KR102658118B1 - Biomarker composition for diagnosing depression with earlier age at onset using the FYB gene, information provision method and diagnostic kit for diagnosing depression with earlier age at onset using the same - Google Patents

Abstract

The present invention relates to a technology for diagnosing depression using biomarkers. More specifically, the present invention relates to a technology for diagnosing depression using biomarkers. More specifically, the present invention relates to a technology for diagnosing depression using an early onset age using the FYB gene, which was developed on the basis that mutations in specific genes increase the risk of developing depression at an earlier onset age. This topic is about a biomarker composition for diagnosing depression, a method of providing information on diagnosing depression with an early onset age using the same, and a diagnostic kit.

Description

Biomarker composition for diagnosing depression with an early age at onset using the FYB gene, information provision method and diagnostic kit for diagnosing depression with an early onset age using the same {Biomarker composition for diagnosing depression with earlier age at onset using the FYB gene, information provision method and diagnostic kit for diagnosing depression with earlier age at onset using the same}

The present invention relates to a technology for diagnosing depression using biomarkers. More specifically, the present invention relates to a technology for diagnosing depression using biomarkers. More specifically, the present invention relates to a technology for diagnosing depression using an early onset age using the FYB gene, which was developed on the basis that mutations in specific genes increase the risk of developing depression at an earlier onset age. This topic is about a biomarker composition for diagnosing depression, a method of providing information on diagnosing depression with an early onset age using the same, and a diagnostic kit.

Depression is a prevalent mental disorder with a high disease burden. Despite substantial evidence for the genetic heritability of depression, the genetic architecture of depression remains unclear (Sullivan et al., 2000). Previous genome-wide association studies (GWAS) that failed to identify genetic markers for major depression (Major Depressive Disorder Working Group of the Psychiatric Gwas Consortium et al., 2013), for example, maximized sample size (Howard et al. , 2019; Hyde et al., 2016; Wray et al., 2018;) or heterogeneity of depression, such as purifying the sample in terms of age of onset (AAO) and relapse (CONVERGE Consortium, 2015; Power et al., 2012). It was suggested that various analysis strategies are needed to overcome this.

Age at onset (AAO) has been considered a potential indicator of underlying genetic risk for depression based on family aggregation and heritability studies showing that early-onset depression is associated with a family history of the disorder (Kendler et al., 2009; Nienberg et al., 2007; Tozzi et al., 2008). Moreover, the risk of early-onset depression in children of disabled parents was 13 times greater than in control children (Wickramarantne and Weissman et al., 1998). A recent GWAS suggested that the importance of genetic factors for depression differs between adult and early-onset depression, with early-onset cases showing greater genetic overlap with schizophrenia and bipolar disorder (Power et al., 2017 ). This finding was supported by a follow-up study showing that adolescent-onset depression has a distinct depression-specific genetic profile (based on polygenic risk scores) for very early-onset depression, with genetic overlap with ADHD and schizophrenia (Kwong et al ., 2019; Rice et al., 2019).

As an example of research results on genetic variants associated with AAO of depression, the TUSC3 gene is associated with early-onset depression in men, while the intergenic region on chromosome 3 is associated with adult-onset depression (Power et al., 2012 and 2017). Moreover, previous GWAS have examined only common, low-impact genetic variants that do not fully capture the genetic background of depression. Additionally, GWAS failed to confirm the association of specific variants with AAO of depression due to lack of repeated validation of specific variants and unknown functions of the identified variants.

Therefore, identifying genetic markers for depression with an early age of onset (AAO) can reduce the disease burden through genetic susceptibility screening, and new genetic markers to predict depression with an early age of onset in adults have been developed. needs to be

Republic of Korea Patent No. 10-1919753

The present inventors completed the present invention by revealing, as a result of numerous studies, a biomarker for predicting depression with an early onset age that is significantly associated with the risk of developing depression before the age of 40.

Therefore, the purpose of the present invention is to confirm that a mutation in a specific region in one or more of the CCL14, FYB, GPRASP1, and CTNND2 genes can predict depression with an early onset age that occurs before the age of 40, and to detect depression in adults over the age of 40. The aim is to provide a biomarker composition for predicting depression with an early onset age that includes a biomarker for predicting depression with an early onset age that can relatively accurately determine the risk of previously developing depression.

Another object of the present invention is to identify depression patients before or in the early stages of depression through genetic susceptibility screening by detecting biomarkers for predicting depression with an early onset age, and to provide a decision-making process for treatment drugs or treatment methods through early intervention. It provides a method of providing information on diagnosing depression with an early age of onset, which can contribute to reducing the burden of the disease.

Another purpose of the present invention is to identify patients with genetic risk factors for early-onset depression by identifying the mutation region of a biomarker for predicting depression with an early onset age from biological samples isolated from an individual, thereby predicting the patient's depression symptoms. The goal is to provide a diagnostic kit and microarray for predicting depression with an early age of onset that can be clinically useful as it can preemptively prevent and/or treat depression before it worsens.

The objects of the present invention are not limited to the objects mentioned above, and other objects not mentioned will be clearly understood by those skilled in the art from the description below.

In order to achieve the object of the present invention described above, the present invention includes a polynucleotide of 10 to 100 nucleotides in length containing the rs35384751 (G>T) region of the FYB gene or a complementary polynucleotide thereof as a control, wherein the rs35384751 If the base in the region is confirmed to be T, a biomarker composition for diagnosing depression with an early onset age is predicted to increase the risk of developing depression by 1.53 times compared to the control group in which the base in the rs35384751 region is G. do.

Additionally, the present invention provides a diagnostic kit for depression with an early onset age, including a detection means capable of detecting the above-described biomarker composition.

In a preferred embodiment, the detection means includes one or more of a probe capable of detecting a polynucleotide included in the biomarker composition, a primer pair capable of amplifying the polynucleotide, and cDNA of the polynucleotide.

Additionally, the present invention provides a microarray for diagnosing depression with an early onset age, comprising the above-described biomarker composition.

In addition, the present invention includes the steps of obtaining DNA or RNA of the FYB gene from a biological sample of a test subject; And a step of confirming the base of the rs35384751 region contained in the above-mentioned biomarker composition from the obtained DNA or RNA. When the base of the rs35384751 region is confirmed to be T, a control group in which the base of the rs35384751 region is G In comparison, we provide a method of providing information on diagnosing depression with an early onset age, which is characterized by a predicted 1.53-fold increase in the risk of developing depression at an earlier age.

In a preferred embodiment, the test subject is an adult over 20 years of age, and the biological sample is selected from body fluids or tissues containing blood.

First, according to the biomarker composition of the present invention, it was confirmed that mutations in specific regions in one or more of the CCL14, FYB, GPRASP1, and CTNND2 genes can be a biomarker for predicting depression with an early onset age of onset before the age of 40. By doing so, it is possible to relatively accurately determine the risk of developing depression in adults before the age of 40.

In addition, according to the method of providing information on diagnosing depression with an early onset age of the present invention, it is possible to identify patients with depression before or in the early stages of the onset of depression through genetic susceptibility screening using biomarkers for predicting depression with an early onset age. Early intervention by doctors can help determine medication or treatment before depression deepens.

In addition, according to the diagnostic kit and microarray of the present invention, patients with genetic risk factors for early-onset depression can be predicted by identifying the mutation site of a biomarker for predicting depression with early onset age from biological samples isolated from individuals. Therefore, it has clinical utility as it can preemptively prevent and/or treat patients' depression symptoms before they deepen.

The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below.

1 is a flow chart (VCF: Variant Call Format File, FDR: False Discovery Rate) outlining the analysis process of the present invention.
Figure 2 is a graph of the results of Kaplan-Meier survival analysis using the log-rank test for the association between the age of onset of depression and CCL14 gene mutation for all depression patients.
Figure 3 is a graph of the results of Kaplan-Meier survival analysis using the log-rank test for the association between the age of onset of depression and FYB gene mutation for all depression patients.
Figure 4 is a graph of the results of Kaplan-Meier survival analysis using the log-rank test for the association between the age of onset of depression and GPRASP1 gene mutation for all depression patients.
Figure 5 is a graph of the results of Kaplan-Meier survival analysis using the log-rank test for the association between the age of onset of depression and CTNND2 gene mutation for all depression patients.

The terms used in the present invention are merely used to describe specific embodiments and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this application, terms such as “comprise” or “have” are intended to designate the presence of features, numbers, steps, operations, components, parts, or combinations thereof described in the description of the invention, but are intended to indicate the presence of one or more other It should be understood that this does not exclude in advance the possibility of the presence or addition of features, numbers, steps, operations, components, parts, or combinations thereof.

Terms such as first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The above terms are used only for the purpose of distinguishing one component from another. For example, a first component may be referred to as a second component, and similarly, the second component may also be referred to as a first component without departing from the scope of the present invention.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the technical field to which the present invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and unless clearly defined in the present invention, should not be interpreted in an idealized or excessively formal sense. No.

When interpreting a component, it is interpreted to include the margin of error even if there is no separate explicit description.

In the case of a description of a temporal relationship, for example, if a temporal relationship is described as ‘after’, ‘after’, ‘after’, ‘before’, etc., ‘immediately’ or ‘directly’ Even non-consecutive cases are included unless ' is used.

The human genome shows more than 0.1% genetic diversity, and these differences can be used as a tool for genetic fingerprint recognition. Single nucleotide polymorphisms (SNPs) can be used as individual genetic fingerprint recognition patterns, and more than 10 million variations are widely known by race and country through the human genome project. These SNPs can be easily detected through sequencing, and this diversity is rare, individual, and specific. However, there may be very frequent variations in some areas. The present inventors discovered that depression with an early onset age can be easily diagnosed by using the characteristics of the single nucleotide polymorphism (SNP) described above.

As used herein, the term “diagnosis” means confirming the presence or characteristics of a pathological condition. For the purposes of the present invention, “diagnosis” means determining the likelihood of developing depression before the age of 40 in an adult patient based on in vitro analysis of body fluids.

In this specification, the term "rs_id" refers to rs-ID, an independent marker assigned to all SNPs initially registered by NCBI, which began accumulating SNP information in 1998. As such, rs_id listed in the table refers to a SNP marker indicating the genetic mutation of the present invention.

The term “biomarker” used in the present invention refers to a substance that can indicate a disease state. In the context of the present invention regarding the diagnosis of depression with an early age of onset, “biomarker” refers to a specific SNP region in any one of the CCL14 gene, FYB gene, GPRASP1 gene and CTNND2 gene. Depending on the type, a “biomarker” may or may not appear in an individual suffering from depression or susceptible to depression with an earlier age of onset compared to a normal healthy state.

The term “blood” as used herein includes whole blood, serum, and plasma.

As used herein, the term “predicting” means discovering that an individual is significantly likely to develop a biological disease.

As used herein, the term “biological sample” includes various sample types obtained from an individual and may also be used in diagnostic or monitoring analysis. Biological fluid samples include blood, cerebrospinal fluid (CSF), urine, and other liquid samples of biological origin. If desired, samples can be pre-processed, for example for concentration and separation.

As used herein, the term “individual” refers to a mammal, preferably a human, and the terms individual or subject may be used interchangeably in the present invention.

The term "early onset age" used in the present invention refers to the age of onset, which refers to the age at which depression occurs, especially in adults aged 20 or older who develop depression before the age of 40, meaning 20 years ≤ early onset age ≤ 40 years. .

Hereinafter, the technical configuration of the present invention will be described in detail with reference to the attached drawings and preferred embodiments.

However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. The same reference numerals used to describe the invention throughout the invention indicate like elements.

The technical feature of the present invention is to confirm that mutations in one or more specific regions of the CCL14, FYB, GPRASP1, and CTNND2 genes can be a biomarker for predicting depression with an early onset age, and based on this, the study was conducted on adults over 40 years of age. A diagnostic kit for diagnosing depression with an early onset age, including a configuration for detecting a specific SNP in any one or more of the CCL14 gene, FYB gene, GPRASP1 gene, and CTNND2 gene for prediction and/or diagnosis of the possibility of previously developing depression; and The goal is to provide a method of providing information on predicting depression with an early age of onset.

Here, the CCCL14 gene is a C-C motif chemokine ligand 14 (CCL14) gene. CCL14, one of the cytokine gene clusters, is known to attract and activate leukocytes to regulate immune cell homeostasis and inflammation. (Moelants et al., 2013). The FYB gene, the FYN binding protein (FYB) gene, encodes an adapter protein involved in T cell signaling and the mTOR signaling pathway, which leads to T cell activation and production of inflammatory cytokines, including IL-2. (da Silva et al., 1997; Griffith et al., 2001; Kotajima-Murakami et al., 2019). In accordance with their primary role in inflammation, CCL14 and FYB have been shown to be associated with systemic lupus erythematosus and diabetes (Addobbati et al., 2013, 2015; Vyshkina et al., 2008; Suchy-Dice et al., 2014). Because inflammation has been identified as one of the pathogenic mechanisms underlying depressive disorders, variants in the CCL14 and FYB genes may increase the risk of depression in early AAO.

The GPRASP1 gene is a G protein-coupled receptor-associated sorting protein 1 (GPRASP1) gene and is a direct interaction partner of the M2-muscarinic acetylcholine receptor (CHRM2), which increases cholinergic system activity. Considering its effect on mood (Furey and Drevets, 2006), it can be considered a genetic risk factor for depressive disorder (Luo et al., 2005; Wang et al. 2004). The CTNND2 gene, the catenin delta 2 (CTNND2) gene, encodes δ-catenin expressed in cortical and hippocampal neurons (Ho et al., 2000) and is involved in early cerebral development and dendrite maintenance processes (Turner et al., 2000). al., 2015) plays an essential role in neuron migration in the brain. Based on its important role in neuronal maintenance and plasticity, the CTNND2 gene has been shown to be associated with anxiety-related traits in rats (Rat Genome Sequencing Consortium, 2013) as well as anxiety and major depressive disorder in a human GWAS (Nivard). et al., 2014).

Therefore, the biomarker composition for diagnosing depression with early onset age of the present invention includes a polynucleotide of 10 to 100 nucleotides in length containing the rs75238886 (G>A) region of the CCL14 gene or a complementary polynucleotide thereof as a control, or A polynucleotide of 10 to 100 nucleotides in length containing the rs35384751 (G>T) region of the FYB gene or its complementary polynucleotide is included as a control, or 10 to 100 nucleotides containing the rs201921260 (T>C) region of the GPRASP1 gene. A polynucleotide with a length of 10 nucleotides or its complementary polynucleotide is included as a control, or a polynucleotide with a length of 10 to 100 nucleotides containing the rs202234398 (G>A) region of the CTNND2 gene or a complementary polynucleotide thereof is included as a control. can do.

As described later, the present invention has clearly confirmed the effect of mutations in specific regions of one or more of the CCL14, FYB, GPRASP1, and CTNND2 genes on increasing the risk of developing depression before the age of 40 in adults.

That is, specific SNP polynucleotides obtained from one or more of the CCL14, FYB, GPRASP1, and CTNND2 genes included in the biomarker composition of the present invention are used to identify genetic markers for depression in consideration of AAO, as described later, in Korean patients with depression. It was obtained by performing full-length exome sequencing on 1,000 samples to investigate whether a single nucleotide polymorphism (SNP) in a specific gene could be a biomarker related to the diagnosis of depression with an early onset age. Sequencing revealed that four variants in the genes CCL14, FYB, GPRASP1 and CTNND2 were associated with an increased risk of depressive disorders early in the onset. Whole-exome sequencing analysis had the advantage of capturing both rare and common variants in the exome (protein-coding region).

Meanwhile, the diagnostic kit for depression with early onset age of the present invention includes the rs75238886 (G>A) region of the CCL14 gene, the rs35384751 (G>T) region of the FYB gene, and the rs201921260 (T>C) region of the GPRASP1 gene contained in the biomarker composition. ) region, and a detection means capable of detecting any one polynucleotide of the rs202234398 (G>A) region of the CTNND2 gene or its complementary polynucleotide. Here, the detection means may include one or more of a probe capable of detecting a polynucleotide included in the biomarker composition, a primer pair capable of amplifying the polynucleotide, and cDNA of the polynucleotide.

Of course, if necessary, it can be implemented as a microarray containing a biomarker composition to diagnose depression with an early onset age.

In addition, the method of providing information on diagnosing depression with an early onset age of the present invention includes DNA or RNA of the CCL14 gene, DNA or RNA of the FYB gene, DNA or RNA of the GPRASP1 gene, and DNA or RNA of the CTNND2 gene from a biological sample of the subject. Obtaining any one of the following; And a step of identifying any one of the bases at the rs75238886 region, the base at the rs35384751 region, the base at the rs201921260 region, and the base at the rs202234398 region contained in the biomarker composition from the obtained DNA or RNA. Here, if the base at the rs75238886 region is confirmed to be A as shown in the results of the investigation described later, the risk of developing depression at an earlier age can be diagnosed to be 2.2 times higher compared to the control group where the base at the rs75238886 region is G, and rs35384751 If the base at the rs35384751 region is confirmed to be T, the risk of developing depression at an earlier age can be diagnosed to be 1.53 times higher compared to the control group where the base at the rs35384751 region is G, and if the base at the rs201921260 region is confirmed to be C, Compared to the control group where the base at the rs201921260 region is T, the risk of developing depression at an earlier age can be diagnosed to be 2.95 times higher. If the base at the rs202234398 region is confirmed to be A, the control group where the base at the rs202234398 region is G. Compared to , the risk of developing depression at an earlier age can be diagnosed as 1.94 times higher.

Example

1. Research overview

An analysis was performed on data from the MAKE BETTER study, which investigated biomarkers of treatment response using a prospective design. Details of the study have been published previously (Kang et al., 2018). The current study analyzed baseline data from all participants who agreed to provide blood samples for genetic testing. Patients who recently visited the Department of Psychiatry at Chonnam National University Hospital and were diagnosed with major depressive disorder, dysthymic disorder, or other depressive disorder were asked to take the Mini-International Neuropsychiatric Interview (MINI, based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition [DSM-IV]). (Sheehan et al., 1998) were recruited consecutively through a structured diagnostic interview. Eligibility criteria are as follows.

<Qualification criteria>

For the MAKE BETTER study, the inclusion criteria were as follows: i) 7 years or older; ii) diagnosed with major depressive disorder, dysthymic disorder, not otherwise specified, or depressive disorder not otherwise classified (NOS), as confirmed using the Mini-International Neuropsychiatric Interview (Sheehan et al., 1998); iii) Hamilton Depression Rating Scale (Hamilton, 1960) score ≥ 14; iv) Complete the questionnaire, understand the purpose of the study, and sign the informed consent form. Exclusion criteria are as follows: i) unstable or uncontrollable medical condition; ii) inability to complete psychiatric evaluation or adhere to medication regimen due to serious physical illness; iii) current or lifetime DSM-IV diagnosis of bipolar disorder, schizophrenia, schizoaffective disorder, schizophrenic disorder, psychotic disorder not elsewhere classified, or other psychotic disorder; iv) History of organic psychosis, epilepsy or seizure disorder; v) history of anticonvulsant treatment; vi) Hospitalization for any psychiatric diagnosis except depressive disorder (e.g. alcohol/drug dependence) vii) Electroconvulsive therapy for current depressive episode; viii) Pregnancy or breastfeeding.

The MAKE BETTER study was approved by the institutional review board of Chonnam National University Hospital, and written consent was obtained from all participants.

2. Full-length exome sequencing

DNA was extracted from venous blood samples of MAKE BETTER participants who consented to genetic testing. After exome capture was performed using the SureSelect Human All Exon V5-UTR kit (Agilent Technologies, Santa Clara, CA, USA), sequencing was performed in paired-end mode (in 100-bp or 150-bp reads, according to the manufacturer's protocol). in paired-end mode) (HiSeq2500; Illumina, San Diego, CA, USA). A bioinformatics pipeline following the best practice recommendations included in the Genome Analysis Toolkit (GATK) version 3.3-0 (van der Auwera et al., 2013) was used to process the captured exome data. BWA 0.7.5a was used to align the reads to the human genome reference sequence (hg19 / GRCh37) (Li et al., 2010). Aligned reads were further processed using InDel (short insertion and deletion) realignment, MarkDuplicates package in Picard software (ver. 1.92; http://picard.sourceforge.net), and sequence score recalibration (BQSR).

Single nucleotide variant and InDel discovery was performed simultaneously in all samples using Haplotype Caller (GATK) (DePristo et al., 2011; McKenna et al., 2010; van der Auwera et al., 2013). Variants were annotated with SnpEff 4.2 (Cingolani et al., 2012a) and SnpSift 4.2 (Cingolani et al., 2012b) software using dbNSFP (ver. 2.9.1) and Ensembl GRCh37.75.

3. Demographic and clinical characteristics of depressed patients

Demographic and clinical characteristics potentially associated with AAO of depressive disorders were assessed. Data were obtained on age, years of education, employment status and number of chronic physical disabilities. Information on depressive disorders includes diagnosis of depressive disorder, number of depressive episodes, AAO of depression, duration of current episode, family history of depressive disorder, history of past suicide attempts, severity of depressive symptoms according to the Hamilton Rating Scale (Hamilton, 1960), and hospital anxiety depression. Severity of anxiety symptoms according to the anxiety subscale of the scale (Zigmond and Snaith, 1983), severity of suicidal ideation according to the suicide-related items of the Brief Psychiatric Rating scale (Overall and Gorham, 1962), and DSM-IV criteria (American Psychiatric Association). , 2000), detailed diagnostic aspects of depression including melancholia, atypical, and psychotic aspects were also obtained.

4. Statistical analysis

Participants were divided into three groups (early onset: 20 < age ≤ 40 years, adult onset: 40 < age < 65 years, late onset: age ≥ 65 years) based on previous studies (Alexopoulos, 2005; Power et al., 2012; Wickramarantne and Weissman et al., 1998). Baseline characteristics, including sociodemographic and clinical factors, were compared between groups through analysis of variance (ANOVA) and Scheff's post hoc test or χ2 test was applied as appropriate. Factors significantly associated with AAO (P <0.05) were entered as covariates in the adjusted analysis performed in the next step.

The analysis process is described in Figure 1. All non-synonymous mutations resulting in amino acid changes were analyzed. To estimate the impact of individual variables on the AAO of depressive disorders, hazard ratios were obtained using the Cox proportional hazards model.

We compared carriers and non-carriers of individual coding variants on the AAO aspect of depressive disorders, with adjustment for sociodemographic and clinical characteristics. Due to multiple comparisons, false discovery rate (FDR) corrected P-values were obtained with the Benjamini-Hochberg method. An FDR-corrected P-value <0.05 was considered statistically significant.

The quality of sequenced nucleic acid fragments (reads) was assessed by checking the bam files using the IGV viewer (http://software.broadinstitute.org /software/igv/) for errors that may have occurred during mechanical filtering during preprocessing. . False positive mutations were removed. A final manual review was also performed by three bioinformatics experts to confirm the results. Kaplan-Meier analysis and log-rank test were used to compare AAO according to candidate variants, and the effect of individual variants on AAO of depressive disorder was graphically depicted.

The clinical impact of candidate variants was assessed by comparing heterozygous or homozygous carriers for each individual variant and for any one of all variants identified using the t test, Wilcoxon rank sum test, χ2 test, or Fisher's exact test. It was decided by comparing the holders.

All statistical analyzes were performed using R software (ver. 3.2.3; http://www.rproject.org/).

5. Results

(1) Target

Among the 1,262 patients who participated in the MAKE BETTER study, 1,000 patients with depressive disorders consented to genetic testing and were included in the study population. Patient characteristics are listed in Table 1 below. Compared to those who did, people who did not consent to having their blood drawn had more depressive episodes (P < 0.001), were more likely to be currently employed (P = 0.003), and had less severe suicidal thoughts (P < 0.001). P = 0.039), and there were many cases showing melancholic features (P = 0.037). The mean (SD) AAO of 1,000 patients was 52.0 (16.4) years (range: 8–84 years).

Patients with early onset depression were younger, more educated, more likely to be currently unemployed, experienced more depressive episodes, had a family history of depressive disorder and a history of suicide attempts, and had more severe anxiety symptoms, suicidal thoughts, and atypical features. . Compared to other AAO groups, they were more likely to be single and less likely to show melancholy (P <0.05). These significant factors were included as covariates in the Cox proportional hazards model.

(2) genetic variants associated with AAO in depressive disorders;

To estimate the impact of individual nonsynonymous variants on AAO in depressive disorders, variant carrier and noncarrier groups were compared using Cox proportional hazards models after adjustment for sociodemographic and clinical characteristics. FDR-corrected P-values were obtained with the Benjamini-Hochberg procedure as mentioned above.

After checking the read quality, candidate genes (FDR corrected P < 0.1) were further analyzed and genes with FDR corrected P < 0.05 were included in the final analysis. Table 2 shows the number of significant variants and associated genes derived by Cox proportional hazards model with P-value corrected for false discovery rate (FDR). Table 3 shows statistically significant genetic variations related to the age of early onset of depressive disorder.

FDR adjusted p-value < 0.1 FDR adjusted p-value < 0.05 N =33
RNPEP,SNAP47,SNAP47,PRSS38,TET1,TBRG1,CIT,FOXA1,TEKT5,GCSH,HS3ST3B1,LIG3,CCL14,ABCA7,DOCK6,PER2,COL6A1,ELFN2,CASR,ADIPOQ,MFSD8,FAM218A,FAM114A1,CTNND2,PCDHB11, FYB,DST,LANCL2,EXT1,HTRA4,OTUD6B,OR2K2,ORM1,GPRASP1 N =4
CCL14, CTNND2, FYB, GPRASP1

Gene Position rsID PP2 SIFT CADD Carrier (homozygotes) Wild Type Cox-PH
FDR-adjusted P HR 95% CI 1KGP 1KGP
EAS CCL14 chr17:34313612 G>A rs75238886 0.773 0 12.72 31(0) 969 0.00008 ^‡ 2.2 1.52-3.18 0.02556 0.0466 FYB chr5:39202689 G>T rs35384751 0.459 0 8.0 111(1) 889 0.00008 ^‡ 1.53 1.25-1.87 0.01637 0.0288 GPRASP1 chrX:101909416 T>C rs201921260 0.999 0.51 7.5 10(0) 990 0.00308 ^† 2.95 1.56-5.55 0.00053 0.001 CTNND2 chr5:11364820 G>A rs202234398 0.919 0.16 24.5 14(0) 986 0.02785* 1.94 1.12-3.38 0.0004 0.002

rsID, reference ID; SIFT, =orting Intolerant From Tolerant; PP2, = PolyPhen2; CADD, Combined Annotation Dependent Depletion; P-value corrected for Cox-PH FDR, P-value obtained using Cox proportional hazards model corrected for false discovery rate (FDR), HR, hazard ratio, CI, confidence interval, 1KGP, allele from 1000 Genome Project Phase 3 frequency; Allele frequencies in East Asia from 1KGP EAS, 1000 Genome Project Phase 3.

*, p value <0.05; †, p-value <0.01'‡, p-value <0.001

As shown in Table 3, there were four variants, namely rs75238886 in the CC motif chemokine ligand 14 (CCL14) gene, rs35384751 in the FYN-binding protein (FYB) gene, rs201921260 in the G protein-coupled receptor-related sorting protein 1 (GPRASP1) gene and catenin. The rs202234398 delta 2 (CTNND2) gene was significantly associated with a higher risk (1.53 - 2.95) of early onset.

AAO for depressive disorders was compared between carriers and non-carriers of each individual variant using Kaplan-Meier survival analysis with the log-rank test and the results are shown in Figures 2 to 5. As shown, patients with one of the four variants were more likely to suffer from depression with early AAO.

(3) Association between clinical characteristics and genetic mutations in patients with depression

To estimate the clinical impact of identified variants, carriers and non-carriers of each variant and one or more of the four variants were compared in terms of clinical characteristics using the t test, Wilcoxon's rank sum test, χ2 test, or Fisher's exact. and the results are shown in Table 4.

HAMD, Hamilton Depression Rating Scale; HADS, Hospital Anxiety Depression Scale; BPRS, Brief Psychiatric Rating Scale.

As shown in Table 4, depressed patients with one or more of the four mutations were younger and more likely to suffer from depression in early AAO. Additionally, they were more likely to experience recurrent and atypical depression. However, no individual variations were associated with any clinical characteristics except early AAO.

The main finding of the above whole-exome sequencing analysis is that four variants of the CCL14, FYB, GPRASP1, and CTNND2 genes are associated with an increased risk of developing depression with an early age of onset. There were no individual variants associated with the clinical characteristics of depressed patients except for AAO, but the presence of any one of the four variants was associated with younger age, early AAO, relapse, and atypical depression. That is, depressed patients with at least one candidate variant were younger and more likely to relapse and experience atypical depression, even though the individual variant was not associated with clinical characteristics.

Consistent with these findings, the age of onset (AAO) of depressive disorders is known to influence symptoms and clinical course. Patients with early-onset depression are more likely to have a chronic course and experience more depressive episodes (Zisook et al., 2007, 2004). Atypical depression, characterized by mood reactivity and opposite physical responses (overeating and oversleeping), is generally a form of early-onset depression (Novik et al., 2004; Stewart et al., 2009).

The present invention is the first study to identify genetic markers of depressive disorder AAO using whole-exome sequencing. Because GWAS could not confirm the association of specific variants with AAO of depression due to the lack of repeated validation of specific variants and the unknown function of validated variants, we used whole-exome sequencing, which can capture both common and rare variants. There was an advantage in being able to do it.

In this way, in terms of clinical implications, using it as a biomarker for depression with an early onset age according to the present invention provides the advantages of non-invasiveness and convenience, and genetic susceptibility screening, that is, genetic screening for depression with an early onset age, By identifying the marker, it will be possible to contribute to the decision-making process regarding therapeutic drugs or treatment methods, which could be of great help as a potential tool to preemptively prevent or treat depression in patients.

Although the present invention has been illustrated and described by way of preferred embodiments as discussed above, it is not limited to the above-described embodiments and is intended to be used by those skilled in the art without departing from the spirit of the invention. Various changes and modifications will be possible.

Claims (6)

A polynucleotide of 10 to 100 nucleotides in length containing the rs35384751 (G>T) region of the FYB gene or a complementary polynucleotide thereof is included as a control,
When the base at the rs35384751 region is confirmed to be T, the risk of developing depression at an earlier age is predicted to increase by 1.53 times compared to the control group where the base at the rs35384751 region is G. A biomarker composition for diagnosing depression with an early onset age. .
A diagnostic kit for depression with an early onset age, comprising a detection means capable of detecting the biomarker composition of claim 1.
According to claim 2,
Early onset age, characterized in that the detection means includes one or more of a probe capable of detecting a polynucleotide included in the biomarker composition, a primer pair capable of amplifying the polynucleotide, and cDNA of the polynucleotide. A depression diagnostic kit with .
A microarray for diagnosing depression with an early onset age comprising the biomarker composition of claim 1.
Obtaining DNA or RNA of the FYB gene from a biological sample of the test subject; and
It includes the step of identifying the base at the rs35384751 region included in the biomarker composition of claim 1 from the obtained DNA or RNA,
If the base at the rs35384751 region is confirmed to be T, the risk of developing depression at an earlier age is predicted to increase by 1.53 times compared to the control group where the base at the rs35384751 region is G. Depression with an early onset age. How to provide information about diagnosis.
According to claim 5,
A method of providing information on the diagnosis of depression with an early onset age, wherein the test subject is an adult over 20 years of age, and the biological sample is selected from body fluids or tissues including blood.