# An ethanol-sensitive, Bmp-PCP dependent pathway regulating endoderm cell behaviors

> **NIH NIH R01** · UNIVERSITY OF LOUISVILLE · 2024 · $480,468

## Abstract

Summary
Fetal Alcohol Spectrum Disorders (FASD) describe a continuum of ethanol-induced developmental defects with
jaw defects being common. Genetic risk factors lie at the heart of FASD and are major drivers of FASD etiology,
providing insight into the cellular and molecular processes potentially disrupted in FASD. However, we know little
of these processes. From our studies in zebrafish, we have established an in vivo model for FASD where
mutations in the Bone Morphogenetic Protein (Bmp) and Wnt/Planar Cell Polarity (PCP) signaling pathways
sensitize embryos to ethanol-induced jaw defects by disrupting the cell behaviors driving tissue morphogenesis
during jaw development. Jaw formation involves complex signaling interactions between the neural crest and
the anterior pharyngeal endoderm (anterior PE). Critical to these interactions, anterior PE morphogenesis
requires Bmp- and PCP-dependent actin-based cell movements, polarity and adhesion. Bmp signaling is
required for endodermal E-cadherin localization and PCP receptor expression while PCP regulates cell polarity
and adhesion. Mutations in the Bmp and PCP pathways sensitize embryos to ethanol-induced malformations of
the anterior PE and jaw. Work in mouse liver indicates that ethanol reduces Bmp signaling at the level of the
receptors. Altogether, this suggests that an ethanol-sensitive, Bmp-PCP signaling cascade contributes to
anterior PE morphogenesis. Yet, how ethanol modulates the Bmp and PCP pathways and their crosstalk,
disrupting anterior PE morphogenesis remain unknown. We hypothesize that ethanol attenuates Bmp signaling
there by decreasing PCP component expression levels and disrupting actin dynamics, cell polarity, adhesion
and migration during anterior PE morphogenesis. Using a unique combination of zebrafish-based approaches,
we will test our hypothesis in the following aims. In Specific Aim 1, we will a) analyze an ethanol dose response
on total and phosphorylated Bmp receptor and Smad proteins, b) use scRNA-seq to examine endoderm-specific
gene expression changes of Bmp and, downstream of Bmp, PCP genes, and c) test the epistasis of the Bmp-
PCP signaling axis on jaw development. In Specific Aim 2, we will use 4D-confocal analysis in control and
ethanol-treated Bmp and PCP mutants to quantify ethanol-induced disruptions to a) cell velocity, coherence, and
persistence of migration and resulting changes to size and shape of the anterior PE and b) actin polarization and
cellular protrusion dynamics. In Specific Aim 3, we will a) quantify shape and polarity and b) analyze E-cadherin
gene expression as well as E-cadherin and PCP protein localization in anterior PE cells of control and ethanol-
treated Bmp, PCP and Bmp-PCP compound mutants. We will also c) quantify ethanol-induced jaw defects in
Bmp- and PCP-E-cadherin compound mutants. Overall, this work will greatly advance our understanding of the
cell behaviors and molecular mechanisms driving ethanol-induced structural malformati...

## Key facts

- **NIH application ID:** 10882648
- **Project number:** 1R01AA031043-01A1
- **Recipient organization:** UNIVERSITY OF LOUISVILLE
- **Principal Investigator:** Charles Benjamin Lovely
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $480,468
- **Award type:** 1
- **Project period:** 2024-05-15 → 2029-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10882648

## Citation

> US National Institutes of Health, RePORTER application 10882648, An ethanol-sensitive, Bmp-PCP dependent pathway regulating endoderm cell behaviors (1R01AA031043-01A1). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10882648. Licensed CC0.

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