Novel LEPR mutations in obese Pakistani children identified by PCR‐based enrichment and next generation sequencing. (9th December 2013)

• Record Type: Journal Article
• Authors: Saeed, Sadia; Bonnefond, Amélie; Manzoor, Jaida; Philippe, Julien; Durand, Emmanuelle; Arshad, Mohsin; Sand, Olivier; Butt, Taeed A; Falchi, Mario; Arslan, Muhammad; Froguel, Philippe
• Is Part Of: Obesity. Volume 22:Number 4(2014:Apr.)
• Page Start: 1112
• Page End: 1117
• Publication Date: 2013-12-09
• Subjects: Obesity -- Periodicals; 616.398005
• DOI: 10.1002/oby.20667
• Languages: English
• ISSNs: 1930-7381

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="oby20667-sec-0001" sec-type="section"> <title>Objective</title> <p>Mutations in leptin receptor gene (<italic>LEPR</italic>) result in early onset extreme adiposity. However, their prevalence in different populations is not known. Indeed, <italic>LEPR</italic> screening by gold standard Sanger sequencing has been limited by its large size and the cost. One‐step PCR‐based targeted enrichment could be an option for rapid and cost effective molecular diagnosis of monogenic forms of obesity.</p> </sec> <sec id="oby20667-sec-0002" sec-type="section"> <title>Methods</title> <p>The study is based on 39 unrelated severely obese Pakistani children, previously shown to be negative for leptin (<italic>LEP</italic>) and melanocortin 4 receptor (<italic>MC4R</italic>) gene mutations, from an initial cohort of 62 probands. Patient samples were analyzed by microdroplet PCR‐enrichment (RainDance technologies) targeting coding exons of 26 obesity‐associated genes combined with next generation sequencing. Hormone levels were analyzed by ELISA.</p> </sec> <sec id="oby20667-sec-0003" sec-type="section"> <title>Results</title> <p>The analysis revealed two novel homozygous <italic>LEPR</italic> mutations, an essential splice site mutation in exon 15 (c.2396‐1 G>T), and a nonsense mutation in exon 10 (c.1675 G>A). Both probands had high leptin levels and were phenotypically indistinguishable from age‐matched leptin‐deficient subjects from the same population.</p> </sec> <sec id="oby20667-sec-0004" sec-type="section"> <title>Conclusions</title> <p>The two subjects carrying homozygous <italic>LEPR</italic> mutations, reported here for the first time in the Pakistani population, constitute 3% of the whole cohort of severely obese children (compared to 17% for <italic>LEP</italic> and 3% for <italic>MC4R</italic>).</p> </sec> </abstract>