BACKGROUND: Pituitary development and GH secretion are orchestrated by multiple genes including GH1, GHRHR, GLI2, HESX1, LHX3, LHX4, PROP1, POU1F1, and SOX3. We aimed to assess their mutation frequency and clinical relevance in children with severe GH deficiency (GHD). METHODS: The Genetics and Neuroendocrinology of Short Stature International Study (GeNeSIS; Clinical Trial Registry Number: NCT01088412) was a prospective, open-label, observational research program for pediatric patients receiving GH treatment, conducted in 30 countries between 1999 and 2015. The study included a sub-study to investigate mutations in the genes listed above. PCR products from genomic blood cell DNA were analyzed by Sanger sequencing. DNA variants were classified as pathogenic according to the recommendations of the American College of Medical Genetics and Genomics. Demographic, auxologic, and endocrine data at baseline and during GH treatment were documented and related to the genotyping results. FINDINGS: The analysis comprised 917 patients. In 92 patients (10%) 33 mutations were found, 16 previously described and 17 novel (52%). Mutation carriers were significantly younger, shorter, and more slowly growing than non-carriers. In general, their peak values in GH stimulation tests were very low; however, in 15/77 (20%) patients with GH1, PROP1, and SOX3 mutations they were only moderately diminished (3-6 μg/L). Two patients with a GH1 mutation developed TSH deficiency and one ADH deficiency. Using logistic multi-regression analysis, significant indicators of a mutation were combined pituitary hormone deficiency, greater patient-parent height difference (SDS), low GH peak, and young age. Final height SDS gain in mutation carriers (mean ± SD 3.4 ± 1.4) was greater than in non-carriers (2.0 ± 1.4; P < .001) and in patients with non-GHD short stature. INTERPRETATION: DNA testing for mutations in children with severe GHD shows a positive finding in approximately 10%. Phenotypes of mutation carriers can be variable. The benefit for clinical practice justifies DNA testing as an important component in the diagnostic work-up of patients with severe GHD. FUND: Eli Lilly and Company, Indianapolis, IN, USA. ClinicalTrials.com registration: NCT01088412.

Department of Pediatrics 2nd Faculty of Medicine Charles University University Hospital Motol 5 Uvalu 84 150 06 Prague 5 Czech Republic

Department of Pediatrics Oregon Health and Science University Portland USA

Division of Pediatric Endocrinology and Diabetes Emory University School of Medicine 2015 Uppergate Dr Atlanta GA 30322 USA

Eli Lilly and Company Indianapolis USA

Eli Lilly and Company Werner Reimers Strasse 2 4 61352 Bad Homburg Germany

Eli Lilly and Company Windlesham GU20 6PH UK

Gordon Cutler Consultancy LLC Deltaville USA

Sorbonne Université Inserm UMR_S933 Département de Génétique Hôpital Trousseau AP HP 75012 Paris France

Sydney Children's Hospital Randwick Australia

University Hospital for Children and Adolescents University of Leipzig Liebigstrasse 20a 04103 Leipzig Germany

University Hospital for Children and Adolescents University of Leipzig Liebigstrasse 20a 04103 Leipzig Germany; Center of Child and Adolescent Medicine Justus Liebig University Feulgenstrasse 12 35392 Giessen Germany

University of Montreal and CHU Ste Justine Montreal Canada

Zobrazit více v PubMed

Wit J.M., Oostdijk W., Losekoot M., van Duyvenvoorde H.A., Ruivenkamp C.A.L., Kant S.G. Mechanisms in endocrinology: novel genetic causes of short stature. Eur J Endocrinol. 2016;174:R145-R173. PubMed

Giordano M. Genetic causes of isolated and combined pituitary hormone deficiency. Best Pract Res Clin Endocrinol Metab. 2016;30:679–691. PubMed

Alatzoglou K.S., Dattani M.T. Genetic causes and treatment of isolated growth hormone deficiency-an update. Nat Rev Endocrinol. 2010;6:562–576. PubMed

Pfaffle R., Klammt J. Pituitary transcription factors in the aetiology of combined pituitary hormone deficiency. Best Pract Res Clin Endocrinol Metab. 2011;25:43–60. PubMed

Fang Q., George A.S., Brinkmeier M.L. Genetics of combined pituitary hormone deficiency: roadmap into the genome era. Endocr Rev. 2016;37:636–675. PubMed PMC

Cerbone M., Dattani M.T. Progression from isolated growth hormone deficiency to combined pituitary hormone deficiency. Growth Horm IGF Res. 2017;37:19–25. PubMed

Dauber A., Rosenfeld R.G., Hirschhorn J.N. Genetic evaluation of short stature. J Clin Endocrinol Metab. 2014;99:3080–3092. PubMed PMC

de Rienzo F., Mellone S., Bellone S. Frequency of genetic defects in combined pituitary hormone deficiency: a systematic review and analysis of a multicentre Italian cohort. Clin Endocrinol (Oxf) 2015;83:849–860. PubMed

Blum W.F., Deal C., Zimmermann A.G. Development of additional pituitary hormone deficiencies in pediatric patients originally diagnosed with idiopathic isolated GH deficiency. Eur J Endocrinol. 2014;170:13–21. PubMed

Deal C., Hasselmann C., Pfäffle R.W. Associations between pituitary imaging abnormalities and clinical and biochemical phenotypes in children with congenital growth hormone deficiency. Data from an international observational study. Horm Res Paediatr. 2013;79:283–292. PubMed

Kamijo T., Phillips J.A., Ogawa M., Yuan L.-F., Y-F Shi, X-L Bao. Screening for growth hormone gene deletions in patients with isolated growth hormone deficiency. J Pediatr. 1991;118:245–248. PubMed

Woods K.S., Cundall M., Turton J. Over- and underdosage of SOX3 is associated with infundibular hypoplasia and hypopituitarism. Am J Hum Genet. 2005;76:833–849. PubMed PMC

Jones A.C., Austin J., Hansen N. Optimal temperature selection for mutation detection by denaturing HPLC and comparison to single-stranded conformation polymorphism and heteroduplex analysis. Clin Chem. 1999;45:1133–1140. PubMed

Richards S., Aziz N., Bale S. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med. 2015;17:405–424. PubMed PMC

Arnhold I.J.P., França M.M., Carvalho L.R., Mendonca B.B., Jorge A.A.L. Role of GLI2 in hypopituitarism phenotype. J Mol Endocrinol. 2015;54:R141–R150. PubMed

Cooper D.N., Youssoufian H. The CpG dinucleotide and human genetic disease. Hum Genet. 1988;78:151–155. PubMed

Lek M., Karczewski K.J., Minikel E.V. Analysis of protein-coding genetic variation in 60,706 humans. Nature. 2016;536:285–291. PubMed PMC

Bertko E., Klammt J., Dusatkova P. Combined pituitary hormone deficiency due to gross deletions in the POU1F1 (PIT-1) and PROP1 genes. J Hum Genet. 2017;62:755–762. PubMed PMC

Hemchand K., Anuradha K., Neeti S. Entire prophet of Pit-1 (PROP-1) gene deletion in an Indian girl with combined pituitary hormone deficiencies. J Pediatr Endocrinol Metab. 2011;24:579–580. PubMed

Ryther R.C., Flynt A.S., Harris B.D., Phillips J.A.I., Patton J.G. GH1 splicing is regulated by multiple enhancers whose mutation produces a dominant-negative GH isoform that can be degraded by allele-specific small interfering RNA (siRNA) Endocrinology. 2004;145:2988–2996. PubMed

Babu D., Mellone S., Fusco I. Novel mutations in the GH gene (GH1) uncover putative splicing regulatory elements. Endocrinology. 2014;155:1786–1792. PubMed

Desmet F.-O., Hamroun D., Lalande M., Collod-Béroud G., Claustres M., Béroud C. Human splicing finder: an online bioinformatics tool to predict splicing signals. Nucleic Acids Res. 2009;37:e67. PubMed PMC

Fofanova O.V., Evgrafov O.V., Polyakov A.V., Poltaraus A.B., Peterkova V.A., Dedov I.I. A novel IVS2 −2A>T splicing mutation in the GH-1 gene in familial isolated growth hormone deficiency Type II in the spectrum of other splicing mutations in the Russian population. J Clin Endocrinol Metab. 2003;88:820–826. PubMed

Gucev Z., Tasic V., Saranac L. A novel GH1 mutation in a family with isolated growth hormone deficiency type II. Horm Res Paediatr. 2012;77:200–204. PubMed

Takahashi I., Takahashi T., Komatsu M., Sato T., Takada G. An exonic mutation of the GH-1 gene causing familial isolated growth hormone deficiency type II. Clin Genet. 2002;61:222–225. PubMed

Alatzoglou K.S., Turton J.P., Kelberman D. Expanding the spectrum of mutations in GH1 and GHRHR: genetic screening in a large cohort of patients with congenital isolated growth hormone deficiency. J Clin Endocrinol Metab. 2009;94:3191–3199. PubMed

Wang Q., Diao Y., Xu Z. Identification of a novel splicing mutation in the growth hormone (GH)-releasing hormone receptor gene in a Chinese family with pituitary dwarfism. Mol Cell Endocrinol. 2009;313:50–56. PubMed

Maheshwari H.G., Silverman B.L., Dupuis J., Baumann G. Phenotype and genetic analysis of a syndrome caused by an inactivating mutation in the growth hormone-releasing hormone receptor: dwarfism of sindh. J Clin Endocrinol Metab. 1998;83:4065–4074. PubMed

Pfaeffle R.W., Savage J.J., Hunter C.S. Four novel mutations of the LHX3 gene cause combined pituitary hormone deficiencies with or without limited neck rotation. J Clin Endocrinol Metab. 2007;92:1909–1919. PubMed

Sobrier M.-L., Netchine I., Heinrichs C. Alu-element insertion in the homeodomain of HESX1 and aplasia of the anterior pituitary. Hum Mutat. 2005;25:503. PubMed

Ohta K., Nobukuni Y., Mitsubuchi H. Mutations in the Pit-1 gene in children with combined pituitary hormone deficiency. Biochem Biophys Res Commun. 1992;189:851–855. PubMed

McLennan K., Jeske Y., Cotterill A. Combined pituitary hormone deficiency in Australian children: clinical and genetic correlates. Clin Endocrinol (Oxf) 2003;58:785–794. PubMed

Dattani M.T., Martinez-Barbera J.-P., Thomas P.Q. Mutations in the homeobox gene HESX1/Hesx1 associated with septo-optic dysplasia in human and mouse. Nat Genet. 1998;19:125–133. PubMed

Durmaz B., Cogulu O., Dizdarer C., Stobbe H., Pfaeffle R., Ozkinay F. A novel homozygous HESX1 mutation causes panhypopituitarism without midline defects and optic nerve anomalies. J Pediatr Endocrinol Metab. 2011;24:779–782. PubMed

McCabe M.J., Alatzoglou K.S., Dattani M.T. Septo-optic dysplasia and other midline defects: the role of transcription factors: HESX1 and beyond. Best Pract Res Clin Endocrinol Metab. 2011;25:115–124. PubMed

Kato Y., Kimoto F., Susa T., Nakayama M., Ishikawa A., Kato T. Pituitary homeodomain transcription factors HESX1 and PROP1 form a heterodimer on the inverted TAAT motif. Mol Cell Endocrinol. 2010;315:168–173. PubMed

Dusatkova P., Pfaffle R., Brown M.R. Genesis of two most prevalent PROP1 gene variants causing combined pituitary hormone deficiency in 21 populations. Eur J Hum Genet. 2015;24:415–420. PubMed PMC

Böttner A., Keller E., Kratzsch J. PROP1 mutations cause progressive deterioration of anterior pituitary function including adrenal insufficiency: a longitudinal analysis. J Clin Endocrinol Metab. 2004;89:5256–5265. PubMed

Deladoey J., Fluck C., Buyukgebiz A. “Hot spot” in the PROP1 gene responsible for combined pituitary hormone deficiency. J Clin Endocrinol Metab. 1999;84:1645–1650. PubMed

Grimberg A., Divall S.A., Polychronakos C. Guidelines for growth hormone and insulin-like growth factor-I treatment in children and adolescents. Growth hormone deficiency, idiopathic short stature, and primary insulin-like growth factor-I deficiency. Horm Res Paediatr. 2016;86:361–397. PubMed

Dattani M.T. Growth hormone deficiency and combined pituitary hormone deficiency. Does the genotype matter? Clin Endocrinol (Oxf) 2005;63:121–130. PubMed

Obermannova B., Pfaeffle R., Zygmunt-Gorska A. Mutations and pituitary morphology in a series of 82 patients with PROP1 gene defects. Horm Res Paediatr. 2011;76:348–354. PubMed

Ranke M.B., Lindberg A. Predicting growth in response to growth hormone treatment. Growth Horm IGF Res. 2009;19:1–11. PubMed

Clayton P.E., Cuneo R.C., Juul A., Monson J.P., Shalet S.M., Tauber M. Consensus statement on the management of the GH-treated adolescent in the transition to adult care. Eur J Endocrinol. 2005;152:165–170. PubMed

Wagner J.K., Eble A., Hindmarsh P.C., Mullis P.E. Prevalence of human GH-1 gene alterations in patients with isolated growth hormone deficiency. Pediatr Res. 1998;43:105–110. PubMed

Kamijo T., Phillips J.A. Detection of molecular heterogeneity in GH-1 gene deletions by analysis of polymerase chain reaction amplification products. J Clin Endocrinol Metab. 1992;74:786–789. PubMed

Cogan J.D., Ramel B., Lehto M. A recurring dominant negative mutation causes autosomal dominant growth hormone deficiency—a clinical research center study. J Clin Endocrinol Metab. 1995;80:3591–3595. PubMed

Binder G., Keller E., Mix M. Isolated GH deficiency with dominant inheritance: new mutations, new insights. J Clin Endocrinol Metab. 2001;86:3877–3881. PubMed

Hayashi Y., Yamamoto M., Ohmori S., Kamijo T., Ogawa M., Seo H. Inhibition of growth hormone (GH) secretion by a mutant GH-I gene product in neuroendocrine cells containing secretory granules. An implication for isolated GH deficiency inherited in an autosomal dominant manner. J Clin Endocrinol Metab. 1999;84:2134–2139. PubMed

Vivenza D., Guazzarotti L., Godi M. A novel deletion in the GH1 gene including the IVS3 branch site responsible for autosomal dominant isolated growth hormone deficiency. J Clin Endocrinol Metab. 2006;91:980–986. PubMed

Wajnrajch M.P., Gertner J.M., Harbison M.D., Chua S.C., Leibel R.L. Nonsense mutation in the human growth hormone-releasing hormone receptor causes growth failure analogous to the little (lit) mouse. Nat Genet. 1996;12:88–90. PubMed

Salvatori R., Fan X., Phillips J.A., Prince M., Levine M.A. Isolated growth hormone (GH) deficiency due to compound heterozygosity for two new mutations in the GH-releasing hormone receptor gene. Clin Endocrinol (Oxf) 2001;54:681–687. PubMed

Sobrier M.-L., Brachet C., Vié-Luton M.-P. Symptomatic heterozygotes and prenatal diagnoses in a nonconsanguineous family with syndromic combined pituitary hormone deficiency resulting from two novel LHX3 mutations. J Clin Endocrinol Metab. 2012;97:E503–E509. PubMed

Riepe F.G., Partsch C.J., Blankenstein O., Monig H., Pfaffle R.W., Sippell W.G. Longitudinal imaging reveals pituitary enlargement preceding hypoplasia in two brothers with combined pituitary hormone deficiency attributable to PROP1 mutation. J Clin Endocrinol Metab. 2001;86:4353–4357. PubMed

Paracchini R., Giordano M., Corrias A. Two new PROP1 gene mutations responsible for compound pituitary hormone deficiency. Clin Genet. 2003;64:142–147. PubMed

Duquesnoy P., Roy A., Dastot F. Human Prop-1: cloning, mapping, genomic structure. Mutations in familial combined pituitary hormone deficiency. FEBS Lett. 1998;437:216–220. PubMed

Vallette-Kasic S., Barlier A., Teinturier C. PROP1 gene screening in patients with multiple pituitary hormone deficiency reveals two sites of hypermutability and a high incidence of corticotroph deficiency. J Clin Endocrinol Metab. 2001;86:4529–4535. PubMed

Wu W., Cogan J.D., Pfaffle R.W. Mutations in PROP1 cause familial combined pituitary hormone deficiency. Nat Genet. 1998;18:147–149. PubMed

Laumonnier F., Ronce N., Hamel B.C.J. Transcription factor SOX3 is involved in X-linked mental retardation with growth hormone deficiency. Am J Hum Genet. 2002;71:1450–1455. PubMed PMC