tests is critical to ensuring a complete understanding of the effect of any promoter mutant on FVII deficiency. Only then can an accurate molecular diagnosis be made and further genetic counselling and prenatal diagnosis be offered.

Keywords

FVII deficiency - promoter mutation - HNF4 - COUP-TF1

^* These authors contributed equally.

• References

• 1 Owens AP, Mackman N. Tissue factor and thrombosis: the clot starts here. Thromb Haemost 2010; 104: 432-439.
• 2 Mariani G, Bernardi F. Factor VII Deficiency. Semin Thromb Hemost 2009; 35: 400-406.
• 3 Erdmann D, Heim J. Orphan nuclear receptor HNF-4 binds to the human coagulation factor VII promoter. J Biol Chem 1995; 270: 22988-22996.
• 4 Giansily-Blaizot M, Aguilar-Martinez P, Briquel ME. et al. Two novel cases of cerebral haemorrhages at the neonatal period associated with inherited factor VII deficiency, one of them revealing a new nonsense mutation (Ser52Stop). Blood Coagul Fibrinolysis 2003; 14: 217-220.
• 5 O'Hara PJ, Grant FJ, Haldeman BA. et al. Nucleotide sequence of the gene coding for human factor VII, a vitamin K-dependent protein participating in blood coagulation. Proc Natl Acad Sci USA 1987; 84: 5158-5162.
• 6 Greenberg D, Miao CH, Ho WT, Chung DW, Davie EW. Liver-specific expression of the human factor VII gene. Proc Natl Acad Sci USA 1995; 92: 12347-12351.
• 7 Carew JA, Jackson AA, Bauer KA. ARP1 interacts with the 5' flanking region of the coagulation factor VII gene. J Thromb Haemost 2003; 01: 1220-1227.
• 8 Arbini AA, Pollak ES, Bayleran JK, High KA, Bauer KA. Severe factor VII deficiency due to a mutation disrupting a hepatocyte nuclear factor 4 binding site in the factor VII promoter. Blood 1997; 89: 176-182.
• 9 Zheng XW, Kudaravalli R, Russell TT. et al. Mutation in the factor VII hepatocyte nuclear factor 4α-binding site contributes to factor VII deficiency. Blood Coagul Fibrinolysis 2011; 22: 624-627.
• 10 Carew JA, Pollak ES, Lopaciuk S, Bauer KA. A new mutation in the HNF4 binding region of the factor VII promoter in a patient with severe factor VII deficiency. Blood 2000; 96: 4370-4372.
• 11 Kavlie A, Hiltunen L, Rasi V, Prydz H. Two novel mutations in the human coagulation factor VII promoter. Thromb Haemost 2003; 90: 194-205.
• 12 Borensztajn K, Chafa O, Alhenc-Gelas M. et al. Characterization of two novel splice site mutations in human factor VII gene causing severe plasma factor VII deficiency and bleeding diathesis. Br J Haematol 2002; 117: 168-171.
• 13 Pollak ES, Hung HL, Godin W, Overton GC, High KA. Functional characterization of the human factor VII 5'-flanking region. J Biol Chem 1996; 271: 1738-1747.
• 14 Kudaravalli R, Tidd T, Pinotti M. et al. Polymorphic changes in the 5' flanking region of factor VII have a combined effect on promoter strength. Thromb Haemost 2002; 88: 763-767.
• 15 Ladias JA, Karathanasis SK. Regulation of the apolipoprotein AI gene by ARP-1, a novel member of the steroid receptor superfamily. Science 1991; 251: 561-565.
• 16 Bernardi F, Marchetti G, Pinotti M. et al. Factor VII gene polymorphisms contribute about one third of the factor VII level variation in plasma. Arterioscler Thromb Vasc Biol 1996; 16: 72-76.
• 17 Carew JA, Basso F, Miller GJ. et al. A functional haplotype in the 5' flanking region of the factor VII gene is associated with an increased risk of coronary heart disease. J Thromb Haemost 2003; 01: 2179-2185.
• 18 Sabater-Lleal M, Chillon M, Howard TE. et al. Functional analysis of the genetic variability in the F7 gene promoter. Atherosclerosis 2007; 195: 262-268.
• 19 Ktistaki E, Talianidis I. Chicken ovalbumin upstream promoter transcription factors act as auxiliary cofactors for hepatocyte nuclear factor 4 and enhance hepatic gene expression. Mol Cell Biol 1997; 17: 2790-2797.
• 20 Marinescu VD, Kohane IS, Riva A. MAPPER: a search engine for the computational identification of putative transcription factor binding sites in multiple genomes. BMC Bioinformatics 2005; 06: 79.
• 21 Gustafsdottir SM, Schlingemann J, Rada-Iglesias A. et al. In vitro analysis of DNA-protein interactions by proximity ligation. Proc Natl Acad Sci USA 2007; 104: 3067-3072.
• 22 Tsai SY, Tsai MJ. Chick ovalbumin upstream promoter-transcription factors (COUP-TFs): coming of age. Endocr Rev 1997; 18: 229-240.
• 23 Santangelo L, Marchetti A, Cicchini C, Conigliaro A, Conti B, Mancone C, Bonzo JA, Gonzalez FJ, Alonzi T, Amicone L, Tripodi M. The stable repression of mesenchymal program is required for hepatocyte identity: a novel role for hepatocyte nuclear factor 4α. Hepatology 2011; 53: 2063-2074.
• 24 Rollini P, Fournier RE. The HNF-4HNF-1 transactivation cascade regulates gene activity and chromatin structure of the human serine protease inhibitor gene cluster at 14q32.1. Proc Natl Acad Sci USA 1999; 96: 10308-10313.
• 25 Ungaro P, Teperino R, Mirra P. et al. Hepatocyte nuclear factor (HNF)-4a-driven epigenetic silencing of the human PED gene. Diabetologia 2010; 53: 1482-1492.
• 26 Schuster-Böckler B, Schultz J, Rahmann S. HMM Logos for visualization of protein families. BMC Bioinformatics 2004; 21: 5-7.