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.
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.
|Huang, Miller; Miller, Matthew L; McHenry, Lauren K et al. (2016) Generating trunk neural crest from human pluripotent stem cells. Sci Rep 6:19727|
|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|
|Uribe, Rosa A; Bronner, Marianne E (2015) Meis3 is required for neural crest invasion of the gut during zebrafish enteric nervous system development. Mol Biol Cell 26:3728-40|
|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|
|SimÃµes-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|
|Kerosuo, Laura; Nie, Shuyi; Bajpai, Ruchi et al. (2015) Crestospheres: Long-Term Maintenance of Multipotent, Premigratory Neural Crest Stem Cells. Stem Cell Reports 5:499-507|
|Hu, Na; Strobl-Mazzulla, Pablo H; Bronner, Marianne E (2014) Epigenetic regulation in neural crest development. Dev Biol 396:159-68|
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