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

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. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS

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OFFSPRING BODY MASS INDEX AND BODY FAT FROM BIRTH TO ADOLESCENCE Article 05 October 2021 MATERNAL HIGH-FAT DIET PROGRAMS WHITE AND BROWN ADIPOSE TISSUE LIPIDOME AND TRANSCRIPTOME IN

OFFSPRING IN A SEX- AND TISSUE-DEPENDENT MANNER IN MICE Article Open access 07 January 2022 WESTERN-STYLE DIET CONSUMPTION IMPAIRS MATERNAL INSULIN SENSITIVITY AND GLUCOSE METABOLISM DURING

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obesity-associated gut microbiome with increased capacity for energy harvest. Nature. 2006;444:1027–31. PubMed  Google Scholar  Download references 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. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * 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 * Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada Meghan B. Azad & 

Allan B. Becker * Max Rady College of Medicine, University of Manitoba, Winnipeg, MB, Canada Alyssa Archibald * Department of Pharmacology and Therapeutics, University of Manitoba, Winnipeg,

MB, Canada Mateusz M. Tomczyk, Alanna Head, Kyle G. Cheung & Vernon W. Dolinsky * Department of Clinical Epidemiology & Biostatistics, McMaster University, Hamilton, ON, Canada

Russell J. de Souza * Department of Nutritional Sciences, University of Toronto, Toronto, ON, Canada Russell J. de Souza * Li Ka Shing Knowledge Institute, St. Michael’s Hospital, Toronto,

ON, Canada Russell J. de Souza * Department of Pediatrics, University of Alberta, Edmonton, AB, Canada Piushkumar J. Mandhane * Department of Pediatrics, BC Children’s Hospital, University

of British Columbia, Vancouver, BC, Canada Stuart E. Turvey * Department of Pediatrics, Hospital for Sick Children, University of Toronto, Toronto, ON, Canada Theo J. Moraes & Padmaja

Subbarao * Department of Medicine, McMaster University, Hamilton, ON, Canada Malcolm R. Sears Authors * Meghan B. Azad View author publications You can also search for this author inPubMed 

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this author inPubMed Google Scholar * Alanna Head View author publications You can also search for this author inPubMed Google Scholar * Kyle G. Cheung View author publications You can also

search for this author inPubMed Google Scholar * Russell J. de Souza View author publications You can also search for this author inPubMed Google Scholar * Allan B. Becker View author

publications You can also search for this author inPubMed Google Scholar * Piushkumar J. Mandhane View author publications You can also search for this author inPubMed Google Scholar *

Stuart E. Turvey View author publications You can also search for this author inPubMed Google Scholar * Theo J. Moraes View author publications You can also search for this author inPubMed 

Google Scholar * Malcolm R. Sears View author publications You can also search for this author inPubMed Google Scholar * Padmaja Subbarao View author publications You can also search for

this author inPubMed Google Scholar * Vernon W. Dolinsky View author publications You can also search for this author inPubMed Google Scholar 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. ETHICS DECLARATIONS

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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 Download citation * Received: 14 November 2019 * Revised: 16 March 2020 * Accepted: 27 March 2020 *

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