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Research project

Studying enteric neuron development in zebrafish

Status: Ongoing project

Using zebrafish, we aim to unravel molecular mechanisms involved in gut colonization by enteric neurons during embryonic development (relevant for Hirschsprung disease).

What we do

About our project

Background information

Modelling Hirschsprung disease

In Hirschsprung disease (HSCR), enteric nervous system (ENS) progenitors have failed to migrate, proliferate, differentiate, or survive within the distal intestine. To gain insight in underlying molecular mechanisms, we will generate zebrafish mutants for genes related to HSCR, enabling to model the disease. Next to the genetic models, we will generate non-genetic HSCR models by treating zebrafish larvae with compounds that inhibit ENS colonization of the gut. 

Validation of candidate genes involved in Hirschsprung disease

HSCR is considered a genetically inherited disease. However, the HSCR genes identified up until today explain only roughly 30% of all cases. In zebrafish, we can relatively easily disrupt genes of interest by injecting CRISPR/Cas9 complexes into fertilized eggs. To visualize gut colonization, we use a reporter fish whose enteric neurons have been labelled with a fluorescent protein. Thereby, we could validate causality of candidate genes identified by DNA sequencing of patients with Hirschsprung disease.

Overall aim?

In these projects we have several aims:

  • Generate additional genetic HSCR zebrafish models.
  • Generate non-genetic HSCR zebrafish models.
  • Identify ’new’ genes that contribute to HSCR.
  • Identify factors involved in ENS colonization.

Research method

We will make use of CRISPR/Cas9 genome editing, in vivo (confocal) microscopy, transcriptomics, immunohistochemistry and bowel motility assays. 

Desirable outcome

These projects will give us insight in the development and functioning of the ENS at a molecular level. Besides, we will increase our understanding of the genetic component of HSCR. By generating genetic and non-genetic HSCR models, we can start searching for small molecules that could stimulate the colonization of the gut by ENS progenitors. This could ultimately lead to the validation of new drugs that could prevent aganglionosis and/or improve HSCR patients’ intestinal innervation.

Publications

Whole exome sequencing coupled with unbiased functional analysis reveals new Hirschsprung disease genes.
Gui H, Schriemer D, Cheng WW, Chauhan RK, Antiňolo G, Berrios C, Bleda M, Brooks AS, Brouwer RW, Burns AJ, Cherny SS, Dopazo J, Eggen BJ, Griseri P, Jalloh B, Le TL, Lui VC, Luzón-Toro B, Matera I, Ngan ES, Pelet A, Ruiz-Ferrer M, Sham PC, Shepherd IT, So MT, Sribudiani Y, Tang CS, van den Hout MC, van der Linde HC, van Ham TJ, van IJcken WF, Verheij JB, Amiel J, Borrego S, Ceccherini I, Chakravarti A, Lyonnet S, Tam PK, Garcia-Barceló MM, Hofstra RM. (2017). Genome Biol. 2017 Mar 8;18(1):48.

Identification of Variants in RET and IHH Pathway Members in a Large Family With History of Hirschsprung Disease.
Sribudiani Y, Chauhan RK, Alves MM, Petrova L, Brosens E, Harrison C, Wabbersen T, de Graaf BM, Rügenbrink T, Burzynski G, Brouwer RWW, van IJcken WFJ, Maas SM, de Klein A, Osinga J, Eggen BJL, Burns AJ, Brooks AS, Shepherd IT, Hofstra RMW. (2018). Gastroenterology. 2018 Jul;155(1):118-129.e6.

Our team

  • Laura Kuil, PhD., Postdoc.
  • Robert Hofstra, Prof.dr., Head of the department.
  • Maria Alves, PhD., Senior scientist.