Analysis of CFTR Mutation Spectrum in Ethnic Russian Cystic Fibrosis Patients

The distribution and frequency of the CFTR gene mutations vary considerably between countries and ethnic groups. Russians are an East Slavic ethnic groups are native to Eastern Europe. Russians, the most numerous people of the Russian Federation (RF), make about 80% of the population. The aim is to reveal the molecular causes of CF in ethnic Russian patients as comprehensively as possible. The analysis of most common CFTR mutations utilized for CF diagnosis in multiethnic RF population accounts for about 83% of all CF-causing mutations in 1384 ethnic Russian patients. Variants c.1521_1523delCTT (F508del), c.54-5940_273+10250del21kb (CFTRdele2,3), c.2012delT (2143delT), c.2052_2053insA (2184insA), and c.3691delT (3821delT) are most typical for CF patients of Russian origin. DNA of 154 CF patients, Russian by origin, in whom at least one mutant allele was not previously identified (164 CF alleles), was analyzed by Sanger sequencing followed by the multiplex ligase-dependent probe amplification (MLPA) method. In addition to the 29 variants identified during the previous test for common mutations, 91 pathogenic CFTR variants were also revealed: 29 missense, 19 nonsense, 14 frame shift in/del, 17 splicing, 1 in frame ins, and 11 copy number variations (CNV). Each of the 61 variants was revealed once, and 17 twice. Each of the variants c.1209G>C (E403D), c.2128A>T (K710X), c.3883delA (4015delA), and c.3884_3885insT (4016insT) were detected for three, c.1766+1G>A (1898+1G>A) and c.2834C>T (S945L) for four, c.1766+1G>C (1898+1G>C) and c.(743+1_744-1)_(1584+1_1585-1)dup (CFTRdup6b-10) for five, c.2353C>T (R785X) and c.4004T>C (L1335P) for six, c.3929G>A (W1310X) for seven, c.580-1G>T (712-1G>T for eight, and c.1240_1244delCAAAA (1365del5) for 11 unrelated patients. A comprehensive analysis of CFTR mutant alleles with sequencing followed by MLPA, allowed not only the identification of 163 of 164 unknown alleles in our patient sample, but also expansion of the mutation spectrum with novel and additional frequent variants for ethnic Russians.

Download Full-text

Fibrillin-1 gene mutations in a Chinese cohort with congenital ectopia lentis: spectrum and genotype–phenotype analysis

British Journal of Ophthalmology ◽

10.1136/bjophthalmol-2021-319084 ◽

2021 ◽

pp. bjophthalmol-2021-319084

Author(s):

Zexu Chen ◽

Tianhui Chen ◽

Min Zhang ◽

Jiahui Chen ◽

Michael Deng ◽

...

Keyword(s):

Calcium Binding ◽

Gene Mutations ◽

Mutation Spectrum ◽

Missense Mutations ◽

Ectopia Lentis ◽

Phenotype Analysis ◽

Chinese Cohort ◽

Fibrillin 1 ◽

Epidermal Growth ◽

Dependent Probe

AimsTo identify the mutation spectrum and genotype–phenotype correlations of fibrillin-1 (FBN1) mutations in a Chinese cohort with congenital ectopia lentis (EL).MethodsPatients clinically suspected of congenital zonulopathy were screened using panel-based next-generation sequencing followed by multiplex ligation-dependent probe amplification. All the probands were subjected to thorough ocular examinations. Molecular and clinical data were integrated in pursuit of genotype–phenotype correlation.ResultsA total of 131 probands of FBN1 mutations from unrelated families were recruited. Around 65% of the probands were children younger than 9 years old. Overall, 110 distinct FBN1 mutations were identified, including 39 novel ones. The most at-risk regions were exons 13, 2, 6, 15, 24 and 33 in descending order of mutation frequency. The most prevalent mutation was c.184C>T (seven, 5.34%) in the coding sequence and c.5788+5G>A (three, 2.29%) in introns. Missense mutations were the most frequent type (103, 78.63%); half of which were distributed in the N-terminal regions (53, 51.46%). The majority of missense mutations were detected in one of the calcium-binding epidermal growth factor-like domains (62, 60.19%), and 39 (62.90%) of them were substitutions of conserved cysteine residues. Microspherophakia (MSP) was found in 15 patients (11.45%). Mutations in the middle region (exons 22–42), especially exon 26, had higher risks of combined MSP (OR, 5.51 (95% CI 1.364 to 22.274), p=0.017).ConclusionsThis study extended the knowledge of the FBN1 mutation spectrum and provided novel insights into its clinical correlation regarding EL and MSP in the Chinese population.

Download Full-text

Ethnic Differences in the Frequency of CFTR Gene Mutations in Populations of the European and North Caucasian Part of the Russian Federation

Frontiers in Genetics ◽

10.3389/fgene.2021.678374 ◽

2021 ◽

Vol 12 ◽

Author(s):

Nika Petrova ◽

Natalia Balinova ◽

Andrey Marakhonov ◽

Tatyana Vasilyeva ◽

Nataliya Kashirskaya ◽

...

Keyword(s):

Ethnic Groups ◽

Russian Federation ◽

Gene Mutations ◽

Ural Region ◽

Monogenic Disease ◽

Healthy Population ◽

The North ◽

Pathogenic Variants ◽

The Russian Federation ◽

Volga Ural Region

Cystic fibrosis (CF) is a common monogenic disease caused by pathogenic variants in the CFTR gene. The distribution and frequency of CFTR variants vary in different countries and ethnic groups. The spectrum of pathogenic variants of the CFTR gene was previously studied in more than 1,500 CF patients from different regions of the European and North Caucasian region of Russia and the spectrum of the most frequent pathogenic variants of the CFTR gene and ethnic features of their distribution were determined. To assess the population frequency of CFTR gene mutations some of the common variants were analyzed in the samples of healthy unrelated individuals from the populations of the European part of the Russian Federation: 1,324 Russians from four European regions (Pskov, Tver, Rostov, and Kirov regions), representatives of five indigenous ethnic groups of the Volga-Ural region [Mari (n = 505), Udmurts (n = 613), Chuvash (n = 780), Tatars (n = 704), Bashkirs (n = 517)], and six ethnic groups of the North Caucasus [Karachay (n = 324), Nogais (n = 118), Circassians (n = 102), Abazins (n = 128), Ossetians (n = 310), and Chechens (n = 100)]. The frequency of common CFTR mutations was established in studied ethnic groups. The frequency of F508del mutation in Russians was found to be 0.0056 on average, varying between four regions, from 0.0027 in the Pskov region to 0.0069 in the Rostov region. Three variants W1282X, 1677delTA, and F508del were identified in the samples from the North Caucasian populations: in Karachay, the frequency of W1282X mutation was 0.0092, 1677delTA mutation – 0.0032; W1282X mutation in the Nogais sample – 0.0127, the frequency of F508del mutations was 0.0098 and 1677delTA – 0.0098 in Circassians; in Abazins F508del (0.0039), W1282X (0.0039) and 1677delTA (0.0117) mutations were found. In the indigenous peoples of the Volga-Ural region, the maximum frequency of the F508del mutation was detected in the Tatar population (0.099), while this mutation was never detected in the Mari and Bashkir populations. The E92K variant was found in Chuvash and Tatar populations. Thus, interethnic differences in the spectra of CFTR gene variants were shown both in CF patients and in healthy population of the European and North Caucasian part of Russia.

Download Full-text

Multiplex Ligation-Dependent Probe Amplification Analysis ofGATA4Gene Copy Number Variations in Patients with Isolated Congenital Heart Disease

Disease Markers ◽

10.1155/2010/530360 ◽

2010 ◽

Vol 28 (5) ◽

pp. 287-292 ◽

Cited By ~ 7

Author(s):

Valentina Guida ◽

Francesca Lepri ◽

Raymon Vijzelaar ◽

Andrea De Zorzi ◽

Paolo Versacci ◽

...

Keyword(s):

Congenital Heart Defects ◽

Copy Number ◽

High Performance ◽

Congenital Heart ◽

Clinical Symptoms ◽

Heart Defects ◽

Gene Mutations ◽

Copy Number Variations ◽

Deletion Syndrome ◽

Dependent Probe

GATA4mutations are found in patients with different isolated congenital heart defects (CHDs), mostly cardiac septal defects and tetralogy of Fallot. In addition,GATA4is supposed to be the responsible gene for the CHDs in the chromosomal 8p23 deletion syndrome, which is recognized as a malformation syndrome with clinical symptoms of facial anomalies, microcephaly, mental retardation, and congenital heart defects. Thus far, no study has been carried out to investigate the role ofGATA4copy number variations (CNVs) in non-syndromic CHDs. To explore the possible occurrence ofGATA4gene CNVs in isolated CHDs, we analyzed by multiplex ligation-dependent probe amplification (MLPA) a cohort of 161 non-syndromic patients with cardiac anomalies previously associated withGATA4gene mutations. The patients were mutation-negative forGATA4,NKX2.5, andFOG2genes after screening with denaturing high performance liquid chromatography. MLPA analysis revealed that normalized MLPA signals were all found within the normal range values for all exons in all patients, excluding a major contribution ofGATA4gene CNVs in CHD pathogenesis.

Download Full-text

Identification of Exonic Copy Number Variations in Dystrophin Gene Using Mlpa / Identificarea Variaţiilor Numărului de Copii în Gena Distrofinei Folosind Metoda Mlpa

Revista Romana de Medicina de Laborator ◽

10.2478/rrlm-2014-0038 ◽

2014 ◽

Vol 22 (4) ◽

Author(s):

Cristina Rusu ◽

Adriana Sireteanu ◽

Lăcrămioara Butnariu ◽

Monica Pânzaru ◽

Elena Braha ◽

...

Keyword(s):

Copy Number ◽

Mutation Screening ◽

Gene Mutations ◽

Dystrophin Gene ◽

Copy Number Variations ◽

Sequencing Analysis ◽

Reading Frame ◽

Muscle Disorders ◽

Dmd Gene ◽

Dependent Probe

AbstractDuchenne and Becker muscular dystrophies (DMD/BMD) are X-linked progressive muscle disorders determined by mutations of the dystrophin (DMD) gene. Multiplex Ligation - Dependent Probe Amplification (MLPA) is a simple, inexpensive and reliable test for molecular diagnosis of DMD gene mutations. It identifies exonic copy number variations in the DMD gene, but the test should be completed with sequencing analysis in case of single exon deletions/duplications. The aim of this study was to evaluate the efficiency of MLPA as a DMD mutation screening tool in affected males and carrier females, as well as to appreciate the frequency of different types of mutations and to check the validity of the “reading frame rule”. We have used MLPA for the detection of deletions/ duplications in DMD gene in 53 individuals (30 affected males and 23 asymptomatic female relatives) referred for evaluation and genetic counseling due to the clinical suspicion of DMD/BMD. In the affected males (21 DMD and 9 BMD) MLPA had a detection rate of 63.5% (53.5% deletions and 10% duplications). The most frequently deleted exon was exon 45 and the most frequent duplication involved exons 3-5, confirming the presence of the two hotspot mutation regions reported in the literature. Mutations detected in our study have a slightly different location compared to literature data. Reading frame rule was valid in 84% of our cases.

Download Full-text

Localization of Ethnic Groups in the Regions as a Factor in Cross-Regional Variations in Voting for United Russia

Russian Politics ◽

10.30965/24518921-00502001 ◽

2020 ◽

Vol 5 (2) ◽

pp. 131-153

Author(s):

Eleonora Minaeva ◽

Petr Panov

Keyword(s):

Ethnic Groups ◽

Russian Federation ◽

Political Mobilization ◽

Structural Factors ◽

Regional Variations ◽

Political Machines ◽

Ethnic Networks ◽

Electoral Authoritarianism ◽

Administrative Unit ◽

The Russian Federation

Abstract In the context of electoral authoritarianism, political mobilization is likely to be a more reasonable explanation of cross-regional variations in voting for the party of power than the diversity of the regions’ policy preferences. In the Russian Federation, the political machines which coordinate various activities aimed at mobilizing people to vote for United Russia demonstrate different degrees of effectiveness. This article examines the structural factors that facilitate machine politics focusing on ethnic networks. Although strong ethnic networks are more likely to arise if the members of an ethnic group live close to each other, and at the same time separately from other ethnic groups, so far researchers have neglected to consider the localization of ethnic groups within the territory of an administrative unit as a factor. In order to fill the gap, we have created an original geo-referenced dataset of the localization of non-Russian ethnic groups within every region of the Russian Federation, and developed special GIS (geographic information systems) techniques and tools to measure them in relation to the Russian population. This has made it possible to include the localization of ethnic groups as a variable in the study of cross-regional differences in voting for United Russia. Our analysis finds that the effect of non-Russians’ share of the population on voting for UR increases significantly if non-Russian groups are at least partially geographically segregated from Russians within a region.

Download Full-text

ON THE NEED TO COLLECT, PRESERVE AND STUDY DISAPPEARED NAMES OF SETTLEMENTS

10.34216/2020-1.onomast.187-192 ◽

2020 ◽

pp. 187-192

Author(s):

S.A. Popov

Keyword(s):

Cultural Heritage ◽

Russian Federation ◽

Historical Memory ◽

Comprehensive Analysis ◽

Local History ◽

The Past ◽

Cultural Systems ◽

The Russian Federation ◽

Time Periods ◽

The Disappeared

The article deals with the problem of collecting, preserving and researching the disappeared names of localities in the subjects of the Russian Federation, which for centuries have become an integral part of the historical and cultural heritage of the peoples of our country. The author believes that only a comprehensive analysis of the past oikonyms in nominational, lexical-semantic, historical-cultural, historical-ethnographic, local history aspects will restore the linguistic and cultural systems of different time periods in different microareals of the Russian Federation. The author comes to the conclusion that in order to preserve the historical memory of the disappeared names of geographical objects, local researchers need the support of regional state authorities and local self-government.

Download Full-text

Detection of ras gene mutations in peripheral blood of carcinoma patients using CD45 immunomagnetic separation and nested mutant allele specific amplification.

International Journal of Oncology ◽

10.3892/ijo.12.6.1333 ◽

1998 ◽

Cited By ~ 1

Author(s):

K Shibata ◽

M Mori ◽

S Kitano ◽

T Akiyoshi

Keyword(s):

Peripheral Blood ◽

Mutant Allele ◽

Gene Mutations ◽

Immunomagnetic Separation ◽

Ras Gene ◽

Allele Specific ◽

Specific Amplification

Download Full-text

A Modified Method to Isolate Circulating Tumor Cells and Identify by a Panel of Gene Mutations in Lung Cancer

Technology in Cancer Research & Treatment ◽

10.1177/1533033821995275 ◽

2021 ◽

Vol 20 ◽

pp. 153303382199527

Author(s):

Helin Wang ◽

Jieqing Wu ◽

Qi Zhang ◽

Jianqing Hao ◽

Ying Wang ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Detection Rate ◽

Target Genes ◽

White Blood Cells ◽

Gene Mutations ◽

Copy Number Variations ◽

Immunomagnetic Beads ◽

Membrane Rupture

The CellSearch system is the only FDA approved and successful used detection technology for circulating tumor cells(CTCs). However, the process for identification of CTCs by CellSearch appear to damage the cells, which may adversely affects subsequent molecular biology assays. We aimed to explore and establish a membrane-preserving method for immunofluorescence identification of CTCs that keeping the isolated cells intact. 98 patients with lung cancer were enrolled, and the efficacy of clinical detection of CTCs was examined. Based on the CellSearch principle, we optimized an anti-EpCAM antibody and improved cell membrane rupture. A 5 ml peripheral blood sample was used to enrich CTCs with EpCAM immunomagnetic beads. Fluorescence signals were amplified with secondary antibodies against anti-EpCAM antibody attached on immunomagnetic beads. After identifying CTCs, single CTCs were isolated by micromanipulation. To confirm CTCs, genomic DNA was extracted and amplified at the single cell level to sequence 72 target genes of lung cancer and analyze the mutation copy number variations (CNVs) and gene mutations. A goat anti-mouse polyclonal antibody conjugated with Dylight 488 was selected to stain tumor cells. We identified CTCs based on EpCAM+ and CD45+ cells to exclude white blood cells. In the 98 lung cancer patients, the detection rate of CTCs (≥1 CTC) per 5 ml blood was 87.76%, the number of detections was 1–36, and the median was 2. By sequencing 72 lung cancer-associated genes, we found a high level of CNVs and gene mutations characteristic of tumor cells. We established a new CTCs staining scheme that significantly improves the detection rate and allows further analysis of CTCs characteristics at the genetic level.

Download Full-text