Related application of phosphorylated Tau protein in delirium prediction

Abstract

The invention discloses related application of phosphorylated Tau protein in delirium prediction, and relates to the field of biology, it is found that increase of the expression level of p-Tau 396 is related to the illness risk and / or prognostic risk of delirium, the p-Tau 396 can be used as an independent predictive factor to predict the illness risk and / or prognostic risk of delirium of a subject, and the application of the p-Tau 396 in delirium prediction is provided. And the patient can be intervened or treated in time.

Description

Technical Field

[0001] The present invention relates to the biological field, and in particular to the application of phosphorylated Tau protein in predicting delirium. Background Art

[0002] Delirium is an acute brain dysfunction that lasts from hours to weeks, characterized by changes or fluctuations in mental state, inattention, cognitive dysfunction, confusion, or changes in consciousness. Delirium can occur in people of all ages, from infancy to old age, and is especially prevalent among the elderly. Elderly people are more likely to have adverse prognoses after developing delirium, including falls, disability, and even increased risk of death, which greatly increases medical expenses and places a heavy medical and economic burden on families and society.

[0003] Delirium is transient, and medical delirium is even more hidden. At present, there is almost no multi-omics research on medical delirium. There is still a lack of systematic understanding of delirium in clinical practice, and the corresponding mechanism research is relatively scarce. Delirium is different from dementia and lacks specific diagnostic biological markers. At present, the gold standard diagnosis of delirium is neuropsychological assessment by specialists.

[0004] The pathogenesis of delirium is diverse. Stress response, neurotransmitter imbalance, neuroinflammatory response, and nerve damage have been proven to be related to the occurrence of delirium. Due to many factors such as different primary diseases, different severity of primary diseases, and different physical fitness, the prognosis of patients with delirium varies greatly. Taking patients with cerebrovascular diseases, cardiorenal and pulmonary encephalopathy as an example, the prognosis of delirium caused by the deterioration of some physical diseases is poor, and the mortality rate within one month can reach 25-33%. The onset of delirium is highly correlated with long-term cognitive impairment of 3 months or more. The lack of research on the mechanism of delirium has directly resulted in the current lack of effective treatment for delirium in clinical practice, and symptoms can only be improved through nursing and symptomatic treatment.

[0005] Therefore, finding markers that can predict delirium has become a key issue that needs to be addressed urgently.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] The purpose of the present invention is to provide relevant applications of phosphorylated Tau protein in predicting delirium.

[0008] The present invention is achieved in that:

[0009] In a first aspect, an embodiment of the present invention provides use of a reagent for detecting the expression level of phosphorylated Tau protein in preparing a product for predicting delirium, wherein the phosphorylated Tau protein includes p-Tau 396.

[0010] In a second aspect, an embodiment of the present invention provides the use of an agent for reducing the phosphorylation level of the S396 site of the Tau protein in the preparation of a product for preventing, treating or assisting in the treatment of delirium.

[0011] In a third aspect, an embodiment of the present invention provides a method for training a prediction model, which includes:

[0012] Obtaining the detection results of the phosphorylated Tau protein expression level in the training sample and the corresponding annotation results; the phosphorylated Tau protein includes p-Tau 396, and the annotation results include the disease risk and / or prognostic risk of delirium representing the sample;

[0013] Input the obtained test results into the pre-built machine learning model to obtain the prediction results of the samples;

[0014] According to the labeling results and the prediction results, the parameters of the machine learning model are updated to obtain a prediction model.

[0015] In a fourth aspect, an embodiment of the present invention provides a prediction device, which includes:

[0016] An acquisition module, used to obtain the detection result of the phosphorylated Tau protein expression level of the sample to be tested, wherein the phosphorylated Tau protein includes p-Tau 396;

[0017] The prediction module is used to input the detection result of the sample to be tested into the prediction model trained by the training method described in the above embodiment to obtain the prediction result of the sample to be tested.

[0018] In a fifth aspect, an embodiment of the present invention provides an electronic device, comprising a processor and a memory, wherein the memory is used to store a program, and when the program is executed by the processor, the processor implements a method for predicting delirium;

[0019] The method for predicting delirium comprises the following steps: obtaining the detection result of the expression level of phosphorylated Tau protein of the sample to be tested, wherein the phosphorylated Tau protein includes p-Tau 396; inputting the detection result of the sample to be tested into the prediction model trained by the training method described in the above embodiment to obtain the prediction result of the sample to be tested.

[0020] In a sixth aspect, an embodiment of the present invention provides a computer-readable medium having a computer program stored thereon, and when the computer program is executed by a processor, the method for predicting delirium described in the aforementioned embodiment is implemented.

[0021] The present invention has the following beneficial effects:

[0022] The present application found that an increase in the expression level of p-Tau 396 is associated with the risk of developing delirium and / or the risk of prognosis, and can be used as an independent predictor to predict the risk of developing delirium and / or the risk of prognosis in subjects, so that patients can receive timely intervention or treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0024] Figure 1 Plasma tau testing for patients with delirium;

[0025] Figure 2 Comparison of physical function and cognitive function between the delirium group and the control group 30 days after discharge; A is the repeated measurement analysis results of ADL (Barthel index) at different time points of the two groups of patients; B is the repeated measurement analysis results of IADL at different time points of the two groups of patients; C is the repeated measurement analysis results of Frail scores at different time points of the two groups of patients; D is the repeated measurement analysis results of SPMSQ at different time points of the two groups of patients;

[0026] Figure 3 To establish a mouse model of delirium;

[0027] Figure 4 The phosphorylation level of tau protein at serine 396 in the hippocampus of delirium mice was significantly increased;

[0028] Figure 5 Tau knockout mice showed no cognitive impairment after being injected with scopolamine;

[0029] Figure 6 This is a test of cognitive levels in mice by simulating phosphorylation of site 396 (silence A, activation D) by AAV in vivo. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0031] On the one hand, an embodiment of the present invention provides use of a reagent for detecting the expression level of phosphorylated Tau protein in preparing a product for predicting delirium, wherein the phosphorylated Tau protein includes p-Tau 396.

[0032] In some embodiments, the delirium comprises elderly delirium. Elderly may refer to people aged ≥ 60 years old.

[0033] In some embodiments, predicting delirium comprises: predicting the subject's risk of illness and / or risk of prognosis.

[0034] In some embodiments, predicting delirium comprises: predicting the subject's risk of postoperative illness and / or prognosis risk.

[0035] In some embodiments, the product includes: any one of a reagent, a chip, and a kit.

[0036] On the other hand, an embodiment of the present invention provides the use of an agent for reducing the phosphorylation level of the S396 site of the Tau protein in the preparation of a product for preventing, treating or assisting in the treatment of delirium.

[0037] "Treatment" as used herein includes preventing or alleviating a condition, reducing the rate at which a condition develops or progresses, reducing the risk of developing a condition, preventing or delaying the development of symptoms associated with a condition, reducing or terminating symptoms associated with a condition, producing complete or partial reversal of a condition, curing a condition, or a combination of the above.

[0038] In some embodiments, the product includes: any one of a reagent, a chip, and a kit.

[0039] On the other hand, an embodiment of the present invention provides a prediction model training method, which includes:

[0040] Obtaining the detection results of the phosphorylated Tau protein expression level in the training sample and the corresponding annotation results; the phosphorylated Tau protein includes p-Tau 396, and the annotation results include the disease risk and / or prognostic risk of delirium representing the sample;

[0041] Input the obtained test results into the pre-built machine learning model to obtain the prediction results of the samples;

[0042] According to the labeling results and the prediction results, the parameters of the machine learning model are updated to obtain a prediction model.

[0043] It is understandable that the categories and number of training samples can be routinely selected by those skilled in the art, and the total number of training samples and training samples of various categories (such as healthy people and patients) can be ≥ any value among 10, 50, 100, 200, 300, 400 and 500, or a range between any two of them.

[0044] In some embodiments, the label may be a character or a string. The content of the prediction result corresponds to the content of the annotation result.

[0045] The present invention has no particular restrictions on the type of machine learning model, which can be selected from the algorithm model conventionally used in the field for disease risk prediction and / or prognosis risk prediction. In some embodiments, the machine learning model includes any one of logistic regression, SVM, random forest, gradient boosting classifier (GBC), KNN, decision tree and naive Bayes.

[0046] On the other hand, an embodiment of the present invention provides a prediction device, comprising:

[0047] An acquisition module, used to obtain the detection result of the phosphorylated Tau protein expression level of the sample to be tested, wherein the phosphorylated Tau protein includes p-Tau 396;

[0048] The prediction module is used to input the detection result of the sample to be tested into the prediction model trained by the training method described in any of the above embodiments to obtain the prediction result of the sample to be tested.

[0049] The modules described in the embodiments of the present invention may be stored in a memory in the form of software or firmware or solidified in the operating system (OS) of the electronic device provided by the present application, and may be executed by a processor in the electronic device. At the same time, the data and program codes required to execute the above modules may be stored in the memory.

[0050] On the other hand, an embodiment of the present invention provides an electronic device, comprising a processor and a memory, wherein the memory is used to store a program, and when the program is executed by the processor, the processor implements a method for predicting delirium;

[0051] The method for predicting delirium comprises the following steps: obtaining the detection result of the expression level of phosphorylated Tau protein of the sample to be tested, wherein the phosphorylated Tau protein includes p-Tau 396; inputting the detection result of the sample to be tested into the prediction model trained by the training method described in any of the aforementioned embodiments to obtain the prediction result of the sample to be tested.

[0052] An electronic device may include a memory, a processor, a bus, and a communication interface, wherein the memory, the processor, and the communication interface are electrically connected to each other directly or indirectly to achieve data transmission or interaction. For example, these components may be electrically connected to each other via one or more buses or signal lines.

[0053] The memory can be, but is not limited to, random access memory (RAM), read only memory (ROM), programmable read-only memory (PROM), erasable programmable read-only memory (EPROM), electrically erasable read-only memory (EEPROM), etc.

[0054] The processor can be an integrated circuit chip with signal processing capabilities. The processor 120 can be a general-purpose processor, including a central processing unit (CPU), a network processor (NP), etc.; it can also be a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA) or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components.

[0055] The electronic device can be a server, a cloud platform, a mobile phone, a tablet computer, a laptop computer, an ultra-mobile personal computer (UMPC), a handheld computer, a netbook, a personal digital assistant (PDA), a wearable electronic device, a virtual reality device, etc. Therefore, the embodiments of the present application do not limit the type of electronic device.

[0056] In addition, an embodiment of the present invention provides a computer-readable medium having a computer program stored thereon. When the computer program is executed by a processor, the method for predicting delirium described in any of the aforementioned embodiments is implemented.

[0057] In some embodiments, the computer-readable medium may be a general storage medium, such as a mobile disk, a hard disk, etc.

[0058] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0059] Clinical samples used in the following examples: Elderly patients with primary liver cancer admitted to our hospital from March 2020 to January 2023 were retrospectively selected as research subjects. Inclusion criteria: (1) All patients met the diagnostic criteria for primary liver cancer in the "Guidelines for the Diagnosis and Treatment of Primary Liver Cancer (2017 Edition)" and were confirmed by clinical pathology; (2) Age ≥ 60 years old; (3) Clear consciousness before surgery; (4) All underwent radical resection of liver cancer under general anesthesia; (5) No extrahepatic metastasis was found; (6) Child-Pugh liver function grade was A or B; (7) Complete clinical pathological data. Exclusion criteria: (1) Patients with delirium before surgery; (2) Patients with comorbid neuropathic diseases; (3) Patients with comorbid mental illness and unable to communicate normally; (4) Patients taking psychiatric drugs for a long time; (5) Patients with comorbid severe underlying diseases who could not tolerate surgery; (6) Patients with hepatic encephalopathy.

[0060] Diagnostic criteria and assessment methods for delirium:

[0061] The diagnosis of delirium refers to the relevant criteria in the Diagnostic and Statistical Criteria for Mental Disorders (DSM-IV): (1) impaired consciousness (i.e., the patient's clarity of awareness of the environment is reduced), accompanied by reduced concentration, attention or attention shifting ability, and cognitive changes (including disorientation, memory impairment and speech disorders); (2) the occurrence of perceptual disorders that cannot be explained by the current dementia; (3) usually occurring within a short period of time, such as hours or days, and with a certain degree of volatility; (4) discovered through medical history, laboratory tests or physical examination that it is a physiological result directly caused by physical conditions.

[0062] Example 1

[0063] Relying on the National Clinical Research Center for Geriatric Diseases of West China Hospital, 42 patients with delirium were collected from elderly inpatients in the internal medicine department, and 70 inpatients without delirium were matched by age and gender. Blood samples were collected and plasma was separated for tau testing.

[0064] The results showed that the plasma tau protein in the delirium group increased significantly ( Figure 1 Aβ showed no significant difference ( Figure 1 In the middle (B), the increase in tau occurs simultaneously with the onset of delirium and decreases as the delirium ends ( Figure 1 Middle C).

[0065] Example 2

[0066] First, a delirium clinical cohort was established.

[0067] Relying on the National Clinical Medicine Research Center for Geriatric Diseases of West China Hospital, a delirium cohort of elderly patients hospitalized in the internal medicine department was established. The clinical characteristics and baseline blood samples of all patients were collected, and blood samples were collected 1 day after the onset of delirium, 1 week after the onset, and 1 day before discharge (Tables 1 and 2).

[0068] Table 1: General information of patients with medical delirium (quantitative data, N = 830)

[0069]

[0070] Table 2 General information of surgical patients with delirium (n=299)

[0071]

[0072]

[0073] Some discharged patients were followed up one month after discharge. The results showed that the daily living activities, instrumental daily living activities, frailty scores, and cognitive function SPMSQ scores of the patients in the delirium group were lower than those of the patients without delirium ( Figure 2 The establishment of this cohort lays the foundation for further verifying the effect of tau protein on cognitive dysfunction after delirium in the population.

[0074] 2. An animal model of delirium has been established to detect the key role of delirium in the onset of delirium and the signaling pathway related to the hyperphosphorylation of Tau protein.

[0075] A mouse delirium model was established by intraperitoneal injection of scopolamine. Behavioral and protein marker tests showed that scopolamine affected working memory but not long-term memory and motor ability. The cortical marker S100β increased significantly, which was transient and consistent with the clinical characteristics of delirium. Figure 3 .

[0076] 3. The role of specific site hyperphosphorylation in cognitive dysfunction after delirium

[0077] By extracting the proteins from the hippocampus of delirium mice during the onset period, immunoblotting experiments revealed that the phosphorylation level of tau396 in delirium mice was significantly increased compared with that in the control group ( Figure 4 ).

[0078] Using tau knockout transgenic mice and wild-type mice as research subjects, it was found that tau knockout can alleviate cognitive impairment in delirium mice ( Figure 5 ), which indicates that tau protein plays an important mediating role in the onset of delirium in mice and is a necessary condition.

[0079] The work of expressing simulated phosphorylated tau protein in tau knockout mice was achieved through AAV technology. The preparation method of mice with phosphorylated tau at specific sites includes: replacing the serine at the site with alanine (simulating dephosphorylation) and isoleucine (simulating phosphorylation) in vitro through overlap-PCR technology, using AAV8 to construct a site-directed mutation of tau protein to simulate the phosphorylation and dephosphorylation state of a single serine / threonine, cloning it on the AAV packaging vector plasmid, and together with the other two AAV packaging auxiliary plasmids, the corresponding AAV vector was transfected into 293 cells by three plasmids. The high-purity AAV vector was purified by cesium chloride and 2×10 15 / mice dose-localized injection into the CA1 region of the hippocampus of Tau knockout mice.

[0080] The possibility that reduction of phosphorylation levels at a single site could improve the onset of delirium was examined ( Figure 6 ).

[0081] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of a reagent for detecting the expression level of phosphorylated Tau protein in the preparation of a product for predicting delirium, characterized in that: The phosphorylated Tau protein includes p-Tau 396.

2. The use according to claim 1, characterized in that: The delirium includes senile delirium.

3. The use according to claim 1, characterized in that: The predicting of delirium includes: predicting the subject's risk of illness and / or risk of prognosis.

4. The use according to claim 3, characterized in that: The predicting of delirium includes: predicting the risk of illness and / or prognosis of the subject after surgery.

5. Use of an agent for reducing the phosphorylation level of Tau protein S396 site in the preparation of a product for preventing, treating or assisting in the treatment of delirium.

6. A method for training a prediction model, characterized in that: It includes: Obtain the detection results of the phosphorylated Tau protein expression level in the training samples and their corresponding annotation results; The phosphorylated Tau protein includes p-Tau 396, and the annotation result includes the disease risk and / or prognostic risk of delirium representing the sample; Input the obtained test results into the pre-built machine learning model to obtain the prediction results of the samples; According to the labeling results and the prediction results, the parameters of the machine learning model are updated to obtain a prediction model.

7. The training method according to claim 6, characterized in that: The machine learning model includes any one of logistic regression, SVM, random forest, gradient boosting classifier, KNN, decision tree and naive Bayes.

8. A prediction device, characterized in that: It includes: An acquisition module, used to obtain the detection result of the phosphorylated Tau protein expression level of the sample to be tested, wherein the phosphorylated Tau protein includes p-Tau 396; The prediction module is used to input the detection result of the sample to be tested into the prediction model trained by the training method described in claim 6 or 7 to obtain the prediction result of the sample to be tested.

9. An electronic device, characterized in that: It includes a processor and a memory, wherein the memory is used to store a program, and when the program is executed by the processor, the processor implements a method for predicting delirium; The method for predicting delirium comprises the following steps: obtaining a test result of the expression level of phosphorylated Tau protein of a sample to be tested, wherein the phosphorylated Tau protein includes p-Tau 396; The test result of the sample to be tested is input into the prediction model trained by the training method described in claim 6 or 7 to obtain the prediction result of the sample to be tested.

10. A computer-readable medium, characterized in that The computer readable medium stores a computer program, and when the computer program is executed by a processor, the method for predicting delirium as claimed in claim 9 is implemented.

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