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Exome sequencing and functional analysis identifies a novel mutation in EXT1 gene that causes multiple osteochondromas.

Zhang F, Liang J, Guo X, Zhang Y, Wen Y, Li Q, Zhang Z, Ma W, Dai L, Liu X, Yang L, Wang J - PLoS ONE (2013)

Bottom Line: Exome sequencing has high chromosomal coverage and accuracy, and has recently been successfully used to identify pathogenic gene mutations.After filtering the data from the 1000 Genome Project and the dbSNP database (build 132), the detected candidate gene mutations were further validated via Sanger sequencing of four other members of the same MO family and 200 unrelated healthy subjects.Immunohistochemisty and multiple sequence alignment were performed to evaluate the importance of the identified causal mutation.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Environment and Gene Related Diseases of Ministry Education, Faculty of Public Health, College of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, China.

ABSTRACT
Multiple osteochondromas (MO) is an inherited skeletal disorder, and the molecular mechanism of MO remains elusive. Exome sequencing has high chromosomal coverage and accuracy, and has recently been successfully used to identify pathogenic gene mutations. In this study, exome sequencing followed by Sanger sequencing validation was first used to screen gene mutations in two representative MO patients from a Chinese family. After filtering the data from the 1000 Genome Project and the dbSNP database (build 132), the detected candidate gene mutations were further validated via Sanger sequencing of four other members of the same MO family and 200 unrelated healthy subjects. Immunohistochemisty and multiple sequence alignment were performed to evaluate the importance of the identified causal mutation. A novel frameshift mutation, c.1457insG at codon 486 of exon 6 of EXT1 gene, was identified, which truncated the glycosyltransferase domain of EXT1 gene. Multiple sequence alignment showed that codon 486 of EXT1 gene was highly conserved across various vertebrates. Immunohistochemisty demonstrated that the chondrocytes with functional EXT1 in MO were less than those in extragenetic solitary chondromas. The novel c.1457insG deleterious mutation of EXT1 gene reported in this study expands the causal mutation spectrum of MO, and may be helpful for prenatal genetic screening and early diagnosis of MO.

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Immunohistochemisty screening of chondrocytes with functional EXT1 in the superficial layers of cartilage caps of MO(a) and extragenetic solitary chondroma(b) (40×).The chondrocytes with functional EXT1 in MO are less than those in extragenetic solitary chondroma.
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pone-0072316-g003: Immunohistochemisty screening of chondrocytes with functional EXT1 in the superficial layers of cartilage caps of MO(a) and extragenetic solitary chondroma(b) (40×).The chondrocytes with functional EXT1 in MO are less than those in extragenetic solitary chondroma.

Mentions: To assess the functional impact of c.1457insG on EXT1 protein, immunohistochemistry was used to compare EXT1 protein level between a patient with MO and a patient with extragenetic solitary chondromas. In extragenetic solitary chondromas, EXT1 protein was enriched in the cytoplasm of chondrocytes (Fig. 3b). Comparing with extragenetic solitary chondroma, the chondrocytes with functional EXT1 in MO were less than those in extragenetic solitary chondroma (Fig. 3a).


Exome sequencing and functional analysis identifies a novel mutation in EXT1 gene that causes multiple osteochondromas.

Zhang F, Liang J, Guo X, Zhang Y, Wen Y, Li Q, Zhang Z, Ma W, Dai L, Liu X, Yang L, Wang J - PLoS ONE (2013)

Immunohistochemisty screening of chondrocytes with functional EXT1 in the superficial layers of cartilage caps of MO(a) and extragenetic solitary chondroma(b) (40×).The chondrocytes with functional EXT1 in MO are less than those in extragenetic solitary chondroma.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3757002&req=5

pone-0072316-g003: Immunohistochemisty screening of chondrocytes with functional EXT1 in the superficial layers of cartilage caps of MO(a) and extragenetic solitary chondroma(b) (40×).The chondrocytes with functional EXT1 in MO are less than those in extragenetic solitary chondroma.
Mentions: To assess the functional impact of c.1457insG on EXT1 protein, immunohistochemistry was used to compare EXT1 protein level between a patient with MO and a patient with extragenetic solitary chondromas. In extragenetic solitary chondromas, EXT1 protein was enriched in the cytoplasm of chondrocytes (Fig. 3b). Comparing with extragenetic solitary chondroma, the chondrocytes with functional EXT1 in MO were less than those in extragenetic solitary chondroma (Fig. 3a).

Bottom Line: Exome sequencing has high chromosomal coverage and accuracy, and has recently been successfully used to identify pathogenic gene mutations.After filtering the data from the 1000 Genome Project and the dbSNP database (build 132), the detected candidate gene mutations were further validated via Sanger sequencing of four other members of the same MO family and 200 unrelated healthy subjects.Immunohistochemisty and multiple sequence alignment were performed to evaluate the importance of the identified causal mutation.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Environment and Gene Related Diseases of Ministry Education, Faculty of Public Health, College of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, China.

ABSTRACT
Multiple osteochondromas (MO) is an inherited skeletal disorder, and the molecular mechanism of MO remains elusive. Exome sequencing has high chromosomal coverage and accuracy, and has recently been successfully used to identify pathogenic gene mutations. In this study, exome sequencing followed by Sanger sequencing validation was first used to screen gene mutations in two representative MO patients from a Chinese family. After filtering the data from the 1000 Genome Project and the dbSNP database (build 132), the detected candidate gene mutations were further validated via Sanger sequencing of four other members of the same MO family and 200 unrelated healthy subjects. Immunohistochemisty and multiple sequence alignment were performed to evaluate the importance of the identified causal mutation. A novel frameshift mutation, c.1457insG at codon 486 of exon 6 of EXT1 gene, was identified, which truncated the glycosyltransferase domain of EXT1 gene. Multiple sequence alignment showed that codon 486 of EXT1 gene was highly conserved across various vertebrates. Immunohistochemisty demonstrated that the chondrocytes with functional EXT1 in MO were less than those in extragenetic solitary chondromas. The novel c.1457insG deleterious mutation of EXT1 gene reported in this study expands the causal mutation spectrum of MO, and may be helpful for prenatal genetic screening and early diagnosis of MO.

Show MeSH
Related in: MedlinePlus