KR102417668B1 - Salivary gland tissue biomarker for diagnosing xerostomia and use thereof - Google Patents

Abstract

A composition and kit for diagnosing dry mouth disease, and a method for diagnosing dry mouth disease using the same are provided. According to this, it can be used to diagnose dry mouth disease simply and objectively, with high accuracy and specificity.

Description

Salivary gland tissue biomarker for diagnosing xerostomia and use thereof for diagnosing dry mouth

To a salivary gland tissue biomarker for diagnosing dry mouth, a composition and kit for diagnosing dry mouth using the same, and a method for detecting a biomarker using the same.

Salivary gland function is affected by several conditions. Dry mouth is a subjective feeling of dry mouth because there is no saliva in the mouth. When the function of the salivary glands decreases due to dry mouth and the function of lubricating or antibacterial action decreases, dental caries and periodontal disease may appear as secondary diseases in the oral cavity. Dry mouth disease is difficult to recognize in the early stage of decreased salivary function, and it is difficult to distinguish between temporary dry mouth and organic dry mouth. Geriatric dry mouth due to aging is also caused by organic salivary gland dysfunction. Dry mouth caused by salivary gland dysfunction is difficult to cure after the disease has progressed, and treatment focuses on alleviating symptoms rather than completely curing the disease.

Dry mouth can be caused by various causes, such as Sjogren's syndrome, anemia, diabetes, aging, drug use, and nervous system diseases. Recently, a method for diagnosing and treating Sjogren's syndrome by detecting a biomarker in a saliva sample of an individual having dry mouth has been developed (US Patent Publication No. 9,833,519B2). However, a biomarker for diagnosing dry mouth has not been developed.

Therefore, there is a need to develop a biomarker capable of diagnosing dry mouth disease quickly, objectively, simply and inexpensively, and with high diagnostic accuracy and specificity.

Provided is a composition for diagnosing dry mouth.

Provides a kit for diagnosing dry mouth.

A method for diagnosing dry mouth is provided.

One aspect is ANP32E (Acid leucine-rich nuclear phosphoprotein 32 family member E), SLC25A1 (Tricarboxylate transport protein, mitochondrial), CCAR1 (Cell division cycle and apoptosis regulator protein 1), PGRMC1 (Membrane-associated progesterone receptor component 1), MTARC2 (Mitochondrial amidoxime reducing component 2), SORL1 (Sortilin-related receptor), ITGA5 (Integrin alpha-5), SOD2 (Superoxide dismutase [Mn], mitochondrial), ERH (Enhancer of rudimentary homolog), MYH4 (Myosin-4), A gene selected from the group consisting of 40S ribosomal protein S4, X isoform (RPS4X), Regulation of nuclear pre-mRNA domain-containing protein 1B (RPRD1B), Unconventional myosin-VI (MYO6), and Troponin I, slow skeletal muscle (TNNI1). It provides a composition for diagnosing dry mouth, comprising an agent for measuring the expression level of, or the expression level of a protein or fragment thereof.

The term "xerostomia or dry mouth" refers to a symptom of dry mouth due to decreased salivation or various other causes. Dry mouth can also be referred to as salivary gland hypofunction. Dry mouth is caused by Sjogren's syndrome, anemia, diabetes, nutritional deficiencies, aging, drug use (e.g., antihistamines, drugs that act on the nervous system, high blood pressure), nervous system disorders, mental disorders (e.g. depression), and radiation therapy. can be caused by When dry mouth is induced, it is difficult to swallow food due to the dryness itself, and you may feel discomfort that it is difficult to speak, the occurrence of tooth decay (dental caries) and tooth decay (periodontitis) increases and is easy to worsen, fungal infection in the mouth, It can cause tongue pain, bad breath, and taste abnormalities. Dry mouth can also affect the development of oral diseases such as oral ulcers. Dry mouth can be diagnosed by measurement of salivation (salivation volume at rest less than 0.1 mL/min), salivography, biopsy, or nuclear medicine examination. It can also be diagnosed through salivary gland biopsy, and it can be diagnosed with the degree of regression of the salivary gland parenchyma in the salivary gland tissue. In Sjögren's syndrome, the infiltration pattern of immune cells in the salivary gland tissue is measured as an evaluation of salivary gland dysfunction, and it can be classified into a score of 0 to 6 depending on the degree of infiltration of the immune cells.

The composition may be for detection in a salivary gland tissue sample. The expression level may be increased compared to the expression level of the normal control.

Acidic leucine-rich nuclear phosphoprotein 32 family member E (ANP32E) is a histone chaperone that specifically mediates clearance of histones H2A.Z/H2AZ1 from nucleosomes. ANP32E can also be called LANP-L or LANPL. ANP32E may comprise the amino acid sequence of Uniprot number Q9BTT0 in humans. ANP32E may comprise the amino acid sequence of Uniprot number P97822 in a mouse.

SLC25A1 (Tricarboxylate transport protein, mitochondrial) is a citrate transporter that mediates the exchange of mitochondrial citrate to cytosolic malate. SLC25A1 may also be referred to as citrate transport protein (CTP), solute carrier family 25 member 1, or tricarboxylate carrier protein. SLC25A1 may comprise the amino acid sequence of Uniprot number P53007 in humans. SLC25A1 may comprise the amino acid sequence of Uniprot number Q8JZU2 in a mouse.

CCAR1 (Cell division cycle and apoptosis regulator protein 1) is a protein involved in transmitting regulatory signals from an upstream transcriptional activation protein to a downstream transcriptional machinery. CCAR1 may also be referred to as CARP-1 or Death inducer with SAP domain. CCAR1 may comprise the amino acid sequence of Uniprot number Q8IX12 in humans. CCAR1 may comprise the amino acid sequence of Uniprot number Q8CH18 in a mouse.

PGRMC1 (Membrane-associated progesterone receptor component 1) is a component of the progesterone-binding protein complex. PGRMC1 may also be referred to as HPR6.6, MPR, Dap1, or IZA. PGRMC1 may comprise the amino acid sequence of Uniprot No. O00264 or Q6IB11 in humans. PGRMC1 may comprise the amino acid sequence of Uniprot number O55022 in a mouse.

MTARC2 (Mitochondrial amidoxime reducing component 2) can catalyze the reduction of N-oxidized molecules. MTARC2 may also be referred to as MOSC2 or MARC2. MTARC2 may comprise the amino acid sequence of Uniprot number Q969Z3 in humans. MTARC2 may comprise the amino acid sequence of Uniprot number Q922Q1 in a mouse.

SORL1 (Sortilin-related receptor) is a neuronal apolipoprotein E receptor. SORL1 may also be referred to as L (DLR class) A repeats containing, C11orf32, LR11, LRP9, SORLA, SorLA-1, gp250, or sortilin related receptor 1. SORL1 may comprise the amino acid sequence of Uniprot number Q92673 in humans. SORL1 may comprise the amino acid sequence of Uniprot number 088307 in a mouse.

ITGA5 (Integrin alpha-5) belongs to the integrin alpha chain family. ITGA5 may also be referred to as CD49e, FNRA, VLA5A, or VLA-5. ITGA5 may comprise the amino acid sequence of Uniprot number P08648 in humans. ITGA5 may comprise the amino acid sequence of Uniprot number P11688 in a mouse.

SOD2 (superoxide dismutase [Mn], mitochondrial) is a manganese-dependent superoxide dismutase. SOD2 may also be referred to as IPOB, MNSOD, MVCD6, IPO-B, Mn-SOD, or GClnc1. SOD2 may comprise the amino acid sequence of Uniprot number P04179 in humans. SOD2 may comprise the amino acid sequence of Uniprot number P09671 in a mouse.

ERH (Enhancer of rudimentary homolog) is an enhancer of a gene involved in pyrimidine biosynthesis and may play a role in the cell cycle. ERH may also be referred to as DROER, mRNA splicing and mitosis factor, or ERH mRNA splicing and mitosis factor. ERH may comprise the amino acid sequence of Uniprot number P84090 in humans. ERH may comprise the amino acid sequence of Uniprot number P84089 in a mouse.

MYH4 (Myosin-4) is a sarcomeric myosin. MYH4 may also be referred to as MYH2B, MyHC-2B, MyHC-IIb, myosin heavy chain 4, or skeletal muscle myosin main heavy chain 4. MYH4 may comprise the amino acid sequence of Uniprot number Q9Y623 in humans. MYH4 may comprise the amino acid sequence of Uniprot number Q5SX39 in a mouse.

RPS4X (40S ribosomal protein S4, X isoform) is a protein constituting the ribosome. RPS4X may also be referred to as CCG2, DXS306, RPS4, S4, SCAR, SCR10, ribosomal protein S4, X-linked, or ribosomal protein S4 X-linked. RPS4X may comprise the amino acid sequence of Uniprot number P62701 in humans. RPS4X may comprise the amino acid sequence of Uniprot number P62702 in a mouse.

Regulation of nuclear pre-mRNA domain-containing protein 1B (RPRD1B) is a protein that binds to the phosphorylated C-terminal 7-peptide repeat domain of the RNA polymerase II subunit. RPRD1B can also be called cell cycle-related and expression-elevated protein in tumor. RPRD1B may comprise the amino acid sequence of Uniprot number Q9NQG5 in humans. RPRD1B may comprise the amino acid sequence of Uniprot number Q9CSU0 in a mouse.

MYO6 (Unconventional myosin-VI) is a protein involved in intracellular vesicle and organelle transport. MYO6 may also be referred to as DFNA22, DFNB37, myosin VI, Myo6-008, or Myo6-007. MYO6 may comprise the amino acid sequence of Uniprot number Q9UM54 in humans. MYO6 may comprise the amino acid sequence of Uniprot number Q64331 in a mouse.

TNNI1 (Troponin I, slow skeletal muscle) is a tissue-specific subtype of troponin I constituting the troponin complex. TNNI1 may also be referred to as SSTNI, TNN1, or troponin I1, slow skeletal type. TNNI1 may comprise the amino acid sequence of Uniprot number P19237 in humans. TNNI1 may comprise the amino acid sequence of Uniprot number Q9WUZ5 in a mouse.

The fragment is a part of the protein and may be an immunogenic polypeptide.

The gene refers to a nucleic acid encoding the protein. The expression level of the gene may be the expression level of mRNA encoding the protein. The expression level of mRNA may be a relative amount or an absolute amount of mRNA. Measuring the expression level of the gene may be measuring the amount of mRNA.

The expression level of the protein or fragment thereof may be a relative amount or an absolute amount of the protein or fragment thereof. Measuring the expression level of the protein or fragment thereof may be measuring the amount of the protein or fragment thereof.

The agent may be an antibody or antigen-binding fragment thereof that specifically binds to the protein or fragment thereof, or an aptamer. The antibody may be a polyclonal antibody or a monoclonal antibody. The term “antibody” may be used interchangeably with the term “immunoglobulin”. The antibody may be a polyclonal antibody or a monoclonal antibody. The antibody may be a full-length antibody. The antigen-binding fragment refers to a polypeptide comprising an antigen-binding site. The antigen-binding fragment may be a single-domain antibody, Fab, Fab', or scFv. The antibody or antigen-binding fragment may be attached to a solid support. The solid support is, for example, a metal chip, a plate, or the surface of a well. The antibody or antigen-binding fragment may be an antibody or antigen-binding fragment. The aptamer may be a single-stranded nucleic acid (DNA, RNA or modified nucleic acid) or peptide that specifically binds to the protein or fragment thereof.

The agent may be a nucleic acid comprising a polynucleotide identical to or complementary to a polynucleotide encoding the protein or fragment thereof. The nucleic acid may be a primer, a probe, or an antisense oligonucleotide. The primer, probe, or antisense oligonucleotide may be labeled with a fluorescent material, chemiluminescent material, or radioactive isotope at the end or inside thereof.

Another aspect is the expression level of a gene selected from the group consisting of ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ITGA5, SOD2, ERH, MYH4, RPS4X, RPRD1B, MYO6, and TNNI1, or the expression level of a protein or fragment thereof. It provides a kit for diagnosing dry mouth disease including a measuring agent.

ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ITGA5, SOD2, ERH, MYH4, RPS4X, RPRD1B, MYO6, TNNI1, and xerostomia as described above.

The kit may further include a sample necessary for diagnosing dry mouth. The kit may include a solid support, a substrate for immunological detection of an antibody or antigen-binding fragment, a suitable buffer, a chromogenic enzyme, a secondary antibody labeled with a fluorescent material, or a chromogenic substrate. The kit may include a polymerase, a buffer, a nucleic acid, a coenzyme, a fluorescent material, or a combination thereof for nucleic acid detection. The polymerase is, for example, Taq polymerase.

Another aspect is a gene selected from the group consisting of ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ITGA5, SOD2, ERH, MYH4, RPS4X, RPRD1B, MYO6, and TNNI1 in a salivary gland tissue sample isolated from a subject suspected of dry mouth measuring the expression level of, or the expression level of a protein or fragment thereof; comparing the measured expression level with the expression level of a normal control; And when the expression level is increased compared to the expression level of the normal control, it provides a method of detecting a biomarker to provide information necessary for diagnosing dry mouth or dry mouth, comprising the step of determining dry mouth do.

ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ITGA5, SOD2, ERH, MYH4, RPS4X, RPRD1B, MYO6, TNNI1, and xerostomia as described above.

The method comprises a gene selected from the group consisting of ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ITGA5, SOD2, ERH, MYH4, RPS4X, RPRD1B, MYO6, and TNNI1 in a salivary gland tissue sample isolated from a subject suspected of dry mouth. measuring the expression level of, or the expression level of a protein or fragment thereof.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be suffering from a dry mouth disorder or may be an individual suspected of suffering from dry mouth.

The salivary gland tissue sample refers to a salivary gland tissue sample obtained from the subject. The salivary gland tissue sample may be a sample that has been subjected to pretreatment such as centrifugation and filtration. The salivary gland tissue sample includes a protein sample or a nucleic acid sample obtained from the salivary gland tissue sample.

In the measuring step, the salivary gland tissue sample is incubated with an antibody or antigen-binding fragment thereof, or an aptamer that specifically binds to the protein or fragment thereof, or the same as or identical to the polynucleotide encoding the gene. incubating the complementary polynucleotide.

The measuring step includes electrophoresis, immunoblotting, enzyme-linked immunosorbent assay (ELISA), polymerase chain semi-Northern blotting, polymerase chain reaction (PCR). , protein chip, immunoprecipitation, microarray, electron microscopy, or a combination thereof. The electrophoresis may be SDS-PAGE, isoelectric point electrophoresis, two-dimensional electrophoresis, or a combination thereof.

The method comprises comparing the measured expression level to the expression level of a normal control. In the method, the expression level of any one selected from the group consisting of ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ITGA5, SOD2, ERH, MYH4, RPS4X, RPRD1B, MYO6, and TNNI1 is compared with that of a normal control group. It may include the step of checking whether the increase. The normal control group is a group that does not have dry mouth and means a negative control group. In the method, the amount of the measured expression level of the salivary gland tissue sample may be increased compared to the expression level of the normal control. For example, when the amount of the detected biomarker is less than that of a normal control, the subject may be diagnosed as having or having a high probability of having dry mouth. Treatments such as oral care, control of drug administration, administration of saliva substitutes, administration of oral moisturizers, electrical stimulation of the oral cavity or acupuncture may be performed on an individual diagnosed with dry mouth.

According to the composition and kit for diagnosing dry mouth according to an aspect, and a method for diagnosing dry mouth using the same, it can be used to diagnose dry mouth disease simply, objectively, with high accuracy and specificity.

1 is an SDS-PAGE gel image of salivary gland tissue samples from a normal control group and a group of patients with dry mouth (left: whole gel, right: fractionated gel).
2 is a Venn diagram showing the number of proteins identified in a salivary gland tissue sample and proteins having a p-value of less than 0.05 analyzed and statistically significant.

Hereinafter, it will be described in more detail through examples. However, these examples are for illustrative purposes of one or more embodiments, and the scope of the present invention is not limited to these examples.

Example 1. Identification of differentially expressed biomarkers in salivary gland tissue samples from dry mouth patients

1. Preparation of salivary gland tissue samples from dry mouth patients

Salivary gland tissue samples were collected from a normal control group (n=3) and a patient diagnosed with dry mouth from Hanyang University. Dry mouth was diagnosed by evaluating salivary gland function in clinical examinations of salivary gland secretion and oral mucosa condition, and salivary gland scans. Salivary gland function was classified into a score of 1 to 5 depending on the condition of the salivary gland tissue and the infiltration of lymphocytes around the parenchyma of the salivary gland by performing a salivary gland biopsy. Salivary gland tissue sampling was performed by classification into 3 groups of score 1, score 2, and score 3 to 5.

To prevent proteolysis or modification of the salivary gland tissue sample, cOmplete as a protease inhibitor, Mini EDTA-free protease inhibitor cocktail (Roche) and PhosSTOP (Roche) as a phosphatase inhibitor were added to the salivary gland tissue sample obtained immediately after serum was obtained. ) was added. The inhibitor was dissolved in 200 μl of phosphate buffered saline (PBS) according to the manufacturer's protocol to make 50 X, and then added to each sample so that the final concentration of the inhibitor was 1.5 X. In order to increase protein extraction efficiency, using scissors, a tissue sample was cut finely, vortexing and incubation were repeated to remove blood contained in the tissue.

2. Concentration and Quantification of Salivary Gland Tissue Samples

Add lysis buffer (50 mM Tris-HCl (pH 7.5), 7 M urea, 2 M thiourea, 1 mM EDTA, 150 mM NaCl, protease and phosphatase inhibitor cocktails (Roche)) to the tissue sample from which blood was removed in 1. did The reaction was repeatedly subjected to vortexing, incubation, and sonication to efficiently perform protein extraction. The obtained protein was quantified using a Bradford assay kit.

3. Preparation of samples for mass spectrometry and in-gel digestion

For proteome analysis, 90 μg of 6 individual samples from 2 groups obtained in 2. were separated according to molecular weight using NuPAGE gels (Bis-Tris gels from 4% to 12%, Invitrogen, Carlsbad, CA, USA). did The gel was stained with Coomassie Brilliant Blue R-250 (FIG. 1). The stained gel for 6 individual samples was divided into 13 slices, and each gel slice was transferred to a microcentrifuge tube. Proteins contained in each gel piece were reduced by disulfide bonds at 56°C, then alkylated in a dark environment at 25°C, and digested with trypsin overnight. Then, the peptide was extracted from a solution of 67% (v/v) acetonitrile (ACN)/5% (v/v) formic acid (FA) (Wako Pure Chemicals, Osaka, Japan). The extracted peptides were dried in Speedvac and dissolved in 20 μl of 0.4% (v/v) acetic acid.

4. Mass Spectrum Analysis

10 μl of each sample obtained in 3. was injected onto a reversed-phase Magic C18AQ column (15 cm X 75 μm) on an Eksigent MDLC system (Eksigent Technologies Dublin, CA, USA). The column was mixed with 95% Buffer A (100% (v/v) water containing 0.1% (v/v) formic acid) and 5% (v/v) Buffer B (0.1% (v/v) of 100% (v/v) ACN) containing formic acid) was equilibrated with a mixed buffer. The working flow rate was 0.35 μl/min (min) under the following gradient conditions:

100% Buffer A from 0 min to 5 min; 92% Buffer A and 8% Buffer B from 5 min to 85 min; 70% Buffer A and 30% Buffer B from 85 to 90 minutes; 30% Buffer A and 70% Buffer B from 90 min to 95 min; 30% Buffer A and 70% Buffer B from 95 to 100 minutes; and 98% Buffer A and 2% Buffer B from 100 to 110 minutes.

The nano HPLC system was mounted on an LTQ XL-Orbitrap mass spectrometer (Thermo Scientific, San Jose, CA, USA). The spray voltage was set at 2.5 kV, and the temperature of the heated capillary was set at 250°C. Survey full-scan MS spectra (300-1,800 m/z) were obtained with 1 microscan and a resolution of 60,000 to select precursors and charge states A preview mode for measurement was set. MS/MS spectra for the 10 most potent ions from the preview irradiation scan were obtained in the ion trap for the full scan with the following options: isolation width, 2 m/z (mass versus charge); normalized collision energy, 35%; dynamic exclusion duration, 360 seconds (sec). Lead materials with +1 charge and undetermined charge states were discarded during data-dependent acquisition.

Each LC-MS/MS file was analyzed using the Andromeda algorithm of MaxQunat1.5.8.3. MS and MS/MS data were compared against SwissProt human database (January 2019). The investigation was limited to accepting tryptic peptides with two missed cleavages. Carboamidomethylation of cysteine was set as a fixed modification (+57.021 Da) and methionine oxidation as a fluid modification (+15.995 Da). Mass tolerance was set at 0.5 Da for MS/MS data and 15 ppm for MS data. For decoy database research, two target values were applied: a strict false discovery rate (FDR) of 0.01 and a relaxed FDR of 0.05. All proteins were identified using two or more matching peptides.

5. Label-free quantitative analysis

Based on the label-free quantification intensity (LFQ intensity), the relative abundance of proteins present in a total of six samples obtained by collecting three salivary gland tissue samples from two groups of the normal control group and the dry mouth group. was calculated. The intensity was extracted using MaxQunat1.5.8.3. Then, the LFQ intensity for each protein was analyzed for relative quantitative analysis using Perseus. Even when no protein was present or below the detection limit, null values were compensated. In order to calculate the fold difference between the normal control group and the group of patients with dry mouth, the area was downshifted by 0.3 and 1.8 from the normal distribution to fill in the missing values. Statistical significance was performed for the LFQ intensity obtained from the ANOVA test with three separate samples of the normal control group and the patient group with dry mouth, and a p-value of less than 0.05 was considered statistically significant.

6. Identification of salivary gland tissue proteins related to patient groups according to dry mouth and identification of differentially expressed proteins

From the LFQ intensity analysis, a total of 3119 proteins were identified in 6 individual samples of the normal control group and the dry mouth group. Among the identified proteins, 20 proteins with a p-value of less than 0.05 were identified. Fig. 2 shows the results of analyzing the identified proteins and proteins having a p-value of less than 0.05 and showing the number as a Venn diagram.

Table 1 shows proteins whose p-value is less than 0.05 and whose expression level is increased or decreased by two or more times compared to the normal control, among proteins that are statistically significant.

UniProt Accession No. gene name
protein name LFQ strength
(normal control) LFQ strength
(Dry mouth disease patient group) Protein expression rate
(dry mouth disease patient group/normal control group) FDR corrected p-value Q9BTT0 ANP32E Acidic leucine-rich nuclear phosphoprotein 32 family member E 3.3.E+06 3.4.E+05 0.106 0.016 P53007 SLC25A1 Tricarboxylate transport protein, mitochondrial 1.1.E+08 4.2.E+07 0.396 0.017 Q8IX12 CCAR1 Cell division cycle and apoptosis regulator protein 1 4.2.E+06 7.6.E+05 0.182 0.018 O00264 PGRMC1 Membrane-associated progesterone receptor component 1 2.6.E+07 1.4.E+07 0.535 0.020 Q969Z3 MTARC2 Mitochondrial amidoxime reducing component 2 1.8.E+07 9.7.E+06 0.542 0.032 Q92673 SORL1 Sortilin-related receptors 9.6.E+06 1.7.E+06 0.175 0.033 P08648 ITGA5 Integrin alpha-5 0 4.5.E+06 Not detected in normal control group 0.035 P04179 SOD2 Superoxide dismutase [Mn], mitochondrial 1.5.E+08 3.7.E+08 2.514 0.037 P84090 ERH Enhancer of rudimentary homolog 2.8.E+07 1.5.E+07 0.547 0.039 Q9Y623 MYH4 Myosin-4 2.4.E+07 1.3.E+08 5.405 0.046 P62701 RPS4X 40S ribosomal protein S4, X isoform 6.3.E+08 2.3.E+08 0.371 0.047 Q9NQG5 RPRD1B Regulation of nuclear pre-mRNA domain-containing protein 1B 5.6.E+06 0 0
(Not detected in score 3 or higher) 0.047 Q9UM54 MYO6 Unconventional myosin-VI 2.7.E+07 7.9.E+06 0.295 0.048 P19237 TNNI1 Troponin I, slow skeletal muscle 2.4.E+07 1.1.E+08 4.473 0.048

As shown in Table 1, expression levels of ITGA5, SOD2, MYH4 and TNNI1 were significantly increased in dry mouth, and ANP32E, SLC25A1, CCAR1, PGRMC1, MTARC2, SORL1, ERH, RPS4X, RPRD1B and MYO6 were significantly increased in dry mouth. It was confirmed that the expression level was decreased.

Claims (13)

A composition for diagnosing dry mouth, comprising an agent for measuring the expression level of the MYH4 (Myosin-4) gene, or the expression level of a protein or fragment thereof. The composition according to claim 1, for detection in a salivary gland tissue sample. The composition of claim 1, wherein the expression level is increased compared to the expression level of the normal control. The method according to claim 1, wherein the agent is
an antibody or antigen-binding fragment thereof, or an aptamer that specifically binds to the protein or fragment thereof; or
The composition is a nucleic acid comprising a polynucleotide identical to or complementary to the polynucleotide encoding the protein or fragment thereof. The composition of claim 4, wherein the antibody is a polyclonal antibody or a monoclonal antibody. The composition of claim 4, wherein the nucleic acid is a primer, a probe, or an antisense oligonucleotide. The method according to claim 1, ANP32E (Acid leucine-rich nuclear phosphoprotein 32 family member E), SLC25A1 (Tricarboxylate transport protein, mitochondrial), CCAR1 (Cell division cycle and apoptosis regulator protein 1), PGRMC1 (Membrane-associated progesterone receptor component 1) , MTARC2 (Mitochondrial amidoxime reducing component 2), SORL1 (Sortilin-related receptor), ITGA5 (Integrin alpha-5), SOD2 (Superoxide dismutase [Mn], mitochondrial), ERH (Enhancer of rudimentary homolog), RPS4X (40S ribosomal protein) the expression level of a gene selected from the group consisting of S4, X isoform), Regulation of nuclear pre-mRNA domain-containing protein 1B (RPRD1B), Unconventional myosin-VI (MYO6), and Troponin I, slow skeletal muscle (TNNI1), or The composition further comprising an agent for measuring the expression level of the protein or fragment thereof. A kit for diagnosing dry mouth disease, comprising an agent for measuring the expression level of the MYH4 gene, or the expression level of a protein or fragment thereof. The method according to claim 8, ANP32E (Acid leucine-rich nuclear phosphoprotein 32 family member E), SLC25A1 (Tricarboxylate transport protein, mitochondrial), CCAR1 (Cell division cycle and apoptosis regulator protein 1), PGRMC1 (Membrane-associated progesterone receptor component 1) , MTARC2 (Mitochondrial amidoxime reducing component 2), SORL1 (Sortilin-related receptor), ITGA5 (Integrin alpha-5), SOD2 (Superoxide dismutase [Mn], mitochondrial), ERH (Enhancer of rudimentary homolog), RPS4X (40S ribosomal protein) the expression level of a gene selected from the group consisting of S4, X isoform), Regulation of nuclear pre-mRNA domain-containing protein 1B (RPRD1B), Unconventional myosin-VI (MYO6), and Troponin I, slow skeletal muscle (TNNI1), or The kit further comprising an agent for measuring the expression level of the protein or fragment thereof. measuring the expression level of the MYH4 gene or the expression level of a protein or fragment thereof in a salivary gland tissue sample isolated from a subject suspected of dry mouth;
comparing the measured expression level with the expression level of a normal control; and
When the expression level is increased compared to the expression level of the normal control, comprising the step of determining dry mouth,
A method of detecting a biomarker to provide information necessary for the diagnosis of dry mouth. The method according to claim 10, wherein the measuring comprises incubating the salivary gland tissue sample with an antibody or antigen-binding fragment thereof, or an aptamer that specifically binds to the protein or fragment thereof, or a polynucleotide encoding the gene A method comprising incubating a polynucleotide identical to or complementary thereto. The method according to claim 10, wherein the measuring step is electrophoresis, immunoblotting, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction, Northern blotting, polymerase amplification reaction ( polymerase chain reaction: PCR), protein chip, immunoprecipitation, microarray, electron microscopy, or a method that is performed by a combination thereof. The method according to claim 10, ANP32E (Acid leucine-rich nuclear phosphoprotein 32 family member E), SLC25A1 (Tricarboxylate transport protein, mitochondrial), CCAR1 (Cell division cycle and apoptosis regulator protein 1), PGRMC1 (Membrane-associated progesterone receptor component 1) , MTARC2 (Mitochondrial amidoxime reducing component 2), SORL1 (Sortilin-related receptor), ITGA5 (Integrin alpha-5), SOD2 (Superoxide dismutase [Mn], mitochondrial), ERH (Enhancer of rudimentary homolog), RPS4X (40S ribosomal protein) the expression level of a gene selected from the group consisting of S4, X isoform), Regulation of nuclear pre-mRNA domain-containing protein 1B (RPRD1B), Unconventional myosin-VI (MYO6), and Troponin I, slow skeletal muscle (TNNI1), or The method further comprising the step of measuring the expression level of the protein or fragment thereof.

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