C101-05 Targeting Senescence to Mitigate Perinatal E-Cigarette Exposure-induced Lung Dysfunction - The Potential Role of Butyrate

Abstract Rationale Maternal smoking has persistent influence on the developing lungs of offspring, including accelerating cellular senescence—a state of irreversible cell-cycle arrest associated with aging and tissue dysfunction. Here, we test whether perinatal e-cigarette exposure predisposes to lung injury and how senescence contributes to airway inflammation and remodeling in asthma during development. We hypothesize that concomitant administration of butyrate, a key short-chain fatty acid metabolite, attenuates nicotine vaping-induced senescence and lung dysfunction in offspring. Methods Using a well-established E-Cigarette Nicotine Delivery System (ENDS), C57BL/J6 dams were exposed with or without nicotine (JUUL E-cigarette 4.8%) and 1mg/ml butyrate oral supplementation (once daily) until weaning (postnatal day 21). The pups were studied at 8 weeks of age with or without allergic sensitization using intranasal administration of House Dust Mite (HDM). Terminally, the animals (N = 5-8) were sacrificed and analyzed for pulmonary function, lung phenotype markers, senescence-associated secretory phenotype (SASP) and SASP factors were studied via, histology, protein, and gene levels. Total lungs were analyzed through RNA sequencing for the mechanistic insights in the offspring. Results The perinatal nicotine exposure impacts offspring lung function with increased airway resistance and decreased compliance after allergen (HDM) challenge, showing nicotine-driven allergic asthma. Moreover, we observed that perinatal nicotine exposure led to the induction of cellular senescence in murine lungs associated with increased SASP phenotype (p16, p21, p53, SA-β-gal or GLB-1, γH2AX, and Lamin B1) and factors including inflammatory makers (IL-6, TNF- α, and IL-1 β, MMPs), at protein and gene levels. The serum and BAL fluid also showed elevated pro-inflammatory cytokines like IL-6, TNF- α, and IL-1 β. The lung Bulk RNA sequencing highlighted the mechanistic insights into differentially expressed genes (DEGs) related to key senescence markers and asthma. Notably, maternal supplementation with butyrate during pregnancy significantly mitigated the cellular senescence induced by perinatal e-cigarette exposure in offspring, as evidenced by reduced expression of senescence-associated markers and decreased inflammatory cytokine levels. Conclusion Collectively, these findings highlight the impact of perinatal nicotine exposure on offspring asthma susceptibility and increased cellular senescence. Maternal butyrate supplementation attenuates the pro-senescent and detrimental pulmonary effects of perinatal e-cigarette exposure in offspring. These findings enhance our understanding of asthma pathogenesis and targeting senescence could be a novel strategy in developmental lung health. This abstract is funded by: HL151769 (NIH); T32IP5044, T32IR5048, T32IR5365, T291R0737, T34IR8406 (TRDRP); and EDUC4-12837 (CIRM)