# Cardiac Transcription Factor Nkx2.7 is a Novel Regulator of Craniofacial Development

> **NIH NIH F31** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2021 · $46,836

## Abstract

PROJECT SUMMARY
The clinical phenotypes associated with DiGeorge Syndrome, the most common microdeletion condition
(22q11.2) in humans, illustrates the developmental link between cardiovascular and craniofacial morphogenesis.
Recent fate mapping studies in mice and zebrafish further support this notion given the identification of a
multipotent progenitor in the cardiopharyngeal field (CPF) that gives rise to the heart, branchiomeric muscles,
and pharyngeal arch arteries, mediated through the pharyngeal arches (PAs). NKX2-5 and NKX2-6 are
homeobox transcription factors frequently mutated in congenital heart defects. In zebrafish, we have previously
shown that Nkx2.5 and Nkx2.7 play redundant roles in cardiac development. Using a novel loss-of-function
nkx2.7 allele that is homozygous lethal, we demonstrate for the first time that nkx2.7-/- embryos fail to form
craniofacial muscles and cartilage necessary for feeding. This developmental deficiency results from early
disrupted genetic regulation in the PAs. We hypothesize that Nkx2.7 functions as an essential transcription factor
during craniofacial development mediated through the pharyngeal arches. We will investigate our model by
uncovering the tissue-specific roles of the nkx2.7+ CPF progenitors and by evaluating the transcriptional targets
of nkx2.7 and assessing for associated human disease phenotypes. In Aim 1, we will characterize the
developmental trajectories of the tissue-specific cell types in the PAs in nkx2.7-/- embryos, benefitting from an
innovative, high speed, volumetric imaging platform, SCAPE microscopy. Moreover, we will determine cell-
autonomous functions of nkx2.7+ progenitors through cell transplantation experiments employing tissue-specific
transgenes. In Aim 2, we will investigate the temporal and molecular mechanisms mediated by Nkx2.7. Using a
novel heat inducible transgene generated in our lab, we will overexpress nkx2.7 to dissect its requirement during
various developmental windows. We will apply this knowledge to single cell RNA-sequencing in nkx2.7-/- embryos
to generate a robust list of genetic targets in all pharyngeal tissues. We will also compare those putative effectors
with genomic data from patients with congenital heart disease and craniofacial defects. To evaluate for direct or
indirect binding of those targets, we will use chromatin immunoprecipitation and DNA sequencing (ChIP-seq).
Altogether, this proposal will provide a comprehensive understanding of the role of Nkx2.7 in cardiopharyngeal
development and will clarify the relationship between cardiac and craniofacial morphogenesis. Moreover, these
studies have potential to ameliorate stem cell therapy strategies in patients with defects of the head musculature
and cartilage.

## Key facts

- **NIH application ID:** 10156452
- **Project number:** 1F31DE030385-01
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** Caitlin Kelly Ford
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $46,836
- **Award type:** 1
- **Project period:** 2021-07-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10156452

## Citation

> US National Institutes of Health, RePORTER application 10156452, Cardiac Transcription Factor Nkx2.7 is a Novel Regulator of Craniofacial Development (1F31DE030385-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10156452. Licensed CC0.

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