ANGPTL3 Mutations in Unrelated Chinese Han Patients with Familial Hypercholesterolemia

Author(s): Yunyun Yang, Song Yang, Xiaolu Jiao, Juan Li, Miaomiao Zhu, Luya Wang, Yanwen Qin*.

Journal Name: Current Pharmaceutical Design

Volume 25 , Issue 2 , 2019


Abstract:

Background and objective: Familial hypercholesterolemia (FH) is a severe genetic hyperlipidemia characterized by increased levels of low-density lipoprotein cholesterol (LDL-C), leading to premature atherosclerosis. Angiopoietin-like protein (ANGPTL3) is a hepatocyte-specific protein that can be used to lower LDL in FH. However, it was unknown whether ANGPTL3 variants are present in FH patients. This study was performed to identify ANGPTL3 variants in unrelated Chinese Han patients with FH.

Methods and Results: We screened 80 patients with FH (total cholesterol >7.8mmol/L, LDL-cholesterol >4.9mmol/L) and 77 controls using targeted next-generation sequencing (NGS) of six FH candidate genes (LDLR, ApoB100, PCSK9, ABCG5, ABCG8, and ANGPTL3). Candidate pathogenic variants identified by NGS were validated by Sanger sequencing. Mutant and wild-type plasmids containing the variant sequence were constructed and verified by Sanger sequencing. The gene expression profile was analyzed by an expression profile chip in transfected HepG2 cells using quantitative real-time (qRT)-PCR. We identified 41 variants in 28 FH patients, including two ANGPTL3 mutations: one exonic (c.A956G: p.K319R) and one in the untranslated region (c.*249G>A). Gene ontology analyses found that the cholesterol metabolic process and ANGPTL3 expression were significantly up-regulated in the ANGPTL3 K319R mutation group compared with the wild-type group. qRT-PCR findings were consistent with the expression profile analysis.

Conclusion: Rare ANGPTL3 variants were identified in Chinese patients with FH, including ANGPTL3: p.(Lys319Arg) which affected the expression of ANGPTL3 and the cholesterol metabolic process as determined by bioinformatics analysis.

Clinical Trial Registration: Chinese Clinical Trial Registration (ChiCTR-ROC-17011027) http://www.chictr.org.cn/listbycreater.aspx

Keywords: Familial hypercholesterolemia, angiopoietin-like protein 3, genetic mutation, cholesterol metabolism, hypercholesterolemia, LDL-C.

[1]
Al-Allaf FA, Athar M, Abduljaleel Z, et al. Next generation sequencing to identify novel genetic variants causative of autosomal dominant familial hypercholesterolemia associated with increased risk of coronary heart disease. Gene 2015; 565(1): 76-84.
[2]
Asahina M, Mashimo T, Takeyama M, et al. Hypercholesterolemia and atherosclerosis in low density lipoprotein receptor mutant rats. Biochem Biophys Res Commun 2012; 418(3): 553-8.
[3]
Alves A, Etxebarria A, Soutar A, et al. Novel functional APOB mutations outside LDL-binding region causing familial hypercholesterolaemia. Hum Mol Genet 2014; 23(7): 1817-28.
[4]
Wang X, Jiang L, Sun L-Y, et al. Genetically confirmed familial hypercholesterolemia in outpatients with hypercholesterolemia. J Geriatr Cardiol 2018; 15(6): 434-40.
[5]
Xu Y, Redon V, Yu H, et al. Role of angiopoietin-like 3 (ANGPTL3) in regulating plasma level of low-density lipoprotein cholesterol. Atherosclerosis 2018; 268: 196-206.
[6]
Musunuru K, Pirruccello J, Do R, et al. Exome sequencing, ANGPTL3 mutations, and familial combined hypolipidemia. N Engl J Med 2010; 363(23): 2220-7.
[7]
Martin-Campos JM, Roig R, Mayoral C, et al. Identification of a novel mutation in the ANGPTL3 gene in two families diagnosed of familial hypobetalipoproteinemia without APOB mutation. Clin Chim Acta 2012; 413(5-6): 552-5.
[8]
Minicocci I, Montali A, Robciuc MR, et al. Mutations in the ANGPTL3 gene and familial combined hypolipidemia: a clinical and biochemical characterization. J Clin Endocrinol Metab 2012; 97(7): E1266-75.
[9]
Biterova E, Esmaeeli M, Alanen HI, Saaranen M, Ruddock LW. Structures of Angptl3 and Angptl4, modulators of triglyceride levels and coronary artery disease. Sci Rep 2018; 8(1): 6752.
[10]
Dewey FE, Gusarova V, Dunbar RL, et al. Genetic and Pharmacologic Inactivation of ANGPTL3 and Cardiovascular Disease. N Engl J Med 2017; 377(3): 211-21.
[11]
Mardis ER. The impact of next-generation sequencing technology on genetics. Trends Genet 2008; 24(3): 133-41.
[12]
Chiou KR and, Charng MJ. Detection of mutations and large rearrangements of the low-density lipoprotein receptor gene in Taiwanese patients with familial hypercholesterolemia. Am J Cardiol 2010; 105(12): 1752-8.
[13]
Lamiquiz-Moneo I, Baila-Rueda L, Bea AM, et al. ABCG5/G8 gene is associated with hypercholesterolemias without mutation in candidate genes and noncholesterol sterols. J Clin Lipidol 2017; 11(6): 1432-1440 e4..
[14]
Volta A, Hovingh GK, Grefhorst A. Genetics of familial hypercholesterolemia: a tool for development of novel lipid lowering pharmaceuticals? Curr Opin Lipidol 2018; 29(2): 80-6.
[15]
Huijgen R, Sjouke B, Vis K, et al. Genetic variation in APOB, PCSK9, and ANGPTL3 in carriers of pathogenic autosomal dominant hypercholesterolemic mutations with unexpected low LDL-Cl Levels. Hum Mutat 2012; 33(2): 448-55.
[16]
Wang X, Jiang L, Sun LY, et al. Genetically confirmed familial hypercholesterolemia in outpatients with hypercholesterolemia. J Geriatr Cardiol 2018; 15(6): 434-40.
[17]
Ose L. An update on familial hypercholesterolaemia. J Ann Med 1999; 13(Suppl. 1): 13-8.
[18]
Richards CS, Bale S, Bellissimo DB, et al. ACMG recommendations for standards for interpretation and reporting of sequence variations: Revisions 2007. Genet Med 2008; 10(4): 294-300.
[19]
Sjouke B, Kusters D, Kindt I, et al. Homozygous autosomal dominant hypercholesterolaemia in the Netherlands: prevalence, genotype-phenotype relationship, and clinical outcome. Eur Heart J 2015; 36(9): 560-5.
[20]
Benn M, Watts G, Tybjærg-Hansen A, et al. Mutations causative of familial hypercholesterolaemia: screening of 98 098 individuals from the Copenhagen General Population Study estimated a prevalence of 1 in 217. Eur Heart J 2016; 37(17): 1384-94.
[21]
Prajapati R, Agrawal V. Familial hypercholesterolemia supravalvular aortic stenosis and extensive atherosclerosis. Indian Heart J 2018; 70(4): 575-7.
[22]
Li S, Zhang H, Guo Y, et al. Familial hypercholesterolemia in very young myocardial infarction. Sci Rep 2018; 8(1): 8861.
[23]
Harada-Shiba M, Ako J, Arai H, et al. Prevalence of familial hypercholesterolemia in patients with acute coronary syndrome in Japan: Results of the EXPLORE-J study. Atherosclerosis 2018; 277: 362-8.
[24]
Béliard S, Boccara F, Cariou B, et al. High burden of recurrent cardiovascular events in heterozygous familial hypercholesterolemia: The French Familial Hypercholesterolemia Registry. Atherosclerosis 2018; 277: 334-40.
[25]
Sun D, Li S, Zhu C, et al. Prevalence and clinical features of familial hypercholesterolemia in Chinese patients with myocardial infarction. Zhonghua Xin Xue Guan Bing Za Zhi 2018; 46(2): 109-13.
[26]
Reiner Z. Management of patients with familial hypercholesterolaemia. Nat Rev Cardiol 2015; 12(10): 565-75.
[27]
Pijlman A, Huijgen R, Verhagen S, et al. Evaluation of cholesterol lowering treatment of patients with familial hypercholesterolemia: a large cross-sectional study in The Netherlands. Atherosclerosis 2010; 209(1): 189-94.
[28]
Daniel Gaudet DAG. Robert Pordy, Zahid Ahmad, Marina Cuchel, Prediman K. Shah, Kuang-Yuh Chyu, William J. Sasiela, Kuo-Chen Chan, Diane Brisson, Etienne Khoury, ANGPTL3 Inhibition in Homozygous Familial Hypercholesterolemia. N Engl J Med 2017; 377: 3.
[29]
Thompson G. Recommendations for the use of LDL apheresis. Atherosclerosis 2008; 198(2): 247-55.
[30]
Shi Z, Yuan B, Zhao D, et al. Familial hypercholesterolemia in China: prevalence and evidence of underdetection and undertreatment in a community population. Int J Cardiol 2014; 174(3): 834-6.
[31]
Athyros VG, Katsiki N, Dimakopoulou A, et al. Drugs that Mimic the Effect of Gene Mutations for the Prevention or the Treatment of Atherosclerotic Disease: From PCSK9 Inhibition to ANGPTL3 Inactivation. Curr Pharm Des 2018; 24(31): 3638-46.
[32]
Ahmad Z, Adams-Huet B, Chen C, et al. Low prevalence of mutations in known loci for autosomal dominant hypercholesterolemia in a multiethnic patient cohort. Circ Cardiovasc Genet 2012; 5(6): 666-75.
[33]
Lange L, Hu Y, Zhang H, et al. Whole-exome sequencing identifies rare and low-frequency coding variants associated with LDL cholesterol. Am J Hum Genet 2014; 94(2): 233-45.
[34]
Paththinige C, Sirisena N and, Dissanayake V. Genetic determinants of inherited susceptibility to hypercholesterolemia - a comprehensive literature review. Lipids Health Dis 2017; 16(1): 103.
[35]
Hatsuda S, Shoji T, Shinohara K, et al. Association between plasma angiopoietin-like protein 3 and arterial wall thickness in healthy subjects. J Vasc Res 2007; 44(1): 61-6.
[36]
Lupo MG, Ferri N. Angiopoietin-Like 3 (ANGPTL3) and Atherosclerosis: Lipid and Non-Lipid Related Effects. J Cardiovasc Dev Dis 2018; 5(3): : E39.
[37]
Stitziel N, Khera A, Wang X, et al. ANGPTL3 Deficiency and Protection Against Coronary Artery Disease. J Am Coll Cardiol 2017; 69(16): 2054-63.
[38]
Dewey FE, Gusarova V, Dunbar RL, et al. Genetic and Pharmacologic Inactivation of ANGPTL3 and Cardiovascular Disease. New Engl J Med 2017; 377(3): 211-21.
[39]
Tarrytown NY. Regeneron Announces Evinacumab has Received FDA Breakthrough Therapy Designation for Homozygous Familial Hypercholesterolemia (HoFH). Regeneron 2017.


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Article Details

VOLUME: 25
ISSUE: 2
Year: 2019
Page: [190 - 200]
Pages: 11
DOI: 10.2174/1381612825666190228000932
Price: $58

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