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Application of tool for targeted activation of LRP6 in preparation of medicine for preventing and treating Alzheimer's disease

An Alzheimer's disease and tool technology, applied in the field of biomedicine, can solve problems such as the lack of prevention and treatment of Alzheimer's disease, relieve the damage of brain endothelial cell function, repair blood-brain barrier damage, application value and good prospects

Pending Publication Date: 2022-08-02
THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no specific and precise treatment to prevent and treat Alzheimer's disease by repairing and maintaining the blood-brain barrier and reshaping the homeostasis of the brain microenvironment in the course of AD

Method used

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  • Application of tool for targeted activation of LRP6 in preparation of medicine for preventing and treating Alzheimer's disease
  • Application of tool for targeted activation of LRP6 in preparation of medicine for preventing and treating Alzheimer's disease
  • Application of tool for targeted activation of LRP6 in preparation of medicine for preventing and treating Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Preparation of optogenetic tool OptoLRP6

[0032] Preparation steps:

[0033] S1. Construct a light-sensitive control plasmid expressing LRP6 protein, namely OptoLRP6 plasmid (provided by Addgene, whose structure is as follows: figure 1 As shown, the used backbone plasmid is pCS2+);

[0034] S2. Using the third-generation lentivirus preparation system, the plasmid in step S1 was transfected into the HEK293T cell line, and the medium for culturing the HEK293T cell line was replaced every 8 hours after transfection;

[0035] S3. Collect the culture medium of HEK293T cell line 48 hours after transfection in step S2, and filter with a 0.22 μm filter, the obtained filtrate is the plasmid of OptoLRP6.

Embodiment 2

[0036] Example 2 In vitro brain endothelial cell blood-brain barrier injury model

[0037] 1. The purpose of the experiment:

[0038] Preparation of an in vitro model of brain endothelial cell blood-brain barrier injury to test optogenetic tools

[0039] 2. Experimental method:

[0040]Take 10-day-old rat suckling mice, remove the brain tissue after anesthesia, and remove the cerebellum, cerebellum, brain stem and white matter on ice to obtain the remaining cortex; the remaining cortex is homogenized in 10 mL and 1 mL of DMEM, DMEM containing 1 mg / mL of collagenase and 0.1 mg / mL DNase I, and shaken at 250 rpm for 1 h at 37 °C, then centrifuged at 1000 g for 8 min at room temperature; the pellet obtained by centrifugation was suspended in 25 mL of DMEM containing 20% ​​BSA, at room temperature The microvessel pellet was obtained by centrifugation at 1000g for 20 minutes. The microvessel pellet was further homogenized in 5 mL of DMEM containing 0.5 mL of 1 mg / mL collagenase-...

Embodiment 3

[0043] Example 3 Optogenetic tool LRP6 in the treatment of blood-brain barrier damage in vitro caused by Aβ oligomer treatment

[0044] 1. The purpose of the experiment:

[0045] Validation of the therapeutic effect of the optogenetic tool OptoLRP6 on blood-brain barrier damage induced by Aβ oligomer treatment in vitro.

[0046] 2. Experimental method:

[0047] The primary BECs were extracted according to the primary BEC extraction method in Example 2, and the BEC cells were treated with 20 μM Aβ using the blood-brain barrier damage model construction method in Example 2 to establish an in vitro blood-brain barrier damage model. Blue light treatment, specifically blue light (wavelength 450nm) cyclic treatment, that is, turn on the blue light for 20 minutes, and then turn off the blue light for 40 minutes for a total of 8 hours. After the end, collect cell samples to extract RNA and protein samples for relevant indicators and blood-brain barrier function. Detection of protein...

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Abstract

The invention discloses application of a tool for repairing and maintaining a blood-brain barrier function based on targeted activation of LRP6 in preparation of a medicine for preventing and treating Alzheimer's disease, in particular to early intervention and prevention of the Alzheimer's disease. Based on the research of the invention, an optogenetics tool OptoLRP6 is utilized to target LRP6, the blood brain barrier injury can be repaired, the blood brain barrier function can be maintained, and the brain microenvironment steady state in the AD disease course can be remodeled, so that the brain endothelial cell function damage caused by A beta oligomer can be relieved, and the method can be applied to treatment and early intervention prevention of Alzheimer's disease. Therefore, development of a tool for repairing and maintaining the blood-brain barrier function based on targeted activation of the LRP6, such as an optogenetics tool, has a very good application prospect in prevention and / or treatment of the Alzheimer's disease.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. Specifically, it relates to the application of a tool for repairing and maintaining the blood-brain barrier function based on the targeted activation of LRP6 in preventing and treating Alzheimer's disease, and more particularly, it relates to the application of an optogenetic tool OptoLRP6 in preventing and treating Alzheimer's disease. Background technique [0002] Alzheimer's Disease (AD) is a neurodegenerative disease. At present, the incidence of AD in my country and the world is increasing rapidly, and it has become a serious social problem. But so far, AD still lacks effective prevention and treatment methods, and new treatment strategies are urgently needed. Because AD has no effective preventive measures, and the early disease process of AD is slow, the diagnosis is difficult, and drug intervention is usually late. Therefore, the current clinical trial drugs for AD mainly target the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K41/00A61P25/28C12N15/867C12N15/12
CPCA61K45/00A61K41/0057A61P25/28C12N15/86C07K14/47C12N2740/15043
Inventor 易陈菊王琪黄小敏苏一洵
Owner THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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