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Diagnosis of alzheimer's disease

A technology for assisting diagnosis of Alzheimer's disease, applied in the direction of disease diagnosis, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problem that neurons cannot start to redifferentiate

Inactive Publication Date: 2014-04-30
THE UNIV OF BIRMINGHAM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, Alzheimer's disease neurons fail to initiate G1 arrest and subsequent redifferentiation due to defects in the G1 / S regulatory checkpoint

Method used

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  • Diagnosis of alzheimer's disease
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  • Diagnosis of alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Example 1 Gene Expression Microarray Analysis of Lymphocytes Treated with Rapamycin

[0138] Genes differentially expressed in rapamycin-treated lymphocytes compared to untreated lymphocytes were identified using two-color-based microarray gene expression analysis (Agilent Technologies). The microarray used is Agilent Human Genome Microarray (Whole Human Genome Microarray): 4x44K.

[0139] 1.1 Lymphocyte cultures with and without rapamycin

[0140] Two parallel lymphoid cultures were established in RPMI medium supplemented with 10% FCS at a concentration of 1x10 6 cells. Phytohemagglutinin (PHA) was added to the culture to a final concentration of 22 μg / ml to activate lymphocytes. at 5% CO 2 The cultures were incubated at 37°C for 48 hours in a humidified environment. After 48 hours of incubation, one culture was treated with 100 ng / ml rapamycin, while the other untreated culture served as a control. After 24 hours, the cells of each lymphocyte culture system were...

Embodiment 2

[0172] Example 2 Analysis of the expression of rapamycin-sensitive genes in lymphocytes isolated from human subjects

[0173] Lymphocytes were isolated for testing from venous blood samples of human subjects. Venous blood samples were obtained from patients and placed in heparinized vacutainer tubes and sent to the laboratory at room temperature within 48 hours. Lymphocytes are isolated from blood using Lymphoprep, Histopaque, Ficoll or equivalent standard reagents according to standard procedures.

[0174] Alternatively, collect venous blood directly into a BD containing sodium heparin CpT TM In the cell preparation tube, immediately isolate the lymphocytes using one of the techniques described above. A sample of lymphocytes is then sent to a laboratory for analysis.

[0175] For each lymphocyte sample obtained from a human subject, two parallel lymphocyte culture systems were established in RPMI medium supplemented with 10% FCS at a concentration of 1 × 10 6 cells. Ph...

Embodiment 3

[0177] Example 3 Differences in gene expression in response to rapamycin measured in Alzheimer's disease patients.

[0178] 3.1 Blood collection and lymphocyte separation

[0179] Peripheral blood samples from elderly subjects were provided by Oxford Project to Investigate Memory and Aging (OPTIMA) subjects, which obtained ethics approval, and patients were informed and informed consent was obtained. Samples presented were from Alzheimer's patients (n=24) who met the NINCDS-ARDRA criteria for probable AD or from healthy age-matched controls (n=21). Appropriate informed consent and ethics approvals were obtained before the study began.

[0180] Peripheral blood was collected in heparinized vacutainer tubes and shipped at room temperature. Lymphocyte isolation was performed within 24 hours of blood collection using an established procedure (Lymphoprep). Briefly, blood was diluted 1:1 with PBS (Ca and Mg free, Sigma). 10ml diluted blood was carefully layered on top of 4ml Lym...

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Abstract

The present invention relates to diagnosis and monitoring of Alzheimer's disease in the live subject. More particularly, the invention relates to methods involving the measurement of differential gene expression in non-neuronal cells taken from human subjects suspected of having Alzheimer's disease wherein the genes to be measured are genes within the m TOR signalling pathway.

Description

technical field [0001] The present invention relates to the diagnosis and monitoring of Alzheimer's disease in living subjects. In particular, although not exclusively, the invention relates to methods comprising measuring differential gene expression in non-neuronal cells taken from a human subject suspected of having Alzheimer's disease. The invention also relates to methods of monitoring mTOR signaling in human cells. Background technique [0002] Alzheimer's disease is the most common form of dementia among older adults. Due to the aging population worldwide, the incidence of this disease will increase significantly in the next few years. Therefore, there is an urgent need to develop better diagnostic tools and new treatment options for those identified with the disease. [0003] Alzheimer's disease is a chronic neurodegenerative disorder characterized by the selective loss of neurons in the hippocampus and cortical layers within the temporal and frontal lobes of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N2333/91215G01N2800/50C12Q1/6827G01N33/6896G01N2800/2821G01N2800/52C12Q1/6883G01N33/68
Inventor 兹苏桑纳·纳吉
Owner THE UNIV OF BIRMINGHAM
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