A Mutation in VWA1, Encoding von Willebrand Factor A Domain-Containing Protein 1, Is Associated With Hemifacial Microsomia

Yibei Wang; Lu Ping; Xiaodong Luan; Yushan Chen; Xinmiao Fan; Lianyan Li; Yaping Liu; Pu Wang; Shuyang Zhang; Bo Zhang; Xiaowei Chen

doi:10.3389/fcell.2020.571004

A Mutation in VWA1, Encoding von Willebrand Factor A Domain-Containing Protein 1, Is Associated With Hemifacial Microsomia

Front Cell Dev Biol. 2020 Sep 9:8:571004. doi: 10.3389/fcell.2020.571004. eCollection 2020.

Authors

Yibei Wang^{1

2}, Lu Ping³, Xiaodong Luan^{4

5

6}, Yushan Chen⁷, Xinmiao Fan¹, Lianyan Li⁸, Yaping Liu⁹, Pu Wang^{1

10}, Shuyang Zhang^{4

5

6}, Bo Zhang⁸, Xiaowei Chen¹

Affiliations

¹ Department of Otolaryngology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
² Department of Otolaryngology, China-Japan Friendship Hospital, Beijing, China.
³ Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
⁴ School of Medicine, Tsinghua University, Beijing, China.
⁵ Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.
⁶ Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China.
⁷ Department of Otolaryngology, The Ohio State University, Columbus, OH, United States.
⁸ Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, China.
⁹ Department of Medical Genetics and National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
¹⁰ Department of Otolaryngology Head and Neck Surgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.

Abstract

Background: Hemifacial microsomia (HFM) is a type of rare congenital syndrome caused by developmental disorders of the first and second pharyngeal arches that occurs in one out of 5,600 live births. There are significant gaps in our knowledge of the pathogenic genes underlying this syndrome.

Methods: Whole exome sequencing (WES) was performed on five patients, one asymptomatic carrier, and two marry-in members of a five-generation pedigree. Structure of WARP (product of VWA1) was predicted using the Phyre2 web portal. In situ hybridization and vwa1-knockdown/knockout studies in zebrafish using morpholino and CRISPR/Cas9 techniques were performed. Cartilage staining and immunofluorescence were carried out.

Results: Through WES and a set of filtration, we identified a c.G905A:p.R302Q point mutation in a novel candidate pathogenic gene, VWA1. The Phyre2 web portal predicted alterations in secondary and tertiary structures of WARP, indicating changes in its function as well. Predictions of protein-to-protein interactions in five pathways related to craniofacial development revealed possible interactions with four proteins in the FGF pathway. Knockdown/knockout studies of the zebrafish revealed deformities of pharyngeal cartilage. A decrease of the proliferation of cranial neural crest cells (CNCCs) and alteration of the structure of pharyngeal chondrocytes were observed in the morphants as well.

Conclusion: Our data suggest that a mutation in VWA1 is functionally linked to HFM through suppression of CNCC proliferation and disruption of the organization of pharyngeal chondrocytes.

Keywords: VWA1; cranial neural crest cell; hemifacial microsomia; whole exome sequencing; zebrafish.