Clinical and molecular evaluation of SHOX/PAR1 duplications in Léri–Weill dyschondrosteosis (LWD) and Idiopathic Short Stature (ISS)

Department of Pediatrics, Universidad Autónoma de Madrid, Madrid, Madrid, Spain
The Journal of Clinical Endocrinology and Metabolism (Impact Factor: 6.21). 02/2011; 96(2):E404-12. DOI: 10.1210/jc.2010-1689
Source: PubMed


Léri-Weill dyschondrosteosis (LWD) is a skeletal dysplasia characterized by disproportionate short stature and the Madelung deformity of the forearm. SHOX mutations and pseudoautosomal region 1 deletions encompassing SHOX or its enhancers have been identified in approximately 60% of LWD and approximately 15% of idiopathic short stature (ISS) individuals. Recently SHOX duplications have been described in LWD/ISS but also in individuals with other clinical manifestations, thus questioning their pathogenicity.
The objective of the study was to investigate the pathogenicity of SHOX duplications in LWD and ISS.
Multiplex ligation-dependent probe amplification is routinely used in our unit to analyze for SHOX/pseudoautosomal region 1 copy number changes in LWD/ISS referrals. Quantitative PCR, microsatellite marker, and fluorescence in situ hybridization analysis were undertaken to confirm all identified duplications.
During the routine analysis of 122 LWD and 613 ISS referrals, a total of four complete and 10 partial SHOX duplications or multiple copy number (n > 3) as well as one duplication of the SHOX 5' flanking region were identified in nine LWD and six ISS cases. Partial SHOX duplications appeared to have a more deleterious effect on skeletal dysplasia and height gain than complete SHOX duplications. Importantly, no increase in SHOX copy number was identified in 340 individuals with normal stature or 104 overgrowth referrals.
MLPA analysis of SHOX/PAR1 led to the identification of partial and complete SHOX duplications or multiple copies associated with LWD or ISS, suggesting that they may represent an additional class of mutations implicated in the molecular etiology of these clinical entities.

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Available from: Karen E Heath, Jul 02, 2015
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    • "Trisomies of the sex chromosomes (47,XXX; 47, XXY; 47,XYY) are associated with normal to tall stature [Ottesen et al., 2010], but contrasting phenotypes result from chromosomal structural rearrangements leading to microscopic deletions or duplications of the SHOX/PAR1 region [Roos et al., 2009; Thomas et al., 2009; Iughetti et al., 2010; Benito-Sanz et al., 2011]. However, duplications of long-range enhancers may cause SHOX-related phenotypes: an upstream duplication, which includes the CNE-5 and CNE-3 elements has been described in a family referred with ISS [Benito-Sanz et al., 2011] and a downstream duplication of ECS4 was separately reported in four ISS patients [Hirschfeldova et al., 2012; Fukami et al., 2015]. "
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    ABSTRACT: Leri-Weill dyschondrosteosis is a pseudoautosomal dominantly-inherited skeletal dysplasia ascribed to haploinsufficiency of the SHOX gene caused by deletions, point mutations, or partial duplications of the gene, or to heterozygous deletions upstream or downstream of the intact SHOX gene involving conserved non-coding cis-regulatory DNA elements that show enhancer activity. Recently, two SHOX conserved non-coding element duplications, one upstream and one downstream, were reported in patients referred with idiopathic short stature. To further evaluate the role of these duplications in SHOX-related disorders, we describe seven patients (five with Leri-Weill dyschondrosteosis and two with short stature) all of whom have duplications of part of the upstream or downstream conserved non-coding element regions, identified by multiplex ligation-dependent probe amplification. In addition, we show data from 32 patients with an apparently identical downstream duplication that includes a proposed putative regulatory element (identified by multiplex ligation-dependent probe amplification or array comparative genome hybridization), which results in a variable phenotype from normal to mild Leri-Weill dyschondrosteosis. These additional data provide further evidence that duplications of upstream and downstream long range cis-regulatory DNA elements can result in a SHOX-related phenotype. © 2015 Wiley Periodicals, Inc.
    Full-text · Article · Dec 2015 · American Journal of Medical Genetics Part A
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    • "Family histories were also documented, including parental heights. In all cases, the presence alterations in SHOX or its enhancers had been previously excluded [4], [7]–[9], [32], [33]. The control cohort consisted of 95 Spanish individuals with normal heights (Spanish DNA Bank, University of Salamanca). "
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    ABSTRACT: SHOX and SHOX2 transcription factors are highly homologous, with even identical homeodomains. Genetic alterations in SHOX result in two skeletal dysplasias; Léri-Weill dyschondrosteosis (LWD) and Langer mesomelic dysplasia (LMD), while no human genetic disease has been linked to date with SHOX2. SHOX2 is, though, involved in skeletal development, as shown by different knockout mice models. Due to the high homology between SHOX and SHOX2, and their functional redundancy during heart development, we postulated that SHOX2 might have the same transcriptional targets and cofactors as SHOX in limb development. We selected two SHOX transcription targets regulated by different mechanisms: 1) the natriuretic peptide precursor B gene (NPPB) involved in the endochondral ossification signalling and directly activated by SHOX; and 2) Aggrecan (ACAN), a major component of cartilage extracellular matrix, regulated by the cooperation of SHOX with the SOX trio (SOX5, SOX6 and SOX9) via the protein interaction between SOX5/SOX6 and SHOX. Using the luciferase assay we have demonstrated that SHOX2, like SHOX, regulates NPPB directly whilst activates ACAN via its cooperation with the SOX trio. Subsequently, we have identified and characterized the protein domains implicated in the SHOX2 dimerization and also its protein interaction with SOX5/SOX6 and SHOX using the yeast-two hybrid and co-immunoprecipitation assays. Immunohistochemistry of human fetal growth plates from different time points demonstrated that SHOX2 is coexpressed with SHOX and the members of the SOX trio. Despite these findings, no mutation was identified in SHOX2 in a cohort of 83 LWD patients with no known molecular defect, suggesting that SHOX2 alterations do not cause LWD. In conclusion, our work has identified the first cofactors and two new transcription targets of SHOX2 in limb development, and we hypothesize a time- and tissue-specific functional redundancy between SHOX and SHOX2.
    Full-text · Article · Jan 2014 · PLoS ONE
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    • "Such heterogeneity prompted us to investigate other genetic mutations in the causation of the HCH phenotype. Mutations in the SHOX gene have been associated with other related syndromes of short stature including idiopathic short stature, Leri–Weill syndrome dyschondrosteosis, and Turner syndrome [Benito-Sanz et al., 2011], though this possibility of the SHOX gene mutations being associated with a HCH phenotype has not been stressed. On the other hand, FGFR1 and 2 mutations were previously reported in patients with other skeletal dysplasias like craniosynostosis [Grigelioniene et al., 2000]. "
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    ABSTRACT: We applied a comprehensive set of clinical and radiological criteria for the diagnosis of hypochondroplasia (HCH) in 160 patients with short stature 58 of whom were diagnosed to have HCH. Taking into account the genotypic and phenotypic variations in HCH, we conducted a study with these 58 patients and tested them for mutations in the fibroblast growth factor receptor 3 (FGFR3) and the short stature homeobox (SHOX) gene. We characterized the phenotypes by clinical and radiologic findings. In the patients with HCH, 19 were included in Group I (FGFR3 mutations-mutations of definite significance), and 39 were in Group II (6 SHOX mutations and 33 negative for disease-causing FGFR3 mutations). The clinical findings were similar in two groups regardless of the presence or absence of mutations. More than 95% of the patients had mesomelic proportions. In Group I, the radiological findings of mesomelia of upper and lower limbs and, L1/L4 ratio in anterior-posterior and lateral view were more typical than in Group II. This study proposes comprehensive clinico-radiological criteria for the diagnosis of HCH, which would help in detecting the true incidence of this underdiagnosed condition. The presence of SHOX mutations suggest genotypic-phenotypic overlap between HCH and Leri-Weill dyschondrosteosis, though further investigation is needed to effectively elucidate the importance of these mutations. Also, the 56.9% of HCH patients with negative mutations for FGFR3 suggests that there are other undiscovered gene mutations associated with this phenotypic entity. © 2012 Wiley Periodicals, Inc.
    Full-text · Article · Oct 2012 · American Journal of Medical Genetics Part A
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