KR101282269B1 - Polynucleotide for predicting prognosis of atrial fibrillation - Google Patents

Abstract

The present invention provides polynucleotides or their complementary polynucleotides comprising Single-Nucleotide Polymorphisms (SNPs) of genes related to structural remodeling of the left atrium, which affects the recurrence of atrial fibrillation. Since the polynucleotide or its complementary polynucleotide can be used as a predictor of the prognosis of atrial fibrillation, according to the present invention, early detection of a patient that is likely to cause structural remodeling of the left atrium causes the patient to progress to chronic persistent atrial fibrillation. Prevention can improve clinical outcomes.

Description

Polynucleotide for predicting prognosis of atrial fibrillation

The present invention relates to a polynucleotide for prognostic prediction of atrial fibrillation.

Atrial fibrillation (AF), one of the most common arrhythmias in the clinic, is associated with increasing mortality and disability rates (van den Berg MP, et al., Eur J Heart Fail . 2002; 4 (5): 571-575 .; Khairy P, et al., CMAJ . Pathology and physiology of atrial fibrillation are heterogeneous (Nattel S, et al., Lancet . 2006; 367 (9506): 262-272.), And older atrial fibrillation is associated with left atrial fibrillation (2002). left atrial, LA). Atrial fibrillation alters the electrophysical properties of myocytes in the atrium and causes alterations in 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 result in a more vulnerable substrate due to increased atrial fibrillation loads (Wijffels MC, et al., Circulation . 1995; 92 (7): 1954-1968). Structural remodeling is associated with interstitial fibrosis, downregulation of gap junctions, and enlargement of the atrium room 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 the degree of structural remodeling measured by scar formation in the left atrium (Verma A, et al., Circulation . 2005; 112 (5): 627-635.) Affect the clinical outcome of rhythm control in patients with

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 atria 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). Ravn et al ( Ravn LS, et al., Pharmacogenet Genomics . ACE 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, the present invention is to investigate the association between the genetic polymorphism of the angiotensin converting enzyme and the structural remodeling of the atrial fibrillation in patients with atrial fibrillation, and to use this result to predict the prognosis of atrial fibrillation.

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

Therefore,

Polynucleotide for prognostic diagnosis of atrial fibrillation consisting of 10 to 50 consecutive DNA sequences containing the 27th base (polymorphic region) of at least one polynucleotide selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 or Complementary polynucleotides are provided.

In one embodiment of the 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 in which two or more alleles exist in a single locus, and a 'polymorphic site' means a locus in which the allele exists. Among polymorphic sites, only a single base that differs from person to person is called "single nucleotide polymorphism," or SNP (single nucleotide polymorphism).

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

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 the allelic types that may appear are shown in parentheses.

Among the polynucleotides selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4, 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.

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

DNA microarray chip for prognostic diagnosis of atrial fibrillation of the present invention can be produced by methods known to those skilled in the art. More specifically, a micropipette using a piezoelectric method to immobilize the polynucleotide or a complementary polynucleotide thereof for prognostic diagnosis of atrial fibrillation on a substrate of a DNA chip using a probe DNA molecule. A micropipetting method or a method using a pin-shaped 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 (amino-silane), poly-L- lysine (poly-L-lysine) and aldehyde (aldehyde) It is not limited to this. 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 prognostic diagnosis of 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 is automatically executable by a computer, the method comprising: at least one member from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4, indicating a risk of recurrence of atrial fibrillation in a nucleic acid sample obtained from a patient In the selected sequence, 10 to 50 consecutive DNA sequences including the 27th base (polymorphic region) or their complementary sequences were analyzed, and as a result, the 27th base of the gene sequence of the patient was SEQ ID NO: 1 C; T for SEQ ID NO: 2; T for SEQ ID NO: 3; And in case of SEQ ID NO: 4, a method for predicting the prognosis of atrial fibrillation, comprising classifying the patient as a high probability of electroanatomical remodeling of the atrium.

The present invention also provides a computer-readable recording medium having recorded thereon a program for performing prognostic prediction of atrial fibrillation, comprising SEQ ID NO: 1 to SEQ ID NO: 4, which represents a risk of recurrence of atrial fibrillation in a nucleic acid sample obtained from a patient. Analyzing from 10 to 50 contiguous DNA sequences comprising the 27th base (polymorphic region) or complementary sequences thereof in a sequence selected from one or more groups from the group, and analyzing, the 27th base of the gene sequence of said patient Is 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 SEQ ID NO: 4, a computer-readable recording medium having recorded thereon a program for causing the computer to classify the patient into a patient with high possibility of electroanatomical remodeling of the atrium.

In accordance with the present invention, early detection of patients susceptible to structural remodeling of the left atrium can improve clinical results by preventing these patients from progressing to chronic persistent atrial fibrillation.

1 is a representative example of a significantly remodeled left atrium (FIGS. 1A and 1C) and a less remodeled left atrium (FIGS. 1B and 1E) in atrial fibrillation patients, and their ACEs Genotype (Figure 1D) Shows.
Figure 2 illustrates the results of ACE variation analysis related to the degree of structural remodeling of the left atrium.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

[ Example ]

Patient Selection

The protocol of this study was approved by the Institutional Review Board. All patients received written consent. The study included 351 atrial fibrillation patients (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 (PeAF). Exclusion criteria were as follows:

1) permanent atrial fibrillation in response to electrical cardioversion;

2) LA size> 55 mm measured by echocardiography;

3) AF with rheumatic valve disease;

4) associated structural heart disease;

5) previous AF block; And

6) sinus rhythm not maintained for LA voltage mapping before

If sinus rhythm is not maintained for LA voltage mapping prior to RFCA.

Patients with blood serum 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 anatomical condition and pulmonary vein of the left atrium. Transthoracic echocardiography is performed in all patients and left ventricular diastolic as measured by 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 the AF RFCA, we use a 5 mapping catheter and a reversible 3.5-mm tip 7 Fr open irrigation tip ablation catheter (Celsius, Johnson & Johnson Inc, Diamond Bar, CA, USA), and catheter ablation) The procedure was performed using 3-D electroanatomy mapping (NavX system, St. Jude Medical Inc., Minneapolis, MN, USA) in all patients. Prior to catheter dissection, we generated a LA 3D electroanalytical map by obtaining contact bipolar diagrams from 250-350 points of the LA endocardium. Voltage maps were color coded. Bipolar conductivity was filtered at 32-300 Hz. Color coded voltage maps were generated by recording bipolar electrical conductivity and measuring peak-to-peak voltage.

Analysis of Left Atrial Remodeling: 3D- Spiral CT And electroanatomical voltage maps

3D-spiral CT images of the left atrium were analyzed on an imaging processing workstation (Aquarius, Terarecon, Inc, USA). Curved length of the left atrium was measured at the 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 chamber and posterior wall), 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 describe the curve lengths of circumferential pulmonary vein ablation (CPVA), 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).

We analyzed color-coded electroanatomical voltage maps in terms of front-rear (AP) and rear-front (PA). The low voltage region below 0.2 mV was coded in gray and the high voltage region above 5.0 mV colored in purple. Reference distance was measured by the internal-electrode distance of the duodecapolar catheter (St. Jude Medical Inc. Minnetonka, MN, USA.). The voltage region in the pulmonary vein was excluded. To quantify the mean 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 are using the HapMap Japanese (JPT) Data banks (http://www.hapmap.org) and the NCBI SNP database (http://www.ncbi.nlm.nih.gov/projects/SNP/) ACE Haplotype-tagging of genes Single nucleotide polymorphisms (SNPs) were selected. To identify the appropriate tagging SNPs, we performed pilot genotyping on 16 selected SNPs out of 48 Korean patients and performed 7 SNPs ( ACE rs1800764, rs4291, rs4309, rs4362, rs4461142, rs61758684 and ACE I / D) was selected. Genomic DNA was extracted from whole blood samples using commercially available kits (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 degree of structural remodeling (total left atrial volume, local left atrial volume, local curve 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 expressed as mean ± standard deviation (SD). For comparison of baseline characteristics, data between groups were analyzed by Student's unpaired t- test for continuous data and 2 tests for categorical data. For statistical analysis, cut to 0.1 decimal places as the median and verified by receiver operating characteristic (ROC) curve analysis. In single-site analysis, we first compared the allele and genotype frequencies between the case and the control by 2 tests or Fisher's exact test. Statistical significance was defined as P <0.05.

Atrial fibrillation In patients  Associated with structural remodeling of the atria ACE  transition

1 is a representative example of a significantly remodeled left atrium (FIGS. 1A and 1C) and a less remodeled left atrium (FIGS. 1B and 1E) in atrial fibrillation patients, and their ACEs Genotype (Figure 1D) Shows. Patients showing significant electroanatomical remodeling of the left ventricle showed extended left atrial volume, low endocardial voltage (FIG. 1A) and high LA / LVEDD ratio (FIG. 1C). In contrast, patients with less remodeled left atrium showed relatively less left atrial volume, higher endocardial voltage (Figure 1B) and lower LA / LVEDD ratio (Figure 1E).

Of the seven genes we evaluated, ACE Five polymorphisms in the gene were associated with structural remodeling of the left ventricle in 351 patients with non-family non-valve atrial fibrillation. Table 2 summarizes the results of ACE variation 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 enlargement of the left atrium volume for the patients. LA expansion relative to LV size measured by echocardiography (LA / LVEDD) was significantly increased in patients with the ACE 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. ACE rs1800764 and rs4291 were closely related to each other in the linkage disequilibrium block (D ′ = 0.977, r ² = 0.93; FIG. 2).

[Table 2]

Structural of the left atrium Remodel  Related ACE  Left Atrial Volume and Voltage with Variation

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

[Table 3]

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

[Table 4]

ACE  Due to variation AF Clinical Results after Catheter Resection

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

In this study, we report that several ACE polymorphisms are associated with electroanatomical remodeling of atrial fibrillation. These are associated with dilatation of the left atrium, decreased endocardial voltage, and early recurrence after catheter resection. These ACE polymorphisms may be useful for early detection of atrial fibrillation patients who are vulnerable to structural remodeling of atrial fibrillation, that is, electroanatomical remodeling. Thus, these patients can be prevented from developing chronic persistent atrial fibrillation. In addition, treatment upstream with ACE inhibitors or angiotensin II receptor blockers for such patients can prevent atrial remodeling as a tailored care. These variations also justify early intervention of catheter resection and can improve clinical outcome with prognostic value.

Attach an electronic file to a sequence list

Claims (6)

Prognostic prediction of atrial fibrillation in Koreans comprising a polynucleotide for predicting atrial fibrillation consisting of 10 to 50 consecutive DNA sequences comprising the 27th base (polymorphic region) of the polynucleotide of SEQ ID NO: 2 or a complementary polynucleotide thereof Composition.
The method of claim 1,
The allele of the 27th base (polymorphic site) of SEQ ID NO: 2 is C / T composition for predicting atrial fibrillation in Koreans.
A polynucleotide consisting of 10 to 50 consecutive DNA sequences including the 27th base (polymorphic region) of the polynucleotide of SEQ ID NO: 2 or a complementary polynucleotide thereof is integrated, or the 27th base of the polynucleotide of SEQ ID NO: 2 Polymorphic site) allele is C / T, polynucleotide consisting of 10 to 50 consecutive DNA sequences comprising the same or DNA microarray chip for predicting prognosis of atrial fibrillation in Koreans with the complementary polynucleotides thereof.
A polynucleotide consisting of 10 to 50 contiguous DNA sequences comprising the 27th base (polymorphism) of the polynucleotide of SEQ ID NO: 2 or a complementary polynucleotide thereof, or the 27th base of the polynucleotide of SEQ ID NO: 2 Polymorphic site) allele is C / T, a kit for predicting the prognosis of atrial fibrillation in Koreans comprising a polynucleotide consisting of 10 to 50 consecutive DNA sequences or the complementary polynucleotides thereof.
delete A computer-readable recording medium recording a program for performing prognostic prediction of atrial fibrillation, comprising the 27th base (polymorphic site) in SEQ ID NO: 2, which represents a risk of recurrence of atrial fibrillation in a nucleic acid sample obtained from a Korean patient Analyzing 10 to 50 consecutive DNA sequences or complementary sequences thereof ; And if the 27th base of the gene sequence of the Korean patient is T, then classifying the Korean patient as a patient with potential for electroanatomical remodeling of the atrial can be read by computer. Recording media.

Images