An intronic variant in LAMB3 contributes to junctional epidermolysis bullosa and enamel hypoplasia via translational attenuation.

Objectives: This study aimed to investigate the genetic etiology of a family affected by junctional epidermolysis bullosa (JEB) and generalized enamel hypoplasia, and to explore how an intronic variant influenced the 5' untranslated region (5'UTR), thereby affecting LAMB3 expression and contributing to the pathogenesis of the disease.

Design: Whole-exome and whole-genome sequencing were used to screen for genetic defects in the patient. Mutational consequences were characterized through luciferase assays, splice assay, in silico analyses, and verification using the patient's gingival sample.

Results: A nonsense variant (c.2983 C>T; p.Gln995*) and an intronic variant (c.-38+2 T>C) of LAMB3 were identified. In vitro assays demonstrated that the intronic variant activated a cryptic splice site, resulting in a 120 bp intronic inclusion. This splicing alteration significantly reduced the translation efficiency of the downstream coding sequence, while overall mRNA expression remained unaffected. Bioinformatic analysis unveiled the creation of three upstream AUG codons, leading to the presence of two upstream open reading frames (uORFs) and one overlapping ORF. The longer uORF's AUG exhibited a moderate Kozak strength similar to that of the main ORF's AUG. Structural analysis of the mutant 5'UTR sequence revealed a more complex secondary structure, characterized by a large branch loop and a stem-loop preceding the coding sequence's start codon.

Conclusion: This study suggests that variants affecting the 5'UTR may contribute to the genetic etiology of JEB. These findings could help enhance the diagnostic accuracy and efficiency in JEB patients.