ANALYSIS OF GENE COPY NUMBER VARIATIONS IN PATIENTS WITH CARDIAC SEPTAL DEFECTS USING MULTIPLEX LIGATION-DEPENDENT PROBE AMPLIFICATION

Yashvanthi Borkar Yashvanthi Borkar,Rajasekhar Moka Rajasekhar Moka,Ranjan Shetty K Ranjan Shetty K,Krishnananda Nayak Krishnananda Nayak

doi:10.22159/ajpcr.2020.v13i1.36189

Yashvanthi Borkar Yashvanthi Borkar, Rajasekhar Moka Rajasekhar Moka + Show 2 more

Open Access

https://doi.org/10.22159/ajpcr.2020.v13i1.36189

Copy DOI

Abstract

Objective: Cardiac septal defects (CSDs), the most common human congenital heart malformations are complex and heterogeneous. Progress in molecular biology has helped to identify many genes responsible for cardiac morphogenesis. However, etiologic factors in familial as well as isolated syndromes are being identified; the root genetic cause still needs to be resolved and its mechanism is yet to be revealed. The objective of this study is to identify DNA copy number variations (CNVs) and their possible association with septal defects. Methods: Multiplex ligation-dependent probe amplification (MLPA) was used to detect DNA copy number in non-syndromic CSDs using the P311-A1 Kit consisting of probes for the key genes, namely, NKX2-5 (NK2 transcription factor related, locus 5), GATA4 (GATA binding protein 4), TBX5 (T-box transcription factor), bone morphogenetic protein 4, and CRELD1 (cysteine rich with EGF-like domains 1). Results: We studied 124 clinically diagnosed CSD subjects, of which 111 (89.5%) had atrial septal defects and 13 (10.5%) had ventricular septal defects. MLPA assay was carried out in all these patients after a thorough clinical and cytogenetic screening. CNVs were identified in 16 (12.9%) cases, of which heterozygous deletions and heterozygous duplications were detected (8 patients each) with apparent phenotypes. Conclusion: MLPA could be a useful assay for the detection of CNVs and to be adopted as the first line of screening in patients with congenital heart diseases.

Highlights

Cardiac septal defects (CSDs) constitute 1% of all congenital heart diseases (CHDs) [1]
copy number variations (CNVs) were identified in 16 (12.9%) cases, of which heterozygous deletions and heterozygous duplications were detected (8 patients each) with apparent phenotypes
multiplex ligation-dependent probe amplification (MLPA) could be a useful assay for the detection of CNVs and to be adopted as the first line of screening in patients with congenital heart diseases

Summary

Introduction

Cardiac septal defects (CSDs) constitute 1% of all congenital heart diseases (CHDs) [1]. CHDs can either be isolated (nonsyndromic; septal defects) or in combination with other cardiac lesions (syndromic; Holt-Oram syndrome). The septal defects such as atrial septal defects (ASDs) and ventricular septal defects (VSDs) may be asymptomatic, with the appearance of murmur as the primary symptom. Congenital septal defects are the most common of all structural malformations of the human heart and are caused by both genetic and environmental factors [3]. Despite recent molecular genetic studies, efforts to identify genotype-phenotype correlation in patients with septal defects suggest a more complex pattern of inheritance, where copy number variations (CNVs), as well as single nucleotide polymorphisms, contribute to the disease pathogenesis and tune the spectrum of septal malformations. MLPA could be a promising technique for proper diagnosis, prognosis, and management of the patients and affected families

Methods

Results

Discussion

Conclusion