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
Lyons, Deirdre C; Martik, Megan L; Saunders, Lindsay R et al. (2014) Specification to biomineralization: following a single cell type as it constructs a skeleton. Integr Comp Biol 54:723-33
Tu, Qiang; Cameron, R Andrew; Davidson, Eric H (2014) Quantitative developmental transcriptomes of the sea urchin Strongylocentrotus purpuratus. Dev Biol 385:160-7
Warner, Jacob F; McCarthy, Ali M; Morris, Robert L et al. (2014) Hedgehog signaling requires motile cilia in the sea urchin. Mol Biol Evol 31:18-22
Warner, Jacob F; McClay, David R (2014) Perturbations to the hedgehog pathway in sea urchin embryos. Methods Mol Biol 1128:211-21
Kerosuo, Laura; Bronner, Marianne E (2014) Biphasic influence of Miz1 on neural crest development by regulating cell survival and apical adhesion complex formation in the developing neural tube. Mol Biol Cell 25:347-55
Hochgreb-Hägele, Tatiana; Koo, Daniel E S; Das, Neha M et al. (2014) Zebrafish stem/progenitor factor msi2b exhibits two phases of activity mediated by different splice variants. Stem Cells 32:558-71
Cheng, Xianrui; Lyons, Deirdre C; Socolar, Joshua E S et al. (2014) Delayed transition to new cell fates during cellular reprogramming. Dev Biol 391:147-57
Kwon, Seung-Hae; Park, Ok Kyu; Nie, Shuyi et al. (2014) Bioinformatic analysis of nematode migration-associated genes identifies novel vertebrate neural crest markers. PLoS One 9:e103024
Simões-Costa, Marcos; Tan-Cabugao, Joanne; Antoshechkin, Igor et al. (2014) Transcriptome analysis reveals novel players in the cranial neural crest gene regulatory network. Genome Res 24:281-90
Betancur, Paola; Simões-Costa, Marcos; Sauka-Spengler, Tatjana et al. (2014) Expression and function of transcription factor cMyb during cranial neural crest development. Mech Dev 132:38-43

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