Additional file 1: of A 18p11.23-p11.31 microduplication in a boy with psychomotor delay, cerebellar vermis hypoplasia, chorioretinal coloboma, deafness and GH deficiency

Supplementary Methods. (DOCX 22 kb

Effects of Growth Hormone (GH) Therapy Withdrawal on Glucose Metabolism in Not Confirmed GH Deficient Adolescents at Final Height

<div><p>Context, objective</p><p>Growth hormone deficiency (GHD) is associated with insulin resistance and diabetes, in particular after treatment in children and adults with pre-existing metabolic risk factors. Our aims were. i) to evaluate the effect on glucose metabolism of rhGH treatment and withdrawal in not confirmed GHD adolescents at the achievement of adult height; ii) to investigate the impact of GH receptor gene genomic deletion of exon 3 (d3GHR).</p><p>Design, setting</p><p>We performed a longitudinal study (1 year) in a tertiary care center.</p><p>Methods</p><p>23 GHD adolescent were followed in the last year of rhGH treatment (T0), 6 (T6) and 12 (T12) months after rhGH withdrawal with fasting and post-OGTT evaluations. 40 healthy adolescents were used as controls. HOMA-IR, HOMA%β, insulinogenic (INS) and disposition (DI) indexes were calculated. GHR genotypes were determined by multiplex PCR.</p><p>Results</p><p>In the group as a whole, fasting insulin (p<0.05), HOMA-IR (p<0.05), insulin and glucose levels during OGTT (p<0.01) progressively decreased from T0 to T12 becoming similar to controls. During rhGH, a compensatory insulin secretion with a stable DI was recorded, and, then, HOMAβ and INS decreased at T6 and T12 (p<0.05). By evaluating the GHR genotype, nDel GHD showed a decrease from T0 to T12 in HOMA-IR, HOMAβ, INS (p<0.05) and DI. Del GHD showed a gradual increase in DI (p<0.05) and INS with a stable HOMA-IR and higher HDL-cholesterol (p<0.01).</p><p>Conclusions</p><p>In not confirmed GHD adolescents the fasting deterioration in glucose homeostasis during rhGH is efficaciously coupled with a compensatory insulin secretion and activity at OGTT. The presence of at least one d3GHR allele is associated with lower glucose levels and higher HOMA-β and DI after rhGH withdrawal. Screening for the d3GHR in the pediatric age may help physicians to follow and phenotype GHD patients also by a metabolic point of view.</p></div

Glucometabolic parameters of growth hormone deficient (GHD, group 1) children at the end of rhGH therapy (T0) and 6 (T6) and 12 (T12) months after rhGH withdrawal.

<p>Abbreviations. DI, disposition index; INS, insulinogenic index; AUC, area under the curve; ΔG0-30: delta glucose;; ΔI0-30: delta insulin; HDL-c, HDL-cholesterol; LDL-c, LDL-cholesterol; T-c, total-cholesterol; TG, triglycerides. Data are expressed as mean±SEM. The significance among the three measures (T0, T6 and T12) was calculated by Friedman test.</p

HOMA-IR, Insulinogenic (INS) and disposition (DI) index in GH deficient (GHD, group 1) and healthy (CS, group 2) adolescents with (Del, 27 subjects) and without (nDel, 31 subjects) the GH receptor (GHR) exon 3 deletion (d3GHR).

<p>GHD adolescents are evaluated in the last year of therapy (T0) and after six (T6) and twelve (T12) rhGH withdrawal. Data are expressed as mean±SEM. The significance among the three measures (T0, T6 and T12) was calculated by Friedman test. The significance between GHD and CS was calculated by Mann-Whitney U test. *p<0.05.</p

Functional characterization of novel GLI2 mutations identified in patients with Combined Pituitary Hormone Deficiency

<div>In a preliminary experiment, the GLI2ΔN-WT induced up to 10-fold reporter activity 306 (p<0.0001) compared with GLI2FL-WT (Supplementary figure 1) due to the absence of the 307 repressor domain, in line with the previous reports.<br></div><div><br></div><div>Primers used for the amplification of the complete coding region of GLI2.<br></div

Summary statistics of all the subjects included in the Italian GWAS.

<p>Summary statistics of all the subjects included in the Italian GWAS.</p

Manhattan plot of genotyped SNPs from logistic additive model.

<p>A) all samples, B) exposed samples.</p

Regional replication of Italian top signals in the Australian study for 5 out of the 20 regions.

<p>(1-tailed binomial test and meta-analysis).</p>a<p>NCBI36/hg18.</p>b<p>Italian study.</p>c<p>Australian study.</p

eQTL: <i>PVT1</i> and <i>MYC</i> gene-expression levels in blood cells across rs78941347 genotypes.

<p>eQTL: <i>PVT1</i> and <i>MYC</i> gene-expression levels in blood cells across rs78941347 genotypes.</p

Nested multivariate logistic regression models: 1) model 1, without genetic component; 2) model 2, with genetic component.

*<p>adjusted for age, gender and center of recruitment.</p><p>MODEL 1: AIC = 871.3, AUC = 0.76.</p><p>MODEL 2: AIC = 730.27, AUC = 0.86.</p