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Functional characterization of a novel p.Ser76Thr variant in IGFBP4 that associates with body mass index in American Indians

Abstract

Insulin-like growth factor binding protein 4 (IGFBP4) is involved in adipogenesis, and IGFBP4 null mice have decreased body fat through decreased PPAR-γ expression. In the current study, we assessed whether variation in the IGFBP4 coding region influences body mass index (BMI) in American Indians who are disproportionately affected by obesity. Whole exome sequence data from a population-based sample of 6779 American Indians with longitudinal measures of BMI were used to identify variation in IGFBP4 that associated with BMI. A novel variant that predicts a p.Ser76Thr in IGFBP4 (Thr-allele frequency = 0.02) was identified which associated with the maximum BMI measured during adulthood (BMI 39.8 kg/m2 for Thr-allele homozygotes combined with heterozygotes vs. 36.2 kg/m2 for Ser-allele homozygotes, β = 6.7% per Thr-allele, p = 8.0 × 10−5, adjusted for age, sex, birth-year and the first five genetic principal components) and the maximum age- and sex-adjusted BMI z-score measured during childhood/adolescence (z-score 0.70 SD for Thr-allele heterozygotes vs. 0.32 SD for Ser-allele homozygotes, β = 0.37 SD per Thr-allele, p = 8.8 × 10−6). In vitro functional studies showed that IGFBP4 with the Thr-allele (BMI-increasing) had a 55% decrease (p = 0.0007) in FOXO-induced transcriptional activity, reflecting increased activation of the PI3K/AKT pathway mediated through increased IGF signaling. Over-expression and knock-down of IGFBP4 in OP9 cells during differentiation showed that IGFBP4 upregulates adipogenesis through PPARγ, CEBPα, AGPAT2 and SREBP1 expression. We propose that this American Indian specific variant in IGFBP4 affects obesity via an increase of IGF signaling.

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Fig. 1: The p.Ser76Thr variant in IGFBP4 associated with lifetime BMI in American Indians.
Fig. 2: The p.Ser76Thr variant in IGFBP4 altered FOXO-induced transcription through PI3K/AKT signaling.
Fig. 3: Over-expressing and knocking-down IGFBP4 during OP9 cell differentiation affected RNA expression of adipogenic markers.
Fig. 4: Effects of IGFBP4 on adipogenesis.

Data availability

Materials are available upon request. Individual-level data are not publicly available due to privacy concerns, but may be made available upon reasonable request—see dbGAP accession number phs002490.v1.p1 for details.

Code availability

Analyses were conducted using available software applications, as described in the Methods.

References

  1. Laron Z. Lessons from 50 years of study of Laron syndrome. Endocr Pr. 2015;21:1395–402.

    Article  Google Scholar 

  2. Janecka A, Kołodziej-Rzepa M, Biesaga B. Clinical and molecular features of laron syndrome, a genetic disorder protecting from cancer. Vivo. 2016;30:375–81.

    CAS  Google Scholar 

  3. Boucher J, Mori MA, Lee KY, Smyth G, Liew CW, Macotela Y, et al. Impaired thermogenesis and adipose tissue development in mice with fat-specific disruption of insulin and IGF-1 signalling. Nat Commun. 2012;3:902.

    Article  Google Scholar 

  4. Maridas DE, DeMambro VE, Le PT, Mohan S, Rosen CJ. IGFBP4 is required for adipogenesis and influences the distribution of adipose depots. Endocrinology. 2017;158:3488–500.

    CAS  Article  Google Scholar 

  5. Knowler WC, Pettitt DJ, Saad MF, Bennett PH. Diabetes mellitus in the Pima Indians: incidence, risk factors and pathogenesis. Diabetes Metab Rev. 1990;6:1–27.

    CAS  Article  Google Scholar 

  6. Barsh GS, Farooqi IS, O’Rahilly S. Genetics of body-weight regulation. Nature. 2000;404:644–51.

    CAS  Article  Google Scholar 

  7. Muller YL, Thearle MS, Piaggi P, Hanson RL, Hoffman D, Gene B, et al. Common genetic variation in and near the melanocortin 4 receptor gene (MC4R) is associated with body mass index in American Indian adults and children. Hum Genet. 2014;133:1431–41.

    CAS  Article  Google Scholar 

  8. Thearle MS, Muller YL, Hanson RL, Mullins M, Abdussamad M, Tran J, et al. Greater impact of melanocortin-4 receptor deficiency on rates of growth and risk of type 2 diabetes during childhood compared with adulthood in Pima Indians. Diabetes. 2012;61:250–7.

    CAS  Article  Google Scholar 

  9. Kim HI, Ye B, Gosalia N, Regeneron Genetics Center, Köroğlu C, Hanson RL, et al. Characterization of exome variants and their metabolic impact in 6,716 American Indians from the Southwest US. Am J Hum Genet. 2020;107:251–64.

    CAS  Article  Google Scholar 

  10. 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;58:1682–9.

    CAS  Article  Google Scholar 

  11. Traurig MT, Perez JM, Ma L, Bian L, Kobes S, Hanson RL, et al. Variants in the LEPR gene are nominally associated with higher BMI and lower 24-h energy expenditure in Pima Indians. Obesity. 2012;20:2426–30.

    CAS  Article  Google Scholar 

  12. The Expert Committee on the Diagnosis and Classification of Diabetes Mellitus. The Expert Committee on the Diagnosis and Classification of Diabetes Mellitus: report of the Expert Committee on the diagnosis and classification of diabetes mellitus. Diabetes Care. 1997;20:1183–97.

  13. Lillioja S, Mott DM, Spraul M, Ferraro R, Foley JE, Ravussin E, et al. Insulin resistance and insulin secretory dysfunction as precursors of non-insulin-dependent diabetes mellitus. Prospective studies of Pima Indians. N Engl J Med. 1993;329:1988–92.

    CAS  Article  Google Scholar 

  14. McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A, et al. The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res. 2010;20:1297–303.

    CAS  Article  Google Scholar 

  15. Hsueh WC, Nair AK, Kobes S, Chen P, Göring HHH, Pollin TI, et al. Identity-by-descent mapping identifies major locus for serum triglycerides in Amerindians largely explained by an APOC3 founder mutation. Circ Cardiovasc Genet. 2017;10:e001809.

  16. Lane JM, Doyle JR, Fortin JP, Kopin AS, Ordovás JM. Development of an OP9 derived cell line as a robust model to rapidly study adipocyte differentiation. PLoS ONE. 2014;9:e112123.

    Article  Google Scholar 

  17. Albert JS, Yerges-Armstrong LM, Horenstein RB, Pollin TI, Sreenivasan UT, Chai S, et al. Null mutation in hormone-sensitive lipase gene and risk of type 2 diabetes. N Engl J Med. 2014;370:2307–15.

    Article  Google Scholar 

  18. Cautivo KM, Lizama CO, Tapia PJ, Agarwal AK, Garg A, Horton JD, et al. AGPAT2 is essential for postnatal development and maintenance of white and brown adipose tissue. Mol Metab. 2016;5:491–505.

    CAS  Article  Google Scholar 

  19. Siwanowicz I, Popowicz GM, Wisniewska M, Huber R, Kuenkele KP, Lang K, et al. Structural basis for the regulation of insulin-like growth factors by IGF binding proteins. Structure. 2005;13:155–67.

    CAS  Article  Google Scholar 

  20. Guex N, Peitsch MC. SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis. 1997;18:2714–23.

    CAS  Article  Google Scholar 

  21. Pandurangan AP, Ochoa-Montaño B, Ascher DB, Blundell TL. SDM: a server for predicting effects of mutations on protein stability. Nucleic Acids Res. 2017;45:W229–35.

    CAS  Article  Google Scholar 

  22. Parthiban V, Gromiha MM, Schomburg D. CUPSAT: prediction of protein stability upon point mutations. Nucleic Acids Res. 2006;34:W239–42.

    CAS  Article  Google Scholar 

  23. Kircher M, Witten DM, Jain P, O’Roak BJ, Cooper GM, Shendure J. A general framework for estimating the relative pathogenicity of human genetic variants. Nat Genet. 2014;46:310–5.

    CAS  Article  Google Scholar 

  24. Burke TW, Vuk-Pavlović S. Insulin-like growth factor-I is a serum component stimulating growth of human neuroblastoma. Vitr Cell Dev Biol Anim. 1993;29:391–4.

    Article  Google Scholar 

  25. Guo S. Molecular basis of insulin resistance: the role of IRS and Foxo1 in the control of diabetes mellitus and its complications. Drug Discov Today Dis Mech. 2013;10:e27–33.

    Article  Google Scholar 

  26. Moll L, Schubert M. The role of insulin and insulin-like growth factor-1/FoxO-mediated transcription for the pathogenesis of obesity-associated dementia. Curr Gerontol Geriatr Res. 2012. https://doi.org/10.1155/2012/384094.

  27. Berryman DE, Glad CAM, List EO, Johannsson G. The GH/IGF-1 axis in obesity: pathophysiology and therapeutic considerations. Nat Rev Endocrinol.2013;9:346–56.

    CAS  Article  Google Scholar 

  28. Gude MF, Hjortebjerg R, Oxvig C, Thyø AA, Magnusson NE, Bjerre M, et al. PAPP-A, IGFBP-4 and IGF-II are secreted by human adipose tissue cultures in a depot-specific manner. Eur J Endocrinol. 2016;175:509–19.

    CAS  Article  Google Scholar 

  29. Yao Y, Li J, Jin Y, Chen Y, He L. Association between PPAR-γ2 Pro12Ala polymorphism and obesity: a meta-analysis. Mol Biol Rep. 2015;42:1029–38.

    CAS  Article  Google Scholar 

  30. Mansoori A, Amini M, Kolahdooz F, Seyedrezazadeh E. Obesity and Pro12Ala polymorphism of peroxisome proliferator-activated receptor-gamma gene in healthy adults: a systematic review and meta-analysis. Ann Nutr Metab. 2015;67:104–18.

    CAS  Article  Google Scholar 

  31. Moseti D, Regassa A, Kim W. Molecular regulation of adipogenesis and potential anti-adipogenic bioactive molecules. Int J Mol Sci. 2016;17:124.

    Article  Google Scholar 

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Acknowledgements

We thank the study participants as well as the clinical staff of the Phoenix Epidemiology and Clinical Research Branch for collecting phenotypes for the study. The opinions expressed in this paper are those of the authors, and do not necessarily reflect the views of the Indian Health Service.

Funding

This work was supported by the Intramural Research Programme of National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health.

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YLM and LJB contributed to the study design. YLM, MS, SD, KB, WCK, CVVH, Regeneron Genetics Center, ARS, RLH, CB and LJB contributed to the data acquisition. CK, SK, WCK, CVVH and RLH contributed to the data analysis. All authors contributed to data interpretation and manuscript drafting; and approved the final version.

Corresponding author

Correspondence to Yunhua L. Muller.

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The authors declare that there is no conflict of interest. ARS or CVVH is or was full-time employees of the Regeneron Genetics Center from Regeneron Pharmaceuticals Inc. and receive or received stock options and restricted stock units as compensation.

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These participants had provided written consent for DNA sequencing; and informed consent was obtained from all subjects. The consent was approved by the Institutional Review Board of the NIDDK and/or by the Phoenix Area Indian Health Service Institutional Review Board.

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Muller, Y.L., Saporito, M., Day, S. et al. Functional characterization of a novel p.Ser76Thr variant in IGFBP4 that associates with body mass index in American Indians. Eur J Hum Genet (2022). https://doi.org/10.1038/s41431-022-01129-3

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