Genetic analysis of the M2/ANXA5 haplotype in Syrian healthy population

Wael Dib, Chadi Soukkarieh, Marwan Alhalabi


Background: Annexin A5 (ANXA5) is an abundantly and ubiquitously expressed protein showing the highest levels of concentration in kidney, liver and placenta. ANXA5 plays a central role in the machinery of membrane repair by enabling of a protective 2D bandage at membrane damaged site, has properties anti-inflammatory, pro-fibrinolytic and anti-thrombotic. Four polymorphisms have been identified in the ANXA5 promoter, which were transmitted as a joint haplotype (M2). M2 haplotype decrease the ANXA5 gene promoter activity and mRNA expression which causes several troubles.

Methods: The aim of this cross-sectional study is to determine the frequency of M2/ANXA5 haplotype in healthy Syrian individuals and compare the genotype and allele distribution with other populations. In this study 94 (female, 71 and male, 23) unrelated healthy Syrian nationals were involved. 94 DNA samples have been collected in order to determine the spread the genotype M2 haplotype using allele specific polymerase chain reaction (AS-PCR).

Results: Our results indicate that the distribution of the alleles and genotypes of M2 haplotype vary considerably in different populations. In the Syrian population the distribution of M2 and wide type (WT) were M2/M2 9.6%, M2/WT 44.7%, WT/WT 45.7%. The M2 haplotype was found in 45 of women (allele frequency 31%) and in 15 of men (allele frequency 32%). The distribution of the ANXA5 genotypes in Syrian study group conformed to Hardy-Weinberg equilibrium (p=0.92).

Conclusions: No significant differences were found at frequency distribution of different genotypes and M2 allele between women and men within this Syrian cohort. In comparison with the results of other studies, the results of this study demonstrate that the frequency and distribution of the M2 haplotype in Syrian population are different from most other populations worldwide.


Annexin A5, M2 haplotype, SNP, Syrian population

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Mirsaeidi M, Gidfar S, Vu A, Schraufnagel D. Annexins family: insights into their functions and potential role in pathogenesis of sarcoidosis. J Transl Med. 2016;14(1):1-9.

Gerke V, Creutz CE, Moss SE. Annexins: linking Ca 2+ signalling to membrane dynamics. Nat Rev Mol Cell Biol. 2005;6(6):449-61.

Moss SE, Morgan RO. The annexins. Genome Biol. 2004;5(4):1-8.

Raynal P, Pollard HB. Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium-and phospholipid-binding proteins. Biochimica et Biophysica Acta (BBA)-Reviews on Biomembranes. 1994;1197(1):63-93.

Ling Q, Jacovina AT, Deora A, et al. Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo. J Clin Investig. 2004;113(1):38-48.

Funakoshi T, Hendrickson LE, McMullen BA, Fujikawa K. Primary structure of human placental anticoagulant protein. Biochemistry. 1987;26(25):8087-92.

Maurer‐Fogy I, Reutelingsperger CP, Pieters J, Bodo G, Stratowa C, Hauptmann R. Cloning and expression of cDNA for human vascular anticoagulant, a Ca2+‐dependent phospholipid‐binding protein. Eur J Biochem. 1988;174(4):585-92.

Tait J, Gibson D, Fujikawa K. Phospholipid binding properties of human placental anticoagulant protein-I, a member of the lipocortin family. J Biol Chem. 1989;264(14):7944-9.

Morgan RO, Bell DW, Testa JR, Fernandez MP. Genomic Locations ofANX11andANX13and the Evolutionary Genetics of Human Annexins. Genomics. 1998;48(1):100-10.

Carcedo M-T, Iglesias J-M, Bances P, Morgan RO, Fernandez M-P. Functional analysis of the human annexin A5 gene promoter: a downstream DNA element and an upstream long terminal repeat regulate transcription. Biochem J. 2001;356(2):571-9.

Lavorato HL, Markoff A, Altholz V. The relevance of ANXA5 genetic variants on male fertility. Journal of assisted reproduction and genetics. 2019;36(7):1355-9.

Srisomsap C, Sawangareetrakul P, Subhasitanont P. Proteomic studies of cholangiocarcinoma and hepatocellular carcinoma cell secretomes. J Biomed Biotechnol. 2010;437143.

Cookson BT, Engelhardt S, Smith C, Bamford HA, Prochazka M, Tait JF. Organization of the human annexin V (ANX5) gene. Genomics. 1994;20(3):463-7.

Bogdanova N, Horst J, Chlystun M, et al. A common haplotype of the annexin A5 (ANXA5) gene promoter is associated with recurrent pregnancy loss. Human molecular genetics. 2007;16(5):573-8.

Chinni E, Tiscia GL, Colaizzo D, Vergura P, Margaglione M, Grandone E. Annexin V expression in human placenta is influenced by the carriership of the common haplotype M2. Fertility and sterility. 2009;91(3):940-2.

Markoff A, Gerdes S, Feldner S, Bogdanova N, Gerke V, Grandone E. Reduced allele specific annexin A5 mRNA levels in placentas carrying the M2/ANXA5 allele. Placenta. 2010;31(10):937-40.

Tüttelmann F, Ivanov P, Dietzel C. Further insights into the role of the annexin A5 M2 haplotype as recurrent pregnancy loss factor, assessing timing of miscarriage and partner risk. Fertility and sterility. 2013;100(5):1321-5.

Demetriou C, Abu-Amero S, White S. Investigation of the Annexin A5 M2 haplotype in 500 white European couples who have experienced recurrent spontaneous abortion. Reproductive biomedicine online. 2015;31(5):681-8.

Pabinger I, Vormittag R. Thrombophilia and pregnancy outcomes. J Thrombosis and Haemostasis. 2005;3(8):1603-10.

Nagirnaja L, Nõmmemees D, Rull K, Christiansen OB, Nielsen HS, Laan M. Annexin A5 promoter haplotype M2 is not a risk factor for recurrent pregnancy loss in Northern Europe. PloS one. 2015;10(7):e0131606.

Ang K-C, Kathirgamanathan S, Ch’ng ES. Genetic analysis of the M2/ANXA5 haplotype as recurrent pregnancy loss predisposition in the Malay population. Journal of assisted reproduction and genetics. 2017;34(4):517-24.

Passos MD. Genetic analysis of the M2/ANXA5 haplotype as adverse pregnancy outcomes risk factor-Evaluation in a Portuguese population group, Universidade de Coimbra. 2018.

Hiddink L, de Visser MC, van Heerde WL. Polymorphisms in the Annexin A5 gene influence circulating Annexin A5 levels in healthy controls. Thrombosis research. 2012;129(6):815-7.

Rogenhofer N, Engels L, Bogdanova N, Tüttelmann F, Markoff A, Thaler C. Paternal and maternal carriage of the annexin A5 M2 haplotype are equal risk factors for recurrent pregnancy loss: a pilot study. Fertility and sterility. 2012;98(2):383-8.

Ang K-C, Bogdanova N, Markoff A, Ch'ng ES, Tang TH. Association between M2/ANXA5 haplotype and repeated pregnancy loss: a meta-analysis. Fertility and sterility. 2019;111(5):971-81.

RPL EGGo, Bender Atik R, Christiansen OB. ESHRE guideline: recurrent pregnancy loss. Human reproduction open. 2018;2018(2):hoy004.

Rogenhofer N, Engels L, Bogdanova N, Tuttelmann F, Markoff A, Thaler C. Paternal and maternal carriage of the annexin A5 M2 haplotype are equal risk factors for recurrent pregnancy loss: a pilot study. Fertil Steril. Aug 2012;98(2):383-8.

Chinni E, Tiscia GL, Colaizzo D, Vergura P, Margaglione M, Grandone E. Annexin V expression in human placenta is influenced by the carriership of the common haplotype M2. Fertil Steril. 2009;91(3):940-2.