Effects of Developmental Alcohol and Nicotine E-Cigarette Co-Exposure on Physiological Measures and Behavioral Development

PROJECT SUMMARY:
Alcohol and nicotine are the two most commonly consumed licit drugs among pregnant people. Although
the potential consequences of prenatal alcohol exposure at different levels are well understood, the patterns of
nicotine consumption have changed over the years with the popularity of electronic cigarettes (e-cigarettes). E-
cigarettes can increase a drug’s presence in blood levels more than traditional routes, which is particularly
alarming given that alcohol and nicotine are commonly used together among pregnant people. However, the
potential consequences of combined alcohol and nicotine exposure via e-cigarettes during early developmental
on brain and behavioral development are unclear. This project will use a mouse model to investigate whether
co-exposure to these drugs during early development alters drug metabolism, shifts intestinal microbiome
populations, changes gene expression, and/or modulates later-life behaviors. Dams and their neonates will be
exposed to alcohol, nicotine via e-cigarettes, the combination, or a control condition (vehicle or ambient air)
during the brain growth spurt period using a novel co-exposure model that mimics the doses and delivery devices
currently seen in human consumption. Aim 1 of this project will examine whether developmental co-exposure of
alcohol and nicotine via e-cigarettes increases blood levels of each drug, alters drug metabolism enzymes in the
liver, increases serum triglyceride levels, and/or shifts the gut microbiota composition. Plasma will be collected
from the dams to examine blood alcohol and nicotine concentrations during exposure. Additionally, plasma, fecal
and cecal matter, and liver tissue will be collected from both the dams and offspring to measure serum triglyceride
concentrations (colorimetric assay), drug metabolism enzymes (western blot), and microbiota shifts (shotgun
metagenomic sequencing and 16s rRNA sequencing) following acute (dams) and developmental (offspring)
exposure. Aim 2 of this project will determine whether developmental co-exposure to alcohol and nicotine via e-
cigarettes alters motor- and anxiety-related behaviors and/or correlated gene expression in the associated brain
regions. Later in life, sex pairs from each litter will be examined for motor- and anxiety-related behaviors using
open-field activity chambers, a forelimb reaching task, and an elevated plus maze. Critical motor- and anxiety-
related brain regions of subjects will also be analyzed to examine gene expression alterations via reverse
transcription quantitative real-time PCR (RT-qPCR). These data will provide valuable information regarding the
potential consequences of e-cigarette use among pregnant people, particularly when used in combination with
alcohol. In addition to providing information for teratology research, these data will also be valuable to general
research in substance use and pharmacology. Importantly, all data will be collected and analyzed by graduate
and undergraduate students in the Psychology, Biology, and Chemistry departments at West Chester University.
This project will enable students in the project-associated labs to advance their research professional
development and prepare them for future Ph.D. and medical school programs.

Grant Number: 1R15AA031576-01
NIH Institute/Center: NIH
Principal Investigator: Kristen Breit