1. Funded by the EC, FP 6, Contract No. 016181 (FOOD) Percentiles of markers of insulin resistance in pre-pubertal normal-weight European children from the IDEFICS cohort ECOG congress, Salzburg, November 14, 2014 Jenny Peplies Leibniz Institute for Prevention Research and Epidemiology − BIPS on behalf of the IDEFICS Consortium

2. 2 Aim  To present age- and sex-specific reference values of markers of insulin resistance (IR) for preadolescent children: o Insulin o Glucose o HOMA-IR (homeostasis model assessment to quantify IR). o HbA1c (glycosylated haemoglobin)  To provide a basis for early detection of metabolic diseases

3. Study population 3  Children who participated in at least one wave of the IDEFICS surveys  Normal weight and without diabetes (self-reported)  3 to 10.9 years old  Availability of biological markers  7.074 children (of a total of 18.745 IDEFICS children)

4. 4 Blood samples and biomarkers  Blood withdrawal after an overnight fast  Exclusion of some children due to unclear fasting status  Fractioning of blood at local survey centres  Freezing of blood fractions at -80ºC and shipping on dry ice to a central lab  Standardised laboratory measurements o Glucose by point-of-care testing (Cholestech LDX*) o Insulin and HbA1c in central laboratory (accredited according to ISO 15189) * Panz et al.: Cardiovasc J S Afr 2005; 16/2: 112-117.

5. Statistical analysis 5  Percentile curves were calculated as a function of age stratified by sex using the ‘Generalised Additive Model for Location Scale and Shape’ (GAMLSS).  The final models used o Box–Cox t (BCT) distribution for insulin and HOMA-IR (modelling μ as a cubic spline depending on age, log(σ) as a linear function of age, and ν and τ as constants). o Lognormal distribution for glucose (considering μ as a linear function of age and log(σ) as a constant). o BCT distribution for HbA1c (modelling μ as a cubic spline depending on age, log(σ) and ν as a linear function of age and τ as constant).

6. Insulin percentiles 6 Reference limit for adults: <174 pmol/l (<25 mIU/l)* *Roche Diastics. Insulin immunoassay package insert. ms_12017547122

7. HOMA percentiles 7 Reference limit for adults: < 2* * Keskin et al.: Pediatrics. 2005 Apr;115(4)..

8. Glucose percentiles 8 Reference limit for adults: 5.6 mmol/l (100 mg/dl)* * Alberti et al.: Lancet. 2005 Sep 24-30;366(9491):1059-62.

9. HbA1c percentiles 9 Reference range for adults: 20 - 42 mmol/mol (4,0 – 6,0 %) * International Expert Committee. Diabetes Care. 2009. Jul;32(7):1327-34..

10. Results  Levels of insulin, fasting glucose and HOMA-IR show a continuously increasing trend with age while HbA1c shows an upward trend only beyond the age of 8 years.  Insulin and HOMA-IR values are higher in girls of all age groups, whereas glucose values are slightly higher in boys and HbA1c values don’t differ between the sexes.  Reference values for adults are only of limitted use for paediatric practise as most biological markers in children rise with age. 10

11. Strengths and limitations Strengths  Large prospective European study, standardised survey protocol, focus on young children Limitations  No assessment of pubertal stage 11

12. Conclusions  Reference percentiles for markers of insulin resistance were modeled on the basis of the so far largest dataset of healthy preadolescent children. 12

13. 13 Thank you for your attention! Also many thanks to  The IDEFICS children and their parents for their participation  Timm Intemann for his enduring help with the GAMLSS modelling  Ina Alvarez for technical support  My co-authors David Jiménez-Pavón, Savvas C. Savva, Christoph Buck, Kathrin Günther, Arno Fraterman, Paola Russo, Licia Iacoviello, Toomas Veidebaum, Michael Tornaritis, Stefaan De Henauw, Staffan Mårild, Dénes Molnár, Luis A. Moreno and Wolfgang Ahrens  The European Community and the Volkswagen Foundation for funding www.idefics.eu

14. Sensitivity analysis HOMA-IR 14 Children with normal weight (defined by Cole, WHO or CDC) and children of all weight groups

15. 15 Sensitivity analysis glucose Children with normal weight (defined by Cole, WHO or CDC) and children of all weight groups