# Deciphering neural crest-specific TFAP2 pathways in midface development and dysplasia

> **NIH NIH F31** · UNIVERSITY OF IOWA · 2024 · $41,580

## Abstract

PROJECT SUMMARY: Craniofacial anomalies are the 2nd leading structural malformation to appear at birth,
with the most common forms occurring in the mid- and upper-face regions. However, our limited understanding
in the molecular mechanisms controlling their development has hindered strategies for effective treatment and
prevention. Several studies have identified the pathways underlying lower face formation during embryonic
development (e.g., jaw and teeth). Yet, less emphasis has been placed on the midface, and the unique
mechanisms orchestrating the development of this region are poorly understood. The long-term goal of this study
is to dissect the molecular underpinnings governing midface development and how their dysregulation causes
dysplasia. We and others have independently uncovered key midface genes that function within the neural crest,
an embryonic stem cell population that gives rise to facial bone and cartilage. For example, deletion of Alx1/3/4
(Alx), whose loss causes syndromic forms of Frontonasal Dysplasia, lead to a severe midfacial cleft in mice. Our
lab has found that loss of Tfap2 in the neural crest compromises expression of Alx genes and presents the same
midface cleft as the Alx knockout embryos. Thus, our overall objective in this research proposal is to leverage
this new animal model for Frontonasal Dysplasia to uncover how TFAP2 transcription factors operate within the
midface-unique gene regulatory networks (GRN’s). To approach this objective, we test the overall hypothesis
that TFAP2 transcription factors directly regulate Alx gene expression for appropriate neural crest survival,
midface fusion, and skeletal formation. In AIM 1, we will employ genome- and epigenome-wide assays
(CUT&RUN, ATAC-seq) to decipher how TFAP2 targets Alx and shapes the genome landscape to regulate their
expression levels. We will leverage zebrafish reporter strategies to test TFAP2-bound Alx noncoding enhancers
for midface specificity. In AIM 2, we will couple sophisticated mouse genetics with immunofluorescence and
skeletal analyses to characterize how Tfap2 and Alx genetically interact to control neural crest viability, behavior,
and formation of the skeletal elements. Completion of these aims will fill a critical knowledge gap in our
understanding for how these genes and pathways are linked to shape the developing midface. Such knowledge
will contribute to the development of strategies in treating midfacial disorders or even preventing them. The
principal investigator of this study, Mr. Nguyen, has extensive training in mouse genetics, embryology, and gene
expression profiling techniques highlighted in this proposal. With combined mentorship from Drs. Van Otterloo
and Amendt, Mr. Nguyen will develop skills in genome-wide molecular assays, next-generation sequencing
approaches, bioinformatic techniques, and immunofluorescent and confocal microscopy methods; collectively,
greatly advancing his doctoral training and expanding his research to...

## Key facts

- **NIH application ID:** 10906653
- **Project number:** 5F31DE032881-02
- **Recipient organization:** UNIVERSITY OF IOWA
- **Principal Investigator:** Timothy Nguyen
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $41,580
- **Award type:** 5
- **Project period:** 2023-07-01 → 2025-05-16

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10906653

## Citation

> US National Institutes of Health, RePORTER application 10906653, Deciphering neural crest-specific TFAP2 pathways in midface development and dysplasia (5F31DE032881-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10906653. Licensed CC0.

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