Genetic analysis in a compound heterozygote family with familial hypercholesterolemia

Wang,Fang; Fan,Qin; Tao,Rong; Gu,Gang; Zhang,Ruiyan; Xi,Rui

doi:10.3892/mmr.2018.8904

Genetic analysis in a compound heterozygote family with familial hypercholesterolemia

Authors:
- Fang Wang
- Qin Fan
- Rong Tao
- Gang Gu
- Ruiyan Zhang
- Rui Xi
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Affiliations: Department of Cardiology, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China
Published online on: April 20, 2018 https://doi.org/10.3892/mmr.2018.8904
Pages: 8439-8449

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Abstract

Homozygous familial hypercholesterolemia (FH) is rare, with an incidence of ~one in a million and commonly presents with a genetic mutation. The genetic variations of families with FH were clinically analyzed to investigate the association between the phenotype and genotype of patients. Direct sequencing was conducted for the proband and her parents to detect mutations in the fragment of 18 exons of the low‑density lipoprotein receptor (LDLR) and apolipoprotein B100 Q3500R in the peripheral blood genomic DNA. The gene sequences were compared with normal ones to find mutations using GenBank. The QX200 Droplet Digital PCR system was used to detect target DNA copy number variations of the proband and her parents. The functional alterations resulting from the novel mutations were verified by quantitative polymerase chain reaction, western blotting and flow cytometric analyses. The lipid levels of the proband and her parents were all elevated. Genetic testing results indicated that the proband and her mother had a novel heterozygous missense mutation (C377G, 28893T>G) in exon 8 of the LDLR gene, whereas the proband and her father had LDLR gene DNA fragment deletions in exon 18. Clinically, the proband was of a compound heterozygous genotype and her parents were of the simple heterozygous genotype. Furthermore, both mutations led to impaired expression and LDL binding and internalization function of LDLR in vitro. The proband's genotype was confirmed to be compound heterozygous FH, leading to clinical manifestations in line with the homozygous FH phenotype. The phenotype is highly associated with the genotype in this type of compound heterozygous FH.

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1	Raal FJ and Santos RD: Homozygous familial hypercholesterolemia: Current perspectives on diagnosis and treatment. Atherosclerosis. 223:262–268. 2012. View Article : Google Scholar : PubMed/NCBI
2	Goldstein JL, Hobbs HH and Brown MS: Familial hypercholesterolemiaThe metabolic basis of inherited disease. McGraw-Hill; New York: pp. 2863–2913. 2001
3	Wang L, Lin J, Liu S, Cao S, Liu J, Yong Q, Yang Y, Wu B, Pan X, Du L, et al: Mutations in the LDL receptor gene in four Chinese homozygous familial hypercholesterolemia phenotype patients. Nutr Metab Cardiovasc Dis. 19:391–400. 2009. View Article : Google Scholar : PubMed/NCBI
4	Hooper AJ, Nguyen LT, Burnett JR, Bates TR, Bell DA, Redgrave TG, Watts GF and van Bockxmeer FM: Genetic analysis of familial hypercholesterolaemia in Western Australia. Atherosclerosis. 224:430–434. 2012. View Article : Google Scholar : PubMed/NCBI
5	Jiang L, Sun LY, Dai YF, Yang SW, Zhang F and Wang LY: The distribution and characteristics of LDL receptor mutations in China: A systematic review. Sci Rep. 5:172722015. View Article : Google Scholar : PubMed/NCBI
6	Humphries SE, Norbury G, Leigh S, Hadfield SG and Nair D: What is the clinical utility of DNA testing in patients with familial hypercholesterolaemia? Curr Opin Lipidol. 19:362–368. 2008. View Article : Google Scholar : PubMed/NCBI
7	Taylor A, Wang D, Patel K, Whittall R, Wood G, Farrer M, Neely RD, Fairgrieve S, Nair D, Barbir M, et al: Mutation detection rate and spectrum in familial hypercholesterolaemia patients in the UK pilot cascade project. Clin Genet. 77:572–580. 2010. View Article : Google Scholar : PubMed/NCBI
8	Lombardi MP, Redeker EJ, van Gent DH, Smeele KL, Weerdesteijn R and Mannens MM: Molecular genetic testing for familial hypercholesterolemia in the Netherlands: A stepwise screening strategy enhances the mutation detection rate. Genet Test. 10:77–84. 2006. View Article : Google Scholar : PubMed/NCBI
9	Poncz M, Solowiejczyk D, Harpel B, Mory Y, Schwartz E and Surrey S: Construction of human gene libraries from small amounts of peripheral blood: Analysis of beta-like globin genes. Hemoglobin. 6:27–36. 1982. View Article : Google Scholar : PubMed/NCBI
10	Belgrader P, Tanner SC, Regan JF, Koehler R, Hindson BJ and Brown AS: Droplet digital PCR measurement of HER2 copy number alteration in formalin-fixed paraffin-embedded breast carcinoma tissue. Clin Chem. 59:991–994. 2013. View Article : Google Scholar : PubMed/NCBI
11	Hindson BJ, Ness KD, Masquelier DA, Belgrader P, Heredia NJ, Makarewicz AJ, Bright IJ, Lucero MY, Hiddessen AL, Legler TC, et al: High-throughput droplet digital PCR system for absolute quantitation of DNA copy number. Anal Chem. 83:8604–8610. 2011. View Article : Google Scholar : PubMed/NCBI
12	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
13	Hartgers ML, Ray KK and Hovingh GK: New approaches in detection and treatment of familial hypercholesterolemia. Curr Cardiol Rep. 17:1092015. View Article : Google Scholar : PubMed/NCBI
14	Watts GF, Sullivan DR, Poplawski N, van Bockxmeer F, Hamilton-Craig I, Clifton PM, O'Brien R, Bishop W, George P, Barter PJ, et al: Familial hypercholesterolaemia: A model of care for Australasia. Atheroscler Suppl. 12:221–263. 2011. View Article : Google Scholar : PubMed/NCBI
15	Harada-Shiba M, Arai H, Oikawa S, Ohta T, Okada T, Okamura T, Nohara A, Bujo H, Yokote K, Wakatsuki A, et al: Guidelines for the management of familial hypercholesterolemia. J Atheroscler Thromb. 19:1043–1060. 2012. View Article : Google Scholar : PubMed/NCBI
16	Williams RR, Hunt SC, Schumacher MC, Hegele RA, Leppert MF, Ludwig EH and Hopkins PN: Diagnosing heterozygous familial hypercholesterolemia using new practical criteria validated by molecular genetics. Am J Cardiol. 72:171–176. 1993. View Article : Google Scholar : PubMed/NCBI
17	Watts GF, Gidding S, Wierzbicki AS, Toth PP, Alonso R, Brown WV, Bruckert E, Defesche J, Lin KK, Livingston M, et al: Integrated guidance on the care of familial hypercholesterolaemia from the International FH Foundation: Executive summary. J Atheroscler Thromb. 21:368–374. 2014.PubMed/NCBI
18	Nordestgaard BG, Chapman MJ, Humphries SE, Ginsberg HN, Masana L, Descamps OS, Wiklund O, Hegele RA, Raal FJ, Defesche JC, et al: Familial hypercholesterolaemia is underdiagnosed and undertreated in the general population: Guidance for clinicians to prevent coronary heart disease: Consensus statement of the European Atherosclerosis Society. Eur Heart J. 34:a3478–a3490. 2013. View Article : Google Scholar
19	Tybjaerg-Hansen A and Humphries SE: Familial defective apolipoprotein B-100: A single mutation that causes hypercholesterolemia and premature coronary artery disease. Atherosclerosis. 96:91–107. 1992. View Article : Google Scholar : PubMed/NCBI
20	Abifadel M, Varret M, Rabès JP, Allard D, Ouguerram K, Devillers M, Cruaud C, Benjannet S, Wickham L, Erlich D, et al: Mutations in PCSK9 cause autosomal dominant hypercholesterolemia. Nat Genet. 34:154–156. 2003. View Article : Google Scholar : PubMed/NCBI
21	van der Graaf A, Avis HJ, Kusters DM, Vissers MN, Hutten BA, Defesche JC, Huijgen R, Fouchier SW, Wijburg FA, Kastelein JJ and Wiegman A: Molecular basis of autosomal dominant hypercholesterolemia: Assessment in a large cohort of hypercholesterolemic children. Circulation. 123:1167–1173. 2011. View Article : Google Scholar : PubMed/NCBI
22	Lindgren V, Luskey KL, Russell DW and Francke U: Human genes involved in cholesterol metabolism: Chromosomal mapping of the loci for the low density lipoprotein receptor and 3-hydroxy-3-methylglutaryl-coenzyme A reductase with cDNA probes. Proc Natl Acad Sci USA. 82:8567–8571. 1985. View Article : Google Scholar : PubMed/NCBI
23	Yamamoto T, Davis CG, Brown MS, Schneider WJ, Casey ML, Goldstein JL and Russell DW: The human LDL receptor: A cysteine-rich protein with multiple Alu sequences in its mRNA. Cell. 39:27–38. 1984. View Article : Google Scholar : PubMed/NCBI
24	Wu WF, Sun LY, Pan XD, Yang SW and Wang LY: Use of targeted exome sequencing in genetic diagnosis of Chinese familial hypercholesterolemia. PLoS One. 9:e946972014. View Article : Google Scholar : PubMed/NCBI
25	Rødningen OK, Tonstad S, Ose L, Berg K and Leren TP: Effects of a 9.6-kb deletion of the LDL receptor gene (FH Helsinki) on structure and levels of mRNA. Hum Mutat. 12:95–102. 1998. View Article : Google Scholar : PubMed/NCBI
26	Rødningen OK, Røsby O, Tonstad S, Ose L, Berg K and Leren TP: A 9.6 kilobase deletion in the low density lipoprotein receptor gene in Norwegian familial hypercholesterolemia subjects. Clin Genet. 42:288–295. 1992. View Article : Google Scholar : PubMed/NCBI
27	Ooi EM, Barrett PH and Watts GF: The extended abnormalities in lipoprotein metabolism in familial hypercholesterolemia: Developing a new framework for future therapies. Int J Cardiol. 168:1811–1818. 2013. View Article : Google Scholar : PubMed/NCBI
28	Gagné C, Gaudet D and Bruckert E: Ezetimibe Study G: Efficacy and safety of ezetimibe coadministered with atorvastatin or simvastatin in patients with homozygous familial hypercholesterolemia. Circulation. 105:2469–2475. 2002. View Article : Google Scholar : PubMed/NCBI
29	Thompson GR, Catapano A, Saheb S, Atassi-Dumont M, Barbir M, Eriksson M, Paulweber B, Sijbrands E, Stalenhoef AF and Parhofer KG: Severe hypercholesterolaemia: Therapeutic goals and eligibility criteria for LDL apheresis in Europe. Curr Opin Lipidol. 21:492–498. 2010. View Article : Google Scholar : PubMed/NCBI
30	Smilde TJ, van Wissen S, Wollersheim H, Trip MD, Kastelein JJ and Stalenhoef AF: Effect of aggressive versus conventional lipid lowering on atherosclerosis progression in familial hypercholesterolaemia (ASAP): A prospective, randomised, double-blind trial. Lancet. 357:577–581. 2001. View Article : Google Scholar : PubMed/NCBI
31	Versmissen J, Oosterveer DM, Yazdanpanah M, Defesche JC, Basart DC, Liem AH, Heeringa J, Witteman JC, Lansberg PJ, Kastelein JJ and Sijbrands EJ: Efficacy of statins in familial hypercholesterolaemia: A long term cohort study. BMJ. 337:a24232008. View Article : Google Scholar : PubMed/NCBI
32	Raal FJ, Pilcher GJ, Panz VR, van Deventer HE, Brice BC, Blom DJ and Marais AD: Reduction in mortality in subjects with homozygous familial hypercholesterolemia associated with advances in lipid-lowering therapy. Circulation. 124:2202–2207. 2011. View Article : Google Scholar : PubMed/NCBI
33	Thompson GR: The evidence-base for the efficacy of lipoprotein apheresis in combating cardiovascular disease. Atheroscler Suppl. 14:67–70. 2013. View Article : Google Scholar : PubMed/NCBI
34	Wiegman A, Gidding SS, Watts GF, Chapman MJ, Ginsberg HN, Cuchel M, Ose L, Averna M, Boileau C, Borén J, et al: Familial hypercholesterolaemia in children and adolescents: Gaining decades of life by optimizing detection and treatment. Eur Heart J. 36:2425–2437. 2015. View Article : Google Scholar : PubMed/NCBI
35	Cuchel M, Bruckert E, Ginsberg HN, Raal FJ, Santos RD, Hegele RA, Kuivenhoven JA, Nordestgaard BG, Descamps OS, Steinhagen-Thiessen E, et al: Homozygous familial hypercholesterolaemia: New insights and guidance for clinicians to improve detection and clinical management. A position paper from the Consensus Panel on Familial Hypercholesterolaemia of the European Atherosclerosis Society. Eur Heart J. 35:2146–2157. 2014. View Article : Google Scholar : PubMed/NCBI