A complementary study approach unravels novel players in the pathoetiology of Hirschsprung disease

• Published in: PLoS genetics 2020-11, Vol.16 (11), p.e1009106-e1009106
• Main Authors: Mederer, Tanja, Schmitteckert, Stefanie, Volz, Julia, Martínez, Cristina, Röth, Ralph, Thumberger, Thomas, Eckstein, Volker, Scheuerer, Jutta, Thöni, Cornelia, Lasitschka, Felix, Carstensen, Leonie, Günther, Patrick, Holland-Cunz, Stefan, Hofstra, Robert, Brosens, Erwin, Rosenfeld, Jill A, Schaaf, Christian P, Schriemer, Duco, Ceccherini, Isabella, Rusmini, Marta, Tilghman, Joseph, Luzón-Toro, Berta, Torroglosa, Ana, Borrego, Salud, Sze-Man Tang, Clara, Garcia-Barceló, Mercè, Tam, Paul, Paramasivam, Nagarajan, Bewerunge-Hudler, Melanie, De La Torre, Carolina, Gretz, Norbert, Rappold, Gudrun A, Romero, Philipp, Niesler, Beate
• Format: Article
• Language: English
• Subjects: Adenosine triphosphatase; Animals; ATP Binding Cassette Transporter, Subfamily D, Member 1 - genetics; Biology and Life Sciences; Candidates; Cell Differentiation - genetics; Cell Line; Cell Proliferation - genetics; Cell Survival - genetics; Central nervous system; Computer Simulation; Copper-Transporting ATPases - genetics; Disease Models, Animal; DNA microarrays; Embryos; Exome sequencing; Gene expression; Gene Expression Profiling; Gene frequency; Gene Knockout Techniques; Genes; Genetic aspects; Genetic research; Glial cell line-derived neurotrophic factor; Hirschsprung Disease - genetics; Hirschsprung's disease; Humans; Identification and classification; Infant; Male; Medicine and Health Sciences; Mice; Mutation; Nervous system; Neural crest; Patients; Phenotypes; Phox2b protein; Protein Inhibitors of Activated STAT - genetics; Proteins; Research and Analysis Methods; Risk factors; Semaphorins; Sox10 protein; Sterol Regulatory Element Binding Protein 1 - genetics; Sterol regulatory element-binding protein; Transcription; Vagus nerve; Viroids; Whole Exome Sequencing
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1009106

Hirschsprung disease (HSCR, OMIM 142623) involves congenital intestinal obstruction caused by dysfunction of neural crest cells and their progeny during enteric nervous system (ENS) development. HSCR is a multifactorial disorder; pathogenetic variants accounting for disease phenotype are identified only in a minority of cases, and the identification of novel disease-relevant genes remains challenging. In order to identify and to validate a potential disease-causing relevance of novel HSCR candidate genes, we established a complementary study approach, combining whole exome sequencing (WES) with transcriptome analysis of murine embryonic ENS-related tissues, literature and database searches, in silico network analyses, and functional readouts using candidate gene-specific genome-edited cell clones. WES datasets of two patients with HSCR and their non-affected parents were analysed, and four novel HSCR candidate genes could be identified: ATP7A, SREBF1, ABCD1 and PIAS2. Further rare variants in these genes were identified in additional HSCR patients, suggesting disease relevance. Transcriptomics revealed that these genes are expressed in embryonic and fetal gastrointestinal tissues. Knockout of these genes in neuronal cells demonstrated impaired cell differentiation, proliferation and/or survival. Our approach identified and validated candidate HSCR genes and provided further insight into the underlying pathomechanisms of HSCR.