Application of EGFR/ERK/TRIM32 signal path in promoting differentiation of spinal nerve stem cells after spinal cord injury

A signal pathway, spinal cord injury technology, applied in the application field of promoting the differentiation of spinal cord neural stem cells after spinal cord injury, to achieve the effect of promoting functional recovery

Active Publication Date: 2019-07-26
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, how myelin proteins inhibit the differentiat

Method used

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  • Application of EGFR/ERK/TRIM32 signal path in promoting differentiation of spinal nerve stem cells after spinal cord injury
  • Application of EGFR/ERK/TRIM32 signal path in promoting differentiation of spinal nerve stem cells after spinal cord injury
  • Application of EGFR/ERK/TRIM32 signal path in promoting differentiation of spinal nerve stem cells after spinal cord injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Discovery of EGFR / ERK / TRIM32 Signaling Pathway

[0051] 1. Myelin proteins inhibit the differentiation of neural stem cells into neurons by activating the EGFR-ERK pathway

[0052] Taking the neonatal mouse telencephalon neural stem cells 12 hours after birth of SD rats as the experimental object, add myelin protein (final concentration: 10 μg / mL) alone during the in vitro differentiation process, or add EGFR inhibitors respectively while adding myelin protein Gefitinib (final concentration 1μM), ERK inhibitor PD98059 (final concentration 1μM), Akt inhibitor MK2206 2HCl (final concentration 100nM), analyzed by immunofluorescence experiments, the results showed that myelin proteins activate EGFR / ERK signaling Pathways inhibit the differentiation of neural stem cells into neurons (such as figure 1 shown in a and b).

[0053] Taking the neonatal mouse telencephalon neural stem cells 12 hours after birth of SD rats as the experimental object, add myelin protein ...

Embodiment 2

[0057] Example 2 Application of EGFR / ERK / TRIM32 signaling pathway inhibitors in promoting the differentiation of spinal cord neural stem cells after spinal cord injury

[0058] Taking adult SD rats as the experimental subjects, after total spinal cord transection injury, the EGFR inhibitor Gefitinib (concentration of 1 μM, 9-10 μL, meeting the final concentration of 50 μM / kg) or the ERK inhibitor PD98059 (concentration of 1 μM , 3.6-4μL, to meet the final concentration of 20μM / kg), and then continuously intraperitoneally inject EGFR inhibitor Gefitinib (concentration of 30mg / mL, 450-500μL, to meet the final concentration of 75mg / kg) and ERK inhibitor PD98059 (concentration 20mg / mL, 90-100μL, meet the final concentration of 10mg / kg). According to the analysis of BBB score and tissue immunofluorescence staining experiment, the results showed that after total spinal cord transection injury, intraperitoneal injection of EGFR / ERK inhibitor can increase the expression of TRIM32 prot...

Embodiment 3

[0061] Example 3 Application of TRIM32 in promoting the differentiation of spinal cord neural stem cells after spinal cord injury

[0062] Taking the neonatal mouse telencephalon neural stem cells 12 hours after birth of SD rats as the experimental object, through lentivirus-mediated TRIM32 (the amino acid sequence is shown in SEQ ID NO.1, the nucleotide sequence of the coding gene is shown in SEQ ID NO.2 (shown) after overexpression in neural stem cells, set the myelin protein group and no myelin protein group, the results show that overexpression of TRIM32 protein in neural stem cells can antagonize the effect of myelin protein on neural stem cells' directed differentiation to neurons Inhibition, promote the differentiation of neural stem cells into neurons (such as figure 2 as shown in d).

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Abstract

The invention relates to the field of biotechnology, and particularly relates to application of an EGFR/ERK/TRIM32 signal path in promoting differentiation of spinal nerve stem cells after spinal cordinjury. According to the application, inhibitors or blockers of the EGFR/ERK/TRIM32 signal path and TRIM32 can be used for promoting the differentiation of the neural stem cells into neurons during the spinal cord injury repair process. After the spinal cord injury, a myelin protein finally inhibits a downstream target TRIM32 protein by activating the EGFR/ERK signal path, and further inhibitingthe differentiation of the neural stem cells into the neurons. The EGFR/ERK/TRIM32 signal path can be used as a new drug target for the spinal cord injury repair, and provides new ideas for repair andtreatment of the spinal cord injury repair.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of EGFR / ERK / TRIM32 signaling pathway in promoting the differentiation of spinal cord neural stem cells after spinal cord injury. Background technique [0002] Neural stem cells are cells with self-renewal and multi-directional differentiation potential in the central nervous system. Neural stem cells and their differentiation play an important role in nerve injury repair and degenerative diseases, such as spinal cord injury, Alzheimer's disease, stroke, etc. . Neural stem cells are scattered in the brain of adult mammals and can differentiate into glial cells and neurons. At the same time, neural stem cells in the body have the ability to migrate to the injured area, but the number of migration and regeneration ability are limited, so the directional differentiation of neural stem cells is current research hotspot. [0003] In recent years, the incidence of spinal c...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P25/00
CPCA61K45/00A61P25/00
Inventor 戴建武薛伟伟肖志峰陈冰
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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