Two novel mutations in the POU1F1 gene generate null alleles through different mechanisms leading to combined pituitary hormone deficiency

Background Mutations in the POU1F1 gene severely affect the development and function of the anterior pituitary gland and lead to combined pituitary hormone deficiency (CPHD). Patient We describe a male patient with extreme short stature, learning difficulties, anterior pituitary hypoplasia, secondary hypothyroidism and undetectable prolactin, growth hormone (GH) and insulin-like growth factor 1 (IGF1), with normal random cortisol. Design The POU1F1 coding region was amplified by PCR and sequenced; the functional consequence of the mutations was analysed by cell transfection and in vitro assays. Results Genetic analysis revealed compound heterozygosity for two novel putative loss of function mutations in POU1F1: a transition at position + 3 of intron 1 [IVS1+ 3nt(A> G)] and a point mutation in exon 6 resulting in a substitution of arginine by tryptophan (R265W). Functional analysis revealed that IVS1+ 3nt(A> G) results in a reduction in the correctly spliced POU1F1 mRNA, which could be corrected by mutations of the + 4, + 5 and + 6 nucleotides. Analysis of POU1F1 R265W revealed complete loss of function resulting from severely reduced protein stability. Conclusions Combined pituitary hormone deficiency in this patient is caused by loss of POU1F1 function by two novel mechanisms, namely aberrant splicing (IVS1+ 3nt (A> G) and protein instability (R265W). Identification of the genetic basis of CPHD enabled the cessation of hydrocortisone therapy without the need for further assessment for evolving endocrinopathy.