System for predicting recurrence of craniopharyngeal tubuloma based on CDK4 CNV amplification

Abstract

The invention discloses a system for predicting recurrence of craniopharyngeal tubuloma based on CDK4 CNV amplification. In the research on patients with recurrent and ectopic recurrent craniopharyngeal tubuloma, the CDK4 gene is found to have a stable copy number gain (CNV) phenomenon for the first time, the proportion of the phenomenon in multiple recurrent patients is obviously higher than that in primary patients, and the recurrent risk of craniopharyngeal tubuloma can be evaluated by detecting the amplification condition of the CDK4 CNV in vascular endothelial cells of tumor tissues. Therefore, the method, the system and the equipment for predicting the recurrence risk of the craniopharyngeal tubuloma have good repeatability and popularization potential, can be expanded into a postoperative pathological sample auxiliary diagnosis tool, and realize layered evaluation of the recurrence risk of a postoperative patient and follow-up strategy optimization.

Description

Technical Field

[0001] The present invention relates to the field of intelligent medical care, and in particular to a system for predicting craniopharyngioma recurrence based on CDK4 CNV amplification. Background Art

[0002] Craniopharyngiomas (CPs) are common intracranial tumors that primarily occur in children and adolescents. These tumors originate from the epithelial cells of the craniopharyngioma and are typically located in the sellar region, closely associated with important structures such as the hypothalamus, pituitary stalk, and optic nerve. Postoperative recurrence is a key characteristic of CPs, but effective recurrence prediction methods are currently lacking. Therefore, developing a method to effectively predict CP recurrence is crucial for CP patients. Summary of the Invention

[0003] To overcome the deficiencies of the prior art, the present invention provides a system for predicting craniopharyngioma recurrence based on CDK4 CNV amplification.

[0004] To achieve the above object, the present invention adopts the following technical solutions:

[0005] A first aspect of the present invention provides a method for predicting the recurrence risk of craniopharyngioma, comprising the following steps:

[0006] Obtain CNV amplification of genes / proteins in the sample to be tested;

[0007] Extracting CNV amplification of target genes / proteins from CNV amplification of the genes / proteins, wherein the target genes / proteins include CDK4;

[0008] The recurrence risk of craniopharyngioma is predicted based on the CNV amplification of CDK4, and a classification result of high or low recurrence risk of craniopharyngioma in the sample to be tested is obtained; if the CNV amplification ratio of CDK4 is high, a result of high recurrence risk of craniopharyngioma in the sample to be tested is obtained; if the CNV amplification ratio of CDK4 is low, a result of low recurrence risk of craniopharyngioma in the sample to be tested is obtained.

[0009] Furthermore, the CNV amplification ratio of CDK4 is the CNV amplification ratio of CDK4 in vascular endothelial cells.

[0010] Furthermore, the CNV amplification ratio of CDK4 was statistically analyzed by image analysis.

[0011] Furthermore, the CDK4 amplification criteria were CDK4 / CEP12>2.0 or the presence of clustered CDK4 signal clusters.

[0012] Furthermore, the CNV amplification ratio of CDK4 = the number of CDK4 amplification-positive vessels / the total number of CD31-positive vessels.

[0013] Furthermore, the craniopharyngioma recurrence is craniopharyngioma recurrence more than 3 times.

[0014] Furthermore, the threshold of the CNV amplification ratio is 3.7%.

[0015] Furthermore, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

[0016] A second aspect of the present invention provides a craniopharyngioma recurrence risk prediction system, the system comprising:

[0017] Acquisition unit: used to obtain CNV amplification status of genes / proteins of the sample to be tested;

[0018] Extraction unit: used for extracting CNV amplification of target genes / proteins from CNV amplification of the genes / proteins, wherein the target genes / proteins include CDK4;

[0019] Prediction unit: predict the recurrence risk of craniopharyngioma based on the CNV amplification of CDK4, and obtain a classification result of high or low recurrence risk of craniopharyngioma of the sample to be tested; if the CNV amplification ratio of CDK4 is high, the result of high recurrence risk of craniopharyngioma of the sample to be tested is obtained; if the CNV amplification ratio of CDK4 is low, the result of low recurrence risk of craniopharyngioma of the sample to be tested is obtained.

[0020] Furthermore, the CNV amplification ratio of CDK4 is the CNV amplification ratio of CDK4 in vascular endothelial cells.

[0021] Furthermore, the CNV amplification ratio of CDK4 was statistically analyzed by image analysis.

[0022] Furthermore, the CDK4 amplification criteria were CDK4 / CEP12>2.0 or the presence of clustered CDK4 signal clusters.

[0023] Furthermore, the CNV amplification ratio of CDK4 = the number of CDK4 amplification-positive vessels / the total number of CD31-positive vessels.

[0024] Furthermore, the craniopharyngioma recurrence is craniopharyngioma recurrence more than 3 times.

[0025] Furthermore, the threshold of the CNV amplification ratio is 3.7%.

[0026] Furthermore, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

[0027] A third aspect of the present invention provides a device for predicting the risk of recurrence of craniopharyngioma, the device comprising:

[0028] Memory: The memory is used to store program instructions;

[0029] Processor: The processor is used to call program instructions, and when the program instructions are executed, it is used to execute the risk prediction method described in the first aspect of the present invention.

[0030] Furthermore, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

[0031] A fourth aspect of the present invention provides a computer-readable storage medium having a computer program stored thereon, which, when executed by a processor, implements the steps of the method described in the first aspect of the present invention.

[0032] A fifth aspect of the present invention provides a computer program product, comprising a computer program, which, when executed by a processor, implements the steps of the method described in the first aspect of the present invention.

[0033] A sixth aspect of the present invention provides the use of a reagent for detecting CDK4 CNV in the preparation of a product for predicting the recurrence risk of craniopharyngioma.

[0034] Furthermore, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

[0035] Furthermore, the reagents include a CDK4 CNV probe and a CDK4 CNV primer.

[0036] Furthermore, the reagent also includes a detectable label.

[0037] A seventh aspect of the present invention provides a product for predicting the recurrence risk of craniopharyngioma, comprising a reagent for detecting CDK4 CNV.

[0038] Furthermore, the reagents include a CDK4 CNV probe and a CDK4 CNV primer.

[0039] Furthermore, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

[0040] Furthermore, the product includes a test kit and a test paper.

[0041] Furthermore, the kit further comprises a detectable label.

[0042] Furthermore, the detectable labels include magnetic beads, fluorescent dyes, radioactive markers, and enzymes.

[0043] Furthermore, the kit also includes instructions.

[0044] Furthermore, the kit further comprises a buffer solution.

[0045] Advantages and beneficial effects of the present invention:

[0046] In a study of patients with recurrent and ectopic recurrent craniopharyngioma, this application discovered for the first time the presence of stable copy number gain (CNV) in the CDK4 gene. This phenomenon was significantly more common in patients with multiple recurrences than in patients with initial onset. By detecting CDK4 CNV amplification in vascular endothelial cells of tumor tissue, the risk of craniopharyngioma recurrence can be assessed. Therefore, this application provides a method, system, and device for predicting the risk of craniopharyngioma recurrence, which has good repeatability and potential for promotion. This method can be expanded into a diagnostic tool for postoperative pathological samples, enabling stratified assessment of recurrence risk and optimization of follow-up strategies for postoperative patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 This is a schematic diagram of the method for predicting the recurrence risk of craniopharyngioma provided in this application;

[0048] Figure 2 This is a schematic diagram of the craniopharyngioma recurrence risk prediction system provided by this application;

[0049] Figure 3 This is a schematic diagram of a craniopharyngioma recurrence risk prediction device provided by this application;

[0050] Figure 4 Figure 4A is a HE staining image of the tumor and immunohistochemical staining of the vascular endothelial cells around the tumor. Figure 4B is a HE staining image of the vascular endothelial cells around the tumor expressing strong positive markers (CD31+ (purple fluorescence), DAPI (blue fluorescence) marks tumor epithelial cells).

[0051] Figure 5 Figures 5A and 5B show HE staining of the tumor and CDK4-DNA fluorescence in situ hybridization (FISH) staining of the vascular endothelial cells surrounding the tumor. Figure 5A shows HE staining of an amelogenin-type craniopharyngioma, and Figure 5B shows CDK4 amplification in some vascular endothelial cells surrounding the tumor (red fluorescence, marked by yellow arrows), while CDK4 expression in some vascular endothelial cells is not significantly amplified (marked by green arrows).

[0052] Figure 6 It is a recurrence risk assessment chart based on the proportion of CDK4 CNV amplified blood vessels. Among them, 6A is a statistical chart between the primary group and the multiple recurrence group, and the proportion of CDK4 CNV amplified blood vessels is significantly increased compared with the multiple recurrence group. 6B is a ROC curve chart constructed based on the training group and the validation group. DETAILED DESCRIPTION

[0053] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention.

[0054] In some of the processes described in the specification and claims of the present invention and the above-mentioned figures, multiple operations that appear in a specific order are included, but it should be clearly understood that these operations may not be executed in the order in which they appear in this article or may be executed in parallel. The serial numbers of the operations, such as 101, 102, etc., are only used to distinguish between different operations, and the serial numbers themselves do not represent any execution order. In addition, these processes may include more or fewer operations, and these operations may be executed in sequence or in parallel. It should be noted that the descriptions of "first", "second", etc. in this article are used to distinguish different messages, devices, modules, etc., and do not represent the order of precedence, nor do they limit "first" and "second" to be different types.

[0055] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making any creative efforts shall fall within the scope of protection of the present invention.

[0056] Figure 1 Schematic diagram of a method for predicting the recurrence risk of craniopharyngioma provided in this application. Specifically, the method comprises the following steps:

[0057] 101: Obtain CNV amplification status of the gene / protein of the sample to be tested.

[0058] 102: Extracting CNV amplification information of target genes / proteins from the CNV amplification information of the genes / proteins, wherein the target genes / proteins include CDK4.

[0059] 103: Predict the recurrence risk of craniopharyngioma based on the CNV amplification of CDK4, and obtain a classification result of high or low recurrence risk of craniopharyngioma of the tested sample; if the CNV amplification ratio of CDK4 is high, the result of high recurrence risk of craniopharyngioma of the tested sample is obtained; if the CNV amplification ratio of CDK4 is low, the result of low recurrence risk of craniopharyngioma of the tested sample is obtained.

[0060] In some embodiments, CDK4 includes wild-type, mutant forms, or fragments thereof. The term encompasses full-length, unprocessed CDK4, any form of CDK4 derived from processing in cells, and naturally occurring variants of CDK4 (e.g., splice variants or allelic variants). The term encompasses, for example, human CDK4 as well as CDK4 from any other vertebrate source, including mammals such as primates and rodents (e.g., mice and rats), Gene ID: 1019.

[0061] In some embodiments, the craniopharyngioma comprises amelogenin-type craniopharyngioma, squamous papillary craniopharyngioma.

[0062] In a specific embodiment, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

[0063] The screening and verification process of the target gene / protein CDK4 in this application is as follows:

[0064] This application collected a total of 118 paraffin-embedded tissue samples of pathologically confirmed enamel cell craniopharyngioma diagnosed after surgery at the Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, between January 2020 and December 2024. Twenty samples of poor quality or incomplete follow-up data were excluded, resulting in a final analysis of 98 cases, including primary (49 cases) and multiple recurrences (49 cases). The samples were randomly divided into groups at a 1:2 ratio, with 33 patients included in the training group (16 multiple recurrences and 17 primary cases) and 65 patients included in the validation group (33 multiple recurrences and 32 primary cases).

[0065] In each sample, the system counted the number of CDK4 CNV-positive blood vessels through image analysis and calculated their proportion in all blood vessels (blood vessels were marked by CD31+), which was defined as the "CDK4 amplified blood vessel ratio". The samples were then randomly divided into a training group and a validation group (1:2), that is, "primary vs. multiple recurrences (multiple recurrences were defined as ≥3 recurrences)" as the grouping criteria for modeling ( Figure 4 、 Figure 5 ).

[0066] ROC curve analysis was used to determine that the optimal segmentation threshold for predicting recurrence was 0.03691454 (i.e., 3.7%). When the proportion of CDK4 amplified blood vessels in a patient is higher than this threshold, it indicates that the patient has a higher risk of recurrence. This indicator has good stability in the training group (AUC = 0.78, sensitivity 82%, specificity 69%) and also showed excellent predictive performance in the validation group (AUC = 0.90, 95%CI: 0.82–0.97; sensitivity 88%, specificity 85%, accuracy 86%). In addition, the positive predictive value (PPV) and negative predictive value (NPV) were 74% and 79% in the training group, and 85% and 88% in the validation group, respectively. Figure 6 , Table 1).

[0067] Table 1 Detailed data information for prediction model construction and validation

[0068]

[0069] In some embodiments, the experimental sample processing and slice preparation method includes:

[0070] Tissue source: Paraffin-embedded surgical tissue of adamantinomatous craniopharyngioma (ACP) obtained clinically.

[0071] Section thickness: Serial sections, each 4 μm thick, numbered as A (for CDK4 / CEP12 FISH) and B (for CD31 immunofluorescence staining).

[0072] Slide type: positively charged slides (Superfrost Plus), numbered to ensure that serial sections correspond to paired analysis areas.

[0073] In some embodiments, the staining method comprises the following detailed steps:

[0074] 1. CDK4 / CEP12 FISH Staining Procedure (Panel A)

[0075] 1) Dewaxing and hydration: Dewaxing in xylene twice, 10 minutes each time; hydrating in 100%, 95%, 85%, and 75% graded alcohol, 2 minutes each time; and finally rinsing with tap water.

[0076] 2) Pretreatment: Pepsin digestion solution (Ambipin, preheated to 37°C) for 10–15 minutes; wash with pure water and air dry.

[0077] 3) Probe hybridization: Add 10 μL of mixed probes (Anbiping CDK4 (12q14) gene amplification probe) (CDK4: red Cy3; CEP12: green FITC); seal with a coverslip and denature at 95°C for 5 minutes in a hybridizer; then hybridize at 37°C for 16 hours.

[0078] 4) Post-hybridization washes: 0.4× SSC / 0.3% NP-40 at 72°C for 5 minutes; 2× SSC / 0.1% NP-40 at room temperature for 1 minute. After drying, add DAPI (nuclear stain) and mount the slides.

[0079] 2. CD31 Immunofluorescence Staining Procedure (Panel B)

[0080] 1) Deparaffinization, hydration, and antigen retrieval: Deparaffinization and hydration were performed as above. Antigen retrieval solution: EDTA buffer (pH 9.0) was used and microwave heating was performed for 10 minutes.

[0081] 2) Blocking and Antibody Incubation: Block with 5% BSA for 30 minutes; Primary antibody: Anti-CD31 Mouse Monoclonal Antibody (CST# 3528) (1:100, overnight at 4°C); Wash the next day and add fluorescent secondary antibody (Alexa Fluor 620, 1:500, 1 hour at room temperature); Stain nuclei with DAPI, seal the slides, and store in dark.

[0082] In some embodiments, the image acquisition device and parameter method include:

[0083] Microscope platform: Olympus VS200 digital pathology slide scanner or Leica DMi8 confocal system.

[0084] Objective magnification: 40× oil objective or 63× oil objective.

[0085] Acquisition channels: DAPI (blue); FITC (CEP12 green); Cy3 (CDK4 red); Alexa Fluor 620 (CD31 purple).

[0086] Slice scanning format: Export in TIFF format for subsequent image analysis.

[0087] In some embodiments, image analysis and cell counting software (ImageJ) includes:

[0088] The software used was ImageJ Fiji 1.53t; the Cell Counter plug-in was used for manual annotation and counting; the ROIManager was used to distinguish the field of view; and the Bio-Formats Importer plug-in was used to assist in loading high-resolution images.

[0089] Image analysis steps:

[0090] A. CD31 slice image analysis (B):

[0091] 1) Open the CD31 staining image (Alexa Fluor 620 channel).

[0092] 2) Identification of continuous vascular structures (CD31+): purple markers appear as tubular or elliptical closed loop structures.

[0093] 3) Manually count the number of vascular units (each closed-loop structure is considered as one vessel).

[0094] 4) Count multiple fields of view for each slice (5–10 fields of view are recommended) and average the total number of vessels.

[0095] B. CDK4 FISH slide image analysis (Slide A):

[0096] 1) Open the CDK4 / CEP12 FISH image (activate the Cy3 and FITC channels, respectively).

[0097] 2) Identify endothelial cell nuclei in the structural control area (based on their position and morphology in serial sections).

[0098] 3) Count the number of CDK4 signals (red dots) and CEP12 signals (green dots) in each endothelial cell nucleus.

[0099] 4) CDK4 amplification criteria: CDK4 / CEP12 > 2.0; or the presence of clustered CDK4 signal clusters (≥6 compact red signals).

[0100] 5) Count the number of blood vessels that meet the above criteria (CDK4 amplification-positive blood vessels).

[0101] The ratio was calculated as follows: CDK4 amplified blood vessel ratio = number of CDK4 amplified positive blood vessels / total number of CD31 positive blood vessels × 100%.

[0102] In some embodiments, the threshold calculation process includes assigning class labels to samples based on recurrence status (primary vs. multiple recurrences, defined as ≥3 recurrences). A receiver operating characteristic (ROC) curve was constructed using the pROC package in R software to evaluate the predictive performance of the "proportion of CDK4-amplified vessels" in distinguishing between the two groups. To identify the optimal predictive threshold, the coords() function was used, and the optimal discrimination criterion was set as maximizing the Youden index. This optimal threshold was determined to achieve the best balance between sensitivity and specificity. The optimal threshold ultimately determined by this method was 0.03691454 (i.e., 3.7%). When the CDK4-amplified vessel proportion in a patient is above this threshold, it indicates a higher risk of recurrence. This threshold performed consistently in the training cohort (AUC = 0.78, sensitivity 82%, specificity 69%) and demonstrated even better predictive performance in the validation cohort (AUC = 0.90, sensitivity 88%, specificity 85%).

[0103] In some embodiments, the specific calculation process and code include:

[0104] 1) Sample labeling and grouping

[0105] All cases were grouped according to the number of surgeries and assigned category labels (primary group = 0, multiple recurrence group = 1) for subsequent ROC analysis.

[0106] 2) ROC curve construction

[0107] The analysis was performed using R language v4.3.0 and the pROC package. The input variable was the "proportion of CDK4-amplified vessels" (continuous variable) for each patient, and the categorical label was the risk of recurrence (0 or 1). Execute the following code:

[0108] library (pROC)

[0109] roc_obj <- roc(response = group, predictor = cdk4_ratio)

[0110] 3) Threshold determination and optimization

[0111] The coords() function is used to extract the optimal cutoff from the ROC curve. The optimization criterion is set to maximize the Youden Index (i.e., maximize Sensitivity + Specificity – 1) to achieve the best balance between sensitivity and specificity:

[0112] opt_threshold <- coords(roc_obj, x = "best", best.method = "youden").

[0113] The optimal threshold finally obtained was: CDK4 amplified blood vessel ratio = 0.03691454 (i.e. 3.7%).

[0114] Figure 2 The present application provides a craniopharyngioma recurrence risk prediction system, specifically, the system includes:

[0115] Acquisition unit 201: used to obtain CNV amplification status of genes / proteins of the sample to be tested.

[0116] Extraction unit 202: used for extracting CNV amplification status of target genes / proteins from CNV amplification status of the genes / proteins, wherein the target genes / proteins include CDK4.

[0117] Prediction unit 203: Predict the recurrence risk of craniopharyngioma based on the CNV amplification of CDK4, and obtain a classification result of high or low recurrence risk of craniopharyngioma of the sample to be tested; if the CNV amplification ratio of CDK4 is high, the result of high recurrence risk of craniopharyngioma of the sample to be tested is obtained; if the CNV amplification ratio of CDK4 is low, the result of low recurrence risk of craniopharyngioma of the sample to be tested is obtained.

[0118] Figure 3 The present application provides a device for predicting the recurrence risk of craniopharyngioma, specifically comprising:

[0119] Memory: The memory is used to store program instructions.

[0120] Processor: The processor is used to call program instructions, and when the program instructions are executed, it is used to execute the above-mentioned craniopharyngioma recurrence risk prediction method.

[0121] The present invention provides a product for predicting the recurrence risk of craniopharyngioma, which comprises a reagent for detecting CDK4 CNV.

[0122] The reagents include a CDK4 CNV probe and a CDK4 CNV primer.

[0123] In some embodiments, the terms "primer" and "amplification primer" are used interchangeably. A primer refers to an oligonucleotide that hybridizes to a target nucleic acid or its complement and participates in a nucleic acid amplification reaction. An amplification primer hybridizes to a template nucleic acid and has a 3'-OH (3'-hydroxyl) group that is extendable by polymerization. In some embodiments, an amplification primer is 15-60, 15-55, 15-50, 15-45, 15-40, 15-35, or 15-30 bases in length. In some embodiments, an amplification primer is 18-30 bases in length. In some embodiments, an amplification primer is 18-60 bases in length and comprises 18-30 contiguous bases (target hybridization region) that hybridize to a corresponding 18-30 nucleotide oligonucleotide hybridization sequence present in the target nucleic acid sequence. In some embodiments, the 18-30 contiguous bases are located at the 3' end of the amplification primer.

[0124] In some embodiments, a probe refers to an oligonucleotide capable of hybridizing to a target nucleic acid of interest. Depending on the stringency of the hybridization conditions, the probe will typically substantially form a chemical bond between the target sequence lacking complete complementarity and the probe sequence. The probe can be associated with an appropriate tag or reporter moiety so that the probe (and its target) can be detected, visually observed, measured, and / or assayed.

[0125] The products include test kits and test strips.

[0126] The kit also includes a detectable label.

[0127] In some embodiments, suitable detectable labels include, but are not limited to, magnetic beads (e.g., Dynabeads™), fluorescent dyes (e.g., fluorescein isothiocyanate, Texas red, rhodamine, green fluorescent protein, red fluorescent protein, yellow fluorescent protein, etc.), radioactive labels (e.g., 3 H, 125 I, 35 S, 14 C, or 32 P), enzymes (e.g., horseradish peroxidase, alkaline phosphatase, luciferase, and others commonly used in enzyme-linked immunosorbent assays (ELISAs)), and colorimetric labels such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads.

[0128] In some embodiments, a kit refers to any delivery system for delivering materials. In the context of nucleic acid purification systems and reaction assays, such delivery systems include systems that allow for the storage, transport, or delivery of reagents and devices (e.g., chaotropic salts, particles, buffers, denaturants, oligonucleotides, filters, etc. in appropriate containers) and / or support materials (e.g., sample processing or sample storage containers, written instructions for performing the procedure, etc.) from one location to another. For example, a kit includes one or more housings (e.g., boxes) containing the relevant reaction reagents and / or support materials.

[0129] In some embodiments, the kits include fragmentation kits and combination kits. A fragmentation kit refers to a delivery system that includes two or more separate containers, each containing a subset of all the kit components. The containers can be delivered to the intended recipient together or individually. For example, a first container may contain materials and a buffer for sample collection, while a second container contains a capture oligonucleotide and a denaturant. A combination kit refers to a delivery system that contains all the components of a reaction assay in a single container (e.g., a single box containing each required component).

[0130] In the several embodiments provided in this application, it should be understood that the disclosed systems, devices and methods can be implemented in other ways. For example, the device embodiments described above are merely schematic. For example, the division of the units is merely a logical function division. In actual implementation, there may be other division methods, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be an indirect coupling or communication connection through some interfaces, devices or units, which can be electrical, mechanical or other forms.

[0131] The units described as separate components may or may not be physically separate, and the components shown as units may or may not be physical units, that is, they may be located in one place or distributed across multiple network units. Some or all of these units may be selected to achieve the purpose of this embodiment according to actual needs.

[0132] In addition, the functional units in the various embodiments of the present invention may be integrated into a single processing unit, each unit may exist physically separately, or two or more units may be integrated into a single unit. The aforementioned integrated units may be implemented in the form of hardware or software functional units.

[0133] Those skilled in the art will appreciate that all or part of the steps in the various methods of the above embodiments can be completed by instructing related hardware through a program, and the program can be stored in a computer-readable storage medium, which may include: a read-only memory (ROM), a random access memory (RAM), a disk or an optical disk, etc.

[0134] Those skilled in the art will understand that all or part of the steps in the above-mentioned embodiment method can be implemented by instructing the relevant hardware through a program, and the program can be stored in a computer-readable storage medium. The above-mentioned storage medium can be a read-only memory, a disk or an optical disk, etc.

[0135] The above embodiments are only provided for understanding the method and core concept of the present invention. It should be noted that, without departing from the principles of the present invention, a number of improvements and modifications may be made to the present invention by a person skilled in the art, and such improvements and modifications shall fall within the scope of protection of the claims of the present invention.

Claims

1. A method for predicting the recurrence risk of craniopharyngioma, characterized in that: The method comprises the following steps: Obtain CNV amplification of genes / proteins in the sample to be tested; Extracting CNV amplification of target genes / proteins from CNV amplification of the genes / proteins, wherein the target genes / proteins include CDK4; Predicting the recurrence risk of craniopharyngioma based on the CNV amplification of CDK4, and obtaining a classification result of the high or low recurrence risk of craniopharyngioma in the tested sample; If the CNV amplification ratio of CDK4 is high, a result is obtained that the risk of craniopharyngioma recurrence of the tested sample is high. If the CNV amplification ratio of CDK4 is low, a result is obtained that the risk of craniopharyngioma recurrence of the tested sample is low.

2. The prediction method according to claim 1, characterized in that The CNV amplification ratio of CDK4 is the CNV amplification ratio of CDK4 in vascular endothelial cells; Preferably, the CNV amplification ratio of CDK4 is statistically analyzed by image analysis; Preferably, the CDK4 amplification criteria are CDK4 / CEP12>2.0 or the presence of clustered CDK4 signal clusters; Preferably, the ratio of CNV amplification of CDK4 = the number of CDK4 amplification-positive blood vessels / the total number of CD31-positive blood vessels; Preferably, the craniopharyngioma recurrence is craniopharyngioma recurrence more than 3 times; Preferably, the threshold value of the CNV amplification ratio is 3.7%; Preferably, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

3. A craniopharyngioma recurrence risk prediction system, characterized in that: The system comprises: Acquisition unit: used to obtain CNV amplification status of genes / proteins of the sample to be tested; Extraction unit: used for extracting CNV amplification of target genes / proteins from CNV amplification of the genes / proteins, wherein the target genes / proteins include CDK4; Prediction unit: predicting the recurrence risk of craniopharyngioma based on the CNV amplification of CDK4, and obtaining a classification result of high or low recurrence risk of craniopharyngioma in the sample to be tested; if the CNV amplification ratio of CDK4 is high, a result of high recurrence risk of craniopharyngioma in the sample to be tested is obtained; if the CNV amplification ratio of CDK4 is low, a result of low recurrence risk of craniopharyngioma in the sample to be tested is obtained; Preferably, the CNV amplification ratio of CDK4 is the CNV amplification ratio of CDK4 in vascular endothelial cells; Preferably, the CNV amplification ratio of CDK4 is statistically analyzed by image analysis; Preferably, the CDK4 amplification criteria are CDK4 / CEP12>2.0 or the presence of clustered CDK4 signal clusters; Preferably, the ratio of CNV amplification of CDK4 = the number of CDK4 amplification-positive blood vessels / the total number of CD31-positive blood vessels; Preferably, the craniopharyngioma recurrence is craniopharyngioma recurrence more than 3 times; Preferably, the threshold value of the CNV amplification ratio is 3.7%; Preferably, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

4. A device for predicting the risk of recurrence of craniopharyngioma, characterized in that: The device comprises: Memory: The memory is used to store program instructions; Processor: The processor is used to call program instructions, and when the program instructions are executed, it is used to execute the risk prediction method according to claim 1 or 2; Preferably, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

5. A computer-readable storage medium, characterized in that A computer program is stored thereon, and when the computer program is executed by a processor, the steps of the method according to claim 1 or 2 are implemented.

6. A computer program product comprising a computer program, characterized in that When the computer program is executed by a processor, the steps of the method according to claim 1 or 2 are implemented.

7. Application of CDK4 CNV detection reagents in the preparation of products for predicting the recurrence risk of craniopharyngioma; Preferably, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

8. The use according to claim 7, characterized in that The reagents include a CDK4 CNV probe and a CDK4 CNV primer; Preferably, the reagent further comprises a detectable label.

9. A product for predicting the recurrence risk of craniopharyngioma, characterized in that: The product includes reagents for detecting CDK4 CNV; Preferably, the reagents include a CDK4 CNV probe and a CDK4 CNV primer; Preferably, the craniopharyngioma is selected from amelogenin-type craniopharyngioma.

10. The product according to claim 9, characterized in that The products include test kits and test strips; Preferably, the kit further comprises a detectable label; Preferably, the detectable label comprises magnetic beads, fluorescent dyes, radioactive markers, enzymes; Preferably, the kit further comprises instructions; Preferably, the kit further comprises a buffer.

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