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A kind of adenovirus vector and application thereof for specifically inhibiting proliferation and migration of smooth muscle cells

A smooth muscle cell and adenovirus technology, applied in the direction of extracellular fluid disease, virus/bacteriophage, virus, etc., can solve the problems of weakening anti-restenosis performance, increasing the risk of thrombosis, and acute injury of blood vessels, so as to reduce the risk of thrombosis, Reduced use and bleeding, less bleeding effect

Active Publication Date: 2019-11-01
姚陈 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are two problems in the current endoluminal therapy based on balloon and stent: ① Postoperative restenosis (restenosis), which is caused by acute injury to blood vessels after balloon expansion or stent implantation, stimulates the proliferation of smooth muscle cells in the vascular intimal and migrate to the intima, leading to pathological thickening of the intima and ultimately postoperative restenosis according to
Its main problem is: inhibiting the repair of endothelium, increasing the risk of thrombosis and weakening its anti-restenotic properties

Method used

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  • A kind of adenovirus vector and application thereof for specifically inhibiting proliferation and migration of smooth muscle cells
  • A kind of adenovirus vector and application thereof for specifically inhibiting proliferation and migration of smooth muscle cells
  • A kind of adenovirus vector and application thereof for specifically inhibiting proliferation and migration of smooth muscle cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Preparation method of adenoviral vector that specifically inhibits smooth muscle cell proliferation and migration

[0076] (1) Construction of the shuttle plasmid pShuttle-TPM1-miR-126-3pTS

[0077] 1) Amplification of TPM1 sequence

[0078] Primer TPM1-F: 5'- agatct GCCACCATGGACGCCATCAAGAAG-3' (SEQ ID NO: 13) and TPM1-R: 5'- gtcgac TTACTATGGAAGTCATATCGTTGAGAG-3' (SEQ ID NO: 14) cloned the TPM1 sequence from the human cDNA library, and the underlined parts in the primers were Bgl , Sal Restriction sites.

[0079] 2) Construction of TPM1-miR-126-3pTS fragment

[0080] The TPM1-miR-126-3pTS fragment is the regulatory sequence that is connected to the miRNA at the 3' end of the above TPM1 sequence, and the regulatory sequence of the miRNA is the reverse complementary sequence of 4 concatenated miR-126-3p (miR-126 The reverse complement of -3p is shown in SEQ ID NO: 7).

[0081] The specific operation method of the TPM1-miR-126-3pTS fragment is as f...

Embodiment 2

[0094] Example 2 Preparation method of adenoviral vector that specifically inhibits smooth muscle cell proliferation and migration

[0095] The method for preparing the adenoviral vector in this example is the same as in Example 1, except that the target gene used in it is Geminin (replication inhibitor, GMNN), its base sequence is shown in SEQ ID NO: 2, and the miRNA regulatory sequence used is 4 tandem miR-1298 reverse complementary sequences (miR-1298 reverse complementary sequence is shown in SEQ ID NO: 8), other operations are the same as in Example 1, and the obtained adenovirus vector is called ad-GMNN-miR-1298TS for short.

Embodiment 3

[0096] Example 3 Preparation method of adenoviral vector that specifically inhibits smooth muscle cell proliferation and migration

[0097] The method for preparing the adenoviral vector in this example is the same as in Example 1, except that the miRNA regulatory sequence used is 4 concatenated miR-140-3p reverse complementary sequences (miR-140-3p reverse complementary sequences such as SEQ ID NO: 9), other operations are the same as in Example 1, and the obtained adenovirus vector is called ad-TPM1-miR-140-3pTS for short.

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Abstract

The invention discloses an adenovirus vector for specifically inhibiting proliferation and migration of smooth muscle cells and application of the adenovirus vector. Target genes and miRNA regulatory sequences are inserted at multiple clone sites of the adenovirus vector. After atherosclerosis disease interventional therapy, a main principle of restenosis lies in that the smooth muscle cells migrate to an intima and perform pathological proliferation. The adenovirus vector disclosed by the invention is applied, so that the movement of the proliferation, the migration and the like of the smooth muscle cells can be well restrained, and restenosis can be avoided. The adenovirus vector is adjusted and controlled based on miRNA, so that the purpose of specifically inhibiting proliferation and the migration of vascular smooth muscle cells can be realized, and influence on vascular endothelial cells is little. Therefore, the repair of endothelium is not influenced, so that the risk of thrombosis is reduced, and the use of antiplatelets and bleeding can be reduced; the effect of interventional treatment of the atherosclerosis diseases can be improved.

Description

technical field [0001] The invention relates to a recombinant adenovirus vector for specifically inhibiting the proliferation and migration of smooth muscle cells based on miRNA regulation and its application. Background technique [0002] Lower extremity arteriosclerosis obliterans (LEASO) is a chronic lower extremity ischemic disease caused by progressive stenosis or even occlusion of the arterial lumen due to lower extremity atherosclerosis. The incidence of LEASO is high. A large number of epidemiological studies have shown that about 10-20% of the elderly (over 60 years old) suffer from LEASO 【1】 . In China, a cross-sectional study based on a Beijing community suggested that the prevalence of LEASO in the elderly population in my country was 12.9%. 【2】 . With the aging of the population, the incidence of LEASO is still increasing year by year. [0003] The typical symptom of LEASO is intermittent claudication, while the more serious clinical manifestations (critica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/861A61K48/00A61K31/7088A61K38/17A61P9/10A61P7/02
CPCA61K31/7088A61K38/1719A61K48/0008C12N15/86C12N2710/10043
Inventor 姚陈王劲松常光其王深明李勇辉
Owner 姚陈
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