title: Human skin fibroblasts as a model of growth hormone (GH) action in GH receptor-positive Laron's syndrome.
abstract: Congenital GH insensitivity (Laron's syndrome, LS) is often associated with a dysfunctional GH receptor (GHR) causing complete insensitivity to GH and absent serum GH-binding protein (GHBP). However, a proportion of children with LS have normal GHBP levels. We have identified four girls from two families with this condition (height SD score, -3.4 to -6.8) and undertaken studies on 1) their GHR genes and 2) their GH responses in cultured skin fibroblasts to define the etiology of their GH insensitivities. No GHR gene mutations were identified in one family. In the other family, the affected siblings, an unaffected brother, and the father were heterozygous for a point mutation within exon 6 (D152H). In addition, use of intron 9 haplotypes to determine linkage to the GHR gene implied inheritance of different maternal GHR alleles in the two affected girls of the latter family. It is unlikely, therefore, that the D152H mutation alone could account for the LS phenotype. End points of GH action [DNA synthesis, insulin-like growth factor-binding protein-3 (IGFBP-3) messenger RNA (mRNA) and peptide production] in skin fibroblast cultures established from three of the LS subjects and four normal children were examined. Whereas normal fibroblasts incorporated [3H]thymidine dose dependently in response to 10-1000 ng/ml GH (increment at 1000 ng/ml, 77 +/- 19%), LS fibroblasts failed to respond significantly above basal levels (P < 0.01). In normal fibroblasts, IGFBP-3 mRNA and peptide increased maximally at 48 h in response to 200 ng/ml GH, as determined by ribonuclease protection assay, Western ligand blotting, and RIA. In comparison, LS fibroblasts produced significantly less IGFBP-3 peptide than normal fibroblasts in response to GH, whereas IGFBP-3 mRNA failed to increase above basal levels. These studies have shown that 1) cultured human skin fibroblasts can be used to define the end points of GH action; 2) fibroblast cultures from the LS children show absent or reduced responses to GH; and 3) GH insensitivity in these children does not appear to be caused exclusively by GHR mutations, but is probably due to dysfunctional GHR signalling. Such patients may prove particularly important to elucidation of the key events in GH signaling.
authors.affiliation: Department of Medicine, University of Manchester, United Kingdom.
authors.forename: J S
authors.initials: JS
authors.lastname: Freeth
authors.forename: R M
authors.initials: RM
authors.lastname: Ayling
authors.forename: A J
authors.initials: AJ
authors.lastname: Whatmore
authors.forename: P
authors.initials: P
authors.lastname: Towner
authors.forename: D A
authors.initials: DA
authors.lastname: Price
authors.forename: M R
authors.initials: MR
authors.lastname: Norman
authors.forename: P E
authors.initials: PE
authors.lastname: Clayton
mesh_terms: D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000187:drug effects; D006130:Growth Disorders / Q000235:genetics* / Q000378:metabolism; D013006:Growth Hormone / Q000172:deficiency / Q000494:pharmacology*; D006801:Humans; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D012333:RNA, Messenger / Q000032:analysis; D011986:Receptors, Somatotropin / Q000032:analysis* / Q000235:genetics; D013577:Syndrome; D013936:Thymidine / Q000378:metabolism
chemical_list: D018972:Insulin-Like Growth Factor Binding Protein 3; D012333:RNA, Messenger; D011986:Receptors, Somatotropin; D013006:Growth Hormone; D013936:Thymidine