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Application of hoxd8 in the preparation of products promoting osteogenic differentiation of bone marrow mesenchymal stem cells

A bone marrow mesenchymal, HOXD8 technology, applied in the application field of the product, can solve the problem of unclear biological role of lncRNA

Active Publication Date: 2020-11-06
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the biological roles of lncRNAs in the pathogenesis of AIS are still unclear

Method used

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  • Application of hoxd8 in the preparation of products promoting osteogenic differentiation of bone marrow mesenchymal stem cells
  • Application of hoxd8 in the preparation of products promoting osteogenic differentiation of bone marrow mesenchymal stem cells
  • Application of hoxd8 in the preparation of products promoting osteogenic differentiation of bone marrow mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Example 1: Identification of LncAIS as a Significantly Differentially Expressed lncRNA Downregulated in AIS Patient Bone Marrow Mesenchymal Stem Cells (BM-MSCs)

[0095] To identify key lncRNAs involved in adolescent idiopathic scoliosis (AIS), we performed microarray analysis of BM-MSCs from 5 healthy donors and 12 AIS patients. 1483 lncRNAs showed differential expression between normal BM-MSCs and BM-MSCs of patients with AIS, of which 718 were up-regulated and 765 were down-regulated, such as figure 1 as shown in a. Among the most downregulated lncRNAs in BM-MSCs of AIS patients, we focused on an uncharacterized lncRNA we termed lncAIS (gene symbol: ENST00000453347). lncAIS is located on human chromosome 1, contains 4 exons, and is a full-length 476nt transcript. lncAIS was identified as a conserved locus, as figure 1 as shown in b.

[0096] Perform real-time qPCR according to routine operations, analyze lncAIS transcripts in BM-MSCs of normal healthy donors and ...

Embodiment 2

[0102] Example 2: Determination of LncAIS activation of HOXD8 transcription

[0103] Transcriptome microarray analysis was performed on lncAIS silenced (shLncAIS) and shCtrl-treated BM-MSCs (the shlncAIS gene sequence used for lncAIS knockdown is shown in SEQ ID NO:4-5 in Table 2 above), and the results are as follows figure 2 As shown in the heat map of a, the 10 most down-regulated transcription factors (TFs) in BM-MSCs silenced by lncAIS were selected; and their expression levels in normal BM-MSCs were detected. Data are from two independent experiments using BM-MSCs derived from 2 healthy donors. Each matched shCtrl and shLncAIS were from the same healthy BM-MSC donor.

[0104] Such as figure 2 As shown in b, the real-time qPCR analysis of BM-MSCs treated with shCtrl and shLncAIS showed that among the 10 TFs with the largest down-regulation range in BM-MSCs treated with shLncAIS, HOXD8, which is a marker gene of BM-MSCs, was significantly affected in normal BM-MSCs. h...

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Abstract

The invention discloses an application of a transcription factor HOXD8 in the preparation of a product for promoting osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs), and finds that the HOXD8 can promote cell proliferation in the BM-MSCs and increase the expression level of self-updating related genes and osteogenic differentiation genes in the BM-MSCs, therefore, the HOXD8 can be used to prepare products that promote cell proliferation and osteogenic differentiation in BM-MSCs, such as plasmids comprising the HOXD8 genes, recombinant expression vectors, transgenic cell lines, and genetically engineered bacteria, and further used in the treatment of adolescent idiopathic scoliosis (AIS). The invention also discloses a pharmaceutical composition for the preventionand / or treatment of AIS, which comprises an effective amount of HOXD8 and a pharmaceutically acceptable carrier and is used for the treatment of AIS associated with abnormal BM-MSC osteogenic differentiation in a patient.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a transcription factor HOXD8 related to osteogenic differentiation in the preparation of products for promoting osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSC). Background technique [0002] Adolescent idiopathic scoliosis (AIS) is a complex three-dimensional deformity of the spine that occurs mainly in girls aged 10 to 16 years during the pubertal growth period. The progression of scoliosis may lead to appearance deformity, back pain, and functional impairment. In severe cases, physiological function problems such as limited cardiopulmonary function will occur, which will affect social activities to varying degrees. Epidemiological survey results show that the global incidence of AIS among adolescents is about 2-4%, and about 10% of those diagnosed with AIS require treatment. The current treatment of AIS includes bracing therapy ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077C12N5/10C12Q1/6888
CPCC12N5/0654C12N2501/60C12N2501/998C12N2506/1353C12Q1/6888C12Q2600/158
Inventor 庄乾宇仉建国惠尚懿范祖森邱贵兴吴志宏叶步青赵春华李静李娜王升儒林莞峰张延斌杨阳
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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