Prenatal Origins of Metabolic Disease – The Interface Between Maternal Smoking and Adulthood Western Diet

Metabolic disease is on the rise in developed and developing countries around the world. Prenatal insults such as cigarette smoke exposure (CSE) during gestation are correlated with the occurrence of metabolic maladies that appear during adulthood. While maternal smoking is on the decline, it was much more prevalent in past decades, meaning that many current adults were exposed in utero. The Developmental Origins of Health and Disease (DOHaD) hypothesis provides a framework to connect gestational insults such as maternal undernutrition to low birth weight and stereotypical adult diseases such as obesity and diabetes. This phenomenon, combined with lifestyle factors such as diet, could promote disease development and progression faster than either insult alone. In this study, we tested a multi-hit hypothesis to see if the effects of multiple moderate insults would compound to give rise to metabolic abnormalities sooner or more aggressively than any insult alone. C57BL/6 mouse dams were exposed to the smoke generated from one pack of cigarettes (CSE) for 3hrs a day starting 4 days before mating and continuing throughout gestation. The offspring of these mice showed no difference in weight at birth compared to controls exposed to filtered room air (Sham), nor did they display differences in metabolic parameters at postnatal week 10 – 12 when both groups were fed breeder chow (24% fat). At postnatal week 15, offspring were placed on either a low-fat (LF: 10 kcal% from fat) or high-fat/high-fructose (HF: 40 kcal% fat, 20 kcal% fructose) western style diet for 5 weeks, a shorter duration than normally needed to produce fatty liver disease with this diet in mice. Measures of adiposity (DEXA, n=5-6) were taken after 3 weeks on the diet, and blood markers of obesity and liver disease (n=5-6) were taken after 5 weeks, following euthanization. Total RNA and protein were extracted from adipose tissue and sequenced for gene expression or processed for proteomic analysis, respectively. Male offspring exposed to cigarette smoke in utero and fed a western style diet in adulthood (i.e., 2 hits) had higher body fat% (Male CSE HF = 37.21%±1.572, Male sham LF =28.95%±1.424, p=0.0137) and blood markers for metabolic abnormalities (cholesterol, leptin) compared to their Sham, low-fat diet (0-hit) counterparts; the 1-hit males (CSE or western diet) expressed intermediate levels. Female offspring did not show the 2-hit phenotype seen in the males for most measures. Pathway analysis of the adipose transcriptomic outcomes using Metacore reveals differential expression of genes in important metabolic pathways such as insulin and PI3K-AKT signaling when comparing 2-hit to 0-hit males, in alignment with the phenotypic outcomes. Our project shows that prenatal cigarette smoke exposure results in a sensitization of male offspring to the effects of a western-style diet in adulthood, in a DOHaD-predicted manner, an effect not found in female offspring within the truncated, 5-week western diet challenge. These results are novel in that we found an effect of the multiple hits even though we did not observe low birth weight for cigarette-exposed offspring, which is a hallmark of the DOHaD hypothesis. This suggests that prenatal exposures to even moderate insults can increase disease susceptibility when challenged with additional factors in adulthood, though more study is needed to fully understand how and why. This study was supported by NIH R15 grant (RS028440-02A1). This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.