阅读说明：本技术 尿毒症标志物及其应用 (Uremia marker and application thereof ) 是由 戴勇 伍宏伟 汤冬娥 张欣洲 王康 郑凤屏 于 2020-05-11 设计创作，主要内容包括：本发明提供一种尿毒症标志物及其应用。该尿毒症标志物,包括以下至少一种：ESRRG、TEAD2、HOXC10、TEAD1、FOS、GRHL1、PAX7、PAX3、PBX1、MAFG、GCM2、NR3C1、NR3C2、JUN、PBX2、SCRT2、MYBL2、SPDEF、STAT1、NEUROG2、JDP2、FOSB。发明人借助scATAC-seq技术发现尿毒症患者与正常健康对照组人群的样本中,多个转录因子的表达水平存在显著差异,因此,可以将这些差异表达的转录因子作为标志物用于尿毒症的诊断、预后评估或筛药。(The invention provides a uremia marker and application thereof. The uremia marker comprises at least one of the following substances: ESRRG, TEAD2, HOXC10, TEAD1, FOS, GRHL1, PAX7, PAX3, PBX1, MAFG, GCM2, NR3C1, NR3C2, JUN, PBX2, SCRT2, MYBL2, SPDEF, STAT1, NEUROG2, JDP2, FOSB. The inventor finds that the expression levels of a plurality of transcription factors in samples of uremia patients and normal healthy control group populations have significant difference by means of the scATAC-seq technology, so that the transcription factors with different expression levels can be used as markers for diagnosis, prognosis evaluation or drug screening of uremia.)

1. A uremic marker comprising at least one of: ESRRG, TEAD2, HOXC10, TEAD1, FOS, GRHL1, PAX7, PAX3, PBX1, MAFG, GCM2, NR3C1, NR3C2, JUN, PBX2, SCRT2, MYBL2, SPDEF, STAT1, NEUROG2, JDP2, FOSB.

2. Use of the uremic marker according to claim 1 for the preparation of a uremic diagnostic and/or prognostic reagent.

3. Use of the uremic marker of claim 1 for screening a medicament for the prevention and/or treatment of uremia.

4. A kit comprising reagents for quantitatively detecting the uremic marker of claim 1.

5. Use of a reagent for quantitatively detecting the uremic marker of claim 1 in the preparation of a diagnostic kit for uremia.

6. The use according to claim 5, wherein the reagent quantitatively detects the uremic marker according to claim 1 at the gene level and/or protein level.

7. A method of assessing the risk of uremia comprising the steps of:

(1) obtaining a relative abundance of the uremic marker of claim 1 in a sample from a subject;

(2) comparing the relative abundance with a preset threshold value, and judging the uremia risk according to a comparison result;

the method is not suitable for the diagnosis or treatment of diseases.

8. A computer-readable storage medium having stored thereon computer-executable instructions for performing the method of claim 7.

9. A system for assessing the risk of uremia, comprising:

a detection device for determining the relative abundance of the uremic marker of claim 1 in a sample from a subject;

and the comparison device is used for comparing the relative abundance with a preset threshold value and judging the uremia risk according to a comparison result.

Technical Field

The invention relates to the technical field of nephropathy, and particularly relates to a uremia marker and application thereof.

Background

Uremia (uremia) refers to a clinical syndrome consisting of a series of clinical manifestations and metabolic disorders that occur as a result of progressive decline in kidney function from various causes, through to loss of kidney function and eventually to the end stage. The disturbed internal environment is liable to cause the change of epigenetic characteristics of Chronic Kidney Disease (CKD) patients and influence gene expression through epigenetic mechanism, thereby accelerating the development of CKD to uremia (end stage kidney disease). This is also evidenced by the increasing number of studies at this stage. Studies have shown that CKD patients have varying degrees of DNA methylation, which is also affected by the uremic component. The inflammatory response in CKD5 stage patients leads to abnormal DNA hypermethylation and is associated with poor prognosis. When epigenetic detection is carried out on a renal tumor patient undergoing dialysis and a renal tumor patient with normal renal function, the fact that various genes of the former show hypermethylation of DNA proves that CKD and dialysis may promote hypermethylation of DNA. The above experimental results show that the uremic environment changes the epigenetic characteristics of the body, which in turn leads to the change of the transcription level. Unlike gene mutations, epigenetic changes are dynamic and possibly reversible, and thus, it is of great interest to analyze the development of disease from an epigenetic perspective.

ATAC-seq (Assay for Transposase-Access chromosome with high-throughput put sequencing) is an emerging scientific research technology for studying Chromatin openness (or Chromatin accessibility) from an epigenetic point of view based on high-throughput sequencing, and the principle is to utilize the characteristic that Transposase Tn5 is easy to specifically bind with open Chromatin, sequence DNA sequence fragments captured by Tn5 enzyme, compare the sequencing result with human genome information, analyze open regions of DNA, and predict Transcription Factor Binding Sites (TFBS) in the whole genome range through further motif analysis. Compared with the traditional DNase-Seq, FAIRE-Seq and other methods for researching open chromatin, the ATAC-Seq has the advantages of less cell quantity requirement, low cost, short period, large data, no need of antibody enrichment, capability of detecting the open state of chromatin in the whole genome range and the like. At present, the single-cell ATAC-Seq is widely applied to the fields of chromosome open map analysis, apparent modification difference research, embryonic development epigenetic modification, tumorigenesis epigenetic mechanism research, tumor heterogeneity and typing research, disease potential marker prediction and the like.

The mechanism of occurrence and development of uremia and its related complications is still unclear, and its specific biomarkers are not discovered at present. Therefore, there is a need to discover uremia-specific biomarkers by means of ATAC-seq technology.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a uremia marker and application thereof.

In a first aspect, an embodiment of the present invention provides a marker for uremia comprising at least one of: ESRRG, TEAD2, HOXC10, TEAD1, FOS, GRHL1, PAX7, PAX3, PBX1, MAFG, GCM2, NR3C1, NR3C2, JUN, PBX2, SCRT2, MYBL2, SPDEF, STAT1, NEUROG2, JDP2, FOSB.

The intestinal flora marker provided by the embodiment of the invention at least has the following beneficial effects:

the inventor finds that the expression levels of a plurality of transcription factors in samples of uremia patients and normal healthy control group populations have significant difference by means of the scATAC-seq technology, so that the transcription factors with different expression levels can be used as markers for diagnosis, prognosis evaluation or drug screening of uremia.

In a second aspect, an embodiment of the present invention provides a use of the above-mentioned uremia marker in the preparation of a uremia diagnostic and/or prognostic reagent. The uremia marker is differentially expressed in uremia patients and normal healthy control groups, so that the diagnosis of uremia or the evaluation of the prognosis of uremia can be realized by quantitatively detecting the uremia marker in a sample of a subject.

In a third aspect, an embodiment of the present invention provides a use of the above-mentioned uremia marker in screening a drug for preventing and/or treating uremia. The uremia marker has differential expression in uremia patients and normal healthy control groups, so that the evaluation of the drug effect of candidate drugs can be realized by quantitatively detecting the uremia marker in a sample after drug administration, thereby achieving the purpose of screening drugs.

In a fourth aspect, an embodiment of the present invention provides a kit comprising reagents for quantitatively detecting the above-mentioned uremia marker. The reagent realizes the quantitative detection of the uremia marker in a sample of a subject, and after the reagent is compared with a preset threshold value, the obtained result can be effectively used for diagnosing uremia, or evaluating the prognosis of uremia, or screening uremia drugs.

In a fifth aspect, an embodiment of the present invention provides a use of a reagent for quantitatively detecting the above-mentioned uremia marker in preparing a diagnostic kit for uremia. The uremia diagnosis product obtained by the reagent can realize quantitative detection on the uremia marker in a sample of a subject, and the detection result can be accurately used for diagnosing uremia, evaluating the prognosis of uremia or screening uremia drugs after being compared with a preset threshold value.

According to some embodiments of the invention, the reagent quantitatively detects the uremic markers at the gene level and/or the protein level. For example, the content of nucleic acid fragments of the uremic marker in the sample is quantitatively determined by PCR, or the expression level of protein of the uremic marker in the sample is quantitatively determined by enzyme-linked immunoassay, radioimmunoassay, Western blotting, protein chip method, or the like.

In a sixth aspect, an embodiment of the present invention provides a method of assessing the risk of uremia, the method comprising the steps of:

(1) obtaining the relative abundance of the uremic markers in the subject sample;

(2) comparing the relative abundance with a preset threshold value, and judging the uremia risk according to the comparison result;

this method is not suitable for the diagnosis and/or treatment of diseases.

Wherein uremia risk refers to the risk of the subject suffering from uremia, or to the prognosis of the subject; or to the effect of a candidate drug on uremia. The preset threshold is a critical value of the uremia marker of the normal subject, and specifically can be a relative abundance value of the uremia marker measured from a normal healthy control group sample or a normalized relative abundance value.

According to the method of some embodiments of the present invention, when directly determining the risk of uremia according to the relative abundance result of the markers, at least one of the following conditions is satisfied, and it can be determined that the subject has uremia, or the prognosis of the uremia patient is poor, or the candidate drug does not meet the screening condition:

(1) the relative abundance of the ESRRG is higher than a preset threshold value;

(2) the relative abundance of TEAD2 is higher than a preset threshold;

(3) the relative abundance of HOXC10 is higher than a preset threshold;

(4) the relative abundance of TEAD1 is higher than a preset threshold;

(5) the relative abundance of FOS is higher than a preset threshold value;

(6) the relative abundance of GRHL1 is lower than a preset threshold value;

(7) the relative abundance of PAX7 is higher than a preset threshold;

(8) the relative abundance of PAX3 is higher than a preset threshold;

(9) the relative abundance of the PBX1 is higher than a preset threshold;

(10) the relative abundance of MAFG is higher than a preset threshold;

(11) the relative abundance of GCM2 is lower than a preset threshold;

(12) the relative abundance of NR3C1 is higher than a preset threshold;

(13) the relative abundance of NR3C2 is higher than a preset threshold;

(14) the relative abundance of JUN is lower than a preset threshold value;

(15) the relative abundance of PBX2 is lower than a preset threshold;

(16) the relative abundance of the SCRT2 is lower than a preset threshold;

(17) the relative abundance of MYBL2 is lower than a preset threshold;

(18) the relative abundance of the SPDEF is lower than a preset threshold;

(19) the relative abundance of STAT1 is higher than a preset threshold;

(20) relative abundance of NEUROG2 is higher than a preset threshold;

(21) the relative abundance of JDP2 is below a preset threshold;

(22) the relative abundance of FOSBs is below a predetermined threshold.

In a seventh aspect, an embodiment of the present invention provides a computer-readable storage medium storing computer-executable instructions for performing the above-mentioned method. By performing the above method, it is possible to accurately and efficiently determine whether a subject has uremia, or the prognosis of a uremia patient, or the uremia efficacy on a candidate drug.

In an eighth aspect, an embodiment of the present invention provides a system for assessing risk of uremia, the system comprising:

a detection device for determining the relative abundance of a uremic marker in a sample from a subject;

and the comparison device is used for comparing the relative abundance with a preset threshold value and judging the uremia risk according to a comparison result.

Drawings

FIG. 1 shows the results of differential transcription factor motif identification of uremic PBMC subpopulations according to example 1 of the present invention.

FIG. 2 shows the results of the transcription factor function enrichment and inflammatory response analysis of the uremic patient of example 1 of the present invention.

Fig. 3 is a schematic composition diagram of a system for assessing risk of uremia according to example 2 of the present invention.

Detailed Description

The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.