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

Nicolas Chassaing; Erica E. Davis; Kelly L. Mcknight; Adrienne R. Niederriter; Alexandre Causse; Véronique David; Annaick Desmaison; Sophie Lamarre; Catherine Vincent-Delorme; Laurent Pasquier; Christine Coubes; Didier Lacombe; Massimiliano Rossi; Jean-Louis Dufier; Hélène Dollfus; Josseline Kaplan; Nicholas Katsanis; Heather C. Etchevers; Stanislas Faguer; Patrick Calvas

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

Nicolas Chassaing ^{1, 2, 3} Erica E. Davis Kelly L. Mcknight Adrienne R. Niederriter Alexandre Causse ^{2, 1} Véronique David ^{4, 5} Annaick Desmaison ⁶ Sophie Lamarre ⁷ Catherine Vincent-Delorme ⁸ Laurent Pasquier ⁹ Christine Coubes ¹⁰ Didier Lacombe ¹¹ Massimiliano Rossi ¹² Jean-Louis Dufier ¹³ Hélène Dollfus ¹⁴ Josseline Kaplan ^{15, 16} Nicholas Katsanis ¹⁷ Heather C. Etchevers Stanislas Faguer ^{18, 19, 20} Patrick Calvas ^{1, 2, 3}

1 Hôpital Purpan [Toulouse]

2 UPS - Université Toulouse III - Paul Sabatier

3 UDEAR - Unité différenciation épidermique et auto-immunité rhumatoïde

4 IGDR - Institut de Génétique et Développement de Rennes

5 Service de biologie moléculaire

6 ITAV - Institut des Technologies Avancées en sciences du Vivant

7 LISBP - Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés

8 Centre de Maladies Rares

9 CHU Pontchaillou [Rennes]

10 Service de Génétique Clinique

11 Service de génétique médicale

12 CRNL - Centre de recherche en neurosciences de Lyon

13 Service d'ophtalmologie [CHU Necker]

14 Centre de Référence pour les Affections Rares en Génétique Ophtalmologique (CARGO) et Service de Génétique Médicale

15 IMAGINE - U1163 - Imagine - Institut des maladies génétiques

16 UPD5 - Université Paris Descartes - Paris 5

17 Center for Human Disease Modeling

18 Département de Néphrologie et Transplantation d'organes

Hôpital de Rangueil

19 Centre de référence des maladies rénales rares

20 I2MC - Institut des Maladies Métaboliques et Cardiovasculaires

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.

Document type :

Journal articles

Domain :

Life Sciences [q-bio] / Development Biology

Life Sciences [q-bio] / Biochemistry, Molecular Biology / Genomics [q-bio.GN]

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Genome Res.-2016-Chassaing-474...

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

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

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