The neural crest is a multipotent embryonic cell population that contributes to diverse derivatives, including peripheral ganglia, cartilage and bone of the face, and melanocytes. We have proposed and tested a multistep gene regulatory network (GRN), comprised of a logical series of distinct regulatory steps that act in concert to imbue the cranial neural crest with its defining traits. However, there are significant differences in developmental potential and migratory pathways of different neural crest populations arising at different axial levels. Here, we propose to explore GRN differences along the neural axis, focusing on premigratory neural crest cells from two distinct regions: cranial versus trunk. Our preliminary transcriptome analysis reveals many transcription factors and signaling molecules specific to the cranial but not trunk neural crest or vice versa. Our goal is to determine the position of these genes in the cranial versus trunk GRNs. This systems level strategy will provide understanding of why neural crest GRNs produces a particular regulatory state for use in preprogramming these cells to a different state.
The aims are:
Aim 1 : Multiplex perturbation analysis of GRN connections at cranial and trunk levels. With the genome-wide representation of the active transcriptome of premigratory cranial and trunk neural crest in hand, we will perform loss-of-function experiments to perturb gene function and quantitate subsequent global transcriptional changes in putative target genes in single embryos using Nanostring analysis.
Aim 2 : Phylogenomic and functional analysis/dissection of neural crest enhancers. We will identify cis-regulatory elements that mediate expression of key GRN factors in cranial versus trunk neural crest populations. We will perform multidimensional modeling that incorporates results of transcriptome data and active enhancers with functional perturbation results into representational models of neural crest GRNs.
Aim 3 : Reengineering of the trunk neural crest program to test skeletogenic potential. Using GRN information, we will challenge the fate of trunk NC by reengineering their regulatory circuits and observing if misexpression/deletion of key GRN subcircuits affects their identity and ability to contribute to cartilage.

Public Health Relevance

The neural crest is a multipotent stem-cell-like population that forms an amazingly diverse set of derivatives, including neurons and glia of the peripheral nervous system, skin melanocytes, and craniofacial skeleton. Because they are involved in many birth defects and cancers (e.g. melanoma, neuroblastoma), our results on their development will yield important clues regarding mistakes causing abnormal development &tumors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
1R01DE024157-01
Application #
8667751
Study Section
Skeletal Biology Development and Disease Study Section (SBDD)
Program Officer
Scholnick, Steven
Project Start
2014-06-01
Project End
2019-03-31
Budget Start
2014-06-01
Budget End
2015-03-31
Support Year
1
Fiscal Year
2014
Total Cost
$416,250
Indirect Cost
$166,250
Name
California Institute of Technology
Department
None
Type
Schools of Arts and Sciences
DUNS #
009584210
City
Pasadena
State
CA
Country
United States
Zip Code
91125
Hu, Na; Strobl-Mazzulla, Pablo H; Bronner, Marianne E (2014) Epigenetic regulation in neural crest development. Dev Biol 396:159-68
Saxena, Ankur; Bronner, Marianne E (2014) A novel HoxB cluster protein expressed in the hindbrain and pharyngeal arches. Genesis 52:858-63