METHOD FOR DETERMINING MORTALITY RISK OF INFECTIOUS INFLAMMATORY DISEASE ON BASIS OF CONCENTRATION OF WARS AND CYTOKINE

Abstract

The present disclosure relates to a method for determining the mortality risk of an infectious inflammatory disease on the basis of the concentrations of tryptophanyl-tRNA synthetase (WARS) and cytokines. The method for determining the mortality risk of the present disclosure can effectively identify patients who are at high risk of mortality due to an infectious inflammatory disease within a short period of time and, as such, can be very helpful for performing timely therapy.

Claims

1-21. (canceled)

22. A kit for determining the mortality risk of an infectious inflammatory disease, comprising: an agent for measuring the expression level of the WARS protein or a gene encoding the same; and an agent for measuring the expression level of the IL-8 protein or a gene encoding the same.

23. The kit for determining the mortality risk according to claim 22, wherein the kit further comprises an agent for measuring the expression level of the IL-6 protein or a gene encoding the same.

24. The kit for determining the mortality risk according to claim 22, wherein the kit determines the mortality risk of an infectious inflammatory disease by identifying whether the concentration of WARS is 100 ng/mL or higher and the concentration of IL-8 is 100 pg/mL or higher in a sample of a subject.

25. The kit for determining the mortality risk according to claim 22, wherein the kit further comprises an instruction about the determination of the mortality risk depending on the correlation between the concentration of WARS and the concentration of IL-8 in a sample.

26. The kit for determining the mortality risk according to claim 25, wherein the instruction describes the identification of whether the concentration of WARS is 100 ng/mL or higher and lower than 200 ng/mL and the concentration of IL-8 is 400 pg/mL or higher or the identification of whether the concentration of WARS is 100 ng/mL or higher and lower than 200 ng/mL and the concentration of IL-8 is 466.5 pg/mL or higher.

27. A method for providing information necessary for determining the mortality risk of an infectious inflammatory disease, comprising: (a) a step of providing a sample of a subject; (b) a step of measuring the concentrations of WARS (tryptophanyl-tRNA synthetase) and IL-8 or IL-6 in the sample; and (c) a step of identifying whether 1) the concentration of WARS is 100 ng/mL or higher and the concentration of IL-8 is 100 pg/mL or higher or 2) the concentration of WARS is lower than 200 ng/mL and the concentration of IL-6 is 1,000 pg/mL or higher.

28. The method for providing information according to claim 27, wherein the infection is infection by one or more selected from a group consisting of viruses, bacteria and fungi.

29. The method for providing information according to claim 27, wherein the infectious inflammatory disease is one or more disease selected from a group consisting of pneumonia, phthisis, tuberculosis, sepsis and septic shock.

30. The method for providing information according to claim 27, wherein the sample is blood, plasma or serum.

31. The method for providing information according to claim 27, wherein, in the step (c), it is identified whether the concentration of WARS is 100 ng/mL or higher and lower than 200 ng/mL and the concentration of IL-8 is 400 pg/mL or higher.

32. The method for providing information according to claim 27, wherein, in the step (c), it is identified whether the concentration of WARS is 200 ng/mL or higher and the concentration of IL-8 is 100 pg/mL or higher.

33. The method for providing information according to claim 27, wherein, in the step (c), it is identified whether the concentration of WARS is lower than 200 ng/mL and the concentration of IL-6 is 1,600 pg/mL or higher.

34. The method for providing information according to claim 27, which further comprises a step (step (d)) of identifying the concentration of one or more cytokine selected from a group consisting of IFN-γ, IL-6, MIP-1α, IL-8 and TNF-α when the concentration of WARS is lower than 200 ng/mL and the concentration of IL-6 is lower than 1,654.5 pg/mL.

35. The method for providing information according to claim 34, wherein the subject is determined as a patient group with low mortality risk if the concentration of IFN-γ is 2.99 pg/mL or higher and the concentration of IL-6 is lower than 54.96 pg/mL.

36. The method for providing information according to claim 34, wherein the subject is determined as a patient group with low mortality risk if the concentration of IFN-γ is lower than 2.99 pg/mL and the concentration of IL-8 is lower than 22.91 pg/mL.

37. The method for providing information according to claim 34, wherein the subject is determined as a patient group with low mortality risk if the concentration of IFN-γ is lower than 2.99 pg/mL, the concentration of IL-8 is 22.91 pg/mL or higher and the concentration of TNF-α is 37.21 pg/mL or higher.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0073] FIG. 1 shows the cohort design for the present disclosure.

[0074] FIGS. 2A-2K show a result of measuring the concentration of WARS1 and cytokines in the samples of survivors and non-survivors. FIG. 2A shows a result of comparing the concentration of WARS1 in survivors and non-survivors, FIG. 2B shows a result of comparing the concentration between a sepsis group and a septic shock group, FIG. 2C shows an AUC analysis result for the survivors and non-survivors, FIG. 2D shows a decision tree obtained using the R2 software from the concentration of WARS1, proinflammatory cytokines and chemokines, FIG. 2E shows the distribution of the concentration of IL-8 and IL-6 based on WARS1. FIG. 2F shows an AUC analysis result for IL-8 and WARS1, FIG. 2G compares days of survival depending on the concentration of WARS1 and IL-8, and FIG. 2H-2K show the correlation and significance of proinflammatory cytokines and chemokines in non-survivors (WARS1 200 ng/mL, IL-8 115.65 pg/mL or higher).

[0075] FIGS. 3A-3C show a result of analyzing the correlation of WARS1 and IL-8 in survivors and non-survivors. FIG. 3A shows a decision tree obtained from analysis of WARS1 and IL-8 only, FIG. 3B shows the distribution of the concentration of IL-8 based on WARS1, and FIG. 3C shows an AUC analysis result for IL-8 and WARS1.

BEST MODE

[0076] Hereinafter, the present disclosure will be described more specifically through examples. The following examples are provided only to describe the present disclosure more specifically and it will be obvious to those having ordinary knowledge in the art that the scope of the present disclosure is not limited by the examples.

EXAMPLES

Example 1. Cohort Design

[0077] In order to investigate whether the death caused by sepsis is correlated with the high concentration of WARS1, target patients were selected based on the concentration of WARS1. Plasma samples were acquired from Asan Hospital. The samples had been taken from the patients within 24 hours after hospitalization. The cohort consisted of a total of 243 patients (FIG. 1), with 54 patients for an H.C group (normal group), 100 patients for a sepsis group and 89 patients for a septic shock group.

Example 2. Comparison of Concentration of WARS1, Cytokines and Chemokines in Survivors and Non-Survivors

[0078] The difference in the level of WARS1 and proinflammatory cytokines and chemokines such as IL-1β, IL-6, IL-8, IFN-γ, MIP-1α and TNF-α in the plasma of survivors and non-survivors was measured. The level of IL-1β, IL-6, IL-8, IFN-γ, MIP-1a and TNF-α was measured by multiplex assay (Millipore), and the level of WARS1 was measured by ELISA using an anti-WARS1 antibody (Abiotech). The measurement was performed according to the manufactures' instructions.

[0079] WARS1 was significantly increased in the non-survivors as compared to the survivors (FIG. 2a), and was significantly increased in the septic shock group as compared to the sepsis group (FIG. 2b). As a result of AUC analysis for WARS1, procalcitonin (PCT) and C-reactive protein (CRP) as biomarkers, there was no significant difference in the level of PCT (p=0.7112) and CRP (p=0.2380) between the non-survivors and the survivors (FIG. 2c). Only WARS1 showed the significant ability of predicting mortality (p=0.006).

[0080] A decision tree (DT) was obtained by analyzing and classifying the concentration of cytokines and chemokines in the plasma of cohort patients depending on the concentration of WARS1 through “rpart” package analysis of the R software (version 3.5.1). The WARS1 concentration of 200 ng/mL was set as a first classification tree root. In the first group, 17 out of 18 septic patients (94.4%) whose IL-8 level was 115.65 pg/mL or higher died. IL-6 was selected as a second root for the patients whose WARS1 level of 200 ng/mL or smaller. 19 out of 24 patients (79.2%) whose IL-6 level was 1654.5 pg/mL or higher died (FIG. 2d). The DT showed a sensitivity of 77.5% and a specificity of 86.2% (FIG. 2e). Non-survivors could be distinguished significantly from the septic patients (p=0.0003) using the AUC with the WARS1 level of 200 ng/mL or higher and the IL-8 level of 115.65 pg/mL or higher (FIG. 2f).

[0081] Average days of survival was about 4.3 days for the first group, which was remarkably shorter than that of the non-survivors whose WARS1 level was lower than 200 ng/mL and the IL-8 level was lower than 115.65 pg/mL (FIG. 2g). The first group showed positive correlations between WARS1 and MIP-1α (p=0.006), IFN-γ (p=0.011), TNF-α (p=0.003) and IL-8 (p=0.01) (FIGS. 2h-2k). As a result of comparing the level of MIP-1α, INF-γ, TNF-α, IL-6, IL-1β, IL-8 and IL-10 for the normal group, the sepsis group and the septic shock group, it was confirmed that MIP-1α (p<0.0001), TNF-α (p<0.0001), IL-6 (p<0.0001), IL-8 (p<0.0001) and IL-10 (p<0.0001) were increased significantly in the septic shock group. In addition, IFN-γ, TNF-α, IL-6, IL-1β, IL-6 and IL-10, except MIP-1α, were increased relatively in the non-survivors as compared to the survivors, and IL-6 (p<0.0001), IL-8 (p<0.0001) and IL-10 (p<0.0001) were increased significantly.

Example 3. Analysis of Correlation of WARS1 and IL-8 in Survivors and Non-Survivors

[0082] A decision tree (DT) was obtained for the concentration of WARS1 and IL-8 only through “rpart” package analysis of the R software (version 3.5.1). The WARS1 concentration of 200 ng/mL was set as a classification tree root (100 ng/mL≤WARS1<200 ng/mL). In the first group, 17 out of 18 septic patients whose WARS1 level was 200 ng/mL or higher and IL-8 level was 115.65 pg/mL or higher died. For the septic patients with 100 ng/mL≤WARS1<200 ng/mL and IL-8 level of 466.5 ng/mL or higher, all the 6 patients died (FIG. 3a). 24 of the 80 dead patients could be targeted with the DT, and the septic patients could be distinguished specifically as non-survivors with an AUC of 0.80 for WARS1 and IL-8 (p=0.007) (FIGS. 3b and 3c).

[0083] Although the examples of the present disclosure have been described above, those having ordinary knowledge in the art can change and modify the present disclosure variously through addition, change or deletion without departing from the scope of the present disclosure defined by the appended claims.