Mutation and gene copy number analyses of six pituitary transcription factor genes in 71 patients with combined pituitary hormone deficiency: identification of a single patient with LHX4 deletion.
BACKGROUND
Mutations of multiple transcription factor genes involved in pituitary development have been identified in a minor portion of patients with combined pituitary hormone deficiency (CPHD). However, copy number aberrations involving such genes have been poorly investigated in patients with CPHD.


OBJECTIVE
We aimed to report the results of mutation and gene copy number analyses in patients with CPHD.


METHODS
Seventy-one Japanese patients with CPHD were examined for mutations and gene copy number aberrations affecting POU1F1, PROP1, HESX1, LHX3, LHX4, and SOX3 by PCR-direct sequencing and multiplex ligation-dependent probe amplification. When a deletion was indicated, it was further studied by fluorescence in situ hybridization, oligoarray comparative genomic hybridization, and serial sequencing for long PCR products encompassing the deletion junction.


RESULTS
We identified a de novo heterozygous 522,009-bp deletion involving LHX4 in a patient with CPHD (GH, TSH, PRL, LH, and FSH deficiencies), anterior pituitary hypoplasia, ectopic posterior pituitary, and underdeveloped sella turcica. We also identified five novel heterozygous missense substitutions (p.V201I and p.H387P in LHX4, p.T63M and p.A322T in LHX3, and p.V53L in SOX3) that were assessed as rare variants by sequencing analyses for control subjects and available parents and by functional studies and in silico analyses.


CONCLUSIONS
The results imply the rarity of abnormalities affecting the six genes in patients with CPHD and the significance of the gene copy number analysis in such patients.
Department of Endocrinology and Metabolism, National Research Institute for Child Health and Development, 2-10-1 Ohkura, Setagaya, Tokyo 157-8535, Japan.
Sumito
S
Dateki
Maki
M
Fukami
Ayumi
A
Uematsu
Masayuki
M
Kaji
Manami
M
Iso
Makoto
M
Ono
Michiyo
M
Mizota
Susumu
S
Yokoya
Katsuaki
K
Motomura
Eiichi
E
Kinoshita
Hiroyuki
H
Moriuchi
Tsutomu
T
Ogata
D005260:Female; D017353:Gene Deletion; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D007018:Hypopituitarism / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D060850:LIM-Homeodomain Proteins; D008297:Male; D016133:Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics*
D018398:Homeodomain Proteins; C486362:LHX4 protein, human; D060850:LIM-Homeodomain Proteins; D014157:Transcription Factors