CN113957139A - Application of human-derived gene in preparation of product for predicting sensitivity of patient to golimumab - Google Patents
Application of human-derived gene in preparation of product for predicting sensitivity of patient to golimumab Download PDFInfo
- Publication number
- CN113957139A CN113957139A CN202111426959.8A CN202111426959A CN113957139A CN 113957139 A CN113957139 A CN 113957139A CN 202111426959 A CN202111426959 A CN 202111426959A CN 113957139 A CN113957139 A CN 113957139A
- Authority
- CN
- China
- Prior art keywords
- golimumab
- ulcerative colitis
- biomarker
- drug
- patient
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4703—Regulators; Modulating activity
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4727—Calcium binding proteins, e.g. calmodulin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/06—Gastro-intestinal diseases
- G01N2800/065—Bowel diseases, e.g. Crohn, ulcerative colitis, IBS
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Abstract
The invention discloses application of a human gene in preparation of a product for predicting sensitivity of a patient to golimumab, wherein the human gene is CPB1, S100A8 and/or NUPR1, the genes CPB1, S100A8 and NUPR1 in an ulcerative colitis patient sensitive to golimumab drug are respectively up-regulated, down-regulated and up-regulated, the sensitivity of the patient to golimumab can be judged by detecting the expression condition of the genes in the body of the ulcerative colitis patient, a judgment index is provided for a clinician in a treatment process, the product has guiding significance for selection of therapeutic drugs for ulcerative colitis, a drug administration scheme can be timely adjusted, and the product has important practical application value.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to application of a human gene in preparation of a product for predicting sensitivity of a patient to golimumab.
Background
Ulcerative Colitis (UC) is a colonic inflammatory disease with complex pathogenesis, mainly including diarrhea, bloody stool and mucopurulent bloody stool as main clinical manifestations, and severe patients with hyperpyrexia and toxic symptoms (Watanabe Y, Murata T, Amakawa M, et al. KAG-308, a new-identified EP4-selective inflammatory disease for treating acute colitis and chronic inflammatory disease of pathological inflammation [ J ]. European journess of pharmacological science 2015,754: 179. 189. alternate onset, remission and recurrence as diseases, which are frequently seen in 20-40 years old and are common and frequently-occurring and difficult diseases of digestive system. The pathological manifestations of the cancer are infiltration of inflammatory cells of mucous membranes and submucosa, formation of multiple ulcers and occurrence of colon cancer. Ulcerative colitis mainly involves the mucosa and submucosa of colon, and is distributed in a continuous and diffuse manner, usually in sigmoid colon and rectum. It is characterized by prolonged and inconsistent disease course and low quality of life (Leblanc K, Mosli M H, Parker C E, et al. the impact of biological interventions for achieving clinical judgment on health-related quality of life [ J ]. Cochrane Database of Systematic Reviews 2015 (9)).
In recent years, the incidence of ulcerative colitis has been on the rise year by year trend in China and worldwide, seriously threatens the health and life of people, and is one of the difficult diseases listed by the World Health Organization (WHO) (Chen S J, Liu X W, Liu J P, et al. Ulcerative science as a microbial infection syndrome barrier [ J ]. World J outer of gastroenterology: WJG,2014,20(28): 9468.). However, the pathogenesis and mechanism of the ulcerative colitis are complex, and the pathogenesis and mechanism are not clear at present, the intrinsic factors including immune disorder, intestinal micro-ecological disorder, intestinal barrier dysfunction, genetic factors, unhealthy diet, smoking, mental factors and the like are considered to be important factors for inducing the ulcerative colitis, the ulcerative colitis is caused by the action of various factors, some serious complications can occur at any time and the risk of canceration exists, and the serious complications further increase the cure difficulty of the ulcerative colitis and seriously affect the life quality of patients. The pathogenesis of ulcerative colitis is very complex, the diseased region mainly relates to the mucosa layer of colon and is not limited by the age of the disease, generally, the pathogenesis of ulcerative colitis is a result of the combined action of multiple factors, the pathogenesis of ulcerative colitis is continuously disclosed along with the development and further research of scientific technology, and immune factors, infection factors, apoptosis, genetic factors, environmental factors and the like are all important pathogenesis of ulcerative colitis.
Golimumab (golimamab) is a novel anti-Tumor Necrosis Factor (TNF) - α monoclonal antibody drug for ulcerative colitis patients, and in the past decade, anti-TNF- α monoclonal antibodies play an important role in the biotherapy of moderate to severe ulcerative colitis. It can promote clinical remission and mucosa healing of ulcerative colitis, reduce disease aggravation risk and operation rate, and has been widely used clinically. To date a total of 3 anti-TNF-alpha mabs, i.e. infliximab, adalimumab and golimumab, have been approved for the treatment of ulcerative colitis. The golimumab is a new member of a TNF-alpha retarder, is a recombinant fully human IgG1 type anti-TNF-alpha monoclonal antibody, has a structure completely the same as that of human IgG, can be specifically and high-affinity combined with TNF-alpha, and can prevent the TNF-alpha from being combined with a TNF-alpha receptor on the surface of a cell, so that the biological activity of the TNF-alpha is antagonized. Compared with infliximab and adalimumab, golimumab is more stable in structure and stronger in binding capacity with TNF-alpha, although the drug has been subjected to classical induced remission research tests and maintenance remission research tests abroad and is also approved by FDA for treating moderate and severe ulcerative colitis, at present, it is still difficult to predict which patients with ulcerative colitis are sensitive or reactive to golimumab drugs, and how to predict the sensitivity and/or reactivity of patients with ulcerative colitis to golimumab drugs remains to be further studied.
Disclosure of Invention
In view of the above-mentioned shortcomings of the current art, the present invention aims to provide the use of a human-derived gene in the preparation of a product for predicting the sensitivity of a patient to golimumab.
The above object of the present invention is achieved by the following technical solutions:
in a first aspect, the present invention provides a biomarker for predicting the susceptibility of patients with ulcerative colitis to a golimumab drug.
Further, the biomarker is CPB1, S100a8, and/or NUPR 1.
Further, the biomarkers described in the present invention include CPB1, S100a8, NUPR1, and the information in NCBI is as follows:
CPB1(Carboxypeptidase B1) has a Gene ID of 1360 in NCBI;
S100A8(S100 calcium binding protein A8) has a Gene ID of 6279 in NCBI;
the Gene ID of NUPR1(Nuclear protein 1, transactional regulator) in NCBI is 26471.
In a second aspect, the invention provides the use of a reagent for detecting a biomarker in a sample for the manufacture of a product for predicting the susceptibility of a patient with ulcerative colitis to a golimumab drug.
Further, the biomarker is CPB1, S100a8, and/or NUPR 1.
Further, the reagent includes a reagent for detecting the mRNA expression level of the biomarker in the sample, and a reagent for detecting the protein expression level of the biomarker.
Further, the reagents include a probe that specifically recognizes the biomarker, a primer that specifically amplifies the biomarker, or a binding agent that specifically binds to a protein encoded by the biomarker.
Further, the binding agent comprises a peptide, a peptidomimetic, a nucleic acid aptamer, a mirror image L-RNA aptamer, an ankyrin repeat protein, a Kunitz-type domain, an antibody, a single domain antibody, or a monovalent antibody fragment;
preferably, the binding agent is an antibody.
In the present invention, the sample may be analyzed to determine the expression level of the biomarker in the sample using any suitable method, including (but not limited to): nucleic acid sequencing, nucleic acid hybridization, nucleic acid amplification technology, protein immunization technology;
illustrative non-limiting examples of nucleic acid sequencing methods include (but are not limited to): chain terminator (Sanger) sequencing and dye terminator sequencing, those of ordinary skill in the art will recognize that RNA is typically reverse transcribed into DNA prior to sequencing, as it is less stable in cells and more susceptible to nuclease attack in experiments; another illustrative, non-limiting example of a nucleic acid sequencing method includes next generation sequencing (deep sequencing/high throughput sequencing), in which a high throughput sequencing technique is a sequencing-by-synthesis technique based on single-molecule clusters, and based on a proprietary principle of reversible termination chemical reaction, random fragments of DNA of a genome are attached to an optically transparent glass surface during sequencing, after extension and bridge amplification of the DNA fragments, hundreds of millions of clusters are formed on the glass surface, each cluster is a single-molecule cluster with thousands of identical templates, and then four specific deoxyribonucleotides with fluorophores are used to sequence template DNA to be tested by the reversible sequencing-by-synthesis technique;
nucleic acid hybridization methods include (but are not limited to): in Situ Hybridization (ISH), which is a hybridization of specific DNA or RNA sequences in a section or slice of tissue using a labeled complementary DNA or RNA strand as a probe (in situ) or in the whole tissue if the tissue is small enough (whole tissue embedded ISH), microarrays, and Southern or Northern blots, respectively, for detecting specific DNA or RNA sequences;
the nucleic acid amplification method is selected from the group consisting of Polymerase Chain Reaction (PCR), reverse transcription polymerase chain reaction (RT PCR), Transcription Mediated Amplification (TMA), Ligase Chain Reaction (LCR), Strand Displacement Amplification (SDA), and Nucleic Acid Sequence Based Amplification (NASBA), wherein PCR requires reverse transcription of RNA into dna prior to amplification (RT PCR), TMA, and NASBA to directly amplify RNA;
protein immunization methods include sandwich immunoassays, such as sandwich ELISA, in which detection of a biomarker is performed using two antibodies that recognize different epitopes on the biomarker; radioimmunoassay (RIA), direct, indirect or contrast enzyme-linked immunosorbent assay (ELISA), Enzyme Immunoassay (EIA), Fluorescence Immunoassay (FIA), western blot, immunoprecipitation, and any particle-based immunoassay (e.g., using gold, silver or latex particles, magnetic particles, or quantum dots, for example, can be performed in microtiter plate or strip formats.
In a third aspect, the invention provides a product for predicting the susceptibility of patients with ulcerative colitis to a golimumab drug.
Further, the product comprises reagents for detecting the biomarkers CPB1, S100a8, and/or NUPR1 in the sample.
Furthermore, the product comprises a kit, a chip, test paper and a high-throughput sequencing platform.
Further, the kit comprises a qPCR kit, an ELISA kit, an immunoblotting detection kit, an immunochromatography detection kit, an immunohistochemical detection kit, a flow cytometry analysis kit and an electrochemiluminescence detection kit.
Further, the kit also comprises a method for predicting and evaluating the sensitivity of the subjects to the golimumab drug by adopting the kit.
Further, in certain embodiments, the kit is marketed, distributed, or sold as a unit for performing the methods of the present invention. Such kits may comprise carrier means compartmentalized to receive, in close confinement, one or more container means (e.g., vials, tubes, etc.), each container means comprising one of the separate components to be used in the method. For example, one of the container means may comprise a probe that carries or can carry a detectable label. Such probes may be polynucleotides specific for polynucleotides of one or more genes comprising gene expression characteristics. Where the kit utilizes nucleic acid hybridization to detect a target nucleic acid, the kit can also have a container containing one or more nucleic acids for amplifying the target nucleic acid sequence and/or a container containing a reporter means.
Further, the kit will generally comprise the above-described container and one or more additional containers containing commercially and user-desired materials, including buffers, diluents, filters, needles, syringes, and package inserts containing instructions for use. A label may be present on the container to indicate the particular application of the composition, and may also indicate the direction of in vivo or in vitro use, such as those described above. Other optional components of the kit include one or more buffers (e.g., blocking buffer, wash buffer, substrate buffer, etc.), other reagents (e.g., substrate chemically altered by enzymatic labeling), epitope retrieval solutions, control samples (positive and/or negative controls), control sections, and the like.
In a fourth aspect, the present invention provides a system or apparatus for predicting the susceptibility of a patient with ulcerative colitis to a golimumab drug.
Further, the system or apparatus comprises:
(1) a detection unit: the detection unit is used for detecting the expression level of the biomarkers CPB1, S100A8 and/or NUPR 1;
(2) a result judgment unit: and the result judging unit is used for outputting the prediction result of the sensitivity of the ulcerative colitis patient to the golimumab drug according to the expression quantity result detected by the detecting unit.
Further, the detection unit comprises a qPCR kit, an ELISA kit, an immunoblotting detection kit, an immunochromatography detection kit, an immunohistochemical detection kit, a flow cytometry kit, an electrochemiluminescence detection kit, a qPCR instrument, an ELISA detection device, an immunoblotting detection device, an immunochromatography detection device, an immunohistochemical detection device, a flow cytometry instrument and an electrochemiluminescence detection device.
Further, the result judging unit comprises an input unit, an analysis unit and an output unit;
the input unit is used for inputting the expression quantity of the biomarker;
the analysis unit is used for analyzing the prediction result of the sensitivity of the ulcerative colitis patient to the golimumab drug according to the expression quantity of the biomarker;
the output unit is used for outputting the analysis result of the analysis unit.
Compared with the prior art, the invention has the advantages and beneficial effects that:
the invention provides a group of gene markers capable of predicting the sensitivity of an ulcerative colitis patient to a golimumab drug, wherein the markers are CPB1, S100A8 and/or NUPR1, the sensitivity of the test subject to the golimumab drug can be predicted by detecting the expression level of the markers in a test subject sample, the test subject does not depend on primary culture or an animal model, the test period is short, the accuracy is high, the clinical application and scientific research value are wide, and the gene markers can be particularly used for predicting the drug effect of the golimumab drug so as to guide the drug administration of the ulcerative colitis patient.
Drawings
Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:
figure 1 shows a graph of the results of CPB1 differential expression between ulcerative colitis patients in the response to golimumab drug group and the non-response to golimumab drug group;
fig. 2 shows a graph of the results of S100A8 differential expression between ulcerative colitis patients in the response to golimumab drug group and the non-response to golimumab drug group;
figure 3 shows a graph of the results of NUPR1 differential expression between ulcerative colitis patients in the response to golimumab drug and non-response to golimumab drug group;
figure 4 shows a graph of the results of CPB1 on the diagnostic efficacy of predicting the susceptibility of patients with ulcerative colitis to golimumab drug;
fig. 5 shows a graph of the results of S100A8 on the efficacy of a diagnosis predicting the susceptibility of patients with ulcerative colitis to golimumab drug;
figure 6 shows a graph of the results of NUPR1 on predicting the diagnostic efficacy of a patient with ulcerative colitis on the sensitivity of golimumab drug;
figure 7 shows a graph of the results of the diagnostic efficacy of the CPB1+ S100a8 combination for predicting the susceptibility of patients with ulcerative colitis to golimumab drug;
figure 8 shows a graph of the results of the CPB1+ NUPR1 combination on predicting the diagnostic efficacy of a patient with ulcerative colitis on the sensitivity to golimumab drug;
figure 9 shows a graph of the results of the diagnostic efficacy of the S100A8+ NUPR1 combination for predicting the sensitivity of patients with ulcerative colitis to golimumab drug;
figure 10 shows a graph of the results of the diagnostic efficacy of the CPB1+ S100a8+ NUPR1 combination for predicting the sensitivity of ulcerative colitis patients to golimumab drugs.
Detailed Description
The present invention is further illustrated by the following examples, which should be construed as being merely illustrative and not limitative of the remainder of the disclosure. As will be understood by those of ordinary skill in the art: many changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention. The following examples are examples of experimental methods not indicating specific conditions, and the detection is usually carried out according to conventional conditions or according to the conditions recommended by the manufacturers. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, and any methods and materials similar or equivalent to those described herein may be applied.
In the context of the present invention, the term "Golimumab drug", like "Golimumab" and "golimamab", is a Tumor Necrosis Factor (TNF) - α inhibitor, and the European Medicines Agency (EMA) announces the recommendation for approval of Golimumab (Simponi, popsonian) for the treatment of moderate to severe ulcerative colitis in adult patients who are ineffective or unable to receive conventional treatment. Conventional therapeutic agents for ulcerative colitis include glucocorticoids, 6-mercaptopurine, or azathioprine. The golimumab is a monoclonal antibody which can inhibit tumor necrosis factor-alpha, and is a pathogenic factor of inflammatory diseases. The united states Food and Drug Administration (FDA) has approved golimumab for the treatment of ulcerative colitis 5 months in 2013. Three other indications for this drug have been approved in both the united states and europe, including moderate to severe rheumatoid arthritis (with methotrexate), psoriatic arthritis, and ankylosing spondylitis.
In the context of the present invention, the term "marker", as well as "biomarker", "genetic marker", "marker molecule", refers to an indicator of a patient's phenotype, such as a pathological state or a possible reactivity to a therapeutic agent, which can be detected in a biological sample of said patient, biomarkers including, but not limited to, DNA, RNA, proteins, small molecule metabolites, carbohydrates, glycolipid-based molecules, and the like.
In the context of the present invention, the term "sample", as well as "sample", "subject sample", refers to a composition obtained or derived from a subject of interest, which comprises cellular entities and/or other molecular entities to be characterized and/or identified, e.g. on the basis of physical, biochemical, chemical and/or physiological characteristics. The sample may be obtained from the blood of a subject and other fluid samples of biological origin and tissue samples, such as biopsy tissue samples or tissue cultures or cells derived therefrom. The source of the tissue sample may be solid tissue, such as from a fresh, frozen and/or preserved organ or tissue sample, biopsy tissue or aspirate; blood or any blood component; a body fluid; cells from any time of pregnancy or development of the individual; or plasma. The term "sample" includes a biological sample that has been treated in any way after it has been obtained, e.g., by treatment with a reagent, stabilization, or enrichment for certain components (e.g., proteins or polynucleotides), or embedded in a semi-solid or solid matrix for sectioning purposes. Samples described herein include, but are not limited to, whole blood, blood-derived cells, serum, plasma, lymph, synovial fluid, cell extracts, and combinations thereof.
In the context of the present invention, the term "primer", as well as "amplification primer", refers to a nucleic acid fragment comprising 5-100 nucleotides, preferably said primer or amplification primer comprises 15-30 nucleotides capable of initiating an enzymatic reaction (e.g., an enzymatic amplification reaction).
In the context of the present invention, the term "probe" refers to a nucleic acid sequence comprising at least 5 nucleotides, e.g.comprising 5 to 100 nucleotides, which probe is capable of hybridizing under the specified conditions with the expression product of the target gene or with the amplification product of this expression product to form a complex. The hybridization probes may also include labels for detection. Such labels include, but are not limited to, labels for fluorescent quantitative PCR or fluorescent in situ hybridization.
In the context of the present invention, the term "antibody", which is well known in the art, refers to a specific immunoglobulin directed against an antigenic site. The antibody of the present invention refers to an antibody that specifically binds to the biomarker protein of the present invention, and can be produced according to a conventional method in the art. Forms of antibodies include polyclonal or monoclonal antibodies, antibody fragments (such as Fab, Fab ', F (ab') 2, and Fv fragments), single chain Fv (scfv) antibodies, multispecific antibodies (such as bispecific antibodies), monospecific antibodies, monovalent antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising an antigen binding site of an antibody, and any other modified immunoglobulin molecule comprising an antigen binding site, so long as the antibody exhibits the desired biological binding activity.
In the context of the present invention, the term "peptide", as well as "polypeptide", refers to a peptide that has the ability to bind to a target substance to a high degree and that is not denatured during heat/chemical treatment. Also, due to its small size, it can be used as a fusion protein by attaching it to other proteins. In particular, since it can be specifically attached to a high molecular protein chain, it can be used as a diagnostic kit and a drug delivery substance.
In the context of the present invention, the term "sensitive", as well as "sensitive", "reactive" and "reacting", means that a patient/subject suffering from, suspected to suffer from or prone to ulcerative colitis shows a positive response to treatment with golimumab drug in some way. One skilled in the art would readily determine whether a patient/subject using a golimumab drug treatment regimen shows a positive response based on the methods described herein. The sensitivity of different patients/subjects suffering from ulcerative colitis to the golimumab drug has obvious difference, and the sensitivity of the patients suffering from ulcerative colitis to the golimumab drug is difficult to accurately judge only by doctors according to clinical characteristics and clinical experience of the patients, so that the effect of clinically adopting the golimumab drug to treat the patients suffering from ulcerative colitis is not ideal at present. With the rapid development of molecular and cellular biology, researchers and clinicians have shifted the focus on how to use biomarkers to predict drug sensitivity. The discovery of a new biomarker capable of being used for predicting the drug sensitivity of ulcerative colitis patients to golimumab can provide a more effective and accurate drug sensitivity evaluation index for ulcerative colitis, thereby facilitating clinicians to set an individualized treatment scheme for ulcerative colitis subjects, reducing medication blindness, monitoring the curative effect of drugs and improving the quality of life of ulcerative colitis patients.
As the skilled person will be familiar with, the step of correlating the expression level or amount of a biomarker with a certain likelihood or risk may be performed and carried out in different ways. Preferably, the measured concentrations of the protein and one or more other markers are mathematically combined and the combined values are correlated to the underlying prediction problem. The determination of marker values may be combined by any suitable prior art mathematical method.
Preferably, the mathematical algorithm applied in the biomarker combinations is a logarithmic function. Preferably, the result of applying such a mathematical algorithm or such a logarithmic function is a single value. Such values can easily be associated, according to the underlying diagnostic question, for example with the sensitivity of an individual with ulcerative colitis with respect to the drug golimumab or with other deliberate diagnostic uses that help to assess patients with ulcerative colitis. In a preferred manner, such a logarithmic function is obtained as follows: a) classifying the individual into groups, e.g., patients with ulcerative colitis who are responsive to golimumab drugs, patients with ulcerative colitis who are not responsive to golimumab drugs, etc.; b) identifying markers that differ significantly between these groups by univariate analysis; c) logistic regression analysis to assess independent difference values of biomarkers that can be used to assess these different sets; d) a logarithmic function is constructed to combine the independent difference values. In this type of analysis, the markers are no longer independent, but represent a combination of markers.
The logarithmic function used to correlate the marker combinations with the sensitivity to golimumab of patients with ulcerative colitis preferably employs algorithms developed and obtained by applying statistical methods. For example, suitable statistical methods are Discriminant Analysis (DA) (i.e., linear, quadratic, regular DA), Kernel methods (i.e., SVM), nonparametric methods (i.e., k-nearest neighbor classifiers), PLS (partial least squares), tree-based methods (i.e., logistic regression, CART, random forest methods, boosting/bagging methods), generalized linear models (i.e., logistic regression), principal component-based methods (i.e., SIMCA), generalized additive models, fuzzy logic-based methods, neural network-and genetic algorithm-based methods. The skilled person will not have problems in selecting a suitable statistical method to evaluate the marker combinations of the invention and thereby obtain a suitable mathematical algorithm. In one embodiment, the statistical method used to obtain the mathematical algorithm used in assessing diabetic nephropathy is selected from DA (i.e., linear, quadratic, regular discriminant analysis), Kernel method (i.e., SVM), non-parametric method (i.e., k-nearest neighbor classifier), PLS (partial least squares), tree-based method (i.e., logistic regression, CART, random forest method, boosting method), or generalized linear model (i.e., logarithmic regression).
The area under the subject's working characteristic curve (AUC) is an indicator of the performance or accuracy of the diagnostic protocol. The accuracy of a diagnostic method is best described by its Receiver Operating Characteristics (ROC). ROC plots are line graphs of all sensitivity/specificity pairs derived from continuously varying decision thresholds across the entire data range observed.
Example 1 screening of genes associated with susceptibility of ulcerative colitis to golimumab drug
1. Data sources and screening methods
59 patients with ulcerative colitis were collected, and colon mucosal tissue samples were collected from the patients with ulcerative colitis before treatment with Golimumab drug (Golimumab). The ulcerative colitis patient in the study is a moderate-to-severe active ulcerative colitis patient, and the golimumab drug can promote clinical remission and mucosal healing of the ulcerative colitis and reduce disease exacerbation risks and operation rate, and is widely applied clinically. The patients with ulcerative colitis were clinically administered a golimumab drug, which was divided into a response group (R, responder) and a non-response group (NR, non-responder) according to their responsiveness to the golimumab drug, wherein the response group included patients with ulcerative colitis who had Complete Response (CR) and patients with ulcerative colitis who had Partial Response (PR), and the non-response group included patients with stable disease (> 4 months) and progressive disease (PD, progressive disease), and the data of this study was derived from GSE 92415. The proportion of patients with ulcerative colitis that were non-responsive and reactive to golimumab drug was as follows: non-responder: (iii) a reposder of 27: 32, a first step of removing the first layer;
detecting genes which are differentially expressed between ulcerative colitis patients in a golimumab drug non-response group and ulcerative colitis patients in a golimumab drug response group after treatment with golimumab drugs, and comparing the genes which are differentially expressed between the ulcerative colitis patients in the golimumab drug non-response group and the ulcerative colitis patients in the golimumab drug response group; analysis of differentially expressed genes was performed using the "limma" package (version 3.36.5) in the R software, where the screening criteria for differentially expressed genes was p<0.01，|log2FC|>0.5。
2. Results of the experiment
The results show that among the screened differentially expressed genes between ulcerative colitis patients in the response group and the non-response group to the golimumab drug, CPB1, S100a8 and NUPR1 show significant differential expression, and CPB1, S100a8 and NUPR1 show the following expression conditions in the ulcerative colitis patients in the response group to the golimumab drug: the up-regulation, down-regulation and up-regulation (see fig. 1-3) show that CPB1, S100A8 and NUPR1 can be used as potential marker molecules for predicting the sensitivity of patients with ulcerative colitis to the drug of golimumide.
Example 2 use of potential marker molecules to predict the susceptibility of patients with ulcerative colitis to golimumab drug
1. Experimental methods
For the genes screened in example 1 and differentially expressed between the response group and the non-response group to the golimumab drug, using the package R "pROC" (version 1.15.0) to draw a subject working curve (ROC curve), calculating the area under the subject working characteristic curve (AUC) to evaluate the accuracy of predicting the sensitivity of the ulcerative colitis patient to the golimumab drug by using a single differentially expressed gene, any two differentially expressed genes in combination, and three differentially expressed genes in combination;
when the diagnosis effectiveness and accuracy of a single gene for predicting the drug sensitivity of an ulcerative colitis patient to golimumab are judged, the expression quantity of the single gene is directly used for analysis, the expression level corresponding to the point with the largest Youden index is selected as the cutoff value, and the gene with the AUC of 0.5< AUC <0.8 is used for combined analysis;
when the diagnosis efficiency and accuracy of the gene combination on predicting the sensitivity of ulcerative colitis patients to the golimumab drug are judged, Logitics regression analysis is carried out on the expression level of each gene in the gene combination, the probability of each individual on the golimumab drug sensitivity is calculated through a fitted regression curve, different probability division thresholds are determined, and the sensitivity, the specificity, the accuracy and the like of the gene combination diagnosis scheme are calculated according to the determined probability division thresholds.
2. Results of the experiment
The results show that the accuracy of the gene combinations CPB1+ S100A8, CPB1+ NUPR1, S100A8+ NUPR1, CPB1+ S100A8+ NUPR1 on the prediction of the drug sensitivity of ulcerative colitis patients to the golimumab, because of the single gene, the AUC values are all high (see table 1, figure 4-figure 10), and the genes CPB1, S100A8 and/or NUPR1 can be applied to the prediction of the drug sensitivity of ulcerative colitis patients to the golimumab.
TABLE 1 diagnostic efficacy of the genes for predicting the susceptibility of patients with ulcerative colitis to the drug golimumab
| Gene | AUC value |
| CPB1 | 0.753 |
| S100A8 | 0.677 |
| NUPR1 | 0.700 |
| CPB1+S100A8 | 0.784 |
| CPB1+NUPR1 | 0.802 |
| S100A8+NUPR1 | 0.748 |
| CPB1+S100A8+NUPR1 | 0.810 |
The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.
Claims (10)
1. A biomarker for predicting the sensitivity of a patient with ulcerative colitis to a golimumab drug, wherein the biomarker is CPB1, S100a8, and/or NUPR 1.
2. Use of a reagent for detecting a biomarker in a sample for the manufacture of a product for predicting the susceptibility of a patient with ulcerative colitis to a golimumab drug, wherein the biomarker is CPB1, S100a8, and/or NUPR 1.
3. The use of claim 2, wherein the reagents comprise a reagent for measuring the amount of mRNA expression of the biomarker in the sample and a reagent for measuring the amount of protein expression of the biomarker.
4. The use of claim 3, wherein the reagents comprise a probe that specifically recognizes the biomarker, a primer that specifically amplifies the biomarker, or a binding agent that specifically binds to a protein encoded by the biomarker.
5. The use of claim 4, wherein the binding agent comprises a peptide, a peptide mimetic, a nucleic acid aptamer, a mirror image L-RNA aptamer, an ankyrin repeat protein, a Kunitz-type domain, an antibody, a single domain antibody, or a monovalent antibody fragment.
6. A product for predicting the susceptibility of a patient with ulcerative colitis to a golimumab drug, comprising reagents for detecting the biomarkers CPB1, S100a8, and/or NUPR1 in a sample.
7. The product of claim 6, wherein the product comprises a kit, a chip, a strip, a high throughput sequencing platform.
8. A system or apparatus for predicting the susceptibility of a patient with ulcerative colitis to a golimumab drug, characterized in that it comprises:
(1) a detection unit: the detection unit is used for detecting the expression level of the biomarkers CPB1, S100A8 and/or NUPR 1;
(2) a result judgment unit: and the result judging unit is used for outputting the prediction result of the sensitivity of the ulcerative colitis patient to the golimumab drug according to the expression quantity result detected by the detecting unit.
9. The system or device of claim 8, wherein the detection unit comprises a qPCR kit, an ELISA kit, an immunoblot detection kit, an immunochromatographic detection kit, an immunohistochemical detection kit, a flow cytometry kit, an electrochemiluminescence detection kit, a qPCR instrument, an ELISA detection device, an immunoblot detection device, an immunochromatographic detection device, an immunohistochemical detection device, a flow cytometry, an electrochemiluminescence detection device.
10. The system or apparatus of claim 8, wherein the result determination unit comprises an input unit, an analysis unit, and an output unit;
the input unit is used for inputting the expression quantity of the biomarker;
the analysis unit is used for analyzing the prediction result of the sensitivity of the ulcerative colitis patient to the golimumab drug according to the expression quantity of the biomarker;
the output unit is used for outputting the analysis result of the analysis unit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111426959.8A CN113957139A (en) | 2021-11-28 | 2021-11-28 | Application of human-derived gene in preparation of product for predicting sensitivity of patient to golimumab |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111426959.8A CN113957139A (en) | 2021-11-28 | 2021-11-28 | Application of human-derived gene in preparation of product for predicting sensitivity of patient to golimumab |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113957139A true CN113957139A (en) | 2022-01-21 |
Family
ID=79472141
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111426959.8A Withdrawn CN113957139A (en) | 2021-11-28 | 2021-11-28 | Application of human-derived gene in preparation of product for predicting sensitivity of patient to golimumab |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113957139A (en) |
-
2021
- 2021-11-28 CN CN202111426959.8A patent/CN113957139A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107209184B (en) | Marker combinations for diagnosing multiple infections and methods of use thereof | |
| JP6401702B2 (en) | Methods and compositions for diagnosis of inflammatory liver disease | |
| KR101771697B1 (en) | Composition and method for detecting a diagnostic marker for infectious disease or infectious complications using tryptophanyl-tRNA synthetase | |
| CN113234830B (en) | Product for lung cancer diagnosis and application | |
| CN112522413A (en) | Biomarker for evaluating gastric cancer risk and application thereof | |
| CN112522412A (en) | Reagent and product for detecting biomarkers and application of reagent and product in diseases | |
| EP3346270A1 (en) | Composition for diagnosing infectious diseases or infectious complications by using tryptophanyl-trna synthetase and method for detecting diagnostic marker | |
| CN113493829B (en) | Application of biomarker in pulmonary hypertension diagnosis and treatment | |
| CN116287220B (en) | Molecular biomarkers and assay methods for rapid diagnosis of kawasaki disease | |
| CN113846164A (en) | Marker molecule for predicting sensitivity of patient to preoperative radiotherapy and chemotherapy combined total rectal resection and derivative product thereof | |
| CN113817825A (en) | Molecular marker for predicting sensitivity of rectal cancer patient to preoperative chemoradiotherapy combined total rectal resection treatment | |
| CN113969312A (en) | Marker for predicting susceptibility of ulcerative colitis patient to golimumab drug | |
| CN112538531A (en) | Product for detecting gastric cancer | |
| CN112746107A (en) | Gastric cancer related biomarkers and their use in diagnosis | |
| CN112795648A (en) | Gastric cancer diagnostic product | |
| CN113444796B (en) | Biomarkers associated with lung cancer and their use in diagnosing cancer | |
| CN112877420B (en) | Biomarker related to retinopathy and application thereof | |
| CN113957139A (en) | Application of human-derived gene in preparation of product for predicting sensitivity of patient to golimumab | |
| CN113897428A (en) | Application of biomarker in judging sensitivity of ulcerative colitis patient to golimumab | |
| CN113999903A (en) | Multi-molecular marker for clinically evaluating agouti-xylol-resistant sensitivity of ulcerative colitis patient and device thereof | |
| CN113999902A (en) | Gene combination and application thereof in preparation of product for evaluating drug sensitivity of subject to golimumab | |
| CN113862356A (en) | Product for predicting sensitivity of rectal cancer to preoperative chemoradiotherapy combined total rectal resection treatment scheme through marker | |
| US20150376723A1 (en) | Prognostic biomarkers for influenza | |
| CN112575089A (en) | Application of gene in diagnosis of gastric cancer | |
| CN112725443A (en) | Biomarker combination and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220121 |