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Application of yap1 gene in the diagnosis and treatment of Alzheimer's disease

A kind of Alzheimer's disease, gene technology, applied in the treatment, human YAP1 gene in the field of diagnosis of Alzheimer's disease, can solve the problem of inability to organize or reverse the disease progression

Active Publication Date: 2017-11-07
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Clinically diagnosed AD patients are basically in the middle and late stages, and existing treatments can only improve symptoms, but cannot prevent or reverse the progression of the disease.

Method used

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  • Application of yap1 gene in the diagnosis and treatment of Alzheimer's disease
  • Application of yap1 gene in the diagnosis and treatment of Alzheimer's disease
  • Application of yap1 gene in the diagnosis and treatment of Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Screening for Gene Markers Related to Alzheimer's Disease

[0057] 1. Collection of peripheral blood samples

[0058] AD patients came from Beijing 301 Hospital, a total of 60 cases, aged 53-84 years old, all cases were diagnosed as AD, and its diagnostic criteria refer to the third revised edition of the American Diagnostic and Statistical Manual of Mental Disorders. A total of 50 cases were selected as the control group, selected from the routine physical examination population of Beijing Hospital. All the subjects were excluded from blood lipid metabolism and other diseases, aged 60-82 years. All subjects signed the informed consent for the testing project and provided peripheral blood for genetic testing.

[0059] 2. Total RNA extraction from blood

[0060] Extraction of total RNA from blood using Biotech Blood RNA Extraction Kit

[0061] (1) Take 250 μl (or 0.25 g) of whole blood into an RNase-Free filter column, centrifuge at 13,000 rpm for 2 minutes,...

Embodiment 2

[0091] Embodiment 2 interferes with the expression of YAP1 gene

[0092] 1. siRNA design and synthesis

[0093] siRNA sequence against YAP1:

[0094] siRNA1-YAP1:

[0095] The sense strand is 5'-AUGUGAUUUAAGAAGUAUCUC-3' (SEQ ID NO.7);

[0096] The antisense strand is 5'-GAUACUUCUUAAAUCACAUCG-3' (SEQ ID NO.8),

[0097] siRNA2-YAP1:

[0098] The sense strand is 5'-UUAUAUAGUAAAUUUCUCCAU-3' (SEQ ID NO.9);

[0099] The antisense strand is 5'-GGAGAAAUUUACUAUAUAAAC-3' (SEQ ID NO.10),

[0100] siRNA3-YAP1:

[0101] The sense strand is 5'-UGACAUUUUGGAGAAUUUGCU-3' (SEQ ID NO.11);

[0102] The antisense strand is 5'-CAAAUUCUCCAAAAUGUCAGG-3' (SEQ ID NO.12)

[0103] Negative control siRNA sequence (siRNA-NC):

[0104] The sense strand is 5'-CGUACGCGGAAUACUUCGA-3' (SEQ ID NO.13);

[0105] The antisense strand is 5'-UCGAAGUAUUCCGCGUACG-3' (SEQ ID NO.14).

[0106] Neural cell line R2L1 cells were divided into 1×10 4 Inoculate / well into 24-well cell culture plates at 37°C, 5% CO 2...

Embodiment 3

[0116] Example 3 Antagonistic effect of YAP1 gene on neuronal cell death induced by Aβ

[0117] 1. Cell transfection: according to the method in Example 2, the neural cell line R2L1 was transfected with siRNA1-YAP1 and siRNA-NC.

[0118] 2. After 24 hours of transfection, the neural cell line R2L1 was cultured in a 96-well plate with a cell density of 0.5×10 4 cells / well. Divide the cells into the following groups:

[0119] Uninduced group: transfected with siRNA-NC, without adding 20 μM Aβ42;

[0120] Negative control group (siRNA-NC+Aβ42): siRNA-NC was transfected, and 20 μM Aβ42 was added to the medium;

[0121] YAP1 gene interference group (siRNA-YAP1+Aβ42): transfect siRNA-YAP1, and add 20 μM Aβ42 to the medium;

[0122] Three replicate holes were set up for each group. After incubation at 37° C. for 20 h, MTT was added and incubated for 4 h. After adding the dissolving solution, incubate at 37°C for another 12h, and read the optical density value at 600nm. The hig...

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Abstract

The invention discloses a YAP1 gene which can be used as a molecular marker for early diagnosis of Alzheimer's disease. The QPCR (quantitative polymerase chain reaction) experiment proves that compared with common people, the YAP1 gene expression in the blood of an Alzheimer's disease patient is obviously enhanced. The RNA (ribonucleic acid) interfering experiment proves that the YAP1 can influence the Abeta mediated neurotoxicity action. According to the research results, a drug capable of inhibiting YAP1 gene expression or inhibiting YAP1 gene expression product functions can be developed, thereby implementing clinical prevention and treatment of Alzheimer's disease.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of human YAP1 gene in the diagnosis and treatment of Alzheimer's disease. Background technique [0002] The English word for Alzheimer's disease is Alzheimer's disease, usually abbreviated as AD. Dementia is a progressive neurodegenerative disease characterized by dementia, and it is an important disease that endangers the health and life of the elderly. At present, there are about 26 million AD patients in the world, and 5 million new cases are diagnosed every year. It is estimated that there will be more than 100 million AD patients in the world by 2040-2050. AD has become the main cause of death in the current population, and the 4th to 5th cause of death in people over 65 years old. The economic burden brought by AD to the family and society ranks third among all diseases, second only to heart disease and cancer. China is facing a severe situation of population aging a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N33/68A61K48/00A61K31/713A61K39/00A61K45/00A61P25/28
CPCA61K31/713A61K39/0005A61K45/00C12Q1/6883C12Q2600/158G01N33/6896G01N2800/2821
Inventor 杨承刚崔双双
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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