Biomarker for detecting Alzheimer disease and application thereof

Abstract

The invention provides a biomarker for detecting Alzheimer's disease and application thereof, and the biomarker comprises CCL3 protein as a biomarker for detecting Alzheimer's disease, CCL3 protein as a biomarker for evaluating the risk of Alzheimer's disease, CCL3 protein as an intervention target of Alzheimer's disease, and CCL3 protein as a biomarker for evaluating the risk of Alzheimer's disease. The CCL3 protein is used as an intervention target for preventing and / or treating Alzheimer's disease related symptoms, the Alzheimer's disease detection kit containing the reagent for detecting the expression level of the CCL3, and the application of the reagent for detecting the expression level of the CCL3 in preparing the Alzheimer's disease detection reagent. The serum CCL3 serving as a biomarker of the Alzheimer's disease has the advantages of being high in specificity and sensitivity, and serum CCL3 detection further has the advantages of being low in detection cost, simple in detection technology, high in detection universality and high in patient acceptability.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a biomarker for detecting Alzheimer's disease and an application thereof. Background Art

[0002] With the continuous development of science and technology, human life expectancy is constantly increasing, but the quality of life of the elderly is affected by various factors. Alzheimer's disease (AD) is a common central nervous system degenerative disease occurring in the elderly. Its main clinical manifestations are progressive cognitive dysfunction and behavioral impairment. Its characteristic pathological changes are neuritic plaques (formed by nerve axons wrapping around Aβ), neurofibrillary tangles (formed by abnormal phosphorylated tau protein highly spiraled in neurons) and neuronal loss.

[0003] According to the survey, the prevalence of AD is increasing year by year. The pathogenesis of AD is complex and early diagnosis is difficult. At present, the clinical diagnosis of AD is mainly based on clinical manifestations, neuropsychological scales and cranial magnetic resonance imaging, with low specificity and sensitivity. Molecular pathology diagnosis is expensive, not widely used and not widely accepted by patients.

[0004] In recent years, research on Alzheimer's disease has made significant progress. Genetic technology and brain imaging technology have played an important role in the early diagnosis of Alzheimer's disease, and also provided a new perspective for understanding the pathogenesis of the disease. However, the exact pathogenesis of Alzheimer's disease is still unclear, and how to effectively prevent and treat Alzheimer's disease is still a problem that scientists need to explore. Therefore, discovering new and reliable biomarkers as diagnostic targets is of great significance for the early prediction, diagnosis and treatment of AD.

[0005] CCL3, also known as macrophage inflammatory protein α (MIP-1α), is one of the main members of the β-chemokine subfamily. It is a proinflammatory chemokine that can be produced by a variety of leukocytes and tissue cells and is widely expressed in the nervous system and immune system. It can participate in the occurrence and development of immune regulation or neuroinflammation by recruiting immune cells, activating intracellular signaling pathways, and mediating interactions between neurons and glial cells. Studies have found that overactive CCL3 has a direct and harmful effect on the outcome of the disease. The expression of CCL3 is associated with the progression of a variety of inflammatory and immune diseases, such as esophageal squamous cell carcinoma, chronic myeloid leukemia, acute and chronic (fibrotic) liver damage, etc. However, the role of CCL3 in the diagnosis, prevention, and treatment of AD is unclear. Summary of the invention

[0006] In view of the defects of the prior art, the present invention studies the relationship between CCL3 and AD, and proposes that the level of CCL3 in serum can reflect the neuroinflammatory status in the brain and is closely related to the clinical cognitive impairment of AD patients, which has great application potential for early identification and diagnosis of AD in clinical practice.

[0007] A first aspect of the present invention provides a biomarker for detecting Alzheimer's disease, including: the biomarker is CCL3 protein.

[0008] Furthermore, there is a negative correlation between the CCL3 protein level in the serum of Alzheimer's patients and the clinical cognitive ability of Alzheimer's patients.

[0009] Furthermore, CCL3 protein was quantitatively detected by enzyme-linked immunosorbent assay.

[0010] The second aspect of the present invention provides a biomarker for assessing the risk of Alzheimer's disease, including: the biomarker is CCL3 protein.

[0011] Furthermore, there is a negative correlation between the serum CCL3 protein level of people at risk of Alzheimer's disease and their clinical cognitive ability.

[0012] The third aspect of the present invention provides an intervention target for Alzheimer's disease, including: the intervention target is CCL3 protein.

[0013] Furthermore, CCL3 protein serves as an intervention target for preventing and / or treating symptoms associated with Alzheimer's disease.

[0014] A fourth aspect of the present invention provides a use of a reagent for detecting CCL3 levels in the preparation of an Alzheimer's disease detection reagent.

[0015] A fifth aspect of the present invention provides a kit for detecting Alzheimer's disease, characterized in that the kit comprises a reagent for detecting the expression level of CCL3 protein.

[0016] Furthermore, the reagent for detecting the expression level of CCL3 protein is a reagent for detecting the expression level of CCL3 protein in human tissue.

[0017] The embodiments of the present invention provide a biomarker for detecting Alzheimer's disease and its application, including CCL3 protein as a biomarker for detecting Alzheimer's disease, CCL3 protein as a biomarker for assessing the risk of Alzheimer's disease, CCL3 protein as an intervention target for Alzheimer's disease, CCL3 protein as an intervention target for preventing and / or treating symptoms related to Alzheimer's disease, an Alzheimer's disease detection kit containing a reagent for detecting the expression level of CCL3, and the application of the reagent for detecting the level of CCL3 in the preparation of an Alzheimer's disease detection reagent. The technical solution proposed by the present invention is based on the conclusion drawn from the experiment of the expression level of CCL3 in Alzheimer's disease patients. Serum CCL3 as a biomarker for Alzheimer's disease has the characteristics of high specificity and high sensitivity, and serum CCL3 detection also has the advantages of low detection cost, simple detection technology, high detection popularity and high patient acceptance. DETAILED DESCRIPTION

[0018] The present application is further described in detail below in conjunction with experimental results and embodiments. It is to be understood that the specific embodiments described herein are only used to explain the related invention, rather than to limit the invention. It should be noted that, in the absence of conflict, the embodiments in the present application and the features in the embodiments can be combined with each other.

[0019] The present invention provides a biomarker for detecting Alzheimer's disease and a biomarker for assessing the risk of Alzheimer's disease, wherein the biomarker is CCL3 protein.

[0020] Specifically, in the embodiment of the present invention, CCL3 is used as a biomarker for detecting Alzheimer's disease. This scheme has been verified by experiments, including:

[0021] Normal control mice and AD transgenic mice matched in age and sex were selected.

[0022] First, transcriptomic analysis was used to compare the differences in mRNA expression levels in brain tissues of normal control mice and Alzheimer's disease (AD) transgenic mice by high-throughput sequencing technology to identify key molecules associated with AD pathological processes. Analysis of the top 10 differentially expressed mRNAs revealed that CCL3 expression in the brains of AD transgenic mice was significantly increased compared with normal controls (p<0.001). In the preferred embodiment of the present invention, CCL3 protein was quantitatively detected by enzyme-linked immunosorbent assay.

[0023] Secondly, the enzyme-linked immunosorbent assay (ELISA) was used to verify that the expression level of CCL3 protein in the serum of Alzheimer's disease (AD) transgenic mice was significantly increased compared with normal controls (p<0.001). In addition, there was a significant negative correlation between the serum CCL3 level and the clinical cognitive ability of Alzheimer's disease (AD) transgenic mice (p<0.001), that is, the higher the CCL3 level, the worse the cognitive ability. In other words, in the preferred embodiment of the present invention, the relationship between the serum CCL3 protein level of Alzheimer's disease patients and the clinical cognitive ability of Alzheimer's disease patients is a negative correlation.

[0024] The above experiments confirmed that CCL3 is overactivated in the serum of patients with Alzheimer's disease (AD) and plays an important role in cognitive dysfunction and pathological damage in Alzheimer's disease (AD).

[0025] Thus, the first aspect of the present invention provides a CCL3 as a biomarker for detecting Alzheimer's disease, and the second aspect of the present invention provides a CCL3 protein as a biomarker for assessing the risk of Alzheimer's disease.

[0026] The present invention provides an intervention target for Alzheimer's disease, wherein the intervention target is CCL3 protein, and CCL3 protein is used as an intervention target for preventing and / or treating symptoms related to Alzheimer's disease.

[0027] Specifically, in the embodiments of the present invention, CCL3 protein is used as an intervention target for Alzheimer's disease and is verified by experiments, including:

[0028] The following four types of mice were shorthand divided into four groups for the experiment:

[0029] (1) 12-month-old wild-type C57BL / 6 mice (WT);

[0030] (2) 12-month-old wild-type C57BL / 6 mice treated with CCL3 antibody (WT+CCL3 Ab);

[0031] (3) APPswe / PS1 dE9 transgenic mice (APP / PS1);

[0032] (4) 12-month-old APPswe / PS1 dE9 transgenic mice treated with CCL3 antibody (APP / PS1+CCL3 Ab);

[0033] The Morris water maze acquisition experiment was used to evaluate the differences in spatial learning ability of mice between different groups. The experimental results showed that the escape latency of APPswe / PS1 dE9 transgenic mice was significantly longer than that of wild-type C57BL / 6 mice (p<0.001), which means that the experiment confirmed that APPswe / PS1 dE9 transgenic mice had defects in spatial learning ability. The results of the experiment also showed that the escape latency of APPswe / PS1 dE9 transgenic mice treated with CCL3 antibodies was significantly shorter than that of APPswe / PS1 dE9 transgenic mice (p<0.001), which confirmed that blocking the activation of CCL3 can significantly improve the spatial learning ability defects of APPswe / PS1 dE9 transgenic mice.

[0034] The Morris water maze exploration method was used to evaluate the differences in spatial memory ability of mice in different groups. The experimental results showed that the time that APPswe / PS1 dE9 transgenic mice stayed in the target quadrant was significantly shorter than that of wild-type C57BL / 6 mice (p<0.01), which means that the experiment confirmed that APPswe / PS1 dE9 transgenic mice had spatial memory defects. The results of the experiment also showed that the time that APPswe / PS1 dE9 transgenic mice treated with CCL3 antibodies stayed in the target quadrant was significantly longer than that of APPswe / PS1 dE9 transgenic mice (p<0.01), which confirmed that inhibiting CCL3 activity can significantly improve the spatial memory defects of APPswe / PS1 dE9 transgenic mice.

[0035] The above experiments confirmed that inhibiting the activity of CCL3 has the effect of improving AD cognitive dysfunction. Specifically, it is manifested in the following two aspects:

[0036] Spatial learning ability: Inhibition of CCL3 activity can significantly shorten the escape latency of APPswe / PS1 dE9 transgenic mice, indicating that their spatial learning ability is improved.

[0037] Spatial memory ability: Inhibiting CCL3 activity can significantly prolong the time that APPswe / PS1 dE9 transgenic mice stay in the target quadrant, indicating that their spatial memory ability is improved.

[0038] The above experiments confirmed that by inhibiting the activity of CCL3, the cognitive dysfunction of AD model mice can be improved to a certain extent, which provides a new intervention target for the treatment of AD.

[0039] Therefore, the third aspect of the present invention provides a CCL3 protein as an intervention target for Alzheimer's disease, and further proposes CCL3 protein as an intervention target for preventing and / or treating symptoms related to Alzheimer's disease.

[0040] The fourth aspect of the present invention provides an application of a reagent for detecting CCL3 levels in the preparation of an Alzheimer's disease detection reagent, which is mainly reflected in the following aspects:

[0041] Potential tool for early diagnosis: Since CCL3 changes in the early stages of AD, detecting its level may help diagnose AD at an early stage. This is of great significance for early intervention and treatment of AD.

[0042] Monitoring of disease progression: As AD progresses, CCL3 levels will change further. Therefore, regular testing of CCL3 levels can help monitor disease progression, evaluate treatment efficacy, and guide adjustments to treatment regimens.

[0043] Combined application with other biomarkers: In order to improve the accuracy and reliability of diagnosis, CCL3 is usually used in combination with other biomarkers (such as Aβ1-40, Aβ1-42, p-Tau, etc.). This combined detection method can comprehensively consider multiple factors, thereby improving the diagnostic ability of AD.

[0044] Advantages of non-invasive testing: Compared with traditional cerebrospinal fluid testing, blood testing has the advantages of being non-invasive and easy to operate. By testing the level of CCL3 in the blood, the discomfort and risks of cerebrospinal fluid testing can be avoided, while improving patient acceptance.

[0045] Clinical application: CCL3-based detection reagents may become one of the important tools for AD diagnosis and treatment.

[0046] As can be seen from the above, in the embodiments of the present invention, the application of the reagent for detecting CCL3 levels in the preparation of Alzheimer's disease detection reagents is reflected in its role as a biomarker, a potential tool for early diagnosis, a means of monitoring disease progression, and its combined application with other biomarkers.

[0047] A fifth aspect of the present invention provides a kit for detecting Alzheimer's disease, the kit comprising a reagent for detecting the expression level of CCL3 protein, and further the reagent for detecting the expression level of CCL3 protein is a reagent for detecting the expression level of CCL3 protein in human tissue.

[0048] In the embodiment of the present invention, the kit for detecting Alzheimer's disease comprises a plurality of components, such as antibodies, substrates, buffers, etc., for specifically binding and detecting CCL3 protein. The detection process may involve reacting the sample to be tested (such as blood or cerebrospinal fluid) with the reagents in the kit, and then quantifying the expression level of CCL3 protein by a specific detection method (such as enzyme-linked immunosorbent assay ELISA, Western blot, etc.).

[0049] In addition, in order to improve the accuracy and reliability of diagnosis, the kit for detecting Alzheimer's disease provided by the present invention may be used in combination with other biomarkers (such as Aβ1-40, Aβ1-42, p-Tau, etc.). By comprehensively considering multiple factors, it is possible to more accurately assess whether the patient's cognitive impairment is caused by AD and monitor the progression of the disease.

[0050] In conclusion, the Alzheimer's disease detection kit provided by the present invention provides a new tool for early diagnosis and disease monitoring of AD by detecting the expression level of CCL3 protein.

[0051] The embodiments of the present invention provide a biomarker for detecting Alzheimer's disease and its application, including CCL3 protein as a biomarker for detecting Alzheimer's disease, CCL3 protein as a biomarker for assessing the risk of Alzheimer's disease, CCL3 protein as an intervention target for Alzheimer's disease, CCL3 protein as an intervention target for preventing and / or treating symptoms related to Alzheimer's disease, an Alzheimer's disease detection kit containing a reagent for detecting the expression level of CCL3, and the application of the reagent for detecting the level of CCL3 in the preparation of an Alzheimer's disease detection reagent. The technical solution proposed by the present invention is based on the conclusion drawn from the experiment of the expression level of CCL3 in Alzheimer's disease patients. Serum CCL3 as a biomarker for Alzheimer's disease has the characteristics of high specificity and high sensitivity, and serum CCL3 detection also has the advantages of low detection cost, simple detection technology, high detection popularity and high patient acceptance.

[0052] The above specific implementation methods further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above are only specific implementation methods of the present invention and are not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A biomarker for detecting Alzheimer's disease, characterized in that: The biomarker is CCL3 protein.

2. The biomarker according to claim 2, characterized in that The relationship between the CCL3 protein level in the serum of Alzheimer's patients and the clinical cognitive ability of the Alzheimer's patients is a negative correlation.

3. The biomarker according to claim 1 or 2, characterized in that The CCL3 protein is quantitatively detected by enzyme-linked immunosorbent assay.

4. A biomarker for assessing the risk of Alzheimer's disease, characterized in that: The biomarker is CCL3 protein.

5. The biomarker according to claim 4, characterized in that The relationship between the CCL3 protein level in the serum of the person at risk of Alzheimer's disease and the clinical cognitive ability of the person at risk of Alzheimer's disease is a negative correlation.

6. An intervention target for Alzheimer's disease, characterized in that: The intervention target is CCL3 protein.

7. The intervention target according to claim 6, characterized in that: The CCL3 protein serves as an intervention target for preventing and / or treating symptoms associated with Alzheimer's disease.

8. Use of a reagent for detecting CCL3 levels in the preparation of an Alzheimer's disease detection reagent.

9. A kit for detecting Alzheimer's disease, characterized in that: The kit includes reagents for detecting the expression level of CCL3 protein.

10. The kit according to claim 9, characterized in that The reagent for detecting the expression level of CCL3 protein is a reagent for detecting the expression level of CCL3 protein in human tissue.