# Identification of a novel regulatory program underlying the early specification of Neural Crest cells

> **NIH NIH F32** · UNIVERSITY OF CALIFORNIA RIVERSIDE · 2020 · $11,374

## Abstract

Project Summary
The objective of this proposal is to pioneer gene regulatory network studies of early Neural Crest (NC)
specification at previously uncharacterized early stages of development. NC cells are unique to vertebrates,
migrate extensively, and differentiate into multiple derivatives, including neurons, glia, melanocytes,
odontoblasts, and most of the craniofacial skeleton and connective tissue. Their failed development and
homeostasis leads to multiple pathologies (collectively termed “neurocristopathies”) which include prevalent
craniofacial malformations such as cleft lip and cleft palate. Traditionally, NC is thought to be induced from
ectoderm-derived cells at the neural plate border, as a result of inductive interactions involving neural and non-
neural ectoderm and/or underlying mesoderm. However, recent evidence from the Garcia-Castro lab and
others using chick, Xenopus, and human embryonic stem cell models suggest an earlier, pre-gastrula
origin of NC that is independent from post-gastrula tissue contributions. Yet the molecular mechanisms
underlying this early NC-specification events remain uncharacterized, and this proposal precisely aims to
address this issue. The evidence from the Garcia-Castro lab demonstrated the formation of human NC in a
human pluripotent stem cell model, independent of mesodermal or neural contributions, and suggested an earlier
than anticipated stage of specification, preceding the accepted neural plate border stage. Importantly, very
similar conclusions where reached in the chick embryo through specification assays and fate mapping
experiments. This suggests that the NC lineage rapidly departs from a pluripotent state to become specified and
engaged (not committed) in the NC lineage. Our group specifically recognizes this as a novel stage we termed
pre-border. Transcriptomic profiling of hNC supports an early stage, characterized by a set of transcription
factors, whose expression is rapidly initiated, precedes that of commonly used NC markers known as neural
plate border specifiers, further supporting our proposed pre-border stage. Here, I hypothesize that the
transcription factors expressed in the pre-border stage function in the early activation of genes known to operate
at later stages in the NC-gene regulatory network. Therefore, I propose to assess the contributions of a selected
group of pre-border factors to the regulation of the NC lineage, by determining their effects on known NC factors
(knockdown and knockout strategies), and monitoring global changes of expression to identify other regulated
molecules (Aim1) Importantly, comparative efforts in chick embryos will further strengthen these studies. Then I
propose to determine the direct contributions of the pre-border factors through the identification of the cis
regulatory modules to which they bind to regulate the expression of modulated targets (Aim2). This proposal will
reveal for the first time the regulatory interactions between TFs re...

## Key facts

- **NIH application ID:** 9964762
- **Project number:** 5F32DE027862-03
- **Recipient organization:** UNIVERSITY OF CALIFORNIA RIVERSIDE
- **Principal Investigator:** Rebekah Charney
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $11,374
- **Award type:** 5
- **Project period:** 2018-07-01 → 2020-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9964762

## Citation

> US National Institutes of Health, RePORTER application 9964762, Identification of a novel regulatory program underlying the early specification of Neural Crest cells (5F32DE027862-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9964762. Licensed CC0.

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