The 253-kb inversion and deep intronic mutations in UNC13D are present in North American patients with familial hemophagocytic lymphohistiocytosis 3

Pediatr Blood Cancer. 2014 Jun;61(6):1034-40. doi: 10.1002/pbc.24955. Epub 2014 Jan 28.

Abstract

Background: The mutations in UNC13D are responsible for familial hemophagocytic lymphohistiocytosis (FHL) type 3. A 253-kb inversion and two deep intronic mutations, c.118-308C > T and c.118-307G > A, in UNC13D were recently reported in European and Asian FHL3 patients. We sought to determine the prevalence of these three non-coding mutations in North American FHL patients and evaluate the significance of examining these new mutations in genetic testing.

Procedure: We performed DNA sequencing of UNC13D and targeted analysis of these three mutations in 1,709 North American patients with a suspected clinical diagnosis of hemophagocytic lymphohistiocytosis (HLH).

Results: The 253-kb inversion, intronic mutations c.118-308C > T and c.118-307G > A were found in 11, 15, and 4 patients, respectively, in which the genetic basis (bi-allelic mutations) explained 25 additional patients. Taken together with previously diagnosed FHL3 patients in our HLH patient registry, these three non-coding mutations were found in 31.6% (25/79) of the FHL3 patients. The 253-kb inversion, c.118-308C > T and c.118-307G > A accounted for 7.0%, 8.9%, and 1.3% of mutant alleles, respectively. Significantly, eight novel mutations in UNC13D are being reported in this study. To further evaluate the expression level of the newly reported intronic mutation c.118-307G > A, reverse transcription PCR and Western blot analysis revealed a significant reduction of both RNA and protein levels suggesting that the c.118-307G > A mutation affects transcription.

Conclusions: These specified non-coding mutations were found in a significant number of North American patients and inclusion of them in mutation analysis will improve the molecular diagnosis of FHL3.

Keywords: UNC13D; familial hemophagocytic lymphohistiocytosis (FHL); hemophagocytic lymphohistiocytosis (HLH); intronic mutation; inversion.

MeSH terms

  • Adolescent
  • Adult
  • Arabs / genetics
  • Asian / genetics
  • Black or African American / genetics
  • Child
  • Chromosome Inversion
  • Consanguinity
  • DNA Mutational Analysis
  • Female
  • Genetic Testing
  • Hispanic or Latino / genetics
  • Humans
  • Infant
  • Infant, Newborn
  • Introns / genetics
  • Lymphohistiocytosis, Hemophagocytic / ethnology
  • Lymphohistiocytosis, Hemophagocytic / genetics*
  • Male
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics*
  • Membrane Proteins / physiology
  • North America / epidemiology
  • Point Mutation
  • Sequence Analysis, DNA
  • White People / genetics
  • Young Adult

Substances

  • Membrane Proteins
  • UNC13D protein, human

Supplementary concepts

  • Hemophagocytic lymphohistiocytosis, familial, 3