Whole exome sequencing identified two novel homozygous missense variants in the same codon of CLCN7 underlying autosomal recessive infantile malignant osteopetrosis in a Pakistani family.

Autosomal recessive osteopetrosis is a severe fatal disorder with an average incidence of around 1:250,000. It is diagnosed soon after birth or within the 1st year of life with severe symptoms of abnormal bone remodelling. This study was aimed to identify the underlying genetic cause of the disease in a Pakistani family segregating infantile malignant osteopetrosis in autosomal recessive pattern. Whole exome sequencing of the proband was performed using the 51 Mb SureSelect V4 library kit and sequenced using the Illumina HiSeq2500 sequencing system. The reads were analysed using standard bioinformatic data analysis pipeline. The genotype of candidate variants was confirmed in the proband and his normal parents by Sanger sequencing. Two novel homozygous missense variants were found in the same codon 204 of CLCN7 NM_001287.5:c.[610A>T;612C>G] predicting p.(Ser204Trp) variant in the protein. Sanger sequencing and RFLP assay verified that both these variants were heterozygous in the unaffected parents. Moreover, these variants were not detected in the unrelated healthy Pakistani subjects (200 chromosomes), ExAC, dbSNP, or the 1000 Genomes Project data. Multiple bioinformatics tools unanimously predicted the p.(Ser204Trp) variant as deleterious. CLCN7 mutation p.(Ser204Trp) is the likely cause of the osteopetrosis disease in the Pakistani family. This study expands the restricted spectrum of CLCN7 mutations associated with infantile malignant osteopetrosis and indicates clinical significance of whole exome sequencing in the diagnosis of clinically and genetically heterogenous osteopetrosis phenotype. These data should be helpful in the improved genetic counselling, carrier identification and prenatal diagnosis of the affected family.