DHPLC-based mutation analysis of ENG and ALK-1 genes in HHT Italian population

Sabbá, Carlo; Lastella, Patrizia; Giacomo, Marilena Carmela Di; Resta, Nicoletta; Suppressa, Patrizia; Pasculli, Giuseppe; Sabbà, Carlo; Guanti, Ginevra

doi:10.1002/humu.9400

Cited by 35 publications

(31 citation statements)

References 21 publications

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“…A second form of HHT (HHT2) is caused by mutations in the gene coding for the TGF-␤ type I receptor known as activin receptor-like kinase-1 (ACVRL1 or ALK1). Either of the two genes, ENG or ACVRL1, is mutated in more than 90% of patients with HHT (1,19,52,57,94,95). Two additional loci for HHT have been mapped to chromosomes 5 and 7, but the corresponding mutant genes have not been identified yet (10,35).…”

Section: Role Of Endoglin In Vascular Pathologymentioning

confidence: 99%

The physiological role of endoglin in the cardiovascular system

López‐Novoa

Bernabeu

2010

American Journal of Physiology-Heart and Circulatory Physiology

190

200

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Endoglin (CD105) is an integral membrane glycoprotein that serves as a coreceptor for members of the transforming growth factor-β superfamily of proteins. A major role for endoglin in regulating transforming growth factor-β-dependent vascular remodeling and angiogenesis has been postulated based on the following: 1) endoglin is the gene mutated in hereditary hemorrhagic telangiectasia type 1, a disease characterized by vascular malformations; 2) endoglin knockout mice die at midgestation because of defective angiogenesis; 3) endoglin is overexpressed in neoangiogenic vessels, during inflammation, and in solid tumors; and 4) endoglin regulates the expression and activity of endothelial nitric oxide synthase, which is involved in angiogenesis and vascular tone. Besides the predominant form of the endoglin receptor (long endoglin isoform), two additional forms of endoglin have been recently reported to play a role in the vascular pathology and homeostasis: the alternatively spliced short endoglin isoform and a soluble endoglin form that is proteolytically cleaved from membrane-bound endoglin. The purpose of this review is to underline the role that the different forms of endoglin play in regulating angiogenesis, vascular remodeling, and vascular tone, as well as to analyze the molecular and cellular mechanisms supporting these effects.

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Section: Role Of Endoglin In Vascular Pathologymentioning

confidence: 99%

The physiological role of endoglin in the cardiovascular system

López‐Novoa

Bernabeu

2010

American Journal of Physiology-Heart and Circulatory Physiology

190

200

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“…A newer technique that detects heteroduplexes in PCR amplicons by ion-pair reverse-phase high performance liquid chromatography has been developed (Oefner & Underhill 1998, Xiao & Oefner 2001. Denaturing highperformance liquid chromatography (DHPLC) has proved useful for mutational analysis of genes such as MEN1 (Crépin et al 2006), RB1 (Houdayer et al 2004), ENG and ALK-1 (Lenato et al 2006), NPHS2/podocin (He et al 2007), and BRCA1/2 (Gerhardus et al 2007). We (Hendy et al 2003) and others (Waller et al 2004) have successfully utilized the DHPLC method for CASR mutational analysis in a few cases.…”

Section: Introductionmentioning

confidence: 99%

Calcium-sensing receptor mutations and denaturing high performance liquid chromatography

Cole¹,

Yun²,

Wong³

et al. 2009

Journal of Molecular Endocrinology

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The calcium-sensing receptor (CASR), a plasma membrane G-protein-coupled receptor, is expressed in parathyroid gland and kidney, and controls systemic calcium homeostasis. Inactivating CASR mutations are associated with familial hypocalciuric hypercalcemia (FHH) and neonatal severe hyperparathyroidism, and activating mutations cause autosomal dominant hypocalcemia (ADH). CASR mutation identification plays an important role in the clinical management of mineral metabolism disorders. We describe here a high-throughput method using screening with denaturing high performance liquid chromatography (DHPLC) to initially interrogate 12 amplicons covering translated exons and exon/intron boundaries, followed by sequencing of any amplicon with a modified melting curve relative to wild type, and direct sequencing of a 13th amplicon encoding the COOH-terminal tail to distinguish causative mutations from three common missense single nucleotide polymorphisms. A blinded analysis of 32 positive controls representing mutations throughout the CASR sequence, as well as 22 negative controls, yielded a concordance rate of 100%. We report eight novel and five recurrent FHH mutations, along with six novel and two recurrent ADH mutations. Thus, DHPLC provides a rapid and effective means to screen for CASR mutations.

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“…A peculiar distribution of mutations has been reported: ENG mutations are more frequently found in patients from Northern Europe and the Americas, while Mediterranean populations have a majority of ACVRL1 mutations (Olivieri et al 2002;Brusgaard et al 2004;Lesca et al 2004Lesca et al , 2006Abdalla et al 2005;Kjeldsen et al 2005;Letteboer et al 2005Letteboer et al , 2006Bayrak-Toydemir et al 2006b;Fernandez-L et al 2006; Lenato et al 2006;Bossler et al 2006). …”

Section: Introductionmentioning

confidence: 99%

Analysis of ENG and ACVRL1 genes in 137 HHT Italian families identifies 76 different mutations (24 novel). Comparison with other European studies

et al. 2007

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Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disorder causing vascular dysplasias. About 70-80% of HHT patients carries mutations in ENG or ACVRL1 genes, which code for a TGFb receptor type III and I respectively. Molecular data on a large cohort of Italian HHT patients are presented, discussing the significance of missense and splice site mutations. Mutation analysis in ENG and ACVRL1 genes was performed using single strand conformation polymorphisms (SSCP), denaturing high performance liquid chromatography (DHPLC) and subsequent direct sequencing. Overall, 101 mutations were found, with ACVRL1 involved in 71% of cases. The highest number of mutations (28/101 subjects, 14/76 different mutations referring to both genes) was in ACVRL1, exon 3. Mutation analysis was then extended to a total of 356 family members, and 162 proven to carry the mutation. New polymorphisms were identified in both genes, and evidence that ENG P131L change is not a disease-causing mutation was also provided. An in silico analysis was performed in order to characterize splice-site mutations. These results were compared to other European national studies and data from Italy, France and Spain were consistent for an higher incidence of ACVRL1 mutations.

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DHPLC-based mutation analysis of ENG and ALK-1 genes in HHT Italian population

Abstract: Hereditary haemorrhagic telangiectasia (HHT orRendu

Cited by 35 publications

References 21 publications

The physiological role of endoglin in the cardiovascular system

The physiological role of endoglin in the cardiovascular system

Calcium-sensing receptor mutations and denaturing high performance liquid chromatography

Analysis of ENG and ACVRL1 genes in 137 HHT Italian families identifies 76 different mutations (24 novel). Comparison with other European studies

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