S48 Posters / Growth Hormone & IGF Research 22S1 (2014) S25–S52

Results: Plasma PAPP-A increased abruptly upon heparin-admin-istration with mean concentrations pre-PCI, post-PCI, day 1 and day 2 of 13 .0 (11 .2; 15 .2), 14 .8 (13 .1; 16 .8), 1 .03 (0 .902; 1 .18) and 1 .08 (0 .916; 1 .28) mg/l, respectively (p<0 .0001) . Concentrations of IGFBP-4, CT-IGFBP-4 and NT-IGFBP-4 pre-PCI were 154 (142; 166), 53 .1 (47 .2; 59 .7) and 136 (122; 150) mg/l, and levels were unaltered post-PCI . Concentrations increased on day 1 by 63 (43;87)%, 69 (36;110)%, and 47 (21;79)%, respectively (p<0 .0001), i .e . at a time point when PAPP-A levels had normalized .Conclusion: Plasma IGFBP-4-fragments levels are not affected by heparin treatment . Thus, they may possess the potential to serve as biomarkers in ACS .

GP4-3Epigenetic analyses of the insulin-like growth factor binding protein 1 gene in diabetes and diabetic nephropathy

T . Gu1, H .F . Gu1,2, H . Falhammar1,2, A . Hilding1, L .K . Sjöholm3, C .-G . Östenson1,2, T .J . Ekström3, K . Brismar1,2 . 1Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden, 2Endocrinology, Metabolism and Diabetes, Karolinska Hospital, Stockholm, Sweden, 3Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden

Background: High levels of circulating insulin-like growth factor binding protein-1 (IGFBP-1) are associated with type 1 diabetes (T1D), while low serum IGFBP-1 levels are associated with type 2 diabetes (T2D) .Aim: To evaluate the epigenetic effects of the IGFBP1 gene in T1D and T2D .Subjects and methods: A total of 753 Swedish subjects including normal glucose tolerance (NGT), T1D and T2D were included in our studies . IGFBP1 methylation levels in genomic DNA extracted from peripheral blood were analyzed by bisulfite pyrosequenc-ing . Serum IGFBP-1 levels were measured by an in-house radio-immunoassay .Results: The percentages of DNA methylation of the IGFBP1 gene were higher in both newly diagnosed and treated T2D patients with a mean diabetes duration of three years compared with subjects with NGT (19 .8% and 20 .2% vs . 16 .9%, P<0 .001 for both) . Serum levels of IGFBP-1 in newly diagnosed and in treated T2D patients were lower compared with healthy individuals (18 mg/l both vs . 24 mg/l, P=0 .011, P<0 .001) . The IGFBP1 methylation but not serum IGFBP-1 levels in T2D were independent of BMI . Newly diagnosed patients with a family history of diabetes (FHD) had higher IGFBP1 methylation levels than those without FHD (20 .3% vs . 18 .6%, P=0 .017) . Unlike T2D, the percentages of DNA methyla-tion in the IGFBP1 gene were decreased in T1D patients with the mean diabetes duration of 29 years, compared to the subjects with NGT (15 .6% vs . 16 .8%, P<0 .001) . Serum IGFBP-1 levels in T1D patients were higher compared to non-diabetic individuals (31 mg/l vs . 24 mg/l, P=0 .003) .Conclusions: We provide the first evidence that decreased DNA methylation levels in the IGFBP1 gene are associated with T1D, while increased IGFBP1 DNA methylation levels are associated with T2D . Data also implicate that increased circulating IGFBP-1 levels are features of T1D, while decreased IGFBP-1 serum levels are features of T2D with a short duration .

GP4-4SGA short stature bearing with a novel nonsense mutation (p .W1219X) in the IGF1R gene

M . Fujimoto1, Y . Kawashima1, T . Hamajima2, N . Miyahara1, R . Nishimura1, K . Hanaki1, S . Kanzaki1 . 1Pediatrics and Perinatology, Tottori University, Yonago, Japan, 2Pediatric Endocrinology and Metabolism, Aichi Children’s Health and Medical Center, Obu, Japan

Introduction: The insulin-like growth factor (IGF)-I receptor (IGF1R) is widely expressed in various tissues . The IGF plays key roles in fetal and postnatal growth through the IGF1R . Heterozygous IGF1R mutations presenting with intrauterine and postnatal growth retardation have been reported in over 10 families . We previously reported that a heterozygous nonsense mutation (p .Q1220X) led to decrease IGF1R protein expression through endoplasmic-reticulum-associated protein degradation (ERAD) mechanism, resulted in growth failure . Recently, we identified another case with SGA short stature bearing a novel heterozygous nonsense mutation (p .W1219X) in the IGF1R gene .Case: Patient was a 3-year-old Japanese girl, who was born at 40 weeks of gestation, with a birth weight of 2,110 g (-3 .0 SD), birth height of 44 .3 cm (-2 .8 SD) . She had no family history of short stature . At the age of 2 years and 5 months, her basal serum GH level was high (10 .5 ng/ml), but IGF-1 level showed normal [185 ng/ml (normal range: 32-213)] . At the age of 3 years, she presented with missing catch-up growth; her height was 82 .9 cm (-3 .1 SD); head circumference, 46 .0 cm (-1 .6 SD) . Her karyotype was normal (46, XX) . We performed the gene analysis of the IGF1R gene by direct sequencing and MLPA . We identified a novel heterozygous nonsense mutation (p .W1219X) in the IGF1R gene .Discussion and conclusion: This mutation causes premature stop codon within exon 21 of the IGF1R gene, and is considered to results in the absence of the carboxyl terminal fragment of IGF1R after the mutation site . We consider that ERAD might cause a reduced IGF1R protein on cells with the mutated IGF1R . We are going to test whether this mutation results in the degradation of the mutant IGF1R protein through ERAD . Our results provide important new information on the IGF1R mutation in SGA short stature .

GP4-5Circulating levels of insulin-like growth factor-1(IGF-I)/insulin-like growth factor binding protein-3 (IGFBP-3) in obese patients: the state of type 2 diabetes

M .S . Kim, S .-Y . Kim, D .-Y . Lee . Chonbuk National University Medical School, Jeonju-si, Korea, Republic of

Introduction: The aim of this study was to investigate the hypothesis that serum IGF-I and IGFBP-3 levels are possible related with the obesity and the diabetes .Methods: We included 111 adolescents aged 10 to 18 years (63 girls and 48 boys) . They were classified into 3 groups according to the results of oral glucose tolerance test (OGTT) and other clinical/laboratory findings, and; normal glucose tolerance group (NGT), n=48 (non-obese, n=41; obese, n=7); impaired glucose tolerance (IGT) group, n=16 (non-obese, n=10; obese, n=6) ; type 2 diabetes (T2DM) group, n=45 (non-obese, n=23; obese, n=22) . We performed laboratory tests .Results:1) Serum IGF-I and IGFBP-3 levels were significantly higher in

T2DM group than NGT group and IGT group . Serum IGF-I and IGFBP-3 had positive correlation with fasting plasma glucose (FPG), HbA1c, c-peptide, and HOMA-IR . Also serum IGFBP-3 level showed the positive correlation with BMI and lipid profile .

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2) Serum IGF-I level was no significantly different between obese and non-obese groups in all subjects, NGT and T2DM groups . It was correlated with serum IGFBP-3 and age in NGT group . And it was showed positive correlation with HbA1c and FPG, OGTT glucose levels but negative correlation with age in T2DM group .

3) Serum IGFBP-3 level was significantly increased in obese than non-obese groups in all subjects . But it was no significantly different between obese and non-obese groups in NGT and T2DM groups . It had positively associated with age and BMI in NGT group . In T2DM group, it had positive correlation with FPG, HbA1c and lipid profile but no correlation with IGF-I level .

Conclusion: Serum IGF-I and IGFBP-3 levels were significantly elevated in adolescents with type 2 diabetes and correlated with blood glucose related factors . Serum IGFBP-3 level was associ-ated with lipid profile . These findings suggest that IGFBP-3 may related glycemic control and/or obesity in adolescents .

GP4-6Effects of GH/IGF-I axis on retinal vascular morphology: retinal vascular morphology in Laron syndrome

H .T . Sekeroglu1, S . Kadayifcilar1, B . Kasim1, U . Arslan2, Z .A . Ozon3 . 1Department of Ophthalmology, Hacettepe University, Faculty of Medicine, Ankara, Turkey, 2Department of Biostatistics, Hacettepe University, Faculty of Medicine, Ankara, Turkey, 3Department of Pediatrics Division of Endocrinology, Hacettepe University, Faculty of Medicine, Ankara, Turkey

Introduction: IGF-I is suggested to be a critical mediator of reti-nal neovascularization with other growth factors . We aimed to study retinal vascular characteristics of patients with Laron syn-drome (LS) as a genetic model of severe IGF-I deficiency before and after rhIGF-I/IGFBP-3 treatment, and to compare them with retinal vascular morphology of healthy age matched controls .Methods: A total of 28 subjects (11 LS, and 17 age-matched healthy controls) were enrolled . All subjects underwent oph-thalmological examination including digital fundus imaging . Patients with LS received combined rhIGF-I/rhIGFBP-3, 1-2 mg/kg/d in a single dose subcutaneously for one (5 patients) or two (6 patients) years, and digital fundus imaging was repeated at the end of treatment . Branching characteristics and tortuosity index of retinal vessels were studied in all subjects by a blinded retina specialist . Pre- and post-treatment findings were compared with each other and with controls .Results: Number of branching points was significantly lower in LS in comparison to controls (12 .73±3 .41, and 17 .47±5 .82 respectively, p=0 .012) . This difference persisted after treatment (12 .09±2 .66 posttreatment LS vs controls, p=0 .017) . Tortuosity indices of nasal arteries (NA) were significantly less in LS than that of controls (upper NA 1 .07±0 .04 and 1 .12±0 .06 respectively p= 0 .022; lower NA 1 .07±0 .03 and 1 .13±0 .07 respectively, p=0 .004) . This difference also persisted following treatment (p<0 .05) . Remaining vessels didnot differ in tortuosity index . No significant difference was observed pre- and post-treatment in patients with LS, neither in tortuosity index nor number of branching points .Conclusion: Retinal vascular development may be adversely affected in the setting of severe IGF-1 deficiency suggesting a major role for GH/IGF-I axis antenatally . Resolution of IGF defi-ciency after birth using systemic administration of IGF-1 how-ever, may not reverse these changes, suggesting that IGF-I may be necessary but insufficient by itself for postnatal angiogenesis .

PO4-1Alpha-fetoprotein at birth: an early marker of circulating IGF-I in preterm infants

A . Kistner1,2, G . Hellgren3, I . Hansen Pupp4, D . Ley4, C . Löfqvist1, A . Hellström1 . 1The Sahlgrenska Center for Pediatric Ophthalmology Research, Institute of Neuroscience and Physiology, Sahlgrenska Academy, Gothenburg, Sweden, 2Department of Molecular Medicine and Surgery, Karolinska institutet, Stockholm, Sweden, 3Department of Pediatrics, Institute of Clinical Sciences, Sahlgrenska Academy, Gothenburg, Sweden, 4Department of Pediatrics, Institution of Clinical Sciences, Lund University and Skane University Hospital, Lund, Sweden

Background: Alpha-fetoprotein is a serum growth factor involved in liver development in humans .Objectives: We hypothesized that alpha-fetoprotein levels at birth might be of importance for later postnatal IGF-I levels .Methods: This study included 100 preterm infants [born at <30 gestational weeks (GW)] . Alpha-fetoprotein was determined 0-48 hours after birth, and insulin like growth factor-1 (IGF-I) was measured at postmenstrual age (PMA) 30-33 .9 weeks .Results: Boys born below -2 birth-weight standard deviation score (SDS) had lower alpha-fetoprotein levels compared to that of boys born with ≥-2 birth-weight SDS (95% CI) [338 (225; 409) mg/L, n=12, vs . 514 (427; 619) mg/L, n=31, respectively, P=0 .006, adjusted for GW] . In multiple regression analysis with postnatal IGF-I as a dependent variable, the addition of alpha-fetoprotein to IGF-I at birth, birth weight SDS, gestational age, and gender increased the R2 from 27 .5% to 34% (P=0 .003) .Conclusions: We found an association between neonatal levels of alpha-fetoprotein, which may reflect a higher proliferative potential in hepatocytes, and postnatal levels of IGF-I in preterm infants .

PO4-2Variations in growth in relation to IGF-I levels in children born very preterm and target height influences from birth to 3-years of age

A . Kistner1,2, G . Hellgren3, A . Niklasson3, J . Sigurdsson3, C . Löfqvist1, A . Hellström1 . 1The Sahlgrenska Center for Pediatric Ophthalmology Research, Institute of Neuroscience and Physiology, Sahlgrenska Academy, Gothenburg, Sweden, 2Department of Molecular Medicine and Surgery, Karolinska institutet, Stockholm, Sweden, 3Department of Pediatrics, Institute of Clinical Sciences, Sahlgrenska Academy, Gothenburg, Sweden

Background: Insulin-like growth factor-1 (IGF-1) and IGF binding protein-1 (IGFBP-1) in preterms may reflect growth . We aimed to investigate growth in relation to IGF-I and IGFBP-1 as well as target height (TH) standard deviation scores (SDS) influences in preterms from birth to 3 years of age .Design: 52 children born very preterm (GA <32 weeks) were stud-ied from birth to 3 years of age with continuously anthropometry . Weekly blood sampling for IGF-I and IGFBP-1 from birth to 40 postmenstrual weeks (PMA) was performed . IGF-I was measured at corrected age (years) 0 .7 (n=30) and 3 (n=48) . The group was divided according to THSDS ≥0 (high) or <0 (low) .Results: Mean IGF-I and IGFBP-1 at PMA 30-37 weeks and caloric intake correlated with growth increment (gram/day) during this period (r=0 .34, P<0 .05, r=-0 .45, P<0 .001, r=0 .45, P<0 .01, respec-tively) . Children with high THSDS were heavier (in weight SDS) and taller (height SDS) at (years) 1 .5 (p<0 .001) and 3 (p<0 .01) compared with low, not seen at birth or at 40 weeks PMA . IGF-I was similar in the two TH groups at (years) 0 .7 and 3 and cor-related with weight increment (gram/month), at 0 .4-1 .5 years (r=0 .51, P<0 .01) and at 1 .5-3 years (r=0 .44, P=0 .01) . In regression