PUBLICATION
DLX4 is Associated With Orofacial Clefting and Abnormal Jaw Development
- Authors
- Wu, D., Mandal, S., Choi, A., Anderson, A., Prochazkova, M., Perry, H., Gil-da-Silva-Lopes, V.L., Lao, R., Wan, E., Tang, P.L., Kwok, P.Y., Klein, O., Zhuan, B., Slavotinek, A.M.
- ID
- ZDB-PUB-150509-4
- Date
- 2015
- Source
- Human molecular genetics 24(15): 4340-52 (Journal)
- Registered Authors
- Slavotinek, Anne, Wu, Di
- Keywords
- none
- MeSH Terms
-
- Animals
- Bone Morphogenetic Protein 4/genetics
- Brain/abnormalities*
- Brain/pathology
- Cleft Lip/genetics*
- Cleft Lip/pathology
- Cleft Palate/genetics*
- Cleft Palate/pathology
- Exome/genetics
- Gene Expression Regulation, Developmental
- HeLa Cells
- High-Throughput Nucleotide Sequencing
- Homeodomain Proteins/biosynthesis
- Homeodomain Proteins/genetics*
- Humans
- Jaw Abnormalities/genetics*
- Jaw Abnormalities/pathology
- Mesoderm/metabolism
- Mice
- Mice, Knockout
- Morpholinos
- Transcription Factors/biosynthesis
- Transcription Factors/genetics*
- Zebrafish
- Zebrafish Proteins/genetics*
- PubMed
- 25954033 Full text @ Hum. Mol. Genet.
Citation
Wu, D., Mandal, S., Choi, A., Anderson, A., Prochazkova, M., Perry, H., Gil-da-Silva-Lopes, V.L., Lao, R., Wan, E., Tang, P.L., Kwok, P.Y., Klein, O., Zhuan, B., Slavotinek, A.M. (2015) DLX4 is Associated With Orofacial Clefting and Abnormal Jaw Development. Human molecular genetics. 24(15):4340-52.
Abstract
Cleft lip and/or palate (CL/P) are common structural birth defects in humans. We used exome sequencing to study a patient with bilateral CL/P and identified a single nucleotide deletion in the patient and her similarly affected son - c.546_546delG, predicting p.Gln183Argfs*57 in the Distal-less 4 (DLX4) gene. The sequence variant was absent from databases, predicted to be deleterious and was verified by Sanger sequencing. In mammals, there are three Dlx homeobox clusters with closely located gene pairs (Dlx1/Dlx2, Dlx3/Dlx4, Dlx5/Dlx6). In-situ hybridization showed that Dlx4 was expressed in the mesenchyme of the murine palatal shelves at E12.5, prior to palate closure. Wildtype human DLX4, but not mutant DLX4_c.546delG, could activate two murine Dlx conserved regulatory elements, implying that the mutation caused haploinsufficiency. We showed that reduced DLX4 expression after short interfering RNA (siRNA) treatment in a human cell line resulted in significant upregulation of DLX3, DLX5 and DLX6, with reduced expression of DLX2 and significant upregulation of BMP4, although the increased BMP4 expression was demonstrated only in HeLa cells. We used antisense morpholinos (MOs) to target the orthologous Danio rerio gene, dlx4b, and found reduced cranial size and abnormal cartilaginous elements. We sequenced DLX4 in 155 patients with non-syndromic CL/P and CP, but observed no sequence variants. From the published literature, Dlx1/Dlx2 double homozygous null mice and Dlx5 homozygous null mice both have clefts of the secondary palate. This first finding of a DLX4 mutation in a family with CL/P establishes DLX4 as a potential cause of human clefts.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping