PUBLICATION

Targeted resequencing identifies PTCH1 as a major contributor to ocular developmental anomalies and extends the SOX2 regulatory network

Authors
Chassaing, N., Davis, E.E., McKnight, K.L., Niederriter, A.R., Causse, A., David, V., Desmaison, A., Lamarre, S., Vincent-Delorme, C., Pasquier, L., Coubes, C., Lacombe, D., Rossi, M., Dufier, J.L., Dollfus, H., Kaplan, J., Katsanis, N., Etchevers, H.C., Faguer, S., Calvas, P.
ID
ZDB-PUB-160220-2
Date
2016
Source
Genome research   26(4): 474-85 (Journal)
Registered Authors
Davis, Erica, Katsanis, Nicholas
Keywords
none
MeSH Terms
  • Alleles
  • Animals
  • Case-Control Studies
  • Disease Models, Animal
  • Eye Abnormalities/genetics*
  • Eye Abnormalities/metabolism*
  • Gene Expression Regulation*
  • Gene Regulatory Networks*
  • Genetic Loci
  • Heterozygote
  • Humans
  • Mutation
  • Patched-1 Receptor/genetics*
  • Patched-1 Receptor/metabolism
  • Phenotype
  • SOXB1 Transcription Factors/metabolism*
  • Sequence Analysis, DNA
  • Zebrafish
PubMed
26893459 Full text @ Genome Res.
Abstract
Ocular developmental anomalies (ODA) such as Anophthalmia/Microphthalmia (AM) or anterior segment dysgenesis (ASD) have an estimated combined prevalence of 3.7 in 10,000 births. Mutations in SOX2 are the most frequent contributors to severe ODA, yet account for a minority of the genetic drivers. To identify novel ODA loci, we conducted targeted high-throughput sequencing of 407 candidate genes in an initial cohort of 22 sporadic ODA patients. Patched 1 (PTCH1), an inhibitor of sonic hedgehog (SHH) signaling, harbored an enrichment of rare heterozygous variants in comparison to either controls, or to the other candidate genes (four missense and one frameshift); targeted resequencing of PTCH1 in a second cohort of 48 ODA patients identified two additional rare nonsynonymous changes. Using multiple transient models and a CRISPR/Cas9 mutant, we show physiologically relevant phenotypes altering SHH signaling and eye development upon abrogation of ptch1 in zebrafish which in vivo complementation assays using these models showed that all six patient missense mutations affect SHH signaling. Finally, through transcriptomic and ChIP analyses, we show that SOX2 binds to an intronic domain of the PTCH1 locus to regulate PTCH1 expression, findings that were validated both in vitro and in vivo. Together, these results demonstrate that PTCH1 mutations contribute to as much as 10% of ODA, identify the SHH signaling pathway as a novel effector of SOX2 activity during human ocular development, and indicate that ODA is likely the result of overactive SHH signaling in humans harboring mutations in either PTCH1 or SOX2.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping