Next Generation Sequencing For Screening Of Pathogenic Genes Of Coronary Disease Combined With Familial Hypercholesterolemia

Objective:Hyperlipidemia is a part of the risk factors of coronary heart disease.There are numerous ways and types of hyperlipidemia in clinic.For different dyslipidemia the efficacy of lipid-regulating drugs used in the clinic is also different.Familial hypercholesterolemia(FH)is a type of familial hypercholesterolemia with autosomal dominant or recessive inheritance.It has intricate genetic heterogeneity and high risk of cardiovascular disease.There is the absence of specific diagnostic methods in clinical and it is difficult to make a diagnosis based on clinical characteristics.This study conducted a preliminary FH pathogenic gene screening for patients with coronary heart disease in Gannan region by using high-throughput targeted sequencing with the purpose of correctly identifying FH patients and their associated pathogenic gene mutation types in the population of coronary heart disease,and guiding clinicians to adjust reasonable lipid-lowering programs for FH gene mutation types to reduce the risk of cardiovascular disease.Methods:Collect 83 preliminaries considered coronary heart disease patients admitted to the Cardiology Department of the First Affiliated Hospital of Gannan Medical College from October 2020 to October 2022.According to the inclusion criteria and exclusion criteria,78 subjects were finally included in the study.They were divided into two groups according to whether they were combined with hyperlipidemia,45 patients were in the coronary heart disease combined with hyperlipidemia group(CAD-HL group),and 33 patients were in the simple coronary heart disease group(CAD group).The clinical data of the two groups of patients were collected respectively,including gender,age,body mass index(BMI),smoking history,hypertension history,diabetes history,cerebral infarction history,TC,TG,LDL-C,Lp(a),uric acid,coronary artery disease,Dutch lipid score,and the use of lipid regulating drugs.4ml of the study subjects’ peripheral venous blood was taken,DNA samples were extracted,and DNA samples were sent to Guangdong Ruiao Kangchen Biotechnology Co.,Ltd.after passing the test,Hghthroughput targeted sequencing method was used to sequence the eons of low density lipoprotein receptor(LDLR),pro-protein converting enzyme subtilisin 9(PCSK9),apolipoprotein B(APOB),low density lipoprotein receptor ligand protein 1(LDLRAP1),ATP binding cassette transporter G5/8(ABCG5/8),Niman-Pick C1L1(NPC1L1)and other genes.The sequencing results of the two groups of FH pathogenic genes were statistically analyzed,and the differences between the two groups of gene mutations were compared;the related factors of the FH gene were analyzed by binary logistic regression and the ROC curve was drawn.Results:1.Compared with the clinical baseline treatment of the two groups,the CAD-HL group was significantly higher than the CAD group in terms of TC、TG、LDL-C、smoking history、use of lipid-regulating drugs Dutch lipid score and coronary artery disease(P<0.05);There was no significant difference between the two groups in gender、age、BMI、hypertension history、diabetes history、cerebral infarction histoy、LP(a)、uric acid and other factors(P>0.05).2.1n the gene mutation analysis,a total of 30(38.7%)patients carried 37 FH mutation genes,all of which were heterozygous mutations,and the gene mutation categories included LDLR,APOB,PSCK9,LDLRAP1,ABCG5,ABCG8 and other 6 categories.There were 10(22.2%)positive gene mutations in the CAD-HL group and 2(5.7%)positive gene mutations in the CAD group,and the difference in the number of positive gene mutations between the two groups was statistically significant(P=0.04<0.05).The prediction model of FH gene mutation obtained by binary logistic regression analysis suggests that the positive rate of FH gene mutation detection can be improved by comprehensively considering five predictive factors such as sex,smoking history,hypertension history,Dutch lipid score,and lipoprotein a in patients with coronary heart disease.3.The results showed that a total of 30(38.7%)patients carried 37 kinds ofFH mutant genes,all of which were heterozygous mutations,and the gene mutation categories included 6 types such as LDLR,APOB,PSCK9,LDLRAP1,ABCG5,ABCG8,7(18.9%)LDLR gene mutation sites and 14 APOB gene mutation sites(37.8%).There were 2(5.41%)mutation sites in PSCK9 gene,2(5.41%)in LDLRAP1 gene mutation sites,and 6(32.43%)in ABCG5/8 gene mutation sites.This included 1 LDLR/APOB dual gene mutation.Conclusion:In patients with coronary heart disease and hyperlipidemia,comprehensive consideration of factors such as gender,smoking history,hypertension history,Dutch lipid score,and lipoprotein a can increase the positive rate of FH gene testing,reduce medical costs,and reduce waste of medical resources;In terms of the types of FH pathogenic gene mutations,the diseases caused by pathogenic gene mutations are mainly Familial Hypercholesterolemia and Sitosterolemia.Therefore,it is necessary to pay attention to the differential diagnosis between the two in clinical work;The types of FH gene mutations are not limited to single gene mutations,but there is a possibility of multigene and multilocus gene mutations.The synergistic effect of multiple genes can further increase blood lipid levels and cardiovascular disease risk.