Verification of Underlying Genetic Cause in a Cohort of Russian Patients with Familial Hypercholesterolemia Using Targeted Next Generation Sequencing

Russian patients with familial hypercholesterolemia (FH) were screened for pathogenic mutations using targeted next generation sequencing. Genetic testing was performed in 52 probands with definite or probable FH based on the Dutch lipid clinic network criteria (DLCN score ≥ 6). Blood samples were studied by massive parallel sequencing (Illumina HiSeq 1500 platform) using a custom capture library related to dyslipidemia and premature atherosclerosis. Mutations considered to be responsible for monogenic FH were identified in 48% of the probands: 24 with mutations in the LDLR gene and two with a mutation in the APOB gene. There were 22 pathogenic/likely pathogenic mutations in LDLR, eight of which have not been previously described in the literature. Four patients with a clinical picture of homozygous FH had two heterozygous LDLR mutations. Although mutation-negative patients had highly elevated total cholesterol and low-density lipoprotein cholesterol levels, only half of them had a family history of hypercholesterolemia. With respect to heterozygous FH, mutation-positive patients had higher maximum total cholesterol levels (p = 0.01), more severe carotid atherosclerotic lesions, and a higher percentage of premature peripheral artery disease (p = 0.03) than mutation-negative ones. However, the number of patients who suffered from myocardial infarction was similar between the two groups.

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The presence of pathogenic mutations in patients with definite or probable diagnosis of familial hypercholesterolemia defined by targeted next generation sequencing

Atherosclerosis ◽

10.1016/j.atherosclerosis.2017.06.751 ◽

2017 ◽

Vol 263 ◽

pp. e231

Author(s):

Anna Semenova ◽

Igor Sergienko ◽

Marat Ezhov ◽

Diego Garcia-Giustiniani ◽

Lorenzo Monserrat ◽

...

Keyword(s):

Next Generation Sequencing ◽

Familial Hypercholesterolemia ◽

Next Generation ◽

Targeted Next Generation Sequencing ◽

Pathogenic Mutations ◽

Generation Sequencing ◽

Probable Diagnosis

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Targeted next-generation sequencing of the LDLRAP1 gene in patients with familial hypercholesterolemia

Atherosclerosis ◽

10.1016/j.atherosclerosis.2016.07.338 ◽

2016 ◽

Vol 252 ◽

pp. e32-e33

Author(s):

D. Ivanoshchuk ◽

E. Shakhtshneider ◽

K. Makarenkova ◽

P. Orlov ◽

Y. Ragino ◽

...

Keyword(s):

Next Generation Sequencing ◽

Familial Hypercholesterolemia ◽

Next Generation ◽

Targeted Next Generation Sequencing ◽

Generation Sequencing

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Identification of 13 novel USH2A mutations in Chinese retinitis pigmentosa and Usher syndrome patients by targeted next-generation sequencing

Bioscience Reports ◽

10.1042/bsr20193536 ◽

2020 ◽

Vol 40 (1) ◽

Cited By ~ 1

Author(s):

Ling-hui Qu ◽

Xin Jin ◽

Yan-ling Long ◽

Jia-yun Ren ◽

Chuang-huang Weng ◽

...

Keyword(s):

Next Generation Sequencing ◽

Retinitis Pigmentosa ◽

Molecular Mechanisms ◽

Usher Syndrome ◽

Next Generation ◽

Chinese Families ◽

Targeted Next Generation Sequencing ◽

Pathogenic Mutations ◽

Basement Membrane Protein ◽

Generation Sequencing

Abstract Background: The USH2A gene encodes usherin, a basement membrane protein that is involved in the development and homeostasis of the inner ear and retina. Mutations in USH2A are linked to Usher syndrome type II (USH II) and non-syndromic retinitis pigmentosa (RP). Molecular diagnosis can provide insight into the pathogenesis of these diseases, facilitate clinical diagnosis, and identify individuals who can most benefit from gene or cell replacement therapy. Here, we report 21 pathogenic mutations in the USH2A gene identified in 11 Chinese families by using the targeted next-generation sequencing (NGS) technology. Methods: In all, 11 unrelated Chinese families were enrolled, and NGS was performed to identify mutations in the USH2A gene. Variant analysis, Sanger validation, and segregation tests were utilized to validate the disease-causing mutations in these families. Results: We identified 21 pathogenic mutations, of which 13, including 5 associated with non-syndromic RP and 8 with USH II, have not been previously reported. The novel variants segregated with disease phenotype in the affected families and were absent from the control subjects. In general, visual impairment and retinopathy were consistent between the USH II and non-syndromic RP patients with USH2A mutations. Conclusions: These findings provide a basis for investigating genotype–phenotype relationships in Chinese USH II and RP patients and for clarifying the pathophysiology and molecular mechanisms of the diseases associated with USH2A mutations.

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The use of next-generation sequencing in clinical diagnosis of familial hypercholesterolemia

Genetics in Medicine ◽

10.1038/gim.2013.55 ◽

2013 ◽

Vol 15 (12) ◽

pp. 948-957 ◽

Cited By ~ 53

Author(s):

Jana Vandrovcova ◽

Ellen R.A. Thomas ◽

Santosh S Atanur ◽

Penny J. Norsworthy ◽

Clare Neuwirth ◽

...

Keyword(s):

Next Generation Sequencing ◽

Familial Hypercholesterolemia ◽

Molecular Diagnosis ◽

Genetic Diagnosis ◽

Target Enrichment ◽

Next Generation ◽

Large Deletions ◽

Number Of Patients ◽

The Cost ◽

Generation Sequencing

Abstract Purpose: Familial hypercholesterolemia is a common Mendelian disorder associated with early-onset coronary heart disease that can be treated by cholesterol-lowering drugs. The majority of cases in the United Kingdom are currently without a molecular diagnosis, which is partly due to the cost and time associated with standard screening techniques. The main purpose of this study was to test the sensitivity and specificity of two next-generation sequencing protocols for genetic diagnosis of familial hypercholesterolemia. Methods: Libraries were prepared for next-generation sequencing by two target enrichment protocols; one using the SureSelect Target Enrichment System and the other using the PCR-based Access Array platform. Results: In the validation cohort, both protocols showed 100% specificity, whereas the sensitivity for short variant detection was 100% for the SureSelect Target Enrichment and 98% for the Access Array protocol. Large deletions/duplications were only detected using the SureSelect Target Enrichment protocol. In the prospective cohort, the mutation detection rate using the Access Array was highest in patients with clinically definite familial hypercholesterolemia (67%), followed by patients with possible familial hypercholesterolemia (26%). Conclusion: We have shown the potential of target enrichment methods combined with next-generation sequencing for molecular diagnosis of familial hypercholesterolemia. Adopting these assays for patients with suspected familial hypercholesterolemia could improve cost-effectiveness and increase the overall number of patients with a molecular diagnosis. Genet Med 15 12, 948–957.

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