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Application of nf90 in the preparation of biological agents regulating osteogenic differentiation of bone marrow mesenchymal stem cells

A technology for bone marrow mesenchymal and osteogenic differentiation, applied in the field of biomedicine, can solve the problem of unclear biological role of lncRNA

Active Publication Date: 2022-02-22
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • 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 nf90 in the preparation of biological agents regulating osteogenic differentiation of bone marrow mesenchymal stem cells
  • Application of nf90 in the preparation of biological agents regulating osteogenic differentiation of bone marrow mesenchymal stem cells
  • Application of nf90 in the preparation of biological agents regulating osteogenic differentiation of bone marrow mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1: Identification of LncAIS as a significantly differentially expressed lncRNA downregulated in AIS patient bone marrow mesenchymal stem cells (BM-MSCs)

[0091] To identify key lncRNAs involved in adolescent idiopathic scoliosis (AIS), we performed microarray analysis on BM-MSCs from 5 healthy donors and 12 AIS patients. In normal BM-MSCs and BM-MSCs of AIS patients, 1483 lncRNAs showed differential expression, of which 718 were up-regulated and 765 were down-regulated, as shown in figure 1 a shown. Among the most down-regulated lncRNAs in BM-MSCs of AIS patients, we focused on an uncharacterized lncRNA that 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 such as figure 1 b shown.

[0092] Real-time qPCR was performed according to routine operations to analyze lncAIS transcripts in BM-MSCs of normal healthy donors and...

Embodiment 2

[0098] Example 2: Determination of LncAIS interaction with NF90 to enhance mRNA stability of HOXD8

[0099] LncRNAs are often positively correlated with the regulation of their nearby protein-coding genes. However, if figure 2 As shown in a, we found by qRT-PCR assessment that depletion of lncAIS in BM-MSCs did not affect the expression levels of its neighboring genes (the shlncAIS gene sequences used for lncAIS knockdown were as shown in SEQ ID NOs: 4-6 in Table 2 above) shown), where relative gene expression fold changes were calculated as mean ± S.D. NS, not significant. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. This suggests that lncAIS may exert its regulatory role in trans.

[0100] Biotin-labeled RNA knockdown assays were then performed to identify possible lncAIS-related proteins from BM-MSC lysates. Biotin-RNA pulldown was performed with lysates of BM-MSCs using full-length lncAIS transcripts (sense) and antisense s...

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Abstract

The invention discloses the application of NF90 in the preparation of biological preparations for regulating osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSC). We found that the RNA-binding protein NF90 is required for the osteogenic differentiation of BM‑MSCs, and lncAIS enhances the mRNA stability of the transcription factor HOXD8 by binding to NF90 protein to promote BM‑MSCs in bone marrow mesenchymal stem cells (BM‑MSCs). Therefore, NF90 can be used to prepare biological preparations for regulating osteogenic differentiation of BM-MSCs, such as plasmids, recombinant expression vectors, transgenic cell lines, and genetically engineered bacteria that contain overexpressed NF90 genes or can express shNF90. The present invention also discloses a pharmaceutical composition for preventing and / or treating adolescent idiopathic scoliosis (AIS), which comprises an effective amount of NF90 and a pharmaceutically acceptable carrier, and is used for treating patients with BM- AIS associated with abnormal MSC osteogenic differentiation.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of RNA binding protein NF90 in the preparation of a biological preparation for regulating the 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 primarily in girls aged 10 to 16 years during pubertal growth. The progression of scoliosis may cause appearance deformity, back pain, and functional impairment. In severe cases, physiological function problems such as cardiopulmonary function limitation will occur, which will affect social activities to varying degrees. Epidemiological findings suggest that the global incidence of AIS in adolescents is approximately 2-4%, of which approximately 10% of those diagnosed with AIS require treatment. Current treatments for AIS include brace therapy and surgical corre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/10A61P19/08
CPCA61K38/10A61P19/08
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张延斌林莞峰杨阳
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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