Application of CIAPIN1 in diagnosis or prognosis of glioma and related computer readable medium

Abstract

The invention discloses application of a reagent for detecting the expression level of CIAPIN1 in preparation of a kit for diagnosis and / or prognosis of glioma of an object. The invention also discloses a kit, a method and a computer readable medium for diagnosis and / or prognosis of glioma. According to the invention, the diagnosis and / or prognosis of glioma can be efficiently and accurately provided by detecting CIAPIN1.

Description

Technical Field

[0001] The present invention relates to a kit, a method and a computer-readable medium for diagnosis and / or prognosis of glioma, and more particularly, to providing diagnosis and / or prognosis of glioma by detecting CIAPIN1 expression. Background Art

[0002] Glioma is a tumor that occurs in the brain and spinal cord, accounting for about 30% of all primary brain tumors and 80% of all malignant brain tumors. The clinical characteristics of glioma are high morbidity, high postoperative recurrence rate and low cure rate.

[0003] According to the degree of morphological similarity between tumor cells and normal brain glial cells, gliomas can be divided into astrocytomas (astrocytes), oligodendrogliomas (oligodendroglial cells), ependymomas (ependymal cells) and mixed gliomas (such as oligodendrogliomas and astrocytomas, including mixed glial cells). According to the molecular pathological characteristics and biological behavior of the tumor, the malignancy of gliomas can be divided into WHO grades I to IV from low to high. Grade I gliomas are usually benign. The average survival time of grade II is about 7 years. Grade II gliomas can progress to grade III (high-grade gliomas) and eventually grade IV (secondary glioblastomas), of which grade IV glioblastomas account for more than 50%, have the highest degree of malignancy, and have an extremely poor prognosis. Grade III and IV gliomas are collectively referred to as high-grade gliomas (HGG), including anaplastic astrocytomas, anaplastic oligodendrogliomas, and the most malignant glioblastoma (GBM). HGG is the most common invasive primary intracranial tumor. It is often treated with surgery and radiotherapy and chemotherapy in clinical practice, but the treatment effect is still not ideal and recurrence is very likely to occur.

[0004] At present, the auxiliary diagnostic molecular markers for gliomas include IDH, MGMT, EGFR, P53, PTEN, etc. Somatic mutations of IDH1 / 2, genomic amplification of EGFR, and co-deletion of chromosome arms 1p and 19q are usually examined for accurate diagnosis. IDH1 / 2 mutations are common in low-grade gliomas and secondary glioblastomas, and it is well known that it has a profound impact on the extended survival of glioma patients. In addition, about 50% of glioblastomas show focal amplification of EGFR. Furthermore, EGFR gene amplification is known to promote cell proliferation, so clinical trials of EGFR inhibitors are often used for cancer treatment.

[0005] In this context, in order to effectively diagnose gliomas, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) are usually used for pathological diagnosis of target mutations of IDH1, a gene considered essential for glioma proliferation, and focal amplification of EGFR. However, there is an increasing demand for effective diagnosis of gliomas in a simpler manner.

[0006] CIAPIN1 (cytokine induced apoptosis inhibitor 1) is a protein that prevents apoptosis or programmed cell death. A series of reports have shown that CIAPIN1 has no homology with Bcl-2, caspases, the IAP family, or other signal transduction molecules. The expression of CIAPIN1 is dependent on the stimulation of growth factors. It is naturally present in various organs, especially those that can regenerate or proliferate. The gene ID of CIAPIN1 in NCBI is 57019, and the reference number of the encoded protein in UniProt is Q6FI81.

[0007] CIAPIN1 is a regulator and important effector in the non-homologous RAS signaling pathway, and is closely related to apoptosis and multidrug resistance in a variety of malignant tumors, such as gastric cancer and cholangiocarcinoma. CIAPIN1 has also been shown to play a key role in malignant tumors such as gastric cancer, hepatocellular carcinoma, and renal cancer. In addition, CIAPIN1 was found to be ubiquitous in normal fetal and tumor tissues, and highly expressed in active metabolic tissues. Therefore, CIAPIN1 may be involved in important physiological functions of cancer.

[0008] However, the expression and function of CIAPIN1 have never been reported in glioma cells. So far, there has been no research or report on the application of CIAPIN1 in the diagnosis and / or prognosis of glioma. Summary of the invention

[0009] The inventors of the present application unexpectedly discovered that CIAPIN1 is significantly overexpressed in glioma tissues relative to normal tissues, and that overexpression of CIAPIN1 is significantly associated with poor prognosis of gliomas. The inventors used reagents that can bind to the expression product of CIAPIN1 to detect the presence and / or level of CIAPIN1 in brain tissues, thereby efficiently and accurately providing diagnosis and / or prognosis of gliomas.

[0010] In a first aspect, the present invention provides a kit for diagnosis and / or prognosis of glioma in a subject, the kit comprising a reagent for detecting the expression level of CIAPIN1.

[0011] In a second aspect, the present invention provides use of a reagent for detecting the expression level of CIAPIN1 in preparing a kit for diagnosis and / or prognosis of glioma in a subject.

[0012] In a third aspect, the present invention provides a reagent for the expression level of CIAPIN1 for use in the diagnosis and / or prognosis of glioma in a subject.

[0013] In a fourth aspect, the present invention provides a method for diagnosing and / or prognosing a glioma in a subject, the method comprising detecting the expression level of CIAPIN1 in a sample from the subject. In some embodiments, the method comprises contacting a sample from the subject with an agent capable of binding to an expression product of CIAPIN1; detecting the presence and / or level of a complex formed by the agent and the expression product of CIAPIN1 in the sample after contact; and providing a diagnosis and / or prognosis of a glioma in the subject based on the presence and / or level of the complex.

[0014] In a fifth aspect, the present invention provides a computer-readable storage medium having stored thereon computer instructions for being read and executed by a computer, wherein the computer instructions are executed to perform a method for diagnosing and / or prognosing a glioma of a subject, the method comprising: (a) contacting a sample from the subject with a reagent capable of binding to an expression product of CIAPIN1; (b) detecting and reading a signal of the contacted sample to determine whether the reagent forms a complex with the expression product of CIAPIN1 in the sample; and (c) determining whether the signal exceeds a predetermined threshold, and providing a diagnosis and / or prognosis of the glioma of the subject when the signal exceeds the predetermined threshold.

[0015] In a sixth aspect, the present invention provides a method for treating a glioma in a subject, the method comprising: (a) contacting a sample from the subject with an agent capable of binding to an expression product of CIAPIN1; (b) detecting and reading a signal of the sample after contact to determine whether the agent forms a complex with the expression product of CIAPIN1 in the sample; and (c) determining whether the signal exceeds a predetermined threshold, and when the signal exceeds the predetermined threshold, providing a diagnosis and / or prognosis of the glioma in the subject; and (d) administering an anti-tumor therapy to a subject determined to have a glioma. In a preferred embodiment, the anti-tumor therapy is temozolomide.

[0016] In any of the above aspects, the difference between the level of CIAPIN1 in the sample from the subject and the control sample level or control value provides a diagnosis and / or prognosis of a glioma in the subject. In some embodiments, a level of CIAPIN1 in the sample from the subject that is higher than the level of a control without the disease indicates that the subject suffers from a glioma. In some embodiments, a higher expression level of CIAPIN1 in the sample from the subject indicates a poor prognosis for a glioma in the subject.

[0017] In any of the above aspects, the reagent for detecting the expression level of CIAPIN1 includes a reagent capable of binding to an expression product of CIAPIN1 to detect the expression level of CIAPIN1 in the sample. In some embodiments, the reagent capable of binding to an expression product of CIAPIN1 includes a protein, a nucleic acid or a small molecule compound. In some embodiments, the reagent capable of binding to an expression product of CIAPIN1 is an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof, or an anti-CIAPIN1 polyclonal antibody.

[0018] In some embodiments, the agent capable of binding to the expression product of CIAPIN1 is labeled with a detectable marker. In some embodiments, the detectable marker is selected from fluorescent markers, chemiluminescent markers, paramagnetic markers, radioisotope markers and enzyme markers.

[0019] In any of the above aspects, the sample is brain tissue. In any of the above aspects, the glioma is selected from astrocytoma, oligodendroglioma, ependymoma and mixed glioma. In some embodiments, the glioma is selected from anaplastic astrocytoma, anaplastic oligodendroglioma and glioblastoma. In some embodiments, the glioma is glioblastoma.

[0020] Other aspects and advantages of the present invention will be described in detail in the specific embodiments, and those skilled in the art will be able to recognize other aspects and advantages that are not explicitly stated in the present disclosure from the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 : The expression level of CIAPIN1 was jointly analyzed by TCGA database and GTEx database. 1157 normal human cortical tissues in the database were classified as the normal group, and 689 gliomas were classified as the tumor group. The results showed that CIAPIN1 was significantly overexpressed in the tumor group (***: P<0.001).

[0022] Figure 2 :Analysis of CIAPIN protein expression level using HPA database. IHC staining images of CIAPIN1 in human cortex and glioma tissues downloaded from the HPA database showed that CIAPIN1 was significantly overexpressed in glioma tumor tissues.

[0023] Figure 3:Prognostic analysis based on CIAPIN1 expression. According to the expression of CIAPIN1 in glioma patient tissues, the samples were divided into CIAPIN1 high expression group and CIAPIN1 low expression group according to the median expression value of CIAPIN1. The overall survival (OS) and progression-free survival (PFS) survival analysis of the CIAPIN1 high expression and low expression groups were performed using the Kaplan-Meier survival analysis method. The results showed that the expression of CIAPIN1 was positively correlated with the poor prognosis of glioma patients.

[0024] Figure 4 :Analysis of the correlation between CIAPIN1 and immune cells. Analysis of the correlation between CIAPIN1 expression and effector immune cell subsets such as NK cells (natural killer cells), pDC cells (plasmacytoid dendritic cells), T Helper cells (helper T cells), and Th2 cells (helper T cells 2).

[0025] Figure 5 : The ROC curve analysis method was used to evaluate the accuracy of CIAPIN1 expression in predicting glioma occurrence. DETAILED DESCRIPTION definition

[0026] Throughout the specification and claims, unless the context indicates otherwise, the word "comprise" and variations such as "include" and "contain" will be understood to include the stated integers, steps or components but not to exclude any other integers, steps or components. When used herein, the term "comprise" may be replaced with the term "include" or "contain", or the term "have" is sometimes used herein instead.

[0027] In the present invention, "about" means that the value is within the acceptable error range of the specific value determined by a person of ordinary skill in the art, and the value depends in part on how it is measured or determined (i.e., the limits of the measurement system). For example, in each practice in the art, "about" can mean within 1 or more than 1 standard deviation. Alternatively, "about" or "substantially including" can mean a range of up to 20%. In addition, for biological systems or processes, the term can mean up to an order of magnitude or up to 5 times the value. Unless otherwise stated, when a specific value appears in the application and claims, the meaning of "about" or "substantially including" should be assumed to be within the acceptable error range of the specific value.

[0028] As used herein, the term "subject", "patient" or "individual" refers to any object for which diagnosis, prognosis or treatment is desired, particularly a mammalian subject. Mammals include humans, domestic animals, farm animals, zoo animals, competitive animals or pets, such as dogs, cats, pigs, rabbits, rats, mice, horses, cattle, cows, etc. The subject referred to herein is preferably a human. In some embodiments, the subject suffers from or is susceptible to one or more disorders or conditions. The patient may exhibit one or more symptoms of a disorder or condition, or may have been diagnosed with one or more disorders or conditions. In some embodiments, the patient is receiving or has received a therapy for diagnosing and / or treating such diseases, disorders or conditions.

[0029] As used herein, the term "detecting" includes any means of detection, including direct and indirect detection, quantitative and qualitative detection, and means identifying the presence and / or level of a specific molecule (eg, CIAPIN1 mRNA or protein) in a subject or in a sample from a subject.

[0030] As used herein, the term "diagnosis" refers to the identification or classification of a molecular or pathological state, disease or condition. For example, "diagnosis" may refer to the identification of a glioma or the identification of its specific type.

[0031] As used herein, the term "prognosis" refers to the prediction of the possible outcome of a disease and the prospect of recovery from the disease according to the nature and symptoms of the case. Therefore, a negative or poor prognosis is defined as a lower post-treatment survival or survival rate. On the contrary, a positive or good prognosis is defined as an improved post-treatment survival or survival rate. Prognosis is usually provided with progression-free survival (PFS) or overall survival (OS). "Poor prognosis" generally refers to a shorter progression-free survival or overall survival.

[0032] Progression-free survival (PFS) used in the context of the present invention should be defined as the length of time during and after the treatment of a disease (e.g., cancer) that a patient coexists with the disease but the disease does not worsen (i.e., survival without disease progression). PFS should be further defined as the time from random assignment (e.g., in a clinical trial) to disease progression or death from any cause. In clinical trials, measuring PFS is a method of observing the effect of new therapies. The term "overall survival" (OS) used in the context of the present invention should be defined as the duration that a patient is still alive from the date of diagnosis or initiation of treatment of a specific disease. Therefore, in clinical trials, the measurement of overall survival will compare the number of patients who died and did not die.

[0033] The term "expression" or "gene expression" refers to RNA production alone or RNA production and RNA translation into protein or polypeptide. Therefore, the term "expression product" includes (i) polynucleotides, including RNA transcripts and corresponding nucleic acids (e.g., hnRNA, mRNA, siRNA or miRNA), and (ii) proteins encoded by RNA transcripts. Gene expression can be measured at the RNA level or protein level by any method known in the art, including but not limited to Northern blots, RT-PCR, Western blots, or in vitro, in situ or in vivo protein activity assays. In the case of CIAPIN1, the expression product is, for example, CIAPIN1 mRNA (mRNA transcribed from the CIAPIN1 gene) or CIAPIN1 protein (protein translated after transcription of the CIAPIN1 gene).

[0034] The term "expression level" refers to the amount of a gene product present in vivo or in a sample at a specific time point. The expression level can be measured / quantified / detected, for example, by a protein or mRNA expressed by a gene. The expression level can be quantified, for example, as follows: the amount of the target gene product present in a normalized sample of the total amount (total protein or mRNA) of the same type of gene product in the same sample or reference sample (for example, a sample obtained from the same individual at the same time or a portion of the same size (weight, volume) of the same sample), or the sample size (weight, volume, etc.) of the amount / limitation of the target gene product is determined. The expression level can be measured or detected by any method known in the art, such as a method (such as mass spectrometry) for direct detection and quantification of the target gene product, or a method for indirect detection and measurement of the target gene product that is usually combined with one or more different molecules or detection devices (such as primers, probes, antibodies, protein scaffolds) that are specific to the target gene product. It is also known to the skilled person to determine the level of gene copies, which also includes determining the absence or presence of one or more fragments (e.g., by nucleic acid probes or primers, such as quantitative PCR, Multiplex ligation-dependent probe amplification (MLPA) PCR).

[0035] TCGA (The Cancer Genome Atlas) is the largest available dataset for tumor genome analysis. It contains more than 10,000 pre-processed samples of more than 30 cancer types, including RNA sequencing (RNA-seq), DNA methylation, copy number variation and other measurements.

[0036] GTEx (Genotype-Tissue Expression) simultaneously performed transcriptome sequencing and genotyping analysis on samples from multiple combinations and organs of the human body, constructed a tissue-specific database of gene expression and regulation, and studied the relationship between genetic variation and gene expression in various human tissues.

[0037] The Human Protein Atlas (HPA) database is a comprehensive human protein information resource that aims to provide a detailed description of human gene and protein expression patterns. The database integrates proteomic data from a variety of tissues and cell types, including immunohistochemistry, immunohistochemistry, and high-throughput antibody preparation technology.

[0038] According to the present invention, the term "binding" preferably relates to specific binding. "Specific binding" means that an agent binds more strongly to a specific target than to another target. If the dissociation constant (K) of the agent binding to the first target is D ) is less than the dissociation constant for the second target, then it binds more strongly to the first target than to the second target. Preferably, the dissociation constant (K D ) compared to the dissociation constant (K D ) is more than 10 2 times, 10 3 times, 10 4 times, 10 5 times, 10 6 times, 10 7 times, 10 8 times, 10 9 times or 10 10 times lower.

[0039] Preferably, an agent (e.g., a protein or polypeptide) is specific for a predetermined target if it is capable of binding to the predetermined target but it is not capable of binding to other targets, i.e. it has no significant affinity for other targets and does not bind significantly to other targets in standard assays. According to the present invention, an agent is specific for the CIAPIN1 protein if it is capable of binding to the CIAPIN1 protein but is (substantially) not capable of binding to other targets. Preferably, an agent is specific for the CIAPIN1 protein if its K for binding to the predetermined target is D is the K that binds to its nonspecific target D At least 10 2 times, 10 3 times, 10 4 times, 10 5 times, 10 6 times, 10 7 times, 10 8 times, 109 times or 10 10 fold lower, then the agent is specific for the target.

[0040] The binding of an agent to a target can be determined experimentally using any suitable method, which is within the scope of those skilled in the art. Affinity can be readily determined using conventional techniques, such as by equilibrium dialysis; by using surface plasmon resonance analysis using the general procedures outlined by the manufacturer; by radioimmunoassay using a radiolabeled target antigen; or by other methods known to the skilled person. Affinity data can be analyzed, for example, by methods known in the art. The measured affinity of a particular interaction may vary if measured under different conditions (e.g., salt concentration, pH). Therefore, affinity and other binding parameters (e.g., K D 、IC 50 ) is preferably measured using standardized solutions of binding agent and target and a standardized buffer.

[0041] As used herein, the term "antibody" refers to any form of an antibody that exhibits a desired biological activity (e.g., inhibiting the binding of a ligand to its receptor or by inhibiting ligand-induced receptor signal transduction). "Antibody fragment" and "antigen-binding fragment" refer to antigen-binding fragments of antibodies and antibody analogs, which typically include at least a portion of the antigen-binding region or variable region (e.g., one or more CDRs) of the parent antibody. In some embodiments, the antibody is a monoclonal antibody. In other embodiments, the antibody is a polyclonal antibody.

[0042] The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies constituting the population are identical except for possible natural mutants that may be present in small amounts. Monoclonal antibodies are highly specific and can be directed against a single antigenic site. In addition, in contrast to conventional (polyclonal) antibody preparations that typically include a variety of different antibodies directed against a plurality of different determinants (epitopes), each monoclonal antibody is directed against only a single determinant on the antigen. The modifier "monoclonal" indicates the characteristic of an antibody obtained from a substantially homogeneous antibody population and is not to be construed as requiring the preparation of the antibody by any particular method. For example, the monoclonal antibodies used in the present invention can be prepared by hybridoma or recombinant DNA methods.

[0043] Monoclonal antibodies can include "chimeric" antibodies, humanized antibodies, or fully human antibodies. In some embodiments, the antibody constitutes part of a larger biomolecule, such as a fusion protein or an antibody drug conjugate. Antibody fragments retain at least some of the binding specificity of the parent antibody. Typically, when the activity is expressed on a molar basis, the antibody fragment retains at least 10% of the parent binding activity. Preferably, the antibody fragment retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the binding affinity of the parent antibody to the target.

[0044] "Antibody fragment" and "antigen-binding fragment" refer to antigen-binding fragments of antibodies and antibody analogs, which generally include at least a portion of the antigen-binding region or variable region (e.g., one or more CDRs) of the parent antibody. Antibody fragments retain at least some of the binding specificity of the parent antibody. Typically, when activity is expressed on a molar basis, antibody fragments retain at least 10% of the parent binding activity. Preferably, the antibody fragment retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the binding affinity of the parent antibody to the target. Examples of antibody fragments include, but are not limited to: Fab, Fab', F(ab')2 and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, such as scFv (single chain variable fragment); nanobodies; domain antibodies; and multispecific antibodies formed from antibody fragments. Antibodies against CIAPIN1 refer to antibodies that specifically bind to the CIAPIN1 protein, including artificially designed antibodies, and any form of antibodies, such as antibody fragments and antigen-binding fragments as defined above.

[0045] As used herein, the term "nucleic acid" is intended to include deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), such as genomic DNA, cDNA, mRNA, recombinantly produced and chemically synthesized molecules. Nucleic acids can be single-stranded or double-stranded. RNA includes in vitro transcribed RNA or synthetic RNA.

[0046] The term "control", "control sample", "standard control" or "standard" refers to a sample that serves as a reference (usually a known reference) for comparison with a test sample. For example, a test sample may be taken from a patient suspected of having a given disease and compared with a sample from a known disease patient or a known normal (non-disease) individual. A control may also represent an average value collected from a population of similar individuals (e.g., disease patients or healthy individuals with similar medical backgrounds, the same age, weight, etc.). A control value may also be from the same individual, such as from a sample obtained earlier, before the disease, or before treatment. A skilled person will recognize that a control can be designed to assess many parameters. Reagents for detecting the expression level of CIAPIN1

[0047] In one aspect, the present invention provides a reagent for detecting the expression level of CIAPIN1 for use in the diagnosis and / or prognosis of glioma in a subject.

[0048] As used in the present invention, the term "reagent for detecting the expression level of CIAPIN1" refers to any reagent known in the art that can be used to detect CIAPIN1, such as a targeting reagent or affinity reagent for CIAPIN1, and especially includes reagents that can bind (especially specifically bind) to the expression product of CIAPIN1 to form a chemically, physically or biologically detectable complex.

[0049] In some embodiments, the expression product of CIAPIN1 includes CIAPIN1 mRNA and / or CIAPIN1 protein.

[0050] In some embodiments, the agent capable of binding to the expression product of CIAPIN1 includes a protein, a nucleic acid or a small molecule compound, which can, for example, target one or more epitopes of the CIAPIN1 protein.

[0051] In some embodiments, the agent capable of binding to the expression product of CIAPIN1 is an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof, or an anti-CIAPIN1 polyclonal antibody. Anti-CIAPIN1 antibodies are available from commercial sources, such as 12638-1-AP (Proteintech Group), ABIN393217 (antibodies-online), HPA041350 (AtlasAntibodies). For more CIAPIN1 antibodies, see https: / / www.antibodypedia.com / gene / 28857CIAPIN1 (last accessed on October 16, 2024). Alternatively, anti-CIAPIN1 antibodies can be generated de novo using methods known in the art. In some embodiments, the anti-CIAPIN1 antibody is any form of an antibody or an antibody fragment as defined herein.

[0052] In some embodiments, the agent capable of binding to the expression product of CIAPIN1 is a peptide or nucleic acid aptamer. Such an aptamer can be selected from an oligonucleotide or peptide library by any method known in the art. Nucleic acid aptamers can be selected via SELEX (Systematic Evolution of Ligands by Exponential Enrichment). Peptide aptamers can be selected using a yeast or bacterial two-hybrid system.

[0053] In some embodiments, the agent capable of binding to the expression product of CIAPIN1 is labeled with a detectable marker. In some embodiments, the detectable marker is selected from fluorescent markers, chemiluminescent markers, paramagnetic markers, radioisotope markers and enzyme markers.

[0054] The choice of label depends on the means of detection. For example, fluorescent labels (e.g., indocyanine green (ICG), rare earth chelates (e.g., europium chelates)), fluorescein-type labels (e.g., fluorescein, fluorescein isothiocyanate, 5-carboxyfluorescein, 6-carboxyfluorescein, dichlorotriazinylamine fluorescein), rhodamine-type labels (e.g., ALEXA568 (Invitrogen), or dansyl chloride), VIVOTAG 680XL FLUOROCHROME™ (PerkinElmer), phycoerythrin, 7-hydroxycoumarin, Lissamine, cyanine, phycoerythrin, Texas Red, BODIPY (Invitrogen), or the like, are suitable for optical detection.

[0055] Chemiluminescent labels (e.g., luminol, luciferase, luciferin, and aequorin) may also be used. Such diagnosis and detection may also be accomplished by linking a reagent capable of binding to CIAPIN1 to a detectable substance, including but not limited to various enzymes, including but not limited to horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase, or by linking it to prosthetic group complexes, such as but not limited to streptavidin / biotin and avidin / biotin.

[0056] Paramagnetic labels and radioisotope labels may also be used, which are preferably detected using positron emission tomography (PET) or single-photon emission computed tomography (SPECT). Radioactive labels include, but are not limited to, bismuth ( 213 Bi), carbon ( 11 C. 13 C. 14 C), chromium ( 51 Cr), cobalt ( 57 Co. 60 Co、Co、Copper 64 Cu), Dysprosium ( 165 Dy), Erbium (169 Er), fluorine ( 18 F), Gadolinium ( 153 Gd, 159 Gd), Gallium ( 68 Ga, 67 Ga), germanium ( 68 Ge), gold ( 198 Au), holmium ( 166 Ho), hydrogen ( 3 H), Indium ( 111 In, 112 In, 113 In, 115 In、Iodine 121 I. 123 I. 125 I. 131 I), iridium ( 192 Ir、Iron 59 Fe), Krypton ( 81m Kr), lanthanum ( 140 La), Lutetium ( 177 Lu), manganese ( 54 Mn), molybdenum ( 99 Mo), nitrogen ( 13 N. 15 N), oxygen ( 15 O), palladium ( 103 Pd), phosphorus ( 32 P), potassium ( 42 K), praseodymium ( 142 Pr), promethium ( 149 Pm), rhenium ( 186 Re, 188 Re), rhodium ( 105 Rh), Rubidium ( 81 Rb, 82 Rb), ruthenium ( 82 Such as 97 Ru), Samarium ( 153 Sm), Scandium ( 47 Sc), Selenium ( 75 Se), sodium ( 24 Na), Strontium ( 85 Sr. 89 Sr. 92 Sr、Sulphur 35 S), technetium ( 99 Tc), thallium ( 201 Tl), tin ( 113 Sn, 117 Sn), Xenon ( 133 Xe), Ytterbium ( 169 Yb, 175 Yb, 177 Yb), yttrium (90 Y) and zinc ( 65 Zn); various positron-emitting metals and non-radioactive paramagnetic metal ions that can be used for positron emission tomography, such as paramagnetic aluminum (Al) ions, barium (Ba) ions, calcium (Ca) ions, cerium (Ce) ions, dysprosium (Dy) ions, erbium (Er) ions, europium (Eu) ions, gadolinium (Gd) ions, holmium (Ho) ions, iridium (Ir) ions, lithium (Li) ions, magnesium (Mg) ions, manganese ( Mn) ions, molybdenum (M) ions, neodymium (Nd) ions, osmium (Os) ions, oxygen (O) ions, palladium (Pd) ions, platinum (Pt) ions, rhodium (Rh) ions, ruthenium (Ru) ions, samarium (Sm) ions, sodium (Na) ions, strontium (Sr) ions, terbium (Tb) ions, thulium (Tm) ions, tin (Sn) ions, titanium (Ti) ions, tungsten (W) ions and zirconium (Zi) ions, especially Co +2 , CR +2 Cr +3 , Cu +2 , Fe +2 , Fe +3 , Ga +3 , Mn +3 、Ni +2 、Ti +3 、V + and V +4 Methods for preparing radiolabeled amino acids and related peptide derivatives are known in the art. For example, radioisotopes can be conjugated by the chloramine T method. Reagent test kit

[0057] In one aspect, the present invention provides a kit for diagnosis and / or prognosis of glioma in a subject, the kit comprising a reagent for detecting the expression level of CIAPIN1.

[0058] Accordingly, in another aspect, the present invention provides use of a reagent for detecting the expression level of CIAPIN1 in preparing a kit for diagnosis and / or prognosis of glioma in a subject.

[0059] In some embodiments, the reagent for detecting the expression level of CIAPIN1 includes a reagent that can bind to the CIAPIN1 protein (especially specifically bind) to form a chemically, physically or biologically detectable complex, such as a protein, a nucleic acid or a small molecule compound, especially an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof, or an anti-CIAPIN1 polyclonal antibody.

[0060] In some embodiments, the agent capable of binding to CIAPIN1 is labeled with a detectable marker. In some embodiments, the detectable marker is selected from a fluorescent marker, a chemiluminescent marker, a paramagnetic marker, a radioisotope marker, and an enzyme marker.

[0061] In some embodiments, the kit further comprises CIAPIN1 recombinant antigen reference substances of different concentrations, so as to prepare a standard curve for quantitative identification. Diagnostic or prognostic method and computer readable storage medium

[0062] In one aspect, the present invention provides use of CIAPIN1 as a marker in the diagnosis and / or prognosis of glioma in a subject.

[0063] In another aspect, the present invention provides a method for in vitro or ex vivo diagnosis and / or prognosis of glioma in a subject, the method comprising detecting the presence and / or level of CIAPIN1 in a sample (eg, brain tissue) from the subject.

[0064] In another aspect, the present invention provides an agent capable of binding to an expression product of CIAPIN1 for in vivo diagnosis and / or prognosis of glioma in a subject.

[0065] Preferably, in each of the above aspects, the difference between the level of CIAPIN1 in the sample from the object and the control sample level or control value provides a diagnosis (e.g., presence and / or stage) and / or prognosis of a glioma of the object. The control sample can be a sample from a healthy tissue or object, or a sample from a tissue or object with a known disease stage or a known prognosis. The control sample level can be the median level of the control sample. In some embodiments, a level of CIAPIN1 in a sample from the object that is higher than the level of a control without the disease indicates that the object suffers from a glioma. In some embodiments, a higher expression level of CIAPIN1 in a sample from the object indicates a poor prognosis for a glioma of the object.

[0066] Preferably, in each of the above aspects, the agent capable of binding to the expression product of CIAPIN1 is an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof, or an anti-CIAPIN1 polyclonal antibody. In a preferred embodiment, the agent capable of binding to the expression product of CIAPIN1 is an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof.

[0067] Preferably, in each of the above aspects, the agent capable of binding to the expression product of CIAPIN1 is labeled with a detectable marker. In a preferred embodiment, the detectable marker is selected from a fluorescent marker, a chemiluminescent marker, a paramagnetic marker, a radioisotope marker and an enzyme marker. In a preferred embodiment, the detectable marker is selected from a fluorescent marker or a chemiluminescent marker.

[0068] Preferably, in each of the above aspects, the glioma is selected from astrocytoma, oligodendroglioma, ependymoma and mixed glioma. In some embodiments, the glioma is selected from anaplastic astrocytoma, anaplastic oligodendroglioma and glioblastoma. In some embodiments, the glioma is glioblastoma.

[0069] Various immunoassays can be used in the diagnosis or prognosis methods. In some embodiments, such immunoassays include competitive and non-competitive detection systems such as radioimmunoassay, immunochromatography, ELISA, "sandwich" immunoassay, precipitation reaction, immunoblot analysis, gel diffusion precipitation reaction, immunodiffusion detection, agglutination detection, complement fixation detection, immunoradiometric detection, fluorescent immunoassay, etc. Both in vitro and in vivo detection can be used.

[0070] The methods for diagnosis or prognosis allow for a quantitative and / or qualitative assessment, such as an absolute and / or relative measurement of a target molecule, such as measuring the amount of CIAPIN1 in a sample.

[0071] In some embodiments of the methods of the present invention, determining the presence and / or amount of CIAPIN1 in a sample comprises: (i) contacting the sample with an agent capable of binding to an expression product of CIAPIN1, and (ii) detecting the formation of a complex between the agent and the expression product of CIAPIN1 and / or determining the amount of the complex.

[0072] In some embodiments, the diagnosis / prognosis method of the present invention can be used in combination with other methods for detecting / diagnosing gliomas. For example, the diagnosis / prognosis method of the present invention can be used in combination with the detection of other biomarkers of gliomas (e.g., IDH, MGMT, EGFR, P53, PTEN).

[0073] On the other hand, the present invention provides a computer-readable storage medium having computer instructions stored thereon for computer reading and execution, the computer instructions being executed to perform a method for diagnosing and / or prognosing a glioma of a subject, the method comprising: (a) contacting a sample from the subject with a reagent capable of binding to an expression product of CIAPIN1; (b) detecting and reading a signal of the sample after contact to determine whether the reagent forms a complex with the expression product of CIAPIN1 in the sample; and (c) determining whether the signal exceeds a predetermined threshold, and when the signal exceeds a predetermined threshold, providing a diagnosis and / or prognosis of a glioma of the subject. In a preferred embodiment, the threshold is the median level from a subject without the disease. In a preferred embodiment, the threshold is the median level from a subject with a known prognosis.

[0074] On the other hand, the present invention provides a method for treating a glioma in a subject, the method comprising: (a) contacting a sample from the subject with an agent capable of binding to an expression product of CIAPIN1; (b) detecting and reading a signal of the sample after contact to determine whether the agent forms a complex with the expression product of CIAPIN1 in the sample; and (c) determining whether the signal exceeds a predetermined threshold, and when the signal exceeds a predetermined threshold, providing a diagnosis and / or prognosis of a glioma in the subject; and (d) administering an anti-tumor therapy to a subject determined to have a glioma. In a preferred embodiment, the threshold is the median level from subjects without the disease. In a preferred embodiment, the threshold is the median level from subjects with a known prognosis.

[0075] In some embodiments, the reagent capable of binding to the expression product of CIAPIN1 includes a protein, a nucleic acid or a small molecule compound. In a preferred embodiment, the reagent capable of binding to the expression product of CIAPIN1 is an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof, or an anti-CIAPIN1 polyclonal antibody. In some embodiments, the reagent capable of binding to the expression product of CIAPIN1 is labeled with a detectable marker. In some embodiments, the detectable marker is selected from a fluorescent marker, a chemiluminescent marker, a paramagnetic marker, a radioisotope marker and an enzyme marker.

[0076] In a preferred embodiment, the anti-tumor therapy is surgical resection, chemotherapy, radiotherapy or a combination thereof. In a preferred embodiment, chemotherapy is administered to an object determined to suffer from high-grade glioma. In a preferred embodiment, the chemotherapy is temozolomide, lomustine, carmustine, nimustine, procarbazine, vinblastine, vincristine, teniposide, etoposide, irinotecan, cisplatin, carboplatin, bevacizumab or a combination thereof. In a preferred embodiment, the chemotherapy is temozolomide.

[0077] In some embodiments, the predetermined threshold value can be a control sample level or control value as described above, wherein the difference in the control sample level or control value provides a diagnosis (e.g., presence and / or stage) and / or prognosis of a glioma of the object. The control sample can be a sample from a healthy tissue or object, or a sample from a tissue or object with a known disease stage or a known prognosis. The control sample level can be the median level of the control sample. In some embodiments, a level of CIAPIN1 in a sample from the object that is higher than the level of a control without the disease indicates that the object suffers from a glioma. In some embodiments, a higher expression level of CIAPIN1 in a sample from the object indicates a poor prognosis for a glioma of the object. Example

[0078] In the following examples, all experiments were repeated 3 times independently, and the results presented in the text are typical representative data. The obtained data are expressed as mean ± SD / SEM, and one-way ANOVA (One-way ANOVA) was performed using SPSS19 and GraphPadPrism 8.0 software. Tukey test was used for comparison of data differences between groups, and Student's t test was used for pairwise comparison. P<0.05 indicated that the difference was statistically significant.

[0079] Example 1: CIAPIN1 is significantly more highly expressed in glioma tissues than in normal tissues

[0080] In order to explore the expression of CIAPIN1 in glioma, this example analyzed the expression of CIAPIN1 in 1157 normal tissues and 689 glioma patient tissues in the TCGA database and GTEx database. The analysis results showed that CIAPIN1 was significantly overexpressed in glioma, P<0.001 ( Figure 1 ).

[0081] In order to further confirm the expression of CIAPIN1 in glioma, the HPA database was used to immunohistochemically analyze the expression of CIAPIN1 in normal human traumatic cortical tissue and glioma tissue. The typical field of view showed that the expression of CIAPIN1 in glioma was significantly higher than that in normal human brain tissue ( Figure 2 ).

[0082] Example 2: High expression of CIAPIN1 is significantly associated with poor prognosis

[0083] According to the expression analysis of CIAPIN1 in glioma samples from the TCGA database, the samples were divided into CIAPIN1 high expression group and CIAPIN1 low expression group based on the median expression of CIAPIN1. Specifically, the median relative expression of CIAPIN1 was 5.798, and the high expression group was defined as greater than or equal to 5.798, and the low expression group was defined as less than 5.798. The Kaplan-Meier survival analysis method was used to perform survival analysis on the CIAPIN1 high expression and low expression groups. The results showed that compared with the low expression group, the high expression group had a relatively poor prognosis in terms of overall survival (OS) (P<0.001) and progression-free survival (PFS) (P=0.002). Figure 3 ).

[0084] Example 3: Expression of CIAPIN1 is associated with the infiltration of various immune cells

[0085] In order to study the relationship between CIAPIN1 and tumor immune cell infiltration, the TCGA database was used to explore the association between CIAPIN1 expression and immune cell infiltration by Spearman correlation analysis. The results showed that CIAPIN1 expression was negatively correlated with NK cells (natural killer cells) and pDC cells (plasmacytoid dendritic cells), and positively correlated with T Helper cells (helper T cells) and Th2 cells (helper T cells 2). Figure 4 ).

[0086] Example 4: Expression of CIAPIN1 can accurately predict glioma

[0087] In order to confirm the value of CIAPIN1 in the diagnosis of glioma, the expression of CIAPIN1 was analyzed in 1157 normal brain tissues and 689 glioma patients in the TCGA database and GTEx database. The sensitivity was used as the vertical axis and 1-specificity was used as the horizontal axis. According to a series of cutoff values ​​divided by the expression values ​​of CIAPIN1 in the normal brain tissue and glioma patient groups, the sensitivity and specificity were calculated respectively. The calculated points were connected into lines to draw the ROC curve. The area under the curve (AUC) was 0.921, and the 95% confidence interval was (0.910-0.933). The analysis results showed that the expression level of CIAPIN1 in the tumor tissue of glioma patients can accurately predict the occurrence of glioma ( Figure 5 ).

[0088] It should be understood that, although the present invention has been specifically disclosed by preferred embodiments and optional features, those skilled in the art may make modifications, improvements and variations to the present invention disclosed herein, and these modifications, improvements and variations are considered to be within the scope of the present invention. The materials, methods and examples provided herein are representative and exemplary of preferred embodiments, and are not intended to be limiting of the scope of the present invention.

Claims

1. Use of a reagent for detecting the expression level of CIAPIN1 in a sample from a subject in the preparation of a kit for diagnosis and / or prognosis of glioma in the subject. 2 . The use according to claim 1 , wherein a level of CIAPIN1 in the sample from the subject that is higher than that of a control without the disease indicates that the subject suffers from glioma. 3 . The use according to claim 1 , wherein a higher expression level of CIAPIN1 in the sample from the subject indicates a poor prognosis of glioma in the subject. 4 . The use according to any one of claims 1 to 3 , wherein the reagent for detecting the expression level of CIAPIN1 comprises a reagent capable of binding to an expression product of CIAPIN1, so as to detect the expression level of CIAPIN1 in the sample. 5 . The use according to claim 4 , wherein the agent capable of binding to the expression product of CIAPIN1 is an anti-CIAPIN1 monoclonal antibody or an antigen-binding fragment thereof, or an anti-CIAPIN1 polyclonal antibody. 6 . The use according to claim 4 , wherein the agent capable of binding to the expression product of CIAPIN1 is labeled with a detectable marker.

7. The use according to claim 6, wherein the detectable label is selected from the group consisting of fluorescent label, chemiluminescent label, paramagnetic label, radioisotope label and enzyme label.

8. The use according to any one of claims 1 to 3, wherein the sample is brain tissue.

9. The use according to any one of claims 1 to 3, wherein the glioma is selected from astrocytoma, oligodendroglioma, ependymoma and mixed glioma.

10. A computer-readable storage medium having stored thereon computer instructions for being read and executed by a computer, wherein the computer instructions are executed to perform a method for diagnosing and / or prognosing a glioma in a subject, the method comprising: (a) contacting a sample from the subject with an agent capable of binding to an expression product of CIAPIN1; (b) detecting and reading the signal of the sample after contact to determine whether the reagent forms a complex with the expression product of CIAPIN1 in the sample; as well as (c) determining whether the signal exceeds a predetermined threshold, and providing a diagnosis and / or prognosis of the subject's glioma when the signal exceeds the predetermined threshold.

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