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 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 NC 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 genomewide representation ofthe 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 funcfional analysis/dissection of neural crest enhancers. We will identify cisregulatory 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 ofthe trunk neural crest program to test skeletogenic potenfial. Using GRN informafion, we will challenge the fate of trunk NC by reengineering their regulatory circuits and observing if misexpression/deletion of key GRN subcircuits affects their identify 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 ofthe 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
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
2P01HD037105-16
Application #
8752121
Study Section
Special Emphasis Panel (ZHD1-DSR-Z (ED))
Project Start
Project End
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
16
Fiscal Year
2014
Total Cost
$312,134
Indirect Cost
$124,666
Name
California Institute of Technology
Department
Type
DUNS #
009584210
City
Pasadena
State
CA
Country
United States
Zip Code
91125
Warner, Jacob F; Miranda, Esther L; McClay, David R (2016) Contribution of hedgehog signaling to the establishment of left-right asymmetry in the sea urchin. Dev Biol 411:314-24
Peter, Isabelle S; Davidson, Eric H (2016) Implications of Developmental Gene Regulatory Networks Inside and Outside Developmental Biology. Curr Top Dev Biol 117:237-51
Simoes-Costa, Marcos; Bronner, Marianne E (2016) Reprogramming of avian neural crest axial identity and cell fate. Science 352:1570-3
Bronner, Marianne E (2016) How inhibitory cues can both constrain and promote cell migration. J Cell Biol 213:505-7
Uribe, Rosa A; Gu, Tiffany; Bronner, Marianne E (2016) A novel subset of enteric neurons revealed by ptf1a:GFP in the developing zebrafish enteric nervous system. Genesis 54:123-8
Hochgreb-Hagele, Tatiana; Koo, Daniel E S; Bronner, Marianne E (2015) Znf385C mediates a novel p53-dependent transcriptional switch to control timing of facial bone formation. Dev Biol 400:23-32
Butler, Samantha J; Bronner, Marianne E (2015) From classical to current: analyzing peripheral nervous system and spinal cord lineage and fate. Dev Biol 398:135-46
Simões-Costa, Marcos; Stone, Michael; Bronner, Marianne E (2015) Axud1 Integrates Wnt Signaling and Transcriptional Inputs to Drive Neural Crest Formation. Dev Cell 34:544-54
Barriga, Elias H; Trainor, Paul A; Bronner, Marianne et al. (2015) Animal models for studying neural crest development: is the mouse different? Development 142:1555-60
Martik, Megan L; McClay, David R (2015) Deployment of a retinal determination gene network drives directed cell migration in the sea urchin embryo. Elife 4:

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