KR20120118845A - Polynucleotide for predicting prognosis of atrial fibrillation - Google Patents

Abstract

PURPOSE: A polynucleotide for predicting atrial fibrillation is provided to polynucleotide or the corresponding polynuclueotide including single-nucleotide polymorphism related to structure remodeling of left atrium. CONSTITUTION: A polynucleotide for predicting atrial fibrillation is composed of 10-50 continuous DNA sequences which include 27th base (polymorphic site) among the polynucleotides of the sequence number 4(SEQ ID NO:4). The sequence number 4 is the allelotype of 27th base (polymorphic site) A / T. The DNA microarray chip for predicting atrial fibrillation is polynucleotide for predicting atrial fibrillation or the corresponding polynucleotide. A kit for predicting atrial fibrillation includes polynucleotide for predicting atrial fibrillation or the corresponding polynucleotide. A prediction method of atrial fibrillation comprises the following steps: analyzing 10-50 continuous DNA sequences which include 27th base (polymorphic site) among the polynucleotides of the sequence number 4(SEQ ID NO:4) or the corresponding sequence from a recording media which can be read by computer; and classifying patients into a group which has potential to be electric anatomical remodeling of atrium if the 27th base of the gene sequence is A.

Description

Polynucleotide for predicting prognosis of atrial fibrillation}

The present invention relates to a polynucleotide for predicting the prognosis of atrial fibrillation.

Atrial fibrillation (AF), one of the most common arrhythmias in the clinic, is associated with an increasing high mortality and disability rate (van den Berg MP, et al., Eur J Heart Fail. 2002;4(5)). :571-575.; Khairy P, et al., CMAJ. 2002;167(9):1012-1020.) The pathophysiology of atrial fibrillation is heterogeneous (Nattel S, et al., Lancet. 2006;367( 9506):262-272.), old atrial fibrillation changes the shape of the left atrial (LA). Atrial fibrillation changes the electrophysical properties of atrial myocytes and causes alteration of atrial myocardial structure (Goette A, et al., Circulation. 1996;94(11):2968-2974.; Nattel S., et al. ., Nature. 2002;415(6868):219-226.). Both electrical and structural remodeling lead to a more susceptible temperament to atrial fibrillation due to an increase in atrial fibrillation load (Wijffels MC, et al., Circulation. 1995;92(7):1954-1968). Structural remodeling has been associated with interstitial fibrosis, downregulation of interstitial binding, and enlargement of atrial chamber size (critical mass) (van der Velden HM, et al., Cardiovasc Res. 2000;46(3):476-486). .; Zou R, et al., Am J Physiol Heart Circ Physiol. 2005;289(3):H1002-1012.). Left atrial size (Shin SH, et al., J Am Soc Echocardiogr. 2008;21(6):697-702.) and left atrial scar formation (Verma A, et al., Circulation. 2005;112(5):627) The degree of structural remodeling measured by -635.) affects the clinical outcome of rhythm control in patients with atrial fibrillation.

The reninangiotensin system (RAS) is associated with many cardiovascular diseases, and has been shown to be associated with fibrosis and hypertrophy of the myocardium. In addition, angiotensin converting enzyme (ACE) stimulates fibroblast proliferation, collagen synthesis, and structural remodeling of the atrium in patients with atrial fibrillation (Goette A, et al., J Am Coll Cardiol . 2000;35(6):1669-1677.; Rosenkranz S., Cardiovasc Res . 2004;63(3):423-432). ACE in Ravn et al ( Ravn LS, et al., Pharmacogenet Genomics . 2008;18(6):525-533) The angiotensinogen (angiotensinogen, AGT) in combination with I / D genotype A-20C genotype but reported that predict increased risk for atrial fibrillation, investigated for the genetic predisposition of the left atrium of the structural renovation of patients with atrial fibrillation There is little research.

Therefore, in the present invention, the relationship between the genetic polymorphism of angiotensin converting enzyme and the structural remodeling of atrial atrial fibrillation patients is investigated, and the results are used to predict the prognosis of atrial fibrillation.

The present inventors assume that the genetic polymorphism of angiotensin converting enzyme is related to the degree of structural remodeling of the atrium of patients with atrial fibrillation, and the genetic polymorphism of angiotensin converting enzyme and measured by 3D-spiral computed tomography (CT) The correlation between left atrial volume or left atrial voltage calculated by 3D-electroanatomical mapping was investigated in Korean non-membrane atrial fibrillation who underwent radiofrequency catheter ablation (RFCA). As a result, ACE The rs4362, rs4309, rs1800764, and rs4291 alleles were found to be highly related to the structural remodeling of the atrium.

Therefore, the present invention is a prognosis of atrial fibrillation consisting of 10 to 50 consecutive DNA sequences including the 27th base (polymorphic site) among polynucleotides selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 A diagnostic polynucleotide or a complementary polynucleotide thereof is provided.

In one embodiment of the present invention, the length of the polynucleotide or its complementary polynucleotide is 8 to 52 nucleotides, 10 to 50 nucleotides, or 20 to 40 nucleotides, but is not limited thereto.

Polymorphism of the present invention refers to a case where two or more alleles exist at a single locus, and a'polymorphic site' refers to a locus in which the allele exists. Among the polymorphic sites, the thing that only a single base differs from person to person is called "single nucleotide polymorphism," that is, single nucleotide polymorphism (SNP).

In another embodiment of the present invention, the allele of the 27th base (polymorphic site) of SEQ ID NO: 1 is C/T; The allele of the 27th base (polymorphic site) of SEQ ID NO: 2 is C/T; The allele of the 27th base (polymorphic site) of SEQ ID NO: 3 is C/T; And the allele of the 27th base (polymorphic site) of SEQ ID NO: 4 may be A/T.

The specific base sequences of the SNPs of SEQ ID NOs: 1 to 4 are shown in the table below.

[Table 1]

The position of the SNP in each of the polynucleotides is the 27th base, and alleles that may appear mainly are shown in parentheses.

Among the polynucleotides selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4, a polynucleotide consisting of 10 to 50 consecutive DNA sequences including the 27th base (polymorphic site) or a complementary polynucleotide thereof Can be used to diagnose atrial fibrillation prognosis.

In addition, the present invention provides a DNA microarray chip for prognostic diagnosis of atrial fibrillation in which the polynucleotide or its complementary polynucleotide is integrated.

The DNA microarray chip for prognostic diagnosis of atrial fibrillation of the present invention can be manufactured by a method known to those skilled in the art. More specifically, micropipette using a piezoelectric method to immobilize on a substrate of a DNA chip using the polynucleotide or a prognostic diagnostic probe for atrial fibrillation made of the polynucleotide thereof as a probe DNA molecule A (micropipetting) method or a method using a pin-type spotter may be used, but is not limited thereto.

In addition, the substrate of the DNA microarray chip is preferably coated with one active group selected from the group consisting of amino-silane, poly-L-lysine, and aldehyde. It is not limited thereto. In addition, the substrate may be selected from the group consisting of slide glass, plastic, metal, silicon, nylon film, and nitrocellulose film, but is not limited thereto.

The present invention also provides a kit for prognosing atrial fibrillation comprising the polynucleotide or a complementary polynucleotide thereof.

The present invention also provides a method for predicting the prognosis of atrial fibrillation that can be executed automatically by a computer, indicating the risk of recurrence of atrial fibrillation in a nucleic acid sample obtained from a patient, at least one type from the group consisting of SEQ ID NOs: 1 to 4 In the selected sequence, 10 to 50 consecutive DNA sequences including the 27th base (polymorphic site) or a complementary sequence thereof were analyzed, and as a result of the analysis, the 27th base of the patient's gene sequence is in the case of SEQ ID NO: 1. C; T for SEQ ID NO: 2; T for SEQ ID NO: 3; And in the case of A in the case of SEQ ID NO: 4, it provides a method for predicting the prognosis of atrial fibrillation comprising classifying the patient as a patient with a high possibility of atrial electroanatomical remodeling.

In addition, the present invention is a computer-readable recording medium in which a program for predicting the prognosis of atrial fibrillation is recorded, which indicates the risk of recurrence of atrial fibrillation in a nucleic acid sample obtained from a patient, consisting of SEQ ID NOs: 1 to 4 Analyzing 10 to 50 contiguous DNA sequences including a 27th base (polymorphic site) or a complementary sequence thereof, and as a result of the analysis, the 27th base of the patient's gene sequence A, C for SEQ ID NO: 1; T for SEQ ID NO: 2; T for SEQ ID NO: 3; And in the case of A in the case of SEQ ID NO: 4, a computer-readable recording medium is provided in which a program for executing the step of classifying the patient as a patient with a high possibility of electroanatomical remodeling of the atrium is recorded on a computer.

According to the present invention, it is possible to improve clinical outcomes by early detection of patients susceptible to structural remodeling of the left atrium, thereby preventing these patients from progressing to chronic persistent atrial fibrillation.

Figure 1 is a representative example of significantly remodeled left atrium (Figures 1A and 1C) and less remodeled left atrium (Figures 1B and 1E) in atrial fibrillation patients, and their ACE Genotype (Figure 1D) Show.
2 is a schematic diagram of the results of analysis of ACE mutations related to the degree of structural remodeling of the left atrium.

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely describe the present invention to those of ordinary skill in the art.

[ Example ]

Patient selection

The protocol for this study was approved by the Institutional Review Board. Written consent was obtained from all patients. The study involved 351 patients with atrial fibrillation (male: female = 282:69, mean age = 54.2±11.1 years) who received RFCA. Among them, 235 patients had paroxysmal atrial fibrillation (PAF) and 116 patients had persistent atrial fibrillation (PAF). The exclusion criteria were as follows:

1) permanent atrial fibrillation refractory to electrical cardioversion (permanent AF);

2) LA size >55 mm measured by echocardiography;

3) AF with rheumatic valve disease;

4) associated structural heart disease;

5) previous AF blocking; And

6) sinus rhythm not maintained for LA voltage mapping before

Sinus rhythm was not maintained for LA voltage mapping prior to RFCA.

Patients with blood vessels in the left atrium were excluded by transesophageal echocardiography. We imaged all patients with 3D-spiral CT (64 Channel, Light Speed Volume CT, Philips, Brilliance 63, Netherlands) to visually clarify the left atrium anatomy and pulmonary veins. Transthoracic echocardiography was performed in all patients, and left ventricular diastolic function measured by the anterior posterior (AP) diameter, left ventricular ejection fraction (LVEF), and E/E'. function, LVDF), LV end-systolic dimension (LVESD), and LV diastolic dimension (LVEDD) were measured.

Electrophysiological Mapping

Intracardiac electrograms were recorded using the Prucka CardioLabTM Electrophysiology system (General Electric Health Care System Inc., Milwaukee, WI, USA). For AF RFCA, we used 5 mapping catheters and a reorientable 3.5-mm tip 7 Fr open irrigation tip ablation catheter (Celsius, Johnson & Johnson Inc, Diamond Bar, CA, USA). ablation) procedure was performed in all patients using 3-D electroanatomical mapping (NavX system, St. Jude Medical Inc., Minneapolis, MN, USA). Prior to catheterization, we generated a LA 3D electroanatomical map by obtaining contact bipolar electrical diagrams from 250-350 points of the LA endocardial membrane. The voltage map was color coded. The bipolar electrical diagram was filtered with 32-300 Hz. The color coded voltage map was generated by recording the bipolar electrical diagram and measuring the peak-to-peak voltage.

Analysis of left atrial remodeling: 3D- Spiral CT And electroanatomical voltage map

3D-spiral CT images of the left atrium were analyzed on an imaging processing workstation (Aquarius, Terarecon, Inc, USA). The curve length of the left atrium was measured at the linear ablation lines (bilateral antral ablation lines, roof line, posterior inferior line, left lateral isthmus line, and anterior line). Each left atrial image was divided into sites by the following embryological organs: venous LA (rear LA including the ventral and posterior walls), LA appendages (LAA), and anterior LA (except LAA and venous LA) (Douglas YL , et al., Am J Cardiol . 2006;97(5):662-670.). We also measured the curve lengths of CPVA (circumferential pulmonary vein ablation), roof line, posterior inferior line, anterior linear line, and left lateral isthmus line as described in previous studies (Park JH, et al., J Cardiovasc). Electrophysiol . 2009), was measured.

We analyzed color-coded electroanatomical voltage maps in terms of anterior-posterior (AP) and posterior-anterior (PA). Low voltage regions of 0.2 mV or less were coded in gray, and high voltage regions of more than 5.0 mV were colored purple. The reference distance was measured by the inner-electrode distance of a duodecapolar catheter (St. Jude Medical Inc. Minnetonka, MN, USA.). The voltage region in the pulmonary vein was excluded. To quantify the average voltage of the left atrium, the percent area for each color was analyzed by a commercialized program (Image Pro) with reference to the color scale bar.

Gene polymorphism analysis

We use the HapMap Japanese (JPT) data bank (http://www.hapmap.org) and the NCBI SNP database ( http://www.ncbi.nlm.nih.gov/projects/SNP/ ) to ACE ACE. Haplotype-tagging single nucleotide polymorphisms (SNPs) of the gene were selected. To identify suitable tagged SNPs, we performed pilot genotyping on 16 selected SNPs from 48 Korean patients, and 7 SNPs ( ACE rs1800764, rs4291, rs4309, rs4362, rs4461142, rs61758684 and ACE I/D) was selected. Genomic DNA was extracted from whole blood samples using a commercially available kit (Qiagen, Valencia, CA, USA). Six SNPs were treated by a single-base expansion method using the SNaPShot™ assay kit (Applied Biosystems, Foster City, CA, USA).

Genotyping for I/D polymorphism was performed by polymerase chain reaction (PCR) (Chiang FT, et al., Clin Chem . 1998;44(6 Pt1):1353-1356.)

Data analysis

We selected angiotensin converting enzyme variants related to the extent of structural remodeling (total left atrial volume, local left atrial volume, local curved left atrial length, mean and local left atrial voltage, and LA/LVEDD ratio). Statistical analysis was performed using SPSS statistical package release 17.0.1 (SPSS, Inc., Chicago, IL, USA). Data are presented as mean±standard deviation (SD). For comparison of baseline characteristics, data between groups was analyzed as Student's unpaired t- test for continuous data and 2 tests for categorical data. For statistical analysis, it was decided to cut to 0.1 decimal place as the median, and verified by ROC (receiver operating characteristic) curve analysis. In single locus analysis, we first compared allele and genotype frequencies between case and control with 2 tests or Fisher's exact test. Statistical significance was defined as P <0.05.

Atrial fibrillation In the patient Associated with the structural remodeling of the atrium ACE transition

Figure 1 is a representative example of significantly remodeled left atrium (Figures 1A and 1C) and less remodeled left atrium (Figures 1B and 1E) in atrial fibrillation patients, and their ACE Genotype (Figure 1D) Show. Patients who showed significant electroanatomical remodeling of the left ventricle showed expanded left atrial volume, low endocardial voltage (Figure 1A), and high LA/LVEDD ratio (Figure 1C). In contrast, patients with less remodeled left atrium showed relatively small left atrial volume, high endocardial voltage (Figure 1B), and low LA/LVEDD ratio (Figure 1E).

Of the 7 genes we evaluated, ACE Five polymorphisms of the genes were associated with structural remodeling of the left ventricle in 351 patients with non-familial non-valvular atrial fibrillation. Table 2 summarizes the results of ACE mutation analysis related to the degree of structural remodeling of the left atrium. ACE The rs4362 C (p=0.024), rs4309 T (p=0.030), rs1800764 T (p=0.030), and rs4291 A (p=0.027) alleles tended to have an expansion of left atrial volume in patients. LA dilatation (LA/LVEDD) relative to LV size measured by echocardiography is ACE It was significantly increased in patients with the rs4362 C allele (p=0.039) and the rs4309 T allele (p=0.026). ACE rs4362 T allele (p=0.021) and ACE The D carriers (DD+ID) allele (p=0.021) was the predominant genotype in patients with low mean LA voltage. ACEs rs1800764 and rs4291 were closely related to each other in a linkage disequilibrium block (D'=0.977, r ² =0.93; FIG. 2).

[Table 2]

Structural of the left atrium Department of Remodeling Related ACE Left atrial volume and voltage according to mutation

Table 3 shows ACE It summarizes the area volume and area curve length of LA adjusted by the body surface area (BSA) according to the rs4362 genotype. We also compared mean and local LA voltage and echocardiographic parameters. Generally, ACE Patients with the rs4362 C allele had a greater overall and local LA volume (p<0.01) and a longer local curve length of LA (p<0.05) than those with the rs4362 TT genotype. In contrast, patients with the rs4362 T allele had a lower LA voltage (p<0.05) than those with the rs4362 CC genotype.

[Table 3]

ACE The characteristics of LA remodeling of patients with the rs4309 allele, rs1800764 allele, and rs4291 allele are summarized in Table 4.

[Table 4]

ACE Mutant AF Results after catheterectomy in patients

The clinical recurrence rate of atrial fibrillation after the 3 month blanking period was 20.45% during the 28.29±5.83 month follow-up period. ACE Regarding genotype, we did not find ACE-associated polymorphisms associated with long-term clinical recurrence after catheterectomy. However, ACE associated with low endocardial left atrial voltage The rs4362 T allele is ACE Compared to the rs4362 CC genotype, it was associated with a higher initial recurrence rate within 3 months after RFCA (27.5%, p=0.0217).

Through this study, we report that several ACE polymorphisms are associated with electroanatomical remodeling of atrial fibrillation. They are associated with dilatation of the left atrium, decreased endocardial voltage, and early recurrence after catheterectomy. These ACE polymorphisms will be useful for structural remodeling of atrial fibrillation, that is, early detection of atrial fibrillation patients vulnerable to electroanatomical remodeling. Thus, it is possible to prevent these patients from progressing to chronic persistent atrial fibrillation. In addition, upstream treatment of such patients with ACE inhibitors or angiotensin II receptor blockers can prevent atrial remodeling as a personalized management. These variations also justify early intervention in catheterectomy and may improve clinical outcomes with prognostic value.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Polynucleotide for predicting prognosis of atrial fibrillation <130> P100868 <140> 10-2010-0117541 <141> 2010-11-24 <160> 4 <170> KopatentIn 2.0 <210> 1 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> rs4362 <220> <221> misc_feature <222> (27) <223> The y in position 27 is C or T. <400> 1 acggcctcgc tctgctccag gtacttygtc agcttcatca tccagttcca gt 52 <210> 2 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> rs4309 <220> <221> misc_feature <222> (27) <223> The y in position 27 is C or T. <400> 2 tactacctgc agtacaagga tctgccygtc tccctgcgtc ggggggccaa cc 52 <210> 3 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> rs1800764 <220> <221> misc_feature <222> (27) <223> The y in position 27 is C or T. <400> 3 atgtacccac tgaggataag ggggggytgc tgtacatact ttgcaaatat ct 52 <210> 4 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> rs4291 <220> <221> misc_feature <222> (27) <223> The y in position 27 is A or T. <400> 4 cggcaaactg ccgggtcccc atcttcyaaa gagaggaggc cctttctcca gc 52

Claims (5)

A polynucleotide for prognostic prediction of atrial fibrillation consisting of 10 to 50 consecutive DNA sequences including the 27th base (polymorphic region) of the polynucleotide of SEQ ID NO: 4 or a complementary polynucleotide thereof.
The method of claim 1,
SEQ ID NO: 4 is the allele of the 27th base (polymorphic region) is A / T prognostic predictive polynucleotide or a complementary polynucleotide thereof.
A DNA microarray chip for prognostic prediction of atrial fibrillation in which the polynucleotide according to claim 1 or the complementary polynucleotide thereof is integrated.
A kit for predicting prognosis of atrial fibrillation comprising the polynucleotide according to claim 1 or the complementary polynucleotide thereof.
A computer-readable recording medium having recorded thereon a program for performing prognostic prediction of atrial fibrillation, comprising: a 27th base (polymorphic site) in SEQ ID NO: 4, indicating a risk of recurrence of atrial fibrillation in a nucleic acid sample obtained from a patient Analyzing 10 to 50 consecutive DNA sequences or their complementary sequences, and
A computer-readable recording medium having recorded thereon a program which causes the computer to perform the step of classifying the patient as a potential patient for electroanatomical remodeling of the atrium if the 27th base of the patient's gene sequence is A.

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