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  • Original Article
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Pediatrics

Mutation screen of the SIM1 gene in pediatric patients with early-onset obesity

International Journal of Obesity volume 38, pages 1000–1004 (2014)Cite this article

Subjects

Abstract

Background:

The transcription factor SIM1 (Single-minded 1) is involved in the control of food intake and in the pathogenesis of obesity. In mice, Sim1 is involved in the development of the paraventricular nucleus, and Sim1 deficiency leads to severe obesity and hyperphagia. In humans, chromosomal abnormalities in the SIM1 gene region have been reported in obese individuals. Furthermore, recent data also suggest that loss-of-function point mutations in SIM1 are responsible for SIM1 haplo-insufficiency that is involved in causing human obesity. In this study, we therefore wanted to expand the evidence regarding the involvement of SIM1 mutations in the pathogenesis of severe early-onset obesity.

Methods:

We screened 561 severely overweight and obese children and adolescents and 453 lean adults for mutations in the coding region of the SIM1 gene. Mutation screening in all patients and lean individuals was performed by high-resolution melting curve analysis combined with direct sequencing. To evaluate the effect of the mutations on SIM1 transcriptional activity, luciferase reporter assays were performed.

Results:

Mutation analysis identified four novel nonsynonymous coding variants in SIM1 in four unrelated obese individuals: p.L242V, p.T481K, p.A517V and p.D590E. Five synonymous variants, p.P57P, p.F93F, p.I183I, p.V208V and p.T653T, were also identified. Screening of the lean control population revealed the occurrence of four other rare SIM1 variants: p.G408R, p.R471P, p.S492P and p.S622F. For variants p.T481K and p.A517V, which were found in obese individuals, a decrease in SIM1 transcriptional activity was observed, whereas the transcriptional activity of all variants found in lean individuals resembled wild type.

Conclusions:

In this study, we have demonstrated the presence of rare SIM1 variants in both an obese pediatric population and a population of lean adult controls. Further, we have shown that functional in vitro analysis of SIM1 variants may help in distinguishing benign variants of no pathogenic significance from variants which contribute to the obesity phenotype.

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References

  1. Michaud JL, Rosenquist T, May NR, Fan CM . Development of neuroendocrine lineages requires the bHLH-PAS transcription factor SIM1. Genes Dev 1998; 12: 3264–3275.

    Article  CAS  Google Scholar 

  2. Palkovits M, Mezey E, Eskay RL . Pro-opiomelanocortin-derived peptides (ACTH/beta-endorphin/alpha-MSH) in brainstem baroreceptor areas of the rat. Brain Res 1987; 436: 323–338.

    Article  CAS  Google Scholar 

  3. Mountjoy KG, Mortrud MT, Low MJ, Simerly RB, Cone RD . Localization of the melanocortin-4 receptor (MC4-R) in neuroendocrine and autonomic control circuits in the brain. Mol Endocrinol 1994; 8: 1298–1308.

    CAS  Google Scholar 

  4. Michaud JL, Boucher F, Melnyk A, Gauthier F, Goshu E, Levy E et al. Sim1 haploinsufficiency causes hyperphagia, obesity and reduction of the paraventricular nucleus of the hypothalamus. Hum Mol Genet 2001; 10: 1465–1473.

    Article  CAS  Google Scholar 

  5. Sims JS, Lorden JF . Effect of paraventricular nucleus lesions on body weight, food intake and insulin levels. Behav Brain Res 1986; 22: 265–281.

    Article  CAS  Google Scholar 

  6. Tolson KP, Gemelli T, Gautron L, Elmquist JK, Zinn AR, Kublaoui BM . Postnatal Sim1 deficiency causes hyperphagic obesity and reduced Mc4r and oxytocin expression. J Neurosci 2010; 30: 3803–3812.

    Article  CAS  Google Scholar 

  7. Kublaoui BM, Holder JL Jr, Tolson KP, Gemelli T, Zinn AR . SIM1 overexpression partially rescues agouti yellow and diet-induced obesity by normalizing food intake. Endocrinology 2006; 147: 4542–4549.

    Article  CAS  Google Scholar 

  8. Holder JL Jr, Butte NF, Zinn AR . Profound obesity associated with a balanced translocation that disrupts the SIM1 gene. Hum Mol Genet 2000; 9: 101–108.

    Article  CAS  Google Scholar 

  9. Faivre L, Cormier-Daire V, Lapierre JM, Colleaux L, Jacquemont S, Genevieve D et al. Deletion of the SIM1 gene (6q16.2) in a patient with a Prader-Willi-like phenotype. J Med Genet 2002; 39: 594–596.

    Article  CAS  Google Scholar 

  10. Varela MC, Simoes-Sato AY, Kim CA, Bertola DR, De Castro CI, Koiffmann CP . A new case of interstitial 6q16.2 deletion in a patient with Prader-Willi-like phenotype and investigation of SIM1 gene deletion in 87 patients with syndromic obesity. Eur J Med Genet 2006; 49: 298–305.

    Article  Google Scholar 

  11. Bonaglia MC, Ciccone R, Gimelli G, Gimelli S, Marelli S, Verheij J et al. Detailed phenotype-genotype study in five patients with chromosome 6q16 deletion: narrowing the critical region for Prader-Willi-like phenotype. Eur J Hum Genet 2008; 16: 1443–1449.

    Article  CAS  Google Scholar 

  12. Stutzmann F, Ghoussaini M, Couturier C, Marchand M, Vatin V, Corset L et al. Loss-of-function mutations in SIM1 cause a specific form of Prader-Willi-like syndrome. Diabetologia 2009; 52 (Suppl 1): 104.

    Google Scholar 

  13. Bonnefond A, Raimondo A, Stutzmann F, Ghoussaini M, Ramachandrappa S, Bersten DC et al. Loss-of-function mutations in SIM1 contribute to obesity and Prader-Willi-like features. J Clin Invest 2013; 123: 3037–3041.

    Article  CAS  Google Scholar 

  14. Ramachandrappa S, Raimondo A, Cali AM, Keogh JM, Henning E, Saeed S et al. Rare variants in single-minded 1 (SIM1) are associated with severe obesity. J Clin Invest 2013; 123: 3042–3050.

    Article  CAS  Google Scholar 

  15. Roelants M, Hauspie R, Hoppenbrouwers K . References for growth and pubertal development from birth to 21 years in Flanders, Belgium. Ann Hum Biol 2009; 36: 680–694.

    Article  CAS  Google Scholar 

  16. Miller SA, Dykes DD, Polesky HF . A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988; 16: 1215.

    Article  CAS  Google Scholar 

  17. Wittwer CT, Reed GH, Gundry CN, Vandersteen JG, Pryor RJ . High-resolution genotyping by amplicon melting analysis using LCGreen. Clin Chem 2003; 49: 853–860.

    Article  CAS  Google Scholar 

  18. Zhou L, Wang L, Palais R, Pryor R, Wittwer CT . High-resolution DNA melting analysis for simultaneous mutation scanning and genotyping in solution. Clin Chem 2005; 51: 1770–1777.

    Article  CAS  Google Scholar 

  19. Zegers D, Beckers S, de Freitas F, Peeters AV, Mertens IL, Verhulst SL et al. Identification of three novel genetic variants in the melanocortin-3 receptor of obese children. Obesity 2011; 19: 152–159.

    Article  CAS  Google Scholar 

  20. Ramensky V, Bork P, Sunyaev S . Human non-synonymous SNPs: server and survey. Nucleic Acids Res 2002; 30: 3894–3900.

    Article  CAS  Google Scholar 

  21. Adzhubei IA, Schmidt S, Peshkin L, Ramensky VE, Gerasimova A, Bork P et al. A method and server for predicting damaging missense mutations. Nat Methods 2010; 7: 248–249.

    Article  CAS  Google Scholar 

  22. Ng PC, Henikoff S . Predicting deleterious amino acid substitutions. Genome Res 2001; 11: 863–874.

    Article  CAS  Google Scholar 

  23. Calabrese R, Capriotti E, Fariselli P, Martelli PL, Casadio R . Functional annotations improve the predictive score of human disease-related mutations in proteins. Hum Mutat 2009; 30: 1237–1244.

    Article  CAS  Google Scholar 

  24. Li B, Krishnan VG, Mort ME, Xin F, Kamati KK, Cooper DN et al. Automated inference of molecular mechanisms of disease from amino acid substitutions. Bioinformatics 2009; 25: 2744–2750.

    Article  CAS  Google Scholar 

  25. 1000 Genomes Project Consortium. A map of human genome variation from population-scale sequencing. Nature 2010; 467: 1061–1073.

    Article  Google Scholar 

  26. Exome Variant Server, NHLBI Exome Sequencing Project (ESP), Seattle, WA [database on the Internet] 2012. [cited July 14]. Available from: URL http://evs.gs.washington.edu/EVS/.

  27. Wang JC, Turner L, Lomax B, Eydoux P . A 5-Mb microdeletion at 6q16.1-q16.3 with SIM gene deletion and obesity. Am J Med Genet A 2008; 146A: 2975–2978.

    Article  Google Scholar 

  28. Swarbrick MM, Evans DS, Valle MI, Favre H, Wu SH, Njajou OT et al. Replication and extension of association between common genetic variants in SIM1 and human adiposity. Obesity 2011; 19: 2394–2403.

    Article  CAS  Google Scholar 

  29. Hung CC, Luan J, Sims M, Keogh JM, Hall C, Wareham NJ et al. Studies of the SIM1 gene in relation to human obesity and obesity-related traits. Int J Obes 2007; 31: 429–434.

    Article  CAS  Google Scholar 

  30. Ghoussaini M, Stutzmann F, Couturier C, Vatin V, Durand E, Lecoeur C et al. Analysis of the SIM1 contribution to polygenic obesity in the French population. Obesity 2010; 18: 1670–1675.

    Article  Google Scholar 

  31. Traurig M, Mack J, Hanson RL, Ghoussaini M, Meyre D, Knowler WC et al. Common variation in SIM1 is reproducibly associated with BMI in Pima Indians. Diabetes 2009; 58: 1682–1689.

    Article  CAS  Google Scholar 

  32. Crews ST . PAS Proteins: Regulators and Sensors of Development and Physiology 1 edn Kluwer Academic Publishers, 2003.

    Book  Google Scholar 

  33. Chrast R, Scott HS, Chen H, Kudoh J, Rossier C, Minoshima S et al. Cloning of two human homologs of the Drosophila single-minded gene SIM1 on chromosome 6q and SIM2 on 21q within the Down syndrome chromosomal region. Genome Res 1997; 7: 615–624.

    Article  CAS  Google Scholar 

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Acknowledgements

Research was funded by a PhD grant of the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen) to DZ. SB is a postdoctoral fellow of the Research Foundation—Flanders (FWO Vlaanderen). This work was supported by a grant (G0028.05) from the Research Foundation—Flanders (FWO Vlaanderen) to LVG and WVH and by a TOP-research grant from the University of Antwerp to WVH. This study was supported by an Interuniversity Attraction Pole Project (Phase VII project 43, BELSPO).

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Authors and Affiliations

  1. Department of Medical Genetics, University of Antwerp and Antwerp University Hospital, Antwerp, Belgium

    D Zegers, S Beckers, R Hendrickx, J K Van Camp, V de Craemer & W Van Hul

  2. Department of Endocrinology, Diabetology and Metabolic Diseases, Antwerp University Hospital, Antwerp, Belgium

    A Verrijken & L F Van Gaal

  3. Department of Paediatrics, Antwerp University Hospital, Antwerp, Belgium

    K Van Hoorenbeeck, S L Verhulst, R P Rooman & K N Desager

  4. Department of Paediatrics, Jessa Hospital, Hasselt, Belgium

    G Massa

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  1. D Zegers
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Correspondence to W Van Hul.

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Zegers, D., Beckers, S., Hendrickx, R. et al. Mutation screen of the SIM1 gene in pediatric patients with early-onset obesity. Int J Obes 38, 1000–1004 (2014). https://doi.org/10.1038/ijo.2013.188

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