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Nonnutritive sweetener consumption during pregnancy, adiposity, and adipocyte differentiation in offspring: evidence from humans, mice, and cells

International Journal of Obesity volume 44, pages 2137–2148 (2020)Cite this article

Subjects

Abstract

Background

Obesity often originates in early life, and is linked to excess sugar intake. Nonnutritive sweeteners (NNS) are widely consumed as “healthier” alternatives to sugar, yet recent evidence suggests NNS may adversely influence weight gain and metabolic health. The impact of NNS during critical periods of early development has rarely been studied. We investigated the effect of prenatal NNS exposure on postnatal adiposity and adipocyte development.

Methods

In the CHILD birth cohort (N = 2298), we assessed maternal NNS beverage intake during pregnancy and child body composition at 3 years, controlling for maternal BMI and other potential confounders. To investigate causal mechanisms, we fed NNS to pregnant C57BL6J mice at doses relevant to human consumption (42 mg/kg/day aspartame or 6.3 mg/kg/day sucralose), and assessed offspring until 12 weeks of age for: body weight, adiposity, adipose tissue morphology and gene expression, glucose and insulin tolerance. We also studied the effect of sucralose on lipid accumulation and gene expression in cultured 3T3-L1 pre-adipocyte cells.

Results

In the CHILD cohort, children born to mothers who regularly consumed NNS beverages had elevated body mass index (mean z-score difference +0.23, 95% CI 0.05–0.42 for daily vs. no consumption, adjusted for maternal BMI). In mice, maternal NNS caused elevated body weight, adiposity, and insulin resistance in offspring, especially in males (e.g., 47% and 15% increase in body fat for aspartame and sucralose vs. controls, p < 0.001). In cultured adipocytes, sucralose exposure at early stages of differentiation caused increased lipid accumulation and expression of adipocyte differentiation genes (e.g., C/EBP-α, FABP4, and FASN). These genes were also upregulated in adipose tissue of male mouse offspring born to sucralose-fed dams.

Conclusion

By triangulating evidence from humans, mice, and cultured adipocytes, this study provides new evidence that maternal NNS consumption during pregnancy may program obesity risk in offspring through effects on adiposity and adipocyte differentiation.

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Fig. 1: Maternal consumption of NNS-sweetened beverages and child body mass index (BMI) at 3 years of age in the CHILD cohort.
Fig. 2: Body composition in male and female mouse offspring of dams fed sucrose, aspartame or sucralose during pregnancy and lactation.
Fig. 3: Glucose tolerance and insulin sensitivity in 10-week-old male and female mouse offspring of dams fed sucrose, aspartame or sucralose during pregnancy and lactation.
Fig. 4: Effect of sucralose on 3T3-L1 adipocyte differentiation in vitro.
Fig. 5: Sucralose increases the expression of pro-adipogenic regulators, fat storage, and mobilization genes in 3T3-L1 cells in vitro.
Fig. 6: Sucralose increases the expression of pro-adipogenic regulators and fat storage and mobilization genes in mouse offspring adipose tissue in vivo.

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Acknowledgements

We are grateful to all the families who took part in the CHILD study, and the whole CHILD team, which includes interviewers, nurses, computer and laboratory technicians, clerical workers, research scientists, volunteers, managers, and receptionists. We also acknowledge the excellent technical work of Mario Fonseca and Bo Xiang (University of Manitoba), critical review by Shirin Moossavi (University of Manitoba), and editorial assistance from John Schellenberg (University of Manitoba). A preprint of this work was posted on bioRxiv (https://doi.org/10.1101/713974).

Funding

The Canadian Institutes of Health Research (CIHR) and the Allergy, Genes and Environment Network of Centers of Excellence (AllerGen NCE) provided core support for the CHILD Study. This research was supported, in part, by the Canada Research Chairs program. MBA holds the Tier 2 Canada Research Chair in the Developmental Origins of Chronic Disease, and is a Canadian Institute for Advanced Research Fellow in the Humans and the Microbiome Program. VWD holds the Allen Rouse-Manitoba Medical Services Foundation Basic Scientist Award. MMT is the recipient of a Research Manitoba/CHRIM studentship. This research was supported by a Children’s Hospital Research Institute of Manitoba Grant, a CIHR Operating Grant #151540, and a CIHR Environments, Genes and Chronic Disease Team Grant #144626. These entities had no role in the design and conduct of the study; collection, management, analysis, and interpretation of the data; and preparation, review, or approval of the paper.

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Authors and Affiliations

  1. Developmental Origins of Chronic Diseases in Children Network (DEVOTION) and the Diabetes Research Envisioned and Accomplished in Manitoba (DREAM) Theme of the Children’s Hospital Research Institute of Manitoba, Winnipeg, MB, Canada

    Meghan B. Azad, Alyssa Archibald, Mateusz M. Tomczyk, Alanna Head, Kyle G. Cheung, Allan B. Becker & Vernon W. Dolinsky

  2. Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada

    Meghan B. Azad & Allan B. Becker

  3. Max Rady College of Medicine, University of Manitoba, Winnipeg, MB, Canada

    Alyssa Archibald

  4. Department of Pharmacology and Therapeutics, University of Manitoba, Winnipeg, MB, Canada

    Mateusz M. Tomczyk, Alanna Head, Kyle G. Cheung & Vernon W. Dolinsky

  5. Department of Clinical Epidemiology & Biostatistics, McMaster University, Hamilton, ON, Canada

    Russell J. de Souza

  6. Department of Nutritional Sciences, University of Toronto, Toronto, ON, Canada

    Russell J. de Souza

  7. Li Ka Shing Knowledge Institute, St. Michael’s Hospital, Toronto, ON, Canada

    Russell J. de Souza

  8. Department of Pediatrics, University of Alberta, Edmonton, AB, Canada

    Piushkumar J. Mandhane

  9. Department of Pediatrics, BC Children’s Hospital, University of British Columbia, Vancouver, BC, Canada

    Stuart E. Turvey

  10. Department of Pediatrics, Hospital for Sick Children, University of Toronto, Toronto, ON, Canada

    Theo J. Moraes & Padmaja Subbarao

  11. Department of Medicine, McMaster University, Hamilton, ON, Canada

    Malcolm R. Sears

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  1. Meghan B. Azad
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Contributions

MBA and VWD conceived of the study design, obtained funding for this research, and drafted the paper. MRS, PS, TJM, SET, PJM, and ABB obtained funding for and oversaw recruitment of the CHILD cohort and data collection. AA performed the statistical analysis of clinical data from the CHILD cohort under the supervision of MBA. RJS contributed nutritional expertise. MMT, AH, and KGC performed mouse and cell culture experiments under the supervision of VWD. All authors critically reviewed and approved the paper. MBA had full access to the human data and VWD had full access to the mouse and adipocyte data, and take final responsibility for the decision to submit for publication.

Corresponding authors

Correspondence to Meghan B. Azad or Vernon W. Dolinsky.

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Azad, M.B., Archibald, A., Tomczyk, M.M. et al. Nonnutritive sweetener consumption during pregnancy, adiposity, and adipocyte differentiation in offspring: evidence from humans, mice, and cells. Int J Obes 44, 2137–2148 (2020). https://doi.org/10.1038/s41366-020-0575-x

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