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Atherosclerosis related cell marker molecule and application thereof

A technology for atherosclerosis and smooth muscle cells, applied in the field of biomedicine, can solve the problem of directly responding to the dynamic changes in cell group biology, ignoring tissue structure and intercellular heterogeneity, and directly analyzing the dynamic changes of each cell group Process and other issues

Active Publication Date: 2021-07-23
首都医科大学脑重大疾病研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Multi-omics research can comprehensively reveal the changes of gene expression profiles in the course of AS lesions. It is a mixed average result of various cell groups, and it is difficult to directly reflect the dynamic change process of each cell group.
Traditional sequencing methods ignore tissue structure and intercellular heterogeneity (Bondareva O, et al. (2019) Identification of atheroprone shear stress responsive regulatory elements inendothelial cells. Cardiovascular research 115(10):1487-1499), the obtained omics The data are often mixed average results of various cell groups, and it is difficult to directly analyze the dynamic changes of each cell group in the process of disease occurrence and development

Method used

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  • Atherosclerosis related cell marker molecule and application thereof
  • Atherosclerosis related cell marker molecule and application thereof
  • Atherosclerosis related cell marker molecule and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Example 1 Single-cell sequencing of AS model animals

[0119] 1. Sample collection

[0120] Collection of large animal models of atherosclerosis ApoE - / - Arterial tissue in Beagle and WT dogs, including carotid, subclavian, aortic arch, coronary arteries, thoracic aorta, abdominal aorta, and iliac arteries. The isolated arteries were snap-frozen in liquid nitrogen and stored at -80°C for RNA extraction and Bulk RNA-seq. In addition, take part of the fresh subclavian artery and iliac artery, wash it repeatedly with pre-cooled saline, place it in a cell culture dish on ice, add RPMI 1640 cell culture medium containing 10% FBS, and use it for single cell Dissociation and scRNA-seq.

[0121] 2. RNA extraction and Bulk RNA-seq

[0122] The total tissue RNA was extracted by Trizol method, the purity of RNA was analyzed by NanoDropSpectrophotometer, and the concentration and integrity of RNA were analyzed by Agilent 2100 Bioanalyzer. After the RNA quality inspection was q...

Embodiment 2

[0142] Example 2 Single-cell sequencing of AS patients

[0143] To determine TNC high Whether the SMC3 subpopulation exists in human AS, the scRNA-seq data of human carotid artery plaques (n=3 patients) were further analyzed, and the method was the same as that in Example 1.

[0144] Human carotid plaque cells were divided into 25 clusters by unbiased clustering, and the main cell types were ECs, SMCs, MFBs, Mφ cells, T cells, B cells, and mast cells ( Figure 6 ). It was pointed out that in carotid atherosclerotic tissue specimens obtained by human carotid endarterectomy, most of the SMCs in the arterial media and the FBs in the adventitia may be lost. In human carotid plaques, the gene expression patterns of SMC-associated and MFB-associated genes (ACTA2, MYH11, TAGLN, CNN1, MYL9, TPM2, ELN, DCN, COL3A1, COL1A1, FMOD) were very similar to those in ApoE- / - dogs.

[0145] Trajectory analysis revealed intercluster relationships between SMC3 and MFB cells ( Figure 7 ). TN...

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Abstract

The invention discloses an atherosclerosis related cell marker molecule and application thereof, and the marker molecule which can be used for identifying, characterizing and optionally enriching or separating smooth muscle cell subset is found by utilizing a method of combining single cell sequencing with bioinformatics analysis. The invention also discloses that the smooth muscle cell subset has significant difference in atherosclerosis.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to atherosclerosis-related cell marker molecules and applications thereof. Background technique [0002] Atherosclerosis (AS) is a chronic inflammatory disease characterized by the formation of fibrous fatty lesions under the arterial intima. Worldwide, AS is the most important cause of cardiovascular and peripheral vascular diseases (such as myocardial infarction, ischemic stroke, lower extremity gangrene, etc.). The susceptibility of AS in different parts of the human arterial system is significantly different, among which the abdominal aorta, iliac artery, carotid artery and coronary artery are the four most susceptible parts of AS, while the subclavian artery, aortic arch and thoracic aorta are not prone to AS. Due to the lack of an ideal large animal model that can simulate human AS, the current understanding of the characteristics and pathogenesis of AS susceptibility sites is very l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6883G01N33/68C12N5/077
CPCC12Q1/6888C12Q1/6883G01N33/56966G01N33/6893C12N5/0661C12Q2600/158C12Q2600/136G01N2800/323C12N2509/00
Inventor 石小峰吉训明刘桂友张永彪朱尚明
Owner 首都医科大学脑重大疾病研究中心
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