How to detect rheumatoid arthritis

Abstract

To provide a more useful novel biomarker that can easily and accurately detect rheumatoid arthritis, and specifically a biomarker that can diagnose rheumatoid arthritis in an early stage.SOLUTION: The method for diagnosing rheumatoid arthritis of the present invention measures acrolein in a sample taken from a subject.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for detecting rheumatoid arthritis.

[0002] As used herein, the term "diagnosis" includes the term "early diagnosis." In this specification, "diagnosis (e.g., early diagnosis)" may be described as "(early) diagnosis." In other words, the description "(early) diagnosis" means "diagnosis (e.g., early diagnosis)." [Background technology]

[0003] Rheumatoid arthritis (RA) is a disease of unknown cause in which inflammation of the synovial membrane of joints leads to joint destruction. It has recently been discovered that this joint damage progresses rapidly and irreversibly in the early stages of onset. Therefore, it is important to suppress inflammation before joint destruction occurs. However, early diagnosis is difficult, and even rheumatoid arthritis specialists often struggle with it. Therefore, the development of an early diagnostic tool that allows anyone to easily and accurately diagnose RA would be a major step forward in the treatment of RA. For example, the AUC (areas under curve) values ​​of SAA4, gelsolin, and vitamin D binding protein, which are new biomarkers for rheumatoid arthritis, are 0.6453, 0.5190, and 0.6098, respectively, individually, and 0.7400 for the combination of these three (Non-Patent Document 1). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Mun S, Lee J, Lim MK, Lee YR, Ihm C, Lee SH, et al. Development of a novel diagnostic biomarker set for rheumatoid arthritis using a proteomic approach. Biomed Res Int. 2018; 26: Article ID 7490723. Summary of the Invention [Problem to be solved by the invention]

[0005] Therefore, an object of the present invention is to provide a novel and more useful biomarker that can easily and accurately detect rheumatoid arthritis, particularly a biomarker that can diagnose rheumatoid arthritis at an early stage. [Means for solving the problem]

[0006] The above problem is solved by the present invention. [1] A method for diagnosing rheumatoid arthritis, comprising measuring acrolein in a sample; [2] A method for early diagnosis of rheumatoid arthritis, which comprises measuring acrolein in a sample. [3] measuring acrolein in a sample collected from a subject; and a step of determining that the patient has rheumatoid arthritis when the acrolein level is higher than that of a healthy subject; A method for diagnosing rheumatoid arthritis, comprising: [4] measuring acrolein in a sample collected from a subject; and a step of determining that the patient has rheumatoid arthritis when the acrolein level is higher than that of a healthy subject; A method for early diagnosis of rheumatoid arthritis, [5] Any of the methods [1] to [4], further measuring a biomarker for rheumatoid arthritis; [6] The method according to [5], wherein the biomarker is C-reactive protein. [7] Acrolein, a biomarker for the diagnosis of rheumatoid arthritis, [8] Acrolein, a biomarker for early diagnosis of rheumatoid arthritis, [9] A kit for diagnosing rheumatoid arthritis, comprising an antibody capable of detecting acrolein.

[10] A kit for early diagnosis of rheumatoid arthritis, comprising an antibody capable of detecting acrolein.

[11] The kit according to [9] or

[10] , further comprising standard data showing the correlation between the acrolein level in the sample and rheumatoid arthritis. This can be solved by:

[0007] The present invention also provides: A method for assisting in the diagnosis of rheumatoid arthritis (for example, early diagnosis of rheumatoid arthritis), which comprises measuring acrolein in a sample (hereinafter, sometimes referred to as an (early) diagnosis assisting method); A method for measuring acrolein in a sample for the diagnosis of rheumatoid arthritis (for example, early diagnosis of rheumatoid arthritis) (hereinafter, sometimes referred to as a measurement method for (early) diagnosis), An in vitro method for diagnosing rheumatoid arthritis (for example, an in vitro method for early diagnosis of rheumatoid arthritis), which comprises measuring acrolein in a sample; Use of an antibody capable of detecting acrolein in the manufacture of a diagnostic kit for rheumatoid arthritis (e.g., a kit for early diagnosis of rheumatoid arthritis); A method for measuring acrolein in a sample to provide information necessary for the diagnosis of rheumatoid arthritis (for example, early diagnosis of rheumatoid arthritis) (hereinafter, sometimes referred to as a measurement method for providing information) Includes: [Effects of the Invention]

[0008] According to the present invention, a novel biomarker for diagnosing rheumatoid arthritis that exhibits a high AUC value and further exhibits high sensitivity and specificity can be provided. Furthermore, according to the present invention, rheumatoid arthritis can be diagnosed at an early stage. [Brief explanation of the drawings]

[0009] [Figure 1] 1 is a graph showing the results of acrolein measurement in a rheumatoid arthritis group and a control group. [Figure 2] 1 is a graph showing the results of ROC analysis for the diagnosis of rheumatoid arthritis and early diagnosis of rheumatoid arthritis using PC-Acro and / or C-reactive protein (CRP). DETAILED DESCRIPTION OF THE INVENTION

[0010] <<Method and Biomarker of the Present Invention>> In the methods of the present invention (including the (early) diagnostic method of the present invention, the (early) diagnostic auxiliary method of the present invention, the measurement method for (early) diagnosis of the present invention, the in vitro (early) diagnostic method of the present invention, the use of the present invention, and the measurement method for providing information of the present invention), acrolein is measured in a sample collected from a subject, particularly a patient suspected of having rheumatoid arthritis or a patient with rheumatoid arthritis. Furthermore, in the present invention, acrolein can be used as a biomarker for the (early) diagnosis of rheumatoid arthritis.

[0011] In this specification, "acrolein" refers to a highly reactive unsaturated aldehyde with the rational formula CH2=CHCHO. Acrolein is a metabolite of polyamines and is highly toxic. This toxicity is usually processed within cells and is not a problem, but when cells are subjected to stress, acrolein production increases, causing it to escape from the cells and causing cell damage.

[0012] In blood, acrolein is mostly bound to proteins and exists as protein-conjugated acrolein (PC-Acro). Methods for measuring acrolein are known, and it can be measured by, for example, ELISA (Enzyme-Linked Immunosorbent Assay).

[0013] The sample used in the method of the present invention is not particularly limited as long as it allows for the measurement of acrolein, and examples that can be used include blood samples (e.g., whole blood, plasma, serum), urine, lymph, cerebrospinal fluid, and synovial fluid.

[0014] In the method of the present invention, an increase in the acrolein value (e.g., amount or concentration of acrolein) in a sample is used as an indicator of rheumatoid arthritis. Specifically, a high acrolein value compared to healthy individuals or compared to normal values ​​in an appropriate population corresponding to the subject can be diagnosed as rheumatoid arthritis. A previously prepared "threshold" can also be used as the standard for diagnosis.

[0015] In the method of the present invention, various biomarkers for rheumatoid arthritis can be further measured in addition to acrolein, if desired. By combining multiple biomarkers, rheumatoid arthritis can be diagnosed more accurately. Examples of biomarkers for rheumatoid arthritis include C-reactive protein (CRP), erythrocyte sedimentation rate, rheumatoid factor, anti-CCP antibody, and MMP-3.

[0016] <<Kit of the present invention>> The kit of the present invention can be used to carry out the method of the present invention and comprises an antibody capable of detecting acrolein, and optionally further comprises standard data showing the correlation between the acrolein level in a sample and rheumatoid arthritis.

[0017] The antibody used in the kit of the present invention may be either a monoclonal antibody or a polyclonal antibody. Alternatively, an antibody fragment that retains specific binding ability to an antigen, such as Fab, Fab', F(ab')2, or Fv, may be used in the kit. Furthermore, the antibody can be used in the kit as it is, or in a form appropriate for the immunological technique to be used, for example, immobilized on a latex carrier if latex agglutination immunoassay is used; immobilized on magnetic particles if a highly sensitive measurement method using magnetic particles or the like is used; immobilized on a substrate if a method using a substrate such as immunochromatography is used; or labeled if labeling with a labeling substance (e.g., enzyme, fluorescent substance, chemiluminescent substance, radioisotope, biotin, avidin) is required.

[0018] The standard data included in the kit of the present invention is not particularly limited as long as it indicates a correlation between acrolein, which is the indicator of the present invention, and rheumatoid arthritis, and examples thereof include a determination threshold value, original data for calculating the determination threshold value, or statistically processed data. The standard data may be described in the instruction manual or attached as a separate data sheet. The attached document may be in the form of paper, electronic media such as a CD-ROM, or downloaded from a website, etc. [Example]

[0019] The present invention will be specifically described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0020] Example 1: Measurement of acrolein in patients with rheumatoid arthritis The subjects were 84 patients with a clinical diagnosis of rheumatoid arthritis, and the control group consisted of 298 healthy adults undergoing regular medical checkups. Acrolein was measured in plasma samples by ELISA using a commercially available kit (Acrolein-Lysine Adduct Competitive EIA Kit; Takara Bio).

[0021] The results are shown in Figure 1. Plasma protein-conjugated acrolein (PC-Acro) levels were 80.5 [63.2-105.2] nmol / mL in the rheumatoid arthritis group (median [interquartile range]) and 65.9 [58.9-78.1] nmol / mL in the control group, with the rheumatoid arthritis group showing significantly higher levels than the control group (p<0.0001).

[0022] Example 2: ROC analysis for the diagnosis of rheumatoid arthritis The rheumatoid arthritis groups were divided into a group of patients with normal rheumatoid arthritis, including those with various stages of onset, and a group of patients with early-stage, untreated rheumatoid arthritis (early naive RA group). Receiver operating characteristic (ROC) analysis was performed for the diagnosis of rheumatoid arthritis using PC-Acro and C-reactive protein (CRP), a known biomarker for rheumatoid arthritis. The early-onset, untreated rheumatoid arthritis group consisted of patients from the regular rheumatoid arthritis group, excluding patients with impaired renal function, and further comprising patients who had been in the rheumatoid arthritis for less than a year and who were not receiving treatment for rheumatoid arthritis.

[0023] The results are shown in Figure 2. In Figure 2, (a) to (c) show the results for the combination group of the normal rheumatoid arthritis group and the control group, and (d) to (f) show the results for the combination group of the early-stage, untreated rheumatoid arthritis group and the control group. In addition, (a) and (d) show the results for PC-Acro alone, (b) and (e) show the results for CRP alone, and (c) and (f) show the results for the combined use of PC-Acro and CRP.

[0024] The AUC (areas under the curve) value for PC-Acro alone was lower than that for CRP alone, but the combined use of PC-Acro and CRP showed a higher AUC value than either PC-Acro or CRP alone. In particular, in a group combining early-stage, untreated rheumatoid arthritis and the control group, the AUC value for the combined use of PC-Acro and CRP was 0.903, compared with 0.6453, 0.5190, and 0.6098 for the new biomarkers for rheumatoid arthritis, SAA4, gelsolin, and vitamin D-binding protein, respectively, alone, and 0.7400 for the combination of these three, and 0.894 for the combination of PC-Acro, CRP, and IL-6, which is already being used clinically to diagnose cerebral infarction. These results indicate that the AUC value for the combined use of PC-Acro and CRP is extremely high and can be considered extremely useful for early diagnosis.

[0025] Furthermore, using the Youden Index, a PC-Acro level of 75.4 nmol / mL or higher can detect early rheumatoid arthritis with a sensitivity of 65.2% and a specificity of 71.1%. Furthermore, when PC-Acro and CRP are used in combination, early rheumatoid arthritis can be detected with a sensitivity of 82.6% and a specificity of 88.3% when the cutoff value is 3.187, calculated as CRP x 3.8865 + PC-Acro x 0.0283. [Industrial Applicability]

[0026] The present invention can be used for the diagnosis of rheumatoid arthritis, particularly for early diagnosis.

Claims

1. A method for measuring acrolein and C-reactive protein in a blood sample and combining the acrolein and C-reactive protein measurements to aid in the diagnosis of rheumatoid arthritis.

2. A method for measuring acrolein and C-reactive protein in a blood sample and combining the acrolein and C-reactive protein measurements to aid in the early diagnosis of rheumatoid arthritis.

3. measuring acrolein and C-reactive protein in a blood sample taken from a subject; Acrolein level and C-reactive protein level each exceeding a predetermined threshold indicate that the subject may have rheumatoid arthritis. A method for aiding in the diagnosis of rheumatoid arthritis.

4. measuring acrolein and C-reactive protein in a blood sample taken from a subject; Acrolein level and C-reactive protein level each exceeding a predetermined threshold indicate that the subject may have rheumatoid arthritis. A method for aiding in the early diagnosis of rheumatoid arthritis.

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