Clinical and Genetic Analysis of Nine Suspected Familial Haemophagocytic Lymphohistiocytosis Patients for MUNC13-4 Deficiency and Introducing Four Novel Mutations in UNC13D

  • Maryam Vahidi Department of Molecular Genetics, Islamic Azad University, Tabriz, Iran
  • Mohsen Badalzadeh Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran
  • Masoomeh Jannesar Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  • Marzieh Mazinani Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Reza Fazlollahi Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  • Nazanin Khodayari Namini Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  • Massoud Houshmand Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
  • Amir Ali Hamidieh Department of Pediatric Stem Cell Transplant, Children’s Medical Center, Tehran University of Medical Sciences, Tehran, Iran
  • Leila Moradi Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  • Zahra Pourpak Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  • Mostafa Moin Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
Keywords: FHL3, Haemophagocytic lymphohistiocytosis, UNC13D

Abstract

Familial haemophagocytic lymphohistiocytosis (FHL) is a rare disorder of immune dysregulation. FHL inherited in an autosomal recessive pattern is classified into five subtypes based on underlying genetic defects. Mutations in four genes including PRF1, UNC13D, STX11 and STXBP2 are responsible for FHL2 to FHL5 respectively. The cause of FHL1 is associated with mutations in an unknown gene located at 9q21.3-22. This study aims to report the clinical features and genetic results of nine Iranian patients suffering from -haemophagocytic lymphohistiocytosis. Nine patients (five males and four females) suspected to FHL whose genetic evaluation of PRF1 and STX11 revealed no mutations, were entered the study to investigate UNC13D mutations. Primers were designed to amplify all coding regions and exon-intron boundaries of the gene. PCR products were then sequenced and analyzed by sequence analysis tools including BLAST. The most frequent clinical manifestations observed in the patients were fever and hepatosplenomegaly. In this study, five mutations were detected in UNC13D including four novel mutations (c.1434_1446delACCCATGGTGCAGinsTGGTGCT, c.1933C>T, c.1389+1G>C and c.2091+1G>A) besides to a previously reported deletion (c.627delT). The pathogenicity of the missense mutation was assessed using online prediction tools including SIFT and PolyPhen2. The study results may provide valuable information for genetic counseling especially for those who have a history of immunodeficiency diseases in their family and can be used for prenatal diagnosis.

References

1. Shibata H, Yasumi T, Shimodera S, Hiejima E, Izawa K, Kawai T, et al. Human CTL-based functional analysis shows the reliability of a munc13-4 protein expression assay for FHL3 diagnosis. Blood 2018; 131(18):2016-25.
2. Vick EJ, Patel K, Prouet P, Martin MG. Proliferation through activation: hemophagocytic lymphohistiocytosis in hematologic malignancy. Blood Adv 2017; 1(12):779-91.
3. Malaga-Dieguez L, Ming W, Trachtman H. Direct Reversible Kidney Injury in Familial Hemophagocytic Lymphohistiocytosis Type 3. J Am Soc Nephrol 2015; 26(8):1777-80.
4. Dufourcq-Lagelouse R, Jabado N, Le Deist F, Stephan JL, Souillet G, Bruin M, et al. Linkage of familial hemophagocytic lymphohistiocytosis to 10q21-22 and evidence for heterogeneity. Am J Hum Genet 1999; 64(1):172-9.
5. Feldmann J, Callebaut I, Raposo G, Certain S, Bacq D, Dumont C, et al. Munc13-4 is essential for cytolytic granules fusion and is mutated in a form of familial hemophagocytic lymphohistiocytosis (FHL3). Cell 2003; 115(4):461-73.
6. zur Stadt U, Schmidt S, Kasper B, Beutel K, Diler AS, Henter JI, et al. Linkage of familial hemophagocytic lymphohistiocytosis (FHL) type-4 to chromosome 6q24 and identification of mutations in syntaxin 11. Hum Mol Genet 2005; 14(6):827-34.
7. Cote M, Menager MM, Burgess A, Mahlaoui N, Picard C, Schaffner C, et al. Munc18-2 deficiency causes familial hemophagocytic lymphohistiocytosis type 5 and impairs cytotoxic granule exocytosis in patient NK cells. J Clin Invest 2009; 119(12):3765-73.
8. zur Stadt U, Rohr J, Seifert W, Koch F, Grieve S, Pagel J, et al. Familial hemophagocytic lymphohistiocytosis type 5 (FHL-5) is caused by mutations in Munc18-2 and impaired binding to syntaxin 11. Am J Hum Genet 2009; 85(4):482-92.
9. Ohadi M, Lalloz MR, Sham P, Zhao J, Dearlove AM, Shiach C, et al. Localization of a gene for familial hemophagocytic lymphohistiocytosis at chromosome 9q21.3-22 by homozygosity mapping. Am J Hum Genet 1999; 64(1):165-71.
10. Rohr J, Beutel K, Maul-Pavicic A, Vraetz T, Thiel J, Warnatz K, et al. Atypical familial hemophagocytic lymphohistiocytosis due to mutations in UNC13D and STXBP2 overlaps with primary immunodeficiency diseases. Haematologica 2010; 95(12):2080-7.
11. Zhang L, Zhou J, Sokol L. Hereditary and acquired hemophagocytic lymphohistiocytosis. Cancer Control 2014; 21(4):301-12.
12. Henter JI, Horne A, Arico M, Egeler RM, Filipovich AH, Imashuku S, et al. HLH-2004: Diagnostic and therapeutic guidelines for hemophagocytic lymphohistiocytosis. Pediatr Blood Cancer 2007; 48(2):124-31.
13. Sullivan EK. Stiehm R. Stiehm's Immune Deficiencies. Elsevier 2014. 1st Edition. Chapter 20: 439, 444-5.
14. Sieni E, Cetica V, Santoro A, Beutel K, Mastrodicasa E, Meeths M, et al. Genotype-phenotype study of familial haemophagocytic lymphohistiocytosis type 3. J Med Genet 2011; 48(5):343-52.
15. Janka GE. Hemophagocytic lymphohistiocytosis. Hematology 2005; (10 Suppl 1):104-7.
16. Usmani GN, Woda BA, Newburger PE. Advances in understanding the pathogenesis of HLH. Br J Haematol 2013; 161(5):609-22.
17. Ng PC, Henikoff S. Predicting deleterious amino acid substitutions. Genome Res 2001; 11(5):863-74.
18. Adzhubei IA, Schmidt S, Peshkin L, Ramensky VE, Gerasimova A, Bork P, et al. A method and server for predicting damaging missense mutations. Nat Methods 2010; 7(4):248-9.
19. Zur Stadt U, Beutel K, Kolberg S, Schneppenheim R, Kabisch H, Janka G, et al. Mutation spectrum in children with primary hemophagocytic lymphohistiocytosis: molecular and functional analyses of PRF1, UNC13D, STX11, and RAB27A. Hum Mutat 2006; 27(1):62-8.
20. Neeft M, Wieffer M, de Jong AS, Negroiu G, Metz CH, van Loon A, et al. Munc13-4 is an effector of rab27a and controls secretion of lysosomes in hematopoietic cells. Mol Biol Cell 2005; 16(2):731-41.
Published
2019-10-23
How to Cite
1.
Vahidi M, Badalzadeh M, Jannesar M, Mazinani M, Fazlollahi MR, Khodayari Namini N, Houshmand M, Hamidieh AA, Moradi L, Pourpak Z, Moin M. Clinical and Genetic Analysis of Nine Suspected Familial Haemophagocytic Lymphohistiocytosis Patients for MUNC13-4 Deficiency and Introducing Four Novel Mutations in UNC13D. Iran J Allergy Asthma Immunol. 18(5):487-492.
Section
Original Article(s)