Objective To explore the potential associations among maternal pre-pregnancy overweight/obesity, the miRNA expression profile of human milk exosomes, and gut microbiota characteristics in 3-month-old infants, so as to provide clues for mechanistic hypotheses. Methods Mother-infant dyads were recruited from healthy check-ups at the Department of Child Health Care, the Second Affiliated Hospital of Nanjing Medical University, between August 2021 and June 2023. Infant fecal samples and human milk samples were collected. Dyads were classified by maternal pre-pregnancy body mass index (BMI) into an overweight/obesity group (BMI ≥24.0 kg/m2; 5 pairs) and a normal-weight group (18.5 kg/m2 ≤BMI<24.0 kg/m2, 9 pairs). High-throughput sequencing of exosomal miRNAs in human milk was performed, followed by differential expression analysis and KEGG pathway enrichment. Infant fecal samples underwent 16S rRNA gene sequencing; α- and β-diversity were compared and differential taxa were identified. Correlations between differentially expressed exosomal miRNAs and gut microbiota were examined based on whether the features of differential taxa were consistent with the predicted functions of differential miRNAs. Results Compared with the normal-weight group, 73 exosomal miRNAs were differentially expressed in the overweight/obesity group, including 16 known miRNAs (5 upregulated and 11 downregulated; |log2FC|>1, P<0.05). KEGG analysis showed that target genes of the differential miRNAs were significantly enriched in lysosome, glycerophospholipid metabolism, and the intestinal immune network for IgA production pathways (P<0.02). No statistically significant differences were observed in α- or β-diversity of infant gut microbiota between groups (P>0.05). The linear discriminant analysis effect size (LEfSe) analysis indicated higher relative abundances of Citrobacter, Prevotella, and Campylobacter in infants of the overweight/obesity group (LDA>2.0, P<0.05). Conclusion Maternal pre-pregnancy overweight/obesity may be associated with altered exosomal miRNA profiles in human milk and differences in the abundance of specific infant gut genera, which still needs to be verified in larger sample sizes and longitudinal studies.
Key words
overweight/obesity /
breast milk /
miRNA /
gut microbiome
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] Verduci E, Gianni ML, Vizzari G, et al. The triad mother-breast milk-infant as predictor of future health: A narrative review[J]. Nutrients, 2021,13(2):486.
[2] Bettag J, Goldenberg D, Carter J, et al. Gut microbiota to microglia: Microbiome influences neurodevelopment in the CNS[J]. Children (Basel), 2023,10(11):1767.
[3] Harold N, Pamela AN, Ruben AM, et al . Early life gut microbiome and its impact on childhood health and chronic conditions[J]. Gut Microbes, 2025, 17(1):2463567.
[4] Marina C, Maria L, Athina V, et al. Human breast milk miRNAs: Investigation of association between breastfeeding children and maternal obesity in obesity development in offspring[J]. Genes(Basel), 2025,16(11):1373.
[5] 王银妃,殷勤,曾思洁,等. 早产儿和足月儿母乳外泌体 miRNA 差异表达的研究[J]. 中国儿童保健杂志, 2024,32(4):377-383.
Wang YF, Yin Q, Zeng SJ, et al. Differences in exosome miRNAs in human breast milk between preterm and full-term infants[J]. Chin J Child Health Care, 2024,32(4):377-383. (in Chinese)
[6] Shah KB, Chernausek SD, Garman LD, et al. Human milk exosomal microRNA: Associations with maternal overweight/obesity and infant body composition at 1 month of life[J]. Nutrients, 2021,13(4).
[7] Tingo L, Ahlberg E, Johansson L, et al. Non-coding RNAs in human breast milk: A systematic review[J]. Front Immunol, 2021,12:725323.
[8] Xi Y, Jiang X, Li R, et al. The levels of human milk microRNAs and their association with maternal weight characteristics[J]. Eur J Clin Nutr, 2016,70(4):445-449.
[9] Carreras-Badosa G, Bonmati A, Ortega F, et al. Dysregulation of placental miRNA in maternal obesity is associated with pre- and postnatal growth[J]. J Clin Endocrinol Metab, 2017,102(7):2584-2594.
[10] Zeng YM, Ye MM, Zhang HY, et al. Retrospective review:Single-and multidonor washed microbiota transplantation have equivalent efficacy in the treatment of autism[J]. Front Cell Infect Microbiol, 2025,15:1606417.
[11] Delphine H, Paula MC, Salome K, et al. Transfer of bacteria from mothers to infants through breast milk: A systematic review[J]. Pediatr infect Dis J, 2025,44(8):804-812.
[12] Katherine D, Victoria H, Kate N, et al. Maternal weight status and the composition of the human milk microbiome: A scoping review[J]. PLos One, 2022,17(10):e0274950.
[13] Kee HC, Yoowon K, Payam HK, et al. Infuence of maternal weight dynamics prior to and throughout gestation on early infant gut microbiome colonization[J]. Microb Ecol, 2025,88(1):32.
[14] Ionescu MI, Maria Catrina A, Dogaru IA, et al. Micro-biome: The trials and errors of developing an experimental model to study the impact of maternal gut microbiome disruption on perinatal asphyxia[J]. Reprod Fertil, 2024,5(4):e240050.
[15] Beckers KF, Flanagan JP, Sones JL. Microbiome and pregnancy:Focus on microbial dysbiosis coupled with maternal obesity[J]. Int J Obes (Lond), 2024,48(4):439-448.
[16] Francesco I, Angelo MI, Giulia L, et al. Difference in the intestinal microbiota between breastfeed infants and infants fed with artificial milk:A systematic review[J]. Pathogens, 2024,13(7):533.
[17] Wu S, Luo G, Jiang FL, et al. Early life bifidobacterial mother-infant transmission:Greater contribution from the infant gut to human milk revealed by microbiomic and culture-based methods[J]. mSystems, 2025,10(7):e0048025.
PDF(2686 KB)
