Application of circulating nucleic acid as marker for hypertension and coronary heart diseases

A technology for hypertension and coronary heart disease, applied in the application field of circulating nucleic acid as a marker of hypertension and coronary heart disease, can solve the problems of lack of large-scale population samples, inaccurate miRNA prediction research results, and restrictions on the discovery of new miRNA markers

Active Publication Date: 2020-06-16
国家心血管病中心 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, although there have been several studies on the use of circulating miRNAs as markers of coronary heart disease, there are still common problems that hinder the advancement of miRNA markers to clinical applications
On the one hand, most of the current miRNA studies use the miRNAs reported in the literature as candidate molecules for direct population experiment verification. This strategy limits the discovery of new miRNA markers, and these studies that lack systematic screening lack the effectiveness of miRNA markers. Compare
On the other hand, the current sample size in marker research generally stays at dozens of cases, and some even only target a small sample of a dozen cases, lacking large-scale population samples for long-term follow-up, which is also consistent with the results found by different platforms This is an important reason for the poor performance, which makes the prediction results of miRNAs inaccurate and the clinical prospects are limited.
In addition, most current studies on circulating miRNAs as markers of coronary heart disease focus on circulating miRNAs as diagnostic markers of coronary heart disease, while studies on circulating miRNAs as predictive markers of coronary heart disease are very scarce

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  • Application of circulating nucleic acid as marker for hypertension and coronary heart diseases
  • Application of circulating nucleic acid as marker for hypertension and coronary heart diseases
  • Application of circulating nucleic acid as marker for hypertension and coronary heart diseases

Examples

Experimental program
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Effect test

Embodiment 1

[0099] (Example 1) 48 candidate miRNAs were initially screened out based on miRNA sequencing data

[0100] Serum and PBMC of CHD patients and healthy subjects were sequenced for miRNA at the Sequencing Center of Peking University. Among them, 1022 and 976 miRNAs were detected in serum samples of coronary heart disease patients and healthy people, respectively; meanwhile, 1027 and 918 miRNAs were detected in PBMC samples of coronary heart disease patients and healthy people. The number of reads for each group of miRNAs was normalized to the t-PM value (t-PM=single miRNA Reads / total reads of the sample*10^6). A total of 483 miRNAs with a t-PM value greater than 50 were clustered. Then, the changes of 483 kinds of miRNA contents in serum and blood cells were counted, and the results are shown in Table 4, among which 132 kinds of miRNA contents changed simultaneously in serum and PBMC. It is speculated that there is a certain correlation between the levels of these miRNAs in ser...

Embodiment 2

[0103] (Example 2) Screen out 5 candidate miRNAs based on miRNA detection stability

[0104] For the 48 miRNA candidate molecules screened out initially, their stability and repeatability in the fluorescent quantitative PCR method were tested. Through multiple q-RT-PCR and sequencing experiments, it was found that only 10 of the miRNAs could be detected in serum and their full-length sequences were detected in sequencing verification. Among these 10 miRNAs, 5 miRNAs have good detection stability in serum, and further repeated detection experiments were carried out on these 5 miRNAs. Five healthy human sera were selected, and each sample was subjected to three replicate experiments starting from the extraction of miRNA. figure 1 It is the repeated experiment result of 5 candidate miRNAs and internal reference U6. From figure 1 It can be seen that the CT value of miRNA is between 15-35, and the standard deviation of repeated experiments for each sample is less than 2.

[010...

Embodiment 3

[0106] (Example 3) miRNA candidate molecules to the diagnostic efficiency of coronary heart disease crowd

[0107] In order to test whether the 5 circulating miRNA molecules screened based on the sequencing results can effectively distinguish patients with coronary heart disease from healthy people, 39 cases of patients with coronary heart disease and 39 cases of healthy people with sex and age matching from the northern region were selected as the discovery set for detection ( The age of patients with coronary heart disease was 63.1±7.41 years old; the age of healthy people was 59±7.29 years old).

[0108] The q-RT-PCR method was used to detect the content of 5 miRNA candidate molecules in serum samples, and the two-tailed T-test was carried out on the data after two groups of normalization, and the analysis results were shown in Table 5 and figure 2 As can be seen from Table 5, 4 of the 5 miRNAs are significantly different between patients and healthy people, namely miR-10a-5...

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Abstract

According to the invention, small RNA sequencing is carried out on serum and blood cells of patients with coronary heart diseases respectively, and based on parameters such as difference change of miRNA and serum abundance in sequencing data and detection of methodological stability, applicable specific candidate miRNA markers are systematically screened out. Furthermore, through cooperation witha prevention and treatment professional team for cardiovascular populations, researches are conducted on application of the candidate miRNA markers to diagnosis and prediction of atherosclerosis-related diseases during large-scale follow-up visit of the population. Research results in the invention provide important theoretical and methodological basis for clinical application of circulating miRNA.

Description

technical field [0001] The invention relates to the application of circulating nucleic acid as a marker for hypertension and coronary heart disease. Specifically, the present invention relates to the application of circulating microRNA (miRNA) in hypertension prediction and coronary heart disease diagnosis and prediction; more specifically, the present invention relates to the application of miR-423-3p in hypertension prediction and coronary heart disease diagnosis and prediction The application of miR-423-3p includes the preparation of a test kit for hypertension prediction and coronary heart disease diagnosis and prediction, a microarray for hypertension prediction and coronary heart disease diagnosis and prediction based on miR-423-3p, and the use of the kit and the Microarray methods for hypertension prediction and coronary heart disease diagnosis and prediction. Background technique [0002] Coronary atherosclerotic heart disease (also known as coronary heart disease) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178C12Q2600/166
Inventor 王增武曹慧青董莹王馨方甜王晓霞
Owner 国家心血管病中心
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