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WO2006115047A1 - Gene whose expression changes in conjunction with diabetic conditions and utilization thereof - Google Patents

Gene whose expression changes in conjunction with diabetic conditions and utilization thereof Download PDF

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Publication number
WO2006115047A1
WO2006115047A1 PCT/JP2006/307700 JP2006307700W WO2006115047A1 WO 2006115047 A1 WO2006115047 A1 WO 2006115047A1 JP 2006307700 W JP2006307700 W JP 2006307700W WO 2006115047 A1 WO2006115047 A1 WO 2006115047A1
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chromosome
reading frame
open reading
gene
type
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Japanese (ja)
Inventor
Shuichi Kaneko
Toshinari Takamura
Hirofumi Misu
Taro Yamashita
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Kanazawa University NUC
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Kanazawa University NUC
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    • 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
    • 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

Definitions

  • the present invention relates to the use of genes that can be key molecules for the pathogenesis of sugar'lipid metabolism, diabetic complications, arteriosclerosis, etc., and the present invention further varies in expression in relation to the pathology of diabetes. Further, the present invention relates to the use of the gene for diagnosis or risk assessment of diabetes or obesity.
  • Type 2 diabetes 'obesity' continues to increase on a global scale and promotes arteriosclerosis such as retinal 'renal' nerve complications and ischemic heart disease, threatening human QOL and life.
  • the liver is an important organ that plays a central role in glucose and fat metabolism.
  • insulin resistance the action of insulin to suppress the release of sugar by hepatic strength is attenuated, and this phenomenon is called insulin resistance (see Non-Patent Documents 1 and 2).
  • Insulin resistance leads to hyperglycemia due to increased sugar release by liver strength and hyperlipidemia due to increased lipid production, and both promote arteriosclerosis.
  • the liver is the largest organ in the body of various physiologically active substances including angiogenic factors that lead to the risk of arteriosclerosis.
  • Non-Patent Document 1 Michael MD. Et al., Mol. Cell 6: 87-97, 2000
  • Non-Patent Document 2 Saltiel AR. Et al., Nature 414: 799-806, 2001
  • the present invention relates to a gene that can be a key molecule for producing a pathological condition such as sugar'lipid metabolism, diabetic complications, arteriosclerosis, and the sugar'lipid metabolism, diabetic complications, arteriosclerosis of the gene.
  • the method of use to elucidate the cause of pathological conditions such as is provided.
  • the present invention relates to a reagent for measuring the expression of a gene whose expression varies in relation to diabetes or obesity using a gene whose expression varies in relation to the pathological condition of diabetes, and diabetes using the reagent,
  • the purpose is to provide methods for detecting obesity, diabetic complications, arteriosclerosis, and the like.
  • the present inventor has constructed the world's largest liver-expressed gene database that exceeds the 600,000-expressed genes obtained from SAGE of normal liver and various liver diseases and DNA chip analysis so far, and analyzes liver disease. I have recommended.
  • a cDNA microarray originally developed in the process, the profile of the gene expressed in the liver of type 2 diabetic patients was analyzed, and it was thought that there was no abnormality in the conventional research method.
  • the expression of various known genes related to vascular complications such as angiogenic factors that are not only directly related to glucose metabolism was increased (Takamura T. et al. al., Diabetologia 47: 638-647, 2004). These known genes may shape the pathology of type 2 diabetes.
  • liver-derived secretory proteins may have unknown secreted proteins that shape the pathology of diabetes, including vascular complications, in livers with type 2 diabetes and obesity. . Therefore, this original research was further developed, and the gene produced by the liver of type 2 diabetic patients, including unknown genes with unknown functions, was analyzed using the serial analysis of gene expression method (hereinafter referred to as SAGE method).
  • SAGE method serial analysis of gene expression method
  • a gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic or obese patient compared to the liver tissue of a normal human subject, which encodes a secreted protein In order to measure the expression of type 2 diabetes-related genes and Z or obesity-related genes containing nucleotides consisting of the base sequence of at least one selected gene or nucleotides including a partial sequence thereof Reagent.
  • nucleobindin 2 nucleobindin 2
  • matrix metalloproteinase 11 matrix metalloproteinase 11 (stromelysin 3)
  • DST Homo sapiens dystonin
  • glycosylphosphatidylinositol specific phospholipase Dl (32) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15
  • ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1
  • a gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is selected from the group consisting of the following genes whose up- or down-expression is correlated with HbAlc levels in human blood
  • matrix metalloproteinase 11 (stromelysin 3)
  • a gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is at least 1 selected from the group consisting of the following genes whose up-regulation or attenuation is correlated with HOMA-R
  • Phospholipase A2 group VII (platelet-activating factor acetylhydrolase, plasma)
  • a gene whose expression is up- or down-regulated in the liver tissue of human type 2 diabetics is correlated with increased or decreased expression.
  • ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1
  • a gene whose expression is up- or down-regulated in liver thread and tissue of human obesity patients is selected from the group consisting of the following genes whose up-regulation or attenuation is correlated with BMI: A reagent for measuring the expression of genes related to type 2 diabetes and Z or obesity related genes of [1].
  • DST Homo sapiens dystonin
  • a reagent for measuring the expression of a type 2 diabetes-related gene and a Z- or obesity-related gene according to any one of [1] to [5], comprising at least 5 genes.
  • a nucleotide comprising a nucleotide sequence or a partial sequence thereof that also has a base sequence ability of a gene whose expression is increased or decreased in the liver tissue of a human type 2 diabetic patient compared to a liver tissue of a normal human subject
  • a nucleotide comprising a nucleotide sequence or a partial sequence thereof, which also has a base sequence ability of a gene whose expression is increased or decreased in the liver tissue of a human type 2 diabetic patient compared to the liver tissue of a normal human subject
  • the expression in a subject is measured, and if the expression of the gene is increased or decreased compared to a normal person, the subject is type 2 diabetes, How to determine that you have or are at risk of suffering from obesity, diabetic complications or arteriosclerosis.
  • matrix metalloproteinase 11 (stromelysin 3)
  • DST Homo sapiens dystonin
  • ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1
  • a gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is a group consisting of the following genes whose up- or down-expression is correlated with HbAlc levels in human blood: The method of [9], which is a single gene.
  • matrix metalloproteinase 11 (stromelysin 3)
  • a gene whose expression is enhanced or attenuated in the liver tissue of a human type 2 diabetic patient is selected as a group force with the following gene strength in which increased or attenuated expression is correlated with HOMA-R: at least 1 selected The method of [9], which is one gene. [0018] (42) thrombospondin 1
  • Phospholipase A2 group VII (platelet-activating factor acetylhydrolase, plasma)
  • group VII platelet-activating factor acetylhydrolase, plasma
  • ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1
  • chromosome 5 open reading frame 13 (61) chromosome 6 open reading frame 79
  • a gene whose expression is enhanced or attenuated in the liver tissue of a human type 2 diabetes patient is at least one gene selected from the group consisting of the following genes whose expression is correlated with BMI: There is a method of [9].
  • DST Homo sapiens dystonin
  • a nucleotide comprising a nucleotide sequence or a partial sequence thereof, which also has a base sequence ability of a gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic patient compared to the liver tissue of a normal human subject [9] to [14] !, either method.
  • a nucleotide comprising a nucleotide sequence or a partial sequence thereof that also has a base sequence ability of a gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic patient compared to the liver tissue of a normal human subject [9] to [14], any of the methods including analyzing the expression of the gene by real-time PCR targeting at [9].
  • Type 2 diabetes, obesity, diabetic comprising a type 2 diabetes-related gene according to any one of [1] to [8] and a reagent for measuring the expression of Z or an obesity-related gene Kit for detection or risk assessment of complications and arteriosclerosis.
  • the gene group can be a key molecule for the pathogenesis of sugar and lipid metabolism, diabetic complications, arteriosclerosis, etc. By using this gene, sugar / lipid metabolism, diabetic complications, arteriosclerosis, etc. It is possible to elucidate the mechanism of the pathophysiology.
  • the gene whose expression changes in relation to the pathological condition of obesity or obesity of the present invention is a gene whose expression is different between the liver of a type 2 diabetes patient or obese patient and the liver of a normal person, and the liver of a normal person In comparison with the above, it is a gene whose expression in the liver of type 2 diabetic patients or obese patients is increased or decreased. Furthermore, the gene of the present invention is a gene encoding a secretory protein that is secreted extracellularly.
  • type 2 diabetes is a type of diabetes that develops due to a decrease in insulin secretion and a decrease in sensitivity to insulin.
  • a normal person means a person who does not exhibit type 2 diabetes, specifically, a person with normal insulin resistance in the liver or skeletal muscle, that is, a person with normal glucose tolerance.
  • blood HbAlc (hemoglobin Ale) concentration increases, and insulin resistance (insulin resistance in the liver and insulin resistance in skeletal muscle) power S is increased.
  • Insulin resistance in the liver can be determined by measuring HOMA-R (homeostasis model assessment), and the higher the value of HOMA-R, the stronger the insulin resistance in the liver.
  • Insulin resistance in skeletal muscle can be determined by measuring the glucose metabolic clearance rate (MCR). The larger the MCR value, the stronger the insulin resistance in skeletal muscle.
  • MCR glucose metabolic clearance rate
  • HOM AR values fasting blood glucose ⁇ / (11) fasting Insurin concentration (11 1?) (U / ml) / 405 indicated by (M atthews DR et al, Diabetologia 28:. 412-419, 1985), This value is around 1 for normal people.
  • the MCR value can be measured by, for example, the method described in DeFronzo RA et al., Am J Physiol 237: E214-223, 1979.
  • Obese patients are those whose BMI (Body Mass index) is high, and BMI is expressed as body weight (kg) Z height (m) 2 . If the BMI is 25 or greater, it is determined to be obese.
  • the gene whose expression changes in association with the pathological condition or obesity of type 2 diabetes of the present invention is a gene that is significantly correlated with any one of blood HbAlc level, HOMA-R, MCR and BMI.
  • a significant correlation is observed when, for example, the relationship between the measured values of blood HbAlc, HOM AR, MCR and BMI and the expression level of each gene is analyzed by linear correlation analysis. It means being done.
  • genes shown in Table 3 as genes that are significantly correlated with HbAlc levels in blood, and 29 genes shown in Table 4 as genes that are significantly correlated with BMI are significantly correlated with HOMA-R.
  • the 6 genes shown in Table 5 are recognized genes, and the 21 genes shown in Table 6 are examples of genes that have a significant correlation with MCR.
  • genes that have a significant correlation with the HbAlc value are found to have a significant correlation with the HbAlc disease-related gene group, genes that have a significant correlation with the HOMA-R, a HOMA-R disease-related gene group, and MCR.
  • Genes may be referred to as MCR pathology-related genes, and genes that are significantly correlated with BMI may be referred to as BMI pathology-related genes.
  • BMI pathology-related genes genes that are significantly correlated with BMI
  • the HbAlc disease-related gene group, the HOM A-R disease-related gene group, and the MCR disease-related gene group can be referred to as a type 2 diabetes-related gene group
  • the BMI disease-related gene group can be referred to as an obesity-related gene group. Since each gene group has some overlap, there are a total of 62 types of genes.
  • genes include genes whose functions have not been elucidated (in the column “Symbol” in Tables 3 to 6, c (n) ori (m) ((n) and (m) are integers)
  • a protein encoded by such a gene whose function is unknown is also known to be a secreted protein having a signal peptide from amino acid sequence analysis. was first found to be associated with diabetes and obesity.
  • SEQ ID NO: 1 PVR
  • SEQ ID NO: 2 LGALS8
  • SEQ ID NO: 3 FCN3
  • SEQ ID NO: 4 MMP24
  • SEQ ID NO: 5 MDK
  • No. 6 NUCB2
  • SEQ ID NO: 7 MMP11
  • SEQ ID NO: 8 GPX3
  • SEQ ID NO: 9 C14ori2
  • SEQ ID NO: 10 C 14orf4
  • SEQ ID NO: ll ClOorflO
  • SEQ ID NO: 12 C6orf74
  • Sequence It is represented by number 13 (C9or! 83) and SEQ ID NO: 14 (C8orfl).
  • SEQ ID NO: 15 BMP2
  • SEQ ID NO: 16 LECT2
  • SEQ ID NO: 17 ADFP
  • SEQ ID NO: 18 BPAG1
  • SEQ ID NO: 19 SEPINF1
  • SEQ ID NO: 20 GNAS
  • SEQ ID NO: 21 AHSG
  • SEQ ID NO: 22 CCL16
  • SEQ ID NO: 23 AFM
  • SEQ ID NO: 24 LTBP1
  • SEQ ID NO: 25 C4BPA
  • SEQ ID NO: 26 LYZ
  • SEQ ID NO: 4 MMP 24
  • SEQ ID NO: 27 PSAP
  • SEQ ID NO: 28 HRG
  • SEQ ID NO: 29 FST
  • SEQ ID NO: 30 GCC3
  • SEQ ID NO: 42 THBS1
  • SEQ ID NO: 43 CSS
  • SEQ ID NO: 44 PON1
  • SEQ ID NO: 45 DEFB1
  • SEQ ID NO: 46 MA2K2
  • PAG7 PAG7
  • nucleotide sequences of 21 genes shown in Table 6 are represented by SEQ ID NO: 36 (THBS1), SEQ ID NO: 43 (CTSS), SEQ ID NO: 48 (IGFBP1), SEQ ID NO: 49 (SULF2), SEQ ID NO: 50 (WNT5B).
  • SEQ ID NO: 51 (ADAMTS1), SEQ ID NO: 52 (SEPP1), SEQ ID NO: 53 (CHAD), SEQ ID NO: 54 (DCN), SEQ ID NO: 56 (LBP), SEQ ID NO: 4 (MMP24), SEQ ID NO: 57 (SPARCL1), SEQ ID NO: 58 (LRG1), SEQ ID NO: 59 (cHorflOO), SEQ ID NO: 35 (cl4orfl23), SEQ ID NO: 9 (C14orl2), SEQ ID NO: 60 (C18orfl0), SEQ ID NO: 37 (C5orfl3), SEQ ID NO: 61 (C6orf79), SEQ ID NO: It is represented by number 62 (C6ori80) and SEQ ID NO: 38 (C9orfl0).
  • the indication in parentheses indicates “Symbol” in the table.
  • Nucleotides used in the present invention include nucleotides containing the above sequences and fragments thereof.
  • the nucleotides used in the present invention also include nucleotides and fragments thereof that are hybridized under stringent conditions with the nucleotides shown in SEQ ID NOs.
  • Such nucleotides include, for example, a base sequence that is about 80% or more, preferably about 90% or more, more preferably about 95% or more on average in terms of the degree of homology with the above base sequence.
  • a nucleotide etc. can be mentioned. Hybridization is well known in the art, including the method described in Current protocols in molecular biology (edited by Frederick M. Ausubel et al., 1987).
  • ⁇ stringent conditions '' are, for example, conditions of ⁇ 1XSSC, 0.1% SDS, 37 ° C '', and more severe conditions are ⁇ 0.5XSSC, 0.1% SDS, 42 ° C '' The more severe condition is “0.2XSSC, 0.1% SDS, 65 ° C”. As the hybridization conditions become more severe in this way, it can be expected to isolate a nucleotide having a high homology with the probe sequence.
  • the combinations of the above SSC, SDS and temperature conditions are exemplary, and those skilled in the art will recognize the above or other factors (eg, probe concentration, It is possible to achieve the same stringency as described above by appropriately combining the length of the probe and the reaction time of the hybridization.
  • some of the above genes (LGAL S8, FCN3, MDK, GNAS, LTBP1, FST, GPLD1) have a Norint, and the nucleotide of the present invention includes a nucleotide containing the nucleotide sequence of the Norint or a nucleotide thereof. Includes fragments.
  • nucleotide used in the present invention it is also possible to use a shift between a nucleotide comprising the sense strand of the above gene and a nucleotide comprising the antisense strand.
  • genes shown in Tables 3-6 those with a value of “DM (type 2 diabetic patients) / NGT (normal glucose tolerance patients) by SAGE” greater than 1 are pathological conditions of type 2 diabetes Or a gene whose expression is increased in relation to obesity, and a gene whose expression is attenuated in relation to the pathology of type 2 diabetes or obesity.
  • a gene with a higher value or a lower value can be said to be a gene having a stronger association with the pathology of type 2 diabetes or obesity. Therefore, among the genes listed in the table, genes with a value represented by “DM (type 2 diabetic patients) / NGT (normal glucose tolerance patients) by SAGE” in the table are 2 or more or 0.5 or less. It can be used more suitably.
  • This SAGE method is a method for determining a 10 base pair DNA sequence (Tag) that characterizes each of the cDNAs that have been prepared for gene expression and expressed in an arbitrary cell.
  • Tag 10 base pair DNA sequence
  • Ma the expression frequency (copy number) of each gene can be determined by calculating the ratio of all the obtained tag powers and individual tags.
  • genes can be key molecules for pathological conditions such as sugar-lipid metabolism, diabetic complications, and arteriosclerosis.
  • these genes are used to study glycolipid metabolism, elucidate the etiology of type 2 diabetes and obesity, detect (diagnose) type 2 diabetes, evaluate the risk of suffering from type 2 diabetes, and become obese It can be used for risk assessment and development of new treatments for diabetes and obesity.
  • these genes are closely related to insulin resistance, and can be used for determination of insulin resistance.
  • these genes are also associated with diabetic complications and can be used to detect (diagnose) diabetic complications or to determine the risk of suffering from diabetic complications.
  • diabetes, obesity, and insulin resistance are risk factors for arteriosclerosis, and these genes are also closely related to arteriosclerosis, which is associated with the detection (diagnosis) of arteriosclerosis and arteriosclerosis. It can also be used to determine risk. Examples of diabetic complications include diabetic retinopathy, diabetic nephropathy, diabetic neuropathy, diabetic macroangiopathy, myocardial infarction, and stroke.
  • type 2 diabetes, obesity, diabetic complications or arteriosclerosis Even if it is possible to measure changes in the expression of genes related to the pathology of the disease and not detectably present the pathology, it may be determined that the patient is suffering from type 2 diabetes, diabetic complications or arteriosclerosis is there.
  • a condition is a condition that is likely to cause detection of type 2 diabetes, obesity, diabetic complications or arteriosclerotic pathology in the future, and this is a type 2 diabetes, obesity, diabetic complication. Can assess the risk of developing a pathology of atherosclerosis or arteriosclerosis.
  • At least one of the genes belonging to the HbAlc disease state-related gene group, the HOMA-R disease state-related gene group, the MCR disease state-related gene group, and the BMI disease state-related gene group is used.
  • At least one of the genes belonging to each of the HbAlc disease state-related gene group, the HOMA-R disease state-related gene group, the MCR disease state-related gene group, and the BMI disease state-related gene group preferably at least 2, more preferably 5 may be used, more preferably at least 10, more preferably all.
  • the BMI condition-related genes are predicted to be associated with obesity and can be used to assess the risk of becoming obese.
  • the degree of gene expression may be measured using nucleotides containing all or part of the base sequences of the above genes as probes or primers.
  • the level of gene expression can be measured by the Northern plot method, RT-PCR method, real-time PCR method, in situ hybridization method, microarray (microchip) method, etc. Any of the methods mentioned can be carried out by known methods.
  • the expression of a gene can be measured by using a nucleotide probe or primer that hybridizes to the mRNA as long as the amount of messenger RNA (mRNA) transcribed from a part of the gene is measured.
  • mRNA messenger RNA
  • the base length of the probe or primer is 10 to 50 bp, preferably 15 to 25 bp.
  • a liver biopsy sample, serum, plasma or the like may be used as a sample sample for evaluating whether or not there is a risk of suffering from sclerosis.
  • the expression of one or more genes belonging to the HbAlc disease-related gene group, the HOMA-R disease-related gene group, the MCR disease-related gene group, and the BMI disease-related gene group is increased or attenuated compared to normal individuals.
  • the subject can be diagnosed as having type 2 diabetes, obesity, diabetic complications or arteriosclerosis, and the subject has type 2 diabetes, hypertrophy, diabetic complications or arteries. Can be assessed as having a risk of suffering from sclerosis.
  • the quantification of the extent of the attenuation makes the subject more accurate, such as type 2 diabetes, obesity, Can be diagnosed as suffering from diabetic complications or arteriosclerosis and assessed that the subject is at risk of suffering from type 2 diabetes, obesity, diabetic complications or arteriosclerosis can do.
  • the expression of a gene is more enhanced or attenuated, it can be determined that type 2 diabetes, obesity, diabetic complications, or arteriosclerosis is more serious! In addition, if the gene expression is more enhanced or attenuated, it can be evaluated that the risk of developing type 2 diabetes, obesity, diabetic complications or arteriosclerosis is high.
  • gene expression products may be measured as well as directly measuring gene expression.
  • the gene expression product can be measured by using an antibody against the protein encoded by the gene.
  • the present invention also includes a protein encoded by a gene whose expression varies depending on the pathological condition between the liver of a type 2 diabetic or obese patient and the liver of a normal person, an antibody against the protein, a reagent containing these, and the like. To do.
  • the present invention provides detection (diagnosis) of type 2 diabetes, obesity, diabetic complications or arteriosclerosis for elucidation of the etiology of type 2 diabetes, obesity, diabetic complications or arteriosclerosis.
  • detection for human type 2 diabetes, obesity, diabetic complications or atherosclerosis, compared to liver tissue of normal humans, human type 2 diabetes, obesity, Increased or attenuated expression in liver tissue of patients with diabetic complications or arteriosclerosis
  • a reagent for measuring the expression of type 2 diabetes-related genes and Z or obesity-related genes which include nucleotides comprising a nucleotide sequence or a partial sequence thereof.
  • the reagent is a reagent comprising a nucleotide comprising the nucleotide sequence of the gene or a nucleotide comprising a partial sequence thereof as a probe or primer, and a nucleotide comprising the nucleotide sequence of the gene or a nucleotide comprising a partial sequence thereof is immobilized.
  • a phased substrate such as a microarray.
  • the microarray can be prepared by immobilizing a nucleotide having the nucleotide sequence of the gene or a nucleotide containing a partial sequence thereof on an appropriate substrate.
  • Examples of the fixed substrate include a glass plate, a quartz plate, and a silicon wafer.
  • a glass plate For example, 3.5mm x 5.5mm, 18mm x 18mm, 22mm x 75mm, etc. as the size of the board This is set according to the number of probe spots on the board and the size of the spots. can do.
  • a method for immobilizing a polynucleotide or a fragment thereof, using a nucleotide charge it can be electrostatically bound to a solid support surface-treated with a polycation such as polylysine, polyethyleneimine or polyalkylamine, Nucleotides introduced with functional groups such as amino group, aldehyde group, SH group, and pyotin can be covalently bonded to the solid phase surface introduced with functional groups such as amino group, aldehyde group, and epoxy group. Fixing may be performed using an array machine.
  • the reagent is at least one of 62 genes belonging to the HbAlc disease state-related gene group, HOMA-R disease state-related gene group, MCR disease state-related gene group and BMI disease state-related gene group, preferably at least two types, Preferably, it contains at least 5, more preferably at least 10, more preferably at least 20, more preferably at least 30, more preferably all. In addition, at least one, preferably at least two of the genes belonging to the HbAlc disease-related gene group, HOMA-R disease-related gene group, MCR disease-related gene group, and BMI disease-related gene group, More preferably, it includes 5 types, more preferably at least 10 types, and even more preferably all.
  • these genes can serve as key molecules for pathological conditions such as glycolipid metabolism, diabetic complications, and arteriosclerosis. Therefore, the gene of the present invention and the reagent containing the gene are key molecules that produce pathologies such as sugar lipid metabolism, diabetic complications, and arteriosclerosis. Use as a tool for analysis, get.
  • detection of type 2 diabetes, obesity, diabetic complications or arteriosclerosis containing the above reagents, or risk of suffering from type 2 diabetes, obesity, diabetic complications or arteriosclerosis Detection agents for assessing are also encompassed by the present invention.
  • expression is increased or decreased in the liver tissue of patients with human type 2 diabetes, obesity, diabetic complications or arteriosclerosis, as compared to liver tissue of normal humans.
  • a drug that increases or decreases the expression of the gene can be selected as a therapeutic or prophylactic agent by administering the candidate drug to a subject or bringing the candidate drug into contact with the gene.
  • Data are mean ⁇ SD, AST, aspartate t raiisHJiiiB se; ALT, al anine auiinot rans feiase;
  • liver tissue was obtained from 21 patients with type 2 diabetes and 11 patients with normal glucose tolerance (Table 2). Liver tissue was obtained by percutaneous ultrasound-guided liver biopsy, and then immediately frozen in liquid nitrogen.
  • RNA extracted from liver tissue was adenylated using FastTrac mRNA extraction kit (Invitrogen, San Diego, CA). For each sample, 2.5 g of RNA was used in the SAGE method. The protocol for the S AGE method followed previous reports (Yamashita T. et al., Biochem Biophys Res Commun 269: 110-116, 2000; Yamashita T. et al., Biochem Biophys Res Comm un 282: 647-654, 2001). The SAGE library was sequenced using the ABI PRISM 377 DNA sequencer and the BigDye terminator single cycle sequencing kit (PE Biosystems, Foster city, CA). Sequenced files were analyzed using SAGE 2000 software (www.sagenet.org).
  • Double-stranded cDNA was used as a cage for quantitative real-time PCR.
  • ABI Prism 7700 Sequence Detection System (Applied Biosystems, Foster City, Calif., USA) was used.
  • Primer and TaqMan probe set is from Applied Biosystems: C (Assays—on—Demand gene expression product).
  • the gene expression of the target sequence was standardized with the expression level of the endogenous control beta-actin RNA (TaqMan Control Reagent Kit, Appleid Biosystems). .
  • PCR conditions were 50 ° C for 2 minutes and 95 ° C for 10 minutes for 1 cycle, and then 95 ° C for 15 seconds and 60 ° C for 1 minute for 40 cycles.
  • HOMA-R is based on previous reports (Matthews DR et al., Diabetologia 28: 412-419, 1985). Fasting blood glucose level (mg / dL) X fasting blood insulin concentration in each case; z U / ml ) Calculated from / 405.
  • the unpaired-t test was used to compare the clinical characteristics of patients with type 2 diabetes and normal glucose tolerance.
  • the relationship between BMI, HOMA-R, MCR and the expression level of each gene was analyzed by linear correlation analysis. For V and deviation, P ⁇ 0.05 was considered significant.
  • the genes of 144,901 tags were identified.
  • Basic local alignment search tool program (BLAST (http://www.ncbi.nlm.nih.gov/BLAST/), and the corresponding gene was analyzed from the tag sequence. Discover and identify.
  • BLAST http://www.ncbi.nlm.nih.gov/BLAST/
  • the genes were classified according to the subcellular localization of transcripts. As a result, 232 genes were genes encoding "extracellular secretory proteins". It was.
  • genes that were upregulated 1.5 times or more in the type 2 diabetes patient group and 114 genes that were expressed 1.5 times or less in comparison with the group with normal glucose tolerance were identified.
  • the amount of gene expression in each case was measured using quantitative real-time PCR, and compared with clinical parameters such as BMI, H0MA-R, and MCR.
  • genes Six genes were identified that showed a significant correlation between expression levels and HOMA-R (Table 5). These genes vary in their expression in relation to insulin resistance in the liver. Among these genes are genes that regulate cell lipid metabolism and energy metabolism, or pathologies related to insulin resistance. It is expected to contain genes that form
  • expression is attenuated or enhanced in the liver of type 2 diabetic patients, and its expression level is glycemic control, obesity, liver 'skeleton Muscle insulin resistance
  • Correlated secretory protein coding genes were identified.
  • the liver-derived secretory protein encoded by these genes is thought to affect intracellular insulin signals, sugar and lipid metabolism, etc., and is related to systemic fat accumulation and the development of insulin hypotonia and arteriosclerosis. It is expected that elucidating the functions of these secreted proteins may lead to elucidation of the etiology of type 2 diabetes and obesity and the development of new therapies.

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Abstract

It is intended to provide a reagent for measuring the expression of a gene whose expression changes in conjunction with diabetes or obesity utilizing a gene whose expression changes in conjunction with diabetic conditions and a method of detecting diabetes or obesity using the reagent. The invention is directed to the reagent for measuring the expression of a type 2 diabetes-associated gene and/or an obesity-associated gene, which contains nucleotides composed of a base sequence of at least one gene selected from the group consisting of genes whose expression is increased or decreased in a liver tissue of a patient with human type 2 diabetes or obesity compared to a liver tissue of a normal human and which encode a secretory protein or nucleotides containing a partial sequence thereof.

Description

明 細 書  Specification

糖尿病の病態と関連して発現変動する遺伝子およびその利用  Genes whose expression varies in relation to the pathology of diabetes and their use

技術分野  Technical field

[0001] 本発明は、糖'脂質代謝、糖尿病性合併症、動脈硬化症などの病態を作るキー分 子となり得る遺伝子の利用に関し、さらに本発明は糖尿病の病態と関連して発現が 変動する遺伝子に関し、さらに該遺伝子の糖尿病や肥満症の診断もしくはリスク評価 等への利用に関する。  [0001] The present invention relates to the use of genes that can be key molecules for the pathogenesis of sugar'lipid metabolism, diabetic complications, arteriosclerosis, etc., and the present invention further varies in expression in relation to the pathology of diabetes. Further, the present invention relates to the use of the gene for diagnosis or risk assessment of diabetes or obesity.

背景技術  Background art

[0002] 世界規模で増加し続ける 2型糖尿病'肥満は網膜 '腎'神経の合併症や虚血性心 疾患などの動脈硬化症を促進し、人類の QOLと生命を脅かしている。肝臓は糖'脂 質代謝に中心的な役割を担う重要な臓器である。 2型糖尿病と肥満では、インスリン による肝臓力ゝらの糖放出抑制作用は減弱しており、この現象はインスリン抵抗性と呼 ばれる (非特許文献 1および 2を参照)。インスリン抵抗性は肝臓力ゝらの糖放出亢進に よる高血糖と、脂質の産生亢進による高脂血症をもたらし、ともに動脈硬化症を促進 する。さらに肝臓は動脈硬化のリスクにつながる血管新生因子をはじめとする各種生 理活性物質の生体内最大の産生臓器である。  [0002] Type 2 diabetes 'obesity' continues to increase on a global scale and promotes arteriosclerosis such as retinal 'renal' nerve complications and ischemic heart disease, threatening human QOL and life. The liver is an important organ that plays a central role in glucose and fat metabolism. In type 2 diabetes and obesity, the action of insulin to suppress the release of sugar by hepatic strength is attenuated, and this phenomenon is called insulin resistance (see Non-Patent Documents 1 and 2). Insulin resistance leads to hyperglycemia due to increased sugar release by liver strength and hyperlipidemia due to increased lipid production, and both promote arteriosclerosis. Furthermore, the liver is the largest organ in the body of various physiologically active substances including angiogenic factors that lead to the risk of arteriosclerosis.

非特許文献 1 : Michael MD. et al.， Mol. Cell 6:87-97, 2000  Non-Patent Document 1: Michael MD. Et al., Mol. Cell 6: 87-97, 2000

非特許文献 2 : Saltiel AR. et al.， Nature 414:799-806, 2001  Non-Patent Document 2: Saltiel AR. Et al., Nature 414: 799-806, 2001

発明の開示  Disclosure of the invention

発明が解決しょうとする課題  Problems to be solved by the invention

[0003] 本発明は、糖'脂質代謝、糖尿病性合併症、動脈硬化症などの病態を作るキー分 子となり得る遺伝子、ならびに該遺伝子の糖'脂質代謝、糖尿病性合併症、動脈硬 化症などの病態を作る原因の解明への使用方法を提供する。 [0003] The present invention relates to a gene that can be a key molecule for producing a pathological condition such as sugar'lipid metabolism, diabetic complications, arteriosclerosis, and the sugar'lipid metabolism, diabetic complications, arteriosclerosis of the gene. The method of use to elucidate the cause of pathological conditions such as is provided.

[0004] 本発明は、糖尿病の病態と関連して発現が変動する遺伝子を利用した、糖尿病や 肥満症に関連して発現が変動する遺伝子の発現を測定する試薬および該試薬を用 いた糖尿病、肥満症、糖尿病性合併症、動脈硬化症などの検出方法の提供を目的 とする。 課題を解決するための手段 [0004] The present invention relates to a reagent for measuring the expression of a gene whose expression varies in relation to diabetes or obesity using a gene whose expression varies in relation to the pathological condition of diabetes, and diabetes using the reagent, The purpose is to provide methods for detecting obesity, diabetic complications, arteriosclerosis, and the like. Means for solving the problem

[0005] 本発明者はこれまで、正常肝と各種肝疾患の SAGE、およびこれまでの DNAチップ 解析から得た 60万発現遺伝子を超える世界最大の肝臓発現遺伝子データベースを 構築し、肝臓病の解析をすすめてきた。その過程で独自に開発した cDNAマイクロア レイを用いて、 2型糖尿病患者の肝臓に発現する遺伝子のプロファイルを解析したと ころ、従来の研究手法では異常がな 、と考えられて 、た 2型糖尿病患者の肝臓にお いて、糖代謝に直接に関連する遺伝子だけでなぐ血管新生因子などの血管合併症 に関連した多彩な既知遺伝子の発現が亢進していることを見出した (Takamura T. et al.， Diabetologia 47:638-647, 2004)。これら既知の遺伝子は、 2型糖尿病の病態を 形作っている可能性がある。これらの知見は、肝由来分泌蛋白のなかに、 2型糖尿病 や肥満症状態の肝臓が、血管合併症をはじめとする糖尿病の病態を形作る未知の 分泌蛋白が存在する可能性を示唆していた。そこで今回、この独創的な研究をさら に発展させ、 Serial analysis of gene expression法（以下 SAGE法）を用いて、 2型糖尿 病患者の肝臓が産生する遺伝子を、機能不明な未知遺伝子を含めて同定し、それら の中でインスリン抵抗性と関連して発現亢進 ·減弱する分泌タンパクをコードする遺 伝子群を同定し、本発明を完成させるに至った。  [0005] The present inventor has constructed the world's largest liver-expressed gene database that exceeds the 600,000-expressed genes obtained from SAGE of normal liver and various liver diseases and DNA chip analysis so far, and analyzes liver disease. I have recommended. Using a cDNA microarray originally developed in the process, the profile of the gene expressed in the liver of type 2 diabetic patients was analyzed, and it was thought that there was no abnormality in the conventional research method. In the liver of diabetic patients, we found that the expression of various known genes related to vascular complications such as angiogenic factors that are not only directly related to glucose metabolism was increased (Takamura T. et al. al., Diabetologia 47: 638-647, 2004). These known genes may shape the pathology of type 2 diabetes. These findings suggest that liver-derived secretory proteins may have unknown secreted proteins that shape the pathology of diabetes, including vascular complications, in livers with type 2 diabetes and obesity. . Therefore, this original research was further developed, and the gene produced by the liver of type 2 diabetic patients, including unknown genes with unknown functions, was analyzed using the serial analysis of gene expression method (hereinafter referred to as SAGE method). The present inventors have identified and identified a group of genes that encode secreted proteins that are up- or down-regulated in relation to insulin resistance, thereby completing the present invention.

[0006] すなわち、本発明の態様は以下の通りである。 [0006] That is, the embodiments of the present invention are as follows.

[0007] [1] ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者または肥満症患者の肝臓 組織で発現が亢進または減弱して 、る遺伝子であって、分泌タンパク質をコードして いる以下の遺伝子力 なる群力 選択される少なくとも 1つの遺伝子の塩基配列から なるヌクレオチドまたはその一部配列を含むヌクレオチドを含む 2型糖尿病関連遺伝 子および Zまたは肥満症関連遺伝子の発現を測定するための試薬。  [0007] [1] A gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic or obese patient compared to the liver tissue of a normal human subject, which encodes a secreted protein In order to measure the expression of type 2 diabetes-related genes and Z or obesity-related genes containing nucleotides consisting of the base sequence of at least one selected gene or nucleotides including a partial sequence thereof Reagent.

[0008] (1) poliovirus receptor  [0008] (1) poliovirus receptor

(2) lectin, galactoside- binding, soluble, 8 (galectin 8)  (2) lectin, galactoside-binding, soluble, 8 (galectin 8)

(3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen)

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2)

(6) nucleobindin 2 (7) matrix metalloproteinase 11 (stromelysin 3) (6) nucleobindin 2 (7) matrix metalloproteinase 11 (stromelysin 3)

(8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma)

(9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2

(10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4

(11) chromosome 10 open reading frame 10  (11) chromosome 10 open reading frame 10

(12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74

(13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA

(14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1

(15) bone morphogenetic protein 2  (15) bone morphogenetic protein 2

(16) leukocyte cell-derived chemotaxin 2  (16) leukocyte cell-derived chemotaxin 2

(17) adipose differentiation- related protein  (17) adipose differentiation- related protein

(18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA

(19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1

(20) GNAS complex locus  (20) GNAS complex locus

(21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein

(22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16

(23) afamin  (23) afamin

(24) LTBPl latent transforming growth factor beta binding protein 1  (24) LTBPl latent transforming growth factor beta binding protein 1

(25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha

(26) lysozyme (renal amyloidosis)  (26) lysozyme (renal amyloidosis)

(27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)  (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)

(28) histidine- rich glycoprotein (28) histidine-rich glycoprotein

(29) FST follistatin  (29) FST follistatin

(30) glypican 3  (30) glypican 3

(31) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15 (31) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15

(33) SPINT2 serine protease inhibitor Kunitz type 2  (33) SPINT2 serine protease inhibitor Kunitz type 2

(34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A  (34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A

(35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123

(36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32

(37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13

(38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10

(39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107

(40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38

(41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58

(42) thrombospondin 1  (42) thrombospondin 1

(43) cathepsin S  (43) cathepsin S

(44) paraoxonase 1  (44) paraoxonase 1

(45) DEFBl defensin beta 1  (45) DEFBl defensin beta 1

(46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2

(47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma)

(48) IGFBPl insulin like growth factor binding protein 1 (48) IGFBPl insulin like growth factor binding protein 1

(49) sulfatase 2  (49) sulfatase 2

(50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B

(51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1

(52) selenoprotein P， plasma, In  (52) selenoprotein P, plasma, In

(53) chondroadherin  (53) chondroadherin

(54) decorin  (54) decorin

(56) lipopolysaccharide binding protein  (56) lipopolysaccharide binding protein

(57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin)

(58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1

(59) chromosome 14 open reading frame 100 (60) C18orfl0 chromosome 18 open reading frame 10 (59) chromosome 14 open reading frame 100 (60) C18orfl0 chromosome 18 open reading frame 10

(61) chromosome 6 open reading frame 79  (61) chromosome 6 open reading frame 79

(62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80

[2] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱している遺伝子が、発 現の亢進または減弱がヒト血中 HbAlc値と相関している以下の遺伝子からなる群から 選択される少なくとも 1つの遺伝子である [1]の 2型糖尿病関連遺伝子および Zまた は肥満症関連遺伝子の発現を測定するための試薬。  [2] A gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is selected from the group consisting of the following genes whose up- or down-expression is correlated with HbAlc levels in human blood A reagent for measuring the expression of at least one gene, type 2 diabetes-related gene and Z or obesity-related gene of [1].

[0009] (1) poliovirus receptor [0009] (1) poliovirus receptor

(2) lectin, galactoside- binding, soluble, 8 (galectin 8)  (2) lectin, galactoside-binding, soluble, 8 (galectin 8)

(3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen)

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2)

(6) nucleobindin 2  (6) nucleobindin 2

(7) matrix metalloproteinase 11 (stromelysin 3)  (7) matrix metalloproteinase 11 (stromelysin 3)

(8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma)

(9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2

(10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4

丄丄) chromosome 10 open reading frame 10  丄 丄) chromosome 10 open reading frame 10

(12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74

(13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA

丄 4) chromosome 8 open reading frame 1  丄 4) chromosome 8 open reading frame 1

[3] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱している遺伝子が、発 現の亢進または減弱が HOMA-Rと相関している以下の遺伝子からなる群から選択さ れる少なくとも 1つの遺伝子である [1]の 2型糖尿病関連遺伝子および Zまたは肥満 症関連遺伝子の発現を測定するための試薬。  [3] A gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is at least 1 selected from the group consisting of the following genes whose up-regulation or attenuation is correlated with HOMA-R A reagent for measuring the expression of type 2 diabetes-related gene and Z or obesity-related gene of [1], which are two genes.

[0010] (42) thrombospondin 1 [0010] (42) thrombospondin 1

(43) cathepsin S (44) paraoxonase 1 (43) cathepsin S (44) paraoxonase 1

(45) DEFB1 defensin beta 1  (45) DEFB1 defensin beta 1

(46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2

(47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) [4] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱している遺伝子が、発 現の亢進または減弱が MCRと相関している以下の遺伝子からなる群から選択される 少なくとも 1つの遺伝子である [1]の 2型糖尿病関連遺伝子および Zまたは肥満症関 連遺伝子の発現を測定するための試薬。  (47) Phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) [4] A gene whose expression is up- or down-regulated in the liver tissue of human type 2 diabetics is correlated with increased or decreased expression. A reagent for measuring the expression of a type 2 diabetes-related gene and a Z- or obesity-related gene of [1], which is at least one gene selected from the group consisting of the following genes:

(42) thrombospondin 1 (42) thrombospondin 1

(43) cathepsin S (43) cathepsin S

(48) IGFBP1 insulin like growth factor binding protein 1  (48) IGFBP1 insulin like growth factor binding protein 1

(49) sulfatase 2  (49) sulfatase 2

(50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B

(51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1

(52) selenoprotein P， plasma, In  (52) selenoprotein P, plasma, In

(53) chondroadherin  (53) chondroadherin

(54) decorin  (54) decorin

(56) lipopolysacchande binding protein  (56) lipopolysacchande binding protein

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin)

(58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1

(59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100

(35) chromosome 14 open reading frame 123 (35) chromosome 14 open reading frame 123

(9) chromosome 14 open reading frame 2 (9) chromosome 14 open reading frame 2

(60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10

(37) chromosome 5 open reading frame 13 (37) chromosome 5 open reading frame 13

(61) chromosome 6 open reading frame 79 (62) C6orf80 chromosome 6 open reading frame 80 (61) chromosome 6 open reading frame 79 (62) C6orf80 chromosome 6 open reading frame 80

(38) chromosome 9 open reading frame 10 (38) chromosome 9 open reading frame 10

[5] ヒト肥満症患者の肝臓糸且織で発現が亢進または減弱している遺伝子が、発現の 亢進または減弱が BMIと相関している以下の遺伝子からなる群から選択される少なく とも 1つの遺伝子である [1]の 2型糖尿病関連遺伝子および Zまたは肥満症関連遺 伝子の発現を測定するための試薬。  [5] A gene whose expression is up- or down-regulated in liver thread and tissue of human obesity patients is selected from the group consisting of the following genes whose up-regulation or attenuation is correlated with BMI: A reagent for measuring the expression of genes related to type 2 diabetes and Z or obesity related genes of [1].

(15) bone morphogenetic protein 2 (15) bone morphogenetic protein 2

uoノ leukocyte cell-derived cnemotaxin 2 uo leukocyte cell-derived cnemotaxin 2

(17) adipose differentiation- related protein  (17) adipose differentiation- related protein

(18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA

(19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1

(20) GNAh complex locus  (20) GNAh complex locus

(21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein

(22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16

(23) afamin  (23) afamin

(24) LTBP1 latent transforming growth factor beta binding protein 1  (24) LTBP1 latent transforming growth factor beta binding protein 1

(25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha

(26) lysozyme (renal amyloidosis;  (26) lysozyme (renal amyloidosis;

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)  (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)

(28) histidine- rich glycoprotein (28) histidine-rich glycoprotein

(29) FST follistatin  (29) FST follistatin

(30) glypican 3  (30) glypican 3

(31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl

(32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15

(33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A (33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A

(35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123

(14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1

(36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32

(37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13

(38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10

(39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107

(40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38

(41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58

[6] 少なくとも 5つの遺伝子を含む [1]〜[5]のいずれかの 2型糖尿病関連遺伝子お よび Zまたは肥満症関連遺伝子の発現を測定するための試薬。  [6] A reagent for measuring the expression of a type 2 diabetes-related gene and a Z- or obesity-related gene according to any one of [1] to [5], comprising at least 5 genes.

[0013] [7] ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進 または減弱している遺伝子の塩基配列力もなるヌクレオチドまたはその一部配列を含 むヌクレオチドを固相化したマイクロアレイを含む [1]〜[6]のいずれかの 2型糖尿病 関連遺伝子および Zまたは肥満症関連遺伝子の発現を測定するための試薬。  [0013] [7] A nucleotide comprising a nucleotide sequence or a partial sequence thereof that also has a base sequence ability of a gene whose expression is increased or decreased in the liver tissue of a human type 2 diabetic patient compared to a liver tissue of a normal human subject A reagent for measuring the expression of type 2 diabetes-related gene and Z or obesity-related gene according to any one of [1] to [6].

[0014] [8] ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進 または減弱している遺伝子の塩基配列力もなるヌクレオチドまたはその一部配列を含 むヌクレオチドをターゲットとしたプローブまたはプライマーを含む [1]〜[6]のいずれ 力の 2型糖尿病関連遺伝子および Zまたは肥満症関連遺伝子の発現を測定するた めの試薬。  [0014] [8] A nucleotide comprising a nucleotide sequence or a partial sequence thereof, which also has a base sequence ability of a gene whose expression is increased or decreased in the liver tissue of a human type 2 diabetic patient compared to the liver tissue of a normal human subject A reagent for measuring the expression of type 2 diabetes-related gene and Z or obesity-related gene according to any one of [1] to [6], which comprises a probe or primer targeted for.

[0015] [9] ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進 または減弱して 、る遺伝子であって、分泌タンパク質をコードして 、る以下の遺伝子 力 なる群力 選択される少なくとも 1つの遺伝子について、被検体における発現を 測定し、該遺伝子の発現が正常人に比較して亢進または減弱して 、る場合に前記 被検体が 2型糖尿病、肥満症、糖尿病性合併症もしくは動脈硬化症に罹患している 力 または罹患するリスクがあると判断する方法。  [0015] [9] A gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic patient as compared with the liver tissue of a normal human subject, which encodes a secreted protein and Gene force is a group force. For at least one selected gene, the expression in a subject is measured, and if the expression of the gene is increased or decreased compared to a normal person, the subject is type 2 diabetes, How to determine that you have or are at risk of suffering from obesity, diabetic complications or arteriosclerosis.

[0016] (1) poliovirus receptor  [0016] (1) poliovirus receptor

(2) lectin, galactoside— binding, soluble, 8 (galectin 8) (3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen) (2) lectin, galactoside— binding, soluble, 8 (galectin 8) (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen)

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2)

(6) nucleobindin 2  (6) nucleobindin 2

(7) matrix metalloproteinase 11 (stromelysin 3)  (7) matrix metalloproteinase 11 (stromelysin 3)

(8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma)

(9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2

(10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4

(11) chromosome 10 open reading frame 10  (11) chromosome 10 open reading frame 10

(12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74

(13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA

(14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1

(15) bone morphogenetic protein 2  (15) bone morphogenetic protein 2

(16) leukocyte cell-derived chemotaxin 2  (16) leukocyte cell-derived chemotaxin 2

(17) adipose differentiation- related protein  (17) adipose differentiation- related protein

(18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA

(19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1

(20) GNAS complex locus  (20) GNAS complex locus

(21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein

(22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16

(23) afamin  (23) afamin

(24) LTBPl latent transforming growth factor beta binding protein 1  (24) LTBPl latent transforming growth factor beta binding protein 1

(25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha

(26) lysozyme (renal amyloidosis)  (26) lysozyme (renal amyloidosis)

(27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (28) histidine- rich glycoprotein (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (28) histidine-rich glycoprotein

(29) FST follistatin  (29) FST follistatin

(30) glypican 3  (30) glypican 3

(31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl

(32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15

(33) SPINT2 serine protease inhibitor Kunitz type 2  (33) SPINT2 serine protease inhibitor Kunitz type 2

(34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A  (34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A

(35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123

(36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32

(37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13

(38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10

(39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107

(40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38

(41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58

(42) thrombospondin 1  (42) thrombospondin 1

(43) cathepsin S  (43) cathepsin S

(44) paraoxonase 1  (44) paraoxonase 1

(45) DEFBl defensin beta 1  (45) DEFBl defensin beta 1

(46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2

(47) phospholipase A2， group VII (platelet-activating factor acetylhydrolase, plasma) (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma)

(48) IGFBPl insulin like growth factor binding protein 1 (48) IGFBPl insulin like growth factor binding protein 1

(49) sulfatase 2  (49) sulfatase 2

(50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B

(51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1

(52) selenoprotein P， plasma, In  (52) selenoprotein P, plasma, In

(53) chondroadherin  (53) chondroadherin

(54) decorin (56) lipopolysaccharide binding protein (54) decorin (56) lipopolysaccharide binding protein

(57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin)

(58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1

(59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100

(60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10

(61) chromosome 6 open reading frame 79  (61) chromosome 6 open reading frame 79

(62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80

[10] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱している遺伝子が、 発現の亢進または減弱がヒト血中 HbAlc値と相関している以下の遺伝子からなる群 力 選択される少なくとも 1つの遺伝子である [9]の方法。  [10] A gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is a group consisting of the following genes whose up- or down-expression is correlated with HbAlc levels in human blood: The method of [9], which is a single gene.

(1) poliovirus receptor (1) poliovirus receptor

(2) lectin, galactoside- binding, soluble, 8 (galectin 8)  (2) lectin, galactoside-binding, soluble, 8 (galectin 8)

(3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen)

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2)

(6) nucleobindin 2  (6) nucleobindin 2

(7) matrix metalloproteinase 11 (stromelysin 3)  (7) matrix metalloproteinase 11 (stromelysin 3)

(8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma)

(9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2

(10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4

丄丄) chromosome 10 open reading frame 10  丄 丄) chromosome 10 open reading frame 10

(12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74

(13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA

丄 4) chromosome 8 open reading frame 1  丄 4) chromosome 8 open reading frame 1

[11] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して ヽる遺伝子が、 発現の亢進または減弱が HOMA-Rと相関している以下の遺伝子力 なる群力 選択 される少なくとも 1つの遺伝子である [9]の方法。 [0018] (42) thrombospondin 1 [11] A gene whose expression is enhanced or attenuated in the liver tissue of a human type 2 diabetic patient is selected as a group force with the following gene strength in which increased or attenuated expression is correlated with HOMA-R: at least 1 selected The method of [9], which is one gene. [0018] (42) thrombospondin 1

(43) cathepsin S  (43) cathepsin S

(44) paraoxonase 1  (44) paraoxonase 1

(45) DEFB1 defensin beta 1  (45) DEFB1 defensin beta 1

(46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2

(47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) [12] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱している遺伝子が、 発現の亢進または減弱が MCRと相関している以下の遺伝子からなる群から選択され る少なくとも 1つの遺伝子である [9]の方法。  (47) Phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) [12] A gene whose expression is up- or down-regulated in the liver tissue of human type 2 diabetic patients, The method according to [9], wherein the method is at least one gene selected from the group consisting of the following genes:

[0019] (42) thrombospondin 1  [0019] (42) thrombospondin 1

(43) cathepsin S  (43) cathepsin S

(48) IGFBP1 insulin like growth factor binding protein 1  (48) IGFBP1 insulin like growth factor binding protein 1

(49) sulfatase 2  (49) sulfatase 2

(50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B

(51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1

(52) selenoprotein P， plasma, In  (52) selenoprotein P, plasma, In

(53) chondroadherin  (53) chondroadherin

(54) decorin  (54) decorin

(56) lipopolysacchande binding protein  (56) lipopolysacchande binding protein

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin)

(58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1

(59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100

(35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123

(9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2

(60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10

(37) chromosome 5 open reading frame 13 (61) chromosome 6 open reading frame 79 (37) chromosome 5 open reading frame 13 (61) chromosome 6 open reading frame 79

(62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80

(38) chromosome 9 open reading frame 10 (38) chromosome 9 open reading frame 10

[13] ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱している遺伝子が、 発現の亢進または減弱が BMIと相関している以下の遺伝子からなる群から選択され る少なくとも 1つの遺伝子である [9]の方法。  [13] A gene whose expression is enhanced or attenuated in the liver tissue of a human type 2 diabetes patient is at least one gene selected from the group consisting of the following genes whose expression is correlated with BMI: There is a method of [9].

(15) bone morphogenetic protein 2 (15) bone morphogenetic protein 2

uoノ leukocyte cell-derived cnemotaxin 2 uo leukocyte cell-derived cnemotaxin 2

(17) adipose differentiation- related protein  (17) adipose differentiation- related protein

(18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA

(19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1

(20) GNAS complex locus  (20) GNAS complex locus

(21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein

(22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16

(23) afamin  (23) afamin

(24) LTBP1 latent transforming growth factor beta binding protein 1  (24) LTBP1 latent transforming growth factor beta binding protein 1

(25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha

(26) lysozyme (renal amyloidosis;  (26) lysozyme (renal amyloidosis;

(4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted)

(27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)  (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)

(28) histidine- rich glycoprotein (28) histidine-rich glycoprotein

(29) FST follistatin  (29) FST follistatin

(30) glypican 3  (30) glypican 3

(31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl

(32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15

(33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A (33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A

(35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123

(14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1

(36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32

(37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13

(38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10

(39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107

(40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38

(41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58

[14] 少なくとも 5つの遺伝子について被検体における発現を測定する [9]〜[13]の いずれかの方法。  [14] The method according to any one of [9] to [13], wherein the expression in the subject is measured for at least 5 genes.

[0021] [15] ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢 進または減弱している遺伝子の塩基配列力もなるヌクレオチドまたはその一部配列を 含むヌクレオチドを固相化したマイクロアレイを用いる [9]〜[14]の!、ずれかの方法。  [15] A nucleotide comprising a nucleotide sequence or a partial sequence thereof, which also has a base sequence ability of a gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic patient compared to the liver tissue of a normal human subject [9] to [14] !, either method.

[0022] [16] ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢 進または減弱している遺伝子の塩基配列力もなるヌクレオチドまたはその一部配列を 含むヌクレオチドをターゲットとしたリアルタイム PCRにより該遺伝子の発現を解析す ることを含む [9]〜[14]の!、ずれかの方法。  [16] [16] A nucleotide comprising a nucleotide sequence or a partial sequence thereof that also has a base sequence ability of a gene whose expression is increased or attenuated in the liver tissue of a human type 2 diabetic patient compared to the liver tissue of a normal human subject [9] to [14], any of the methods including analyzing the expression of the gene by real-time PCR targeting at [9].

[0023] [17] [1]〜[8]のいずれかの 2型糖尿病関連遺伝子および Zまたは肥満症関連遺 伝子の発現を測定するための試薬を含む 2型糖尿病、肥満症、糖尿病性合併症お よび動脈硬化症の検出またはリスク判定のためのキット。  [0023] [17] Type 2 diabetes, obesity, diabetic comprising a type 2 diabetes-related gene according to any one of [1] to [8] and a reagent for measuring the expression of Z or an obesity-related gene Kit for detection or risk assessment of complications and arteriosclerosis.

発明の効果  The invention's effect

[0024] 実施例に示すように、 2型糖尿病患者または肥満症患者の肝臓と正常人の肝臓と の間で、病態ごとに発現が変動する遺伝子群を見出した。該遺伝子群は、糖'脂質 代謝、糖尿病性合併症、動脈硬化症などの病態を作るキー分子となり得、該遺伝子 を利用することにより、糖 ·脂質代謝、糖尿病性合併症、動脈硬化症などの病態を作 る機構を解明することができる。該遺伝子群に属する遺伝子の発現を調べることによ り、 2型糖尿病、糖尿病性合併症、動脈硬化症または肥満症の病因を解明することが でき、また 2型糖尿病、糖尿病性合併症または動脈硬化症または肥満症の診断、 2 型糖尿病、糖尿病性合併症、動脈硬化症または肥満症に罹るリスクを評価すること ができる。さらに、 2型糖尿病や肥満症の新規治療法の開発につながることが期待さ れる [0024] As shown in the Examples, a group of genes whose expression varies depending on the pathological condition between the liver of a type 2 diabetic patient or obese patient and the liver of a normal person was found. The gene group can be a key molecule for the pathogenesis of sugar and lipid metabolism, diabetic complications, arteriosclerosis, etc. By using this gene, sugar / lipid metabolism, diabetic complications, arteriosclerosis, etc. It is possible to elucidate the mechanism of the pathophysiology. By investigating the expression of genes belonging to this gene group, it is possible to elucidate the etiology of type 2 diabetes, diabetic complications, arteriosclerosis or obesity And the risk of developing type 2 diabetes, diabetic complications or arteriosclerosis or obesity, type 2 diabetes, diabetic complications, arteriosclerosis or obesity can be assessed. Furthermore, it is expected to lead to the development of new treatments for type 2 diabetes and obesity

本明細書は本願の優先権の基礎である日本国特許出願 2005-125689号の明細書 および Zまたは図面に記載される内容を包含する。  This specification includes the contents described in the specification and Z or drawings of Japanese Patent Application No. 2005-125689, which is the basis of the priority of the present application.

発明を実施するための最良の形態 BEST MODE FOR CARRYING OUT THE INVENTION

本発明の糖尿病の病態または肥満と関連して発現変動する遺伝子は、 2型糖尿病 患者または肥満症患者の肝臓と正常人の肝臓との間で発現に差がある遺伝子であり 、正常人の肝臓に比較して 2型糖尿病患者または肥満症患者の肝臓における発現 が亢進または減弱している遺伝子である。さらに、本発明の遺伝子は細胞外に分泌 される分泌性タンパク質をコードする遺伝子である。ここで 2型糖尿病とはインスリンの 分泌量の低下やインスリンに対する感受性の低下により発症するタイプの糖尿病で ある。また、正常人とは 2型糖尿病の病態を呈していない者をいい、具体的には肝臓 または骨格筋におけるインスリン抵抗性が正常値である者、すなわち耐糖能が正常 な者をいう。 2型糖尿病の病態として、血液中 HbAlc (ヘモグロビン Ale)濃度が高くな り、インスリン抵抗性 (肝臓におけるインスリン抵抗性および骨格筋におけるインスリン 抵抗性）力 S高まることが挙げられる。肝臓におけるインスリン抵抗性は、 HOMA-R(hom eostasis model assessment)の測定により決定することができ、 HOMA-Rの数値が大き いほど肝臓におけるインスリン抵抗性が強いと判断される。また、骨格筋におけるイン スリン抵抗性は、 MCR(glucose metabolic clearance rate)の測定により決定することが でき、 MCRの数値が大きいほど骨格筋のインスリン抵抗性が強いと判断される。 HOM A-R値は、空腹時血糖 §/(11) 空腹時ィンスリン濃度(11?1)( U/ml)/405で示され (M atthews DR et al.， Diabetologia 28:412-419, 1985)、正常人ではこの値が 1前後であ る。 MCR値は、例えば DeFronzo RA et al.， Am J Physiol 237:E214-223, 1979に記載 の方法で測定することができる。肥満症患者は、 BMI (Body Mass index)が高値を示 す患者であり、 BMIは体重 (kg)Z身長 (m)2で示される。 BMIが 25以上の場合、肥満症 であると判定される。 [0026] 本発明の 2型糖尿病の病態または肥満症と関連して発現変動する遺伝子は、血液 中 HbAlc値、 HOMA-R、 MCRおよび BMIのいずれか一つと有意の相関が認められる 遺伝子である。ここで、有意の相関が認められるとは、例えば血液中 HbAlc値、 HOM A-R、 MCRおよび BMIの各測定値と各遺伝子の発現レベルの関係を線形相関解析 により解析した場合に有意に相関が認められることをいう。 The gene whose expression changes in relation to the pathological condition of obesity or obesity of the present invention is a gene whose expression is different between the liver of a type 2 diabetes patient or obese patient and the liver of a normal person, and the liver of a normal person In comparison with the above, it is a gene whose expression in the liver of type 2 diabetic patients or obese patients is increased or decreased. Furthermore, the gene of the present invention is a gene encoding a secretory protein that is secreted extracellularly. Here, type 2 diabetes is a type of diabetes that develops due to a decrease in insulin secretion and a decrease in sensitivity to insulin. In addition, a normal person means a person who does not exhibit type 2 diabetes, specifically, a person with normal insulin resistance in the liver or skeletal muscle, that is, a person with normal glucose tolerance. As a pathological condition of type 2 diabetes, blood HbAlc (hemoglobin Ale) concentration increases, and insulin resistance (insulin resistance in the liver and insulin resistance in skeletal muscle) power S is increased. Insulin resistance in the liver can be determined by measuring HOMA-R (homeostasis model assessment), and the higher the value of HOMA-R, the stronger the insulin resistance in the liver. Insulin resistance in skeletal muscle can be determined by measuring the glucose metabolic clearance rate (MCR). The larger the MCR value, the stronger the insulin resistance in skeletal muscle. HOM AR values, fasting blood glucose § / (11) fasting Insurin concentration (11 1?) (U / ml) / 405 indicated by (M atthews DR et al, Diabetologia 28:. 412-419, 1985), This value is around 1 for normal people. The MCR value can be measured by, for example, the method described in DeFronzo RA et al., Am J Physiol 237: E214-223, 1979. Obese patients are those whose BMI (Body Mass index) is high, and BMI is expressed as body weight (kg) Z height (m) 2 . If the BMI is 25 or greater, it is determined to be obese. [0026] The gene whose expression changes in association with the pathological condition or obesity of type 2 diabetes of the present invention is a gene that is significantly correlated with any one of blood HbAlc level, HOMA-R, MCR and BMI. . Here, a significant correlation is observed when, for example, the relationship between the measured values of blood HbAlc, HOM AR, MCR and BMI and the expression level of each gene is analyzed by linear correlation analysis. It means being done.

[0027] 血液中 HbAlc値と有意の相関が認められる遺伝子として表 3に示す 14遺伝子が、 BMIと有意の相関が認められる遺伝子として表 4に示す 29遺伝子力 HOMA-Rと有 意の相関が認められる遺伝子として表 5に示す 6遺伝子が、 MCRと有意の相関が認 められる遺伝子として表 6に示す 21遺伝子が挙げられる。本発明において、 HbAlc 値と有意の相関が認められる遺伝子を HbAlc病態関連遺伝子群、 HOMA-Rと有意 の相関が認められる遺伝子を HOMA-R病態関連遺伝子群、 MCRと有意の相関が認 められる遺伝子を MCR病態関連遺伝子群、 BMIと有意の相関が認められる遺伝子を BMI病態関連遺伝子群と呼ぶことがある。このうち、 HbAlc病態関連遺伝子群、 HOM A-R病態関連遺伝子群および MCR病態関連遺伝子群を 2型糖尿病関連遺伝子群と いうことができ、 BMI病態関連遺伝子群を肥満症関連遺伝子群ということができる。な お、それぞれの遺伝子群には一部重複があるので、遺伝子の種類は全部で 62種類 ある。これらの遺伝子には、機能が解明されていない遺伝子も含まれるが（表 3から 6 の「Symbol」の欄において、 c(n)ori(m)((n)および (m)は整数)、このような機能未知とさ れる遺伝子がコードするタンパク質についても、アミノ酸配列解析から、シグナルぺプ チドを有しており分泌タンパク質であることが判明している。なお、これらの遺伝子は 、本発明において初めて、糖尿病や肥満と関連あることが見出された。  [0027] 14 genes shown in Table 3 as genes that are significantly correlated with HbAlc levels in blood, and 29 genes shown in Table 4 as genes that are significantly correlated with BMI are significantly correlated with HOMA-R. The 6 genes shown in Table 5 are recognized genes, and the 21 genes shown in Table 6 are examples of genes that have a significant correlation with MCR. In the present invention, genes that have a significant correlation with the HbAlc value are found to have a significant correlation with the HbAlc disease-related gene group, genes that have a significant correlation with the HOMA-R, a HOMA-R disease-related gene group, and MCR. Genes may be referred to as MCR pathology-related genes, and genes that are significantly correlated with BMI may be referred to as BMI pathology-related genes. Of these, the HbAlc disease-related gene group, the HOM A-R disease-related gene group, and the MCR disease-related gene group can be referred to as a type 2 diabetes-related gene group, and the BMI disease-related gene group can be referred to as an obesity-related gene group. Since each gene group has some overlap, there are a total of 62 types of genes. These genes include genes whose functions have not been elucidated (in the column “Symbol” in Tables 3 to 6, c (n) ori (m) ((n) and (m) are integers) A protein encoded by such a gene whose function is unknown is also known to be a secreted protein having a signal peptide from amino acid sequence analysis. Was first found to be associated with diabetes and obesity.

[0028] 表 3に示す 14遺伝子の塩基配列を配列番号 1 (PVR)、配列番号 2 (LGALS8)、配 列番号 3 (FCN3)、配列番号 4 (MMP24)、配列番号 5 (MDK)、配列番号 6 (NUCB2) 、配列番号 7 (MMP11)、配列番号 8 (GPX3)、配列番号 9 (C14ori2)、配列番号 10 (C 14orf4)、配列番号 l l (ClOorflO)、配列番号 12 (C6orf74)、配列番号 13 (C9or!83) および配列番号 14 (C8orfl)に表す。  [0028] The nucleotide sequences of the 14 genes shown in Table 3 are shown in SEQ ID NO: 1 (PVR), SEQ ID NO: 2 (LGALS8), SEQ ID NO: 3 (FCN3), SEQ ID NO: 4 (MMP24), SEQ ID NO: 5 (MDK), and sequence. No. 6 (NUCB2), SEQ ID NO: 7 (MMP11), SEQ ID NO: 8 (GPX3), SEQ ID NO: 9 (C14ori2), SEQ ID NO: 10 (C 14orf4), SEQ ID NO: ll (ClOorflO), SEQ ID NO: 12 (C6orf74), Sequence It is represented by number 13 (C9or! 83) and SEQ ID NO: 14 (C8orfl).

[0029] 表 4に示す 29遺伝子の塩基配列を配列番号 15 (BMP2)、配列番号 16 (LECT2)、 配列番号 17 (ADFP)、配列番号 18 (BPAG1)、配列番号 19 (SERPINF1)、配列番号 20 (GNAS)、配列番号 21 (AHSG)、配列番号 22 (CCL16)、配列番号 23 (AFM)、配 列番号 24 (LTBP1)、配列番号 25 (C4BPA)、配列番号 26 (LYZ)、配列番号 4 (MMP 24)、配列番号 27 (PSAP)、配列番号 28 (HRG)、配列番号 29 (FST)、配列番号 30 ( GPC3)、配列番号 31 (GPLD1)、配列番号 32 (GDF15)、配列番号 33 (SPINT2)、配 列番号 34 (TNFRSF1A)、配列番号 35 (cl4orfl23)、配列番号 14 (c8orfl)、配列番 号 36 (cl4orf32)、配列番号 37 (c5orH3)、配列番号 38 (c9orfl0)、配列番号 39 (clO orfl07)、配列番号 40(clorB8)および配列番号 41 (cl0orf58)に表す。 [0029] The nucleotide sequences of 29 genes shown in Table 4 are shown in SEQ ID NO: 15 (BMP2), SEQ ID NO: 16 (LECT2), SEQ ID NO: 17 (ADFP), SEQ ID NO: 18 (BPAG1), SEQ ID NO: 19 (SERPINF1), SEQ ID NO: 20 (GNAS), SEQ ID NO: 21 (AHSG), SEQ ID NO: 22 (CCL16), SEQ ID NO: 23 (AFM), SEQ ID NO: 24 (LTBP1), SEQ ID NO: 25 (C4BPA), SEQ ID NO: 26 (LYZ), SEQ ID NO: 4 (MMP 24), SEQ ID NO: 27 (PSAP), SEQ ID NO: 28 (HRG), SEQ ID NO: 29 (FST), SEQ ID NO: 30 (GPC3), SEQ ID NO: 31 (GPLD1), SEQ ID NO: 32 (GDF15), SEQ ID NO: 33 (SPINT2), SEQ ID NO: 34 (TNFRSF1A), SEQ ID NO: 35 (cl4orfl23), SEQ ID NO: 14 (c8orfl), SEQ ID NO: 36 (cl4orf32), SEQ ID NO: 37 (c5orH3), SEQ ID NO: 38 (c9orfl0), SEQ ID NO: It is represented by SEQ ID NO: 39 (clO orfl07), SEQ ID NO: 40 (clorB8) and SEQ ID NO: 41 (cl0orf58).

[0030] 表 5に示す 6遺伝子の塩基配列を配列番号 42 (THBS1)、配列番号 43 (CTSS)、配 列番号 44 (PON1)、配列番号 45 (DEFB1)、配列番号 46 (MAP2K2)および配列番 号 47 (PLA2G7)に示す。  [0030] The nucleotide sequences of the 6 genes shown in Table 5 are represented by SEQ ID NO: 42 (THBS1), SEQ ID NO: 43 (CTSS), SEQ ID NO: 44 (PON1), SEQ ID NO: 45 (DEFB1), SEQ ID NO: 46 (MAP2K2) and the sequence. It is shown in No. 47 (PLA2G7).

[0031] さらに、表 6に示す 21遺伝子の塩基配列を配列番号 36 (THBS1)、配列番号 43 (C TSS)、配列番号 48 (IGFBP1)、配列番号 49 (SULF2)、配列番号 50 (WNT5B)、配列 番号 51 (ADAMTS1)、配列番号 52 (SEPP1)、配列番号 53 (CHAD)、配列番号 54 ( DCN)、配列番号 56 (LBP)、配列番号 4 (MMP24)、配列番号 57 (SPARCL1)、配列 番号 58 (LRG1)、配列番号 59 (cHorflOO)、配列番号 35 (cl4orfl23)、配列番号 9 ( C14orl2)、配列番号 60 (C18orfl0)、配列番号 37 (C5orfl3)、配列番号 61 (C6orf79 )、配列番号 62 (C6ori80)および配列番号 38 (C9orfl0)に表す。括弧内の表示は、 表中の「Symbol」を示す。  [0031] Furthermore, the nucleotide sequences of 21 genes shown in Table 6 are represented by SEQ ID NO: 36 (THBS1), SEQ ID NO: 43 (CTSS), SEQ ID NO: 48 (IGFBP1), SEQ ID NO: 49 (SULF2), SEQ ID NO: 50 (WNT5B). , SEQ ID NO: 51 (ADAMTS1), SEQ ID NO: 52 (SEPP1), SEQ ID NO: 53 (CHAD), SEQ ID NO: 54 (DCN), SEQ ID NO: 56 (LBP), SEQ ID NO: 4 (MMP24), SEQ ID NO: 57 (SPARCL1), SEQ ID NO: 58 (LRG1), SEQ ID NO: 59 (cHorflOO), SEQ ID NO: 35 (cl4orfl23), SEQ ID NO: 9 (C14orl2), SEQ ID NO: 60 (C18orfl0), SEQ ID NO: 37 (C5orfl3), SEQ ID NO: 61 (C6orf79), SEQ ID NO: It is represented by number 62 (C6ori80) and SEQ ID NO: 38 (C9orfl0). The indication in parentheses indicates “Symbol” in the table.

[0032] 本発明で用いるヌクレオチドは上記配列を含むヌクレオチドおよびその断片を含む 。また、本発明に用いるヌクレオチドは、上記配列番号で示されるヌクレオチドとストリ ンジェントな条件下でノヽイブリダィズするヌクレオチドおよびその断片も含まれる。この ようなヌクレオチドとしては、例えば、上記塩基配列との相同性の程度力 全体の平 均で約 80%以上、好ましくは約 90%以上、より好ましくは約 95%以上である塩基配列 を含有するヌクレオチド等を挙げることが出来る。ハイブリダィゼーシヨンは、カレント' プロトコーノレズ 'イン'モレキュラー.ノ ィォロジー (Current protocols in molecular biol ogy (edited by Frederick M. Ausubel et al.， 1987) )に記載の方法等、当業界で公知 の方法あるいはそれに準じる方法に従って行なうことができる。また、市販のライブラ リーを使用する場合、添付の使用説明書に記載の方法に従って行なうことができる。 ここで、「ストリンジェントな条件」とは、例えば、「1XSSC、0.1% SDS、 37°C」程度の条 件であり、より厳しい条件としては「0.5XSSC、 0.1% SDS、 42°C」程度の条件であり、 さらに厳しい条件としては「0.2XSSC、 0.1% SDS、 65°C」程度の条件である。このよう にハイブリダィゼーシヨンの条件が厳しくなるほどプローブ配列と高い相同性を有す るヌクレオチドを単離を期待しうる。ただし、上記の SSC,SDSおよびに温度の条件の組 み合わせは例示であり、当業者であればノヽイブリダィゼーシヨンのストリンジエンシー を決定する上記もしくは他の要素（例えば、プローブ濃度、プローブの長さ、ハイプリ ダイゼーシヨンの反応時間など）を適宜組み合わせることにより、上記と同様のストリン ジエンシーを実現することが可能である。さらに、上記遺伝子のうちのいくつか (LGAL S8、 FCN3、 MDK、 GNAS、 LTBP1、 FST、 GPLD1)には、ノ リアントが存在し、本発明 のヌクレオチドは該ノ リアントの塩基配列を含むヌクレオチドまたはその断片も含む。 [0032] Nucleotides used in the present invention include nucleotides containing the above sequences and fragments thereof. The nucleotides used in the present invention also include nucleotides and fragments thereof that are hybridized under stringent conditions with the nucleotides shown in SEQ ID NOs. Such nucleotides include, for example, a base sequence that is about 80% or more, preferably about 90% or more, more preferably about 95% or more on average in terms of the degree of homology with the above base sequence. A nucleotide etc. can be mentioned. Hybridization is well known in the art, including the method described in Current protocols in molecular biology (edited by Frederick M. Ausubel et al., 1987). It can carry out according to the method of or the method according to it. When using a commercially available library, it can be carried out according to the method described in the attached instruction manual. Here, `` stringent conditions '' are, for example, conditions of `` 1XSSC, 0.1% SDS, 37 ° C '', and more severe conditions are `` 0.5XSSC, 0.1% SDS, 42 ° C '' The more severe condition is “0.2XSSC, 0.1% SDS, 65 ° C”. As the hybridization conditions become more severe in this way, it can be expected to isolate a nucleotide having a high homology with the probe sequence. However, the combinations of the above SSC, SDS and temperature conditions are exemplary, and those skilled in the art will recognize the above or other factors (eg, probe concentration, It is possible to achieve the same stringency as described above by appropriately combining the length of the probe and the reaction time of the hybridization. Furthermore, some of the above genes (LGAL S8, FCN3, MDK, GNAS, LTBP1, FST, GPLD1) have a Norint, and the nucleotide of the present invention includes a nucleotide containing the nucleotide sequence of the Norint or a nucleotide thereof. Includes fragments.

[0033] 本発明で用いるヌクレオチドとしては、上記遺伝子のセンス鎖よりなるヌクレオチド、 アンチセンス鎖よりなるヌクレオチドの 、ずれをも用いることができる。  [0033] As the nucleotide used in the present invention, it is also possible to use a shift between a nucleotide comprising the sense strand of the above gene and a nucleotide comprising the antisense strand.

[0034] 表 3〜6に示す遺伝子のうち、「DM (2型糖尿病患者群) / NGT (耐糖能正常患者群 ) by SAGE」で表される値が 1を超えるものが 2型糖尿病の病態または肥満症と関連 して発現が亢進する遺伝子であり、 1未満のものが 2型糖尿病の病態または肥満症と 関連して発現が減弱する遺伝子である。また、値がより大きいか、もしくは値がより小 さい遺伝子ほど、 2型糖尿病の病態または肥満症との関連がより強い遺伝子であると いえる。従って、表に挙げた遺伝子のうち、表中の「DM (2型糖尿病患者群) / NGT ( 耐糖能正常患者群） by SAGE」で表される値が 2以上のものまたは 0.5以下の遺伝子 をより好適に用いることができる。  [0034] Among the genes shown in Tables 3-6, those with a value of “DM (type 2 diabetic patients) / NGT (normal glucose tolerance patients) by SAGE” greater than 1 are pathological conditions of type 2 diabetes Or a gene whose expression is increased in relation to obesity, and a gene whose expression is attenuated in relation to the pathology of type 2 diabetes or obesity. In addition, a gene with a higher value or a lower value can be said to be a gene having a stronger association with the pathology of type 2 diabetes or obesity. Therefore, among the genes listed in the table, genes with a value represented by “DM (type 2 diabetic patients) / NGT (normal glucose tolerance patients) by SAGE” in the table are 2 or more or 0.5 or less. It can be used more suitably.

[0035] これらの遺伝子群に属する各遺伝子は公知の SAGE： Serial Analysis of GeneExpre ssion法 (Yamashita T. et al., Biochem Biophys Resし ommun 269:110- 11り, 2000; Ya masnita T. et al., Biochem Biophys Res C ommun 282:647-654, 2001; Science 276:1 268, 1997; Cell 88:243, 1997; Science 270:484, 1995; Nature 389:300, 1997;米国 特許第 5,695,937号公報；特表平 10-511002号公報）によって特定されたものである 。この SAGE法は、任意の細胞で発現している遺伝子力 調製した cDNAのそれぞれ について、個々を特徴づける 10塩基対の DNA配列（Tag)を決定する方法である。ま た、得られた全ての Tag力 個々の Tagの割合を算出することによって、各遺伝子の 発現頻度 (コピー数)を決定することもできる。 [0035] Each gene belonging to these gene groups is known in the known SAGE: Serial Analysis of Gene Expression method (Yamashita T. et al., Biochem Biophys Res ommun 269: 110-11, 2000; Ya masnita T. et al ., Biochem Biophys Res Commun 282: 647-654, 2001; Science 276: 1 268, 1997; Cell 88: 243, 1997; Science 270: 484, 1995; Nature 389: 300, 1997; U.S. Patent No. 5,695,937 ; Specified by Japanese National Publication No. 10-511002). This SAGE method is a method for determining a 10 base pair DNA sequence (Tag) that characterizes each of the cDNAs that have been prepared for gene expression and expressed in an arbitrary cell. Ma In addition, the expression frequency (copy number) of each gene can be determined by calculating the ratio of all the obtained tag powers and individual tags.

[0036] ま 7こ、

Figure imgf000020_0001
Center for Biotechnology Information)の Unigene (nttp: [0036] Ma 7
Figure imgf000020_0001
Unigene (nttp: Center for Biotechnology Information)

//www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=unigene)のァクセッシヨン番号 ) さ れ、該番号でデータベースにアクセスすることで、それぞれの遺伝子の配列情報を 容易に得ることが可能である。  //www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=unigene)), and by accessing the database with that number, the sequence information of each gene can be easily obtained. Is possible.

[0037] これらの遺伝子は、糖'脂質代謝、糖尿病性合併症、動脈硬化症などの病態を作る キー分子となり得る。 [0037] These genes can be key molecules for pathological conditions such as sugar-lipid metabolism, diabetic complications, and arteriosclerosis.

[0038] 従って、これらの遺伝子は、糖'脂質代謝の研究、 2型糖尿病や肥満症の病因解明 、 2型糖尿病の検出（診断)、 2型糖尿病に罹患するリスクの評価、肥満症になるリスク 評価、ならびに糖尿病や肥満症の新規な治療法の開発に利用することができる。ま た、これらの遺伝子は、インスリン抵抗性に密接に関連しており、インスリン抵抗性の 判定にも利用することができる。さらに、これらの遺伝子は糖尿病性合併症とも関連し ており、糖尿病性合併症の検出（診断)または糖尿病性合併症に罹患するリスクの判 定に利用することができる。さらに、糖尿病、肥満およびインスリン抵抗性は動脈硬化 症の危険因子であり、これらの遺伝子は動脈硬化症とも密接に関連しており、動脈硬 化症の検出（診断)および動脈硬化症に罹患するリスクの判定に用いることもできる。 なお、糖尿病性合併症としては、例えば、糖尿病性網膜症、糖尿病性腎症、糖尿病 性神経障害、糖尿病性大血管症、心筋梗塞、脳卒中等が挙げられる。  [0038] Therefore, these genes are used to study glycolipid metabolism, elucidate the etiology of type 2 diabetes and obesity, detect (diagnose) type 2 diabetes, evaluate the risk of suffering from type 2 diabetes, and become obese It can be used for risk assessment and development of new treatments for diabetes and obesity. In addition, these genes are closely related to insulin resistance, and can be used for determination of insulin resistance. In addition, these genes are also associated with diabetic complications and can be used to detect (diagnose) diabetic complications or to determine the risk of suffering from diabetic complications. In addition, diabetes, obesity, and insulin resistance are risk factors for arteriosclerosis, and these genes are also closely related to arteriosclerosis, which is associated with the detection (diagnosis) of arteriosclerosis and arteriosclerosis. It can also be used to determine risk. Examples of diabetic complications include diabetic retinopathy, diabetic nephropathy, diabetic neuropathy, diabetic macroangiopathy, myocardial infarction, and stroke.

[0039] 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症の病因解明のためには、 本発明の遺伝子の肝臓での発現状況を調べる必要がある。また、本発明の遺伝子 の肝臓での発現状況を調べることにより、被験体が 2型糖尿病、肥満症、糖尿病性合 併症または動脈硬化症に罹患しているかどうかの検出 (診断)を行うことができ、さら に、被験体が 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症に罹患するリ スクを有しているかどうかの評価をすることができる。なお、 2型糖尿病、肥満症、糖尿 病性合併症または動脈硬化症に罹患して ヽる場合でも、これらの疾患の初期にお ヽ て、 HOMA_R、 HbAlc等の病態を検出可能に呈さない場合がある。本願発明のヌク レオチドを用いることにより、 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化 症の病態に関連した遺伝子の発現の変動を測定でき、病態を検出可能に呈さない 場合であっても、 2型糖尿病、糖尿病性合併症または動脈硬化症に罹患していると 判断できる場合がある。また、このような状態は今後 2型糖尿病、肥満症、糖尿病性 合併症または動脈硬化症の病態を検出可能に呈する可能性のある状態であり、この 点 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症の病態を呈するリスクを 評価することちできる。 [0039] In order to elucidate the etiology of type 2 diabetes, obesity, diabetic complications or arteriosclerosis, it is necessary to examine the expression state of the gene of the present invention in the liver. Further, by examining the expression state of the gene of the present invention in the liver, it is possible to detect (diagnose) whether or not the subject suffers from type 2 diabetes, obesity, diabetic complications or arteriosclerosis. In addition, it can be assessed whether the subject has a risk of suffering from type 2 diabetes, obesity, diabetic complications or arteriosclerosis. Even if people suffer from type 2 diabetes, obesity, diabetic complications or arteriosclerosis, they do not detect detectable pathological conditions such as HOMA_R and HbAlc in the early stages of these diseases. There is. By using the nucleotide of the present invention, type 2 diabetes, obesity, diabetic complications or arteriosclerosis Even if it is possible to measure changes in the expression of genes related to the pathology of the disease and not detectably present the pathology, it may be determined that the patient is suffering from type 2 diabetes, diabetic complications or arteriosclerosis is there. In addition, such a condition is a condition that is likely to cause detection of type 2 diabetes, obesity, diabetic complications or arteriosclerotic pathology in the future, and this is a type 2 diabetes, obesity, diabetic complication. Can assess the risk of developing a pathology of atherosclerosis or arteriosclerosis.

[0040] このような検出または評価のためには、 HbAlc病態関連遺伝子群、 HOMA-R病態 関連遺伝子群、 MCR病態関連遺伝子群および BMI病態関連遺伝子群に属する遺 伝子の中の少なくとも 1種、好ましくは少なくとも 2種、さらに好ましくは少なくとも 5種、 さらに好ましくは少なくとも 10種、さらに好ましくは少なくとも 20種、さらに好ましくは少 なくとも 30種、さらに好ましくは全部を用いて、遺伝子の発現の亢進または減弱を測 定すればよい。また、 HbAlc病態関連遺伝子群、 HOMA-R病態関連遺伝子群、 MC R病態関連遺伝子群および BMI病態関連遺伝子群のそれぞれの群に属する遺伝子 の中力 少なくとも 1種、好ましくは少なくとも 2種、さらに好ましくは 5種、さらに好まし くは少なくとも 10種、さらに好ましくは全部を用いてもよい。特に BMI病態関連遺伝子 群は肥満症と関連していると予測されるので、肥満になるリスクの評価のために用い ることがでさる。  [0040] For such detection or evaluation, at least one of the genes belonging to the HbAlc disease state-related gene group, the HOMA-R disease state-related gene group, the MCR disease state-related gene group, and the BMI disease state-related gene group is used. , Preferably at least 2, more preferably at least 5, more preferably at least 10, more preferably at least 20, more preferably at least 30, more preferably all, to enhance gene expression Or just measure the attenuation. Further, at least one of the genes belonging to each of the HbAlc disease state-related gene group, the HOMA-R disease state-related gene group, the MCR disease state-related gene group, and the BMI disease state-related gene group, preferably at least 2, more preferably 5 may be used, more preferably at least 10, more preferably all. In particular, the BMI condition-related genes are predicted to be associated with obesity and can be used to assess the risk of becoming obese.

[0041] これらの遺伝子の発現の亢進または減弱の測定のためには、上記の遺伝子の塩基 配列の全部または一部を含むヌクレオチドをプローブまたはプライマーとして用いて 遺伝子発現の程度を測定すればよい。遺伝子発現の程度は、ノーザンプロット法、 R T- PCR法、リアルタイム PCR法、 in situハイブリダィゼーシヨン法、マイクロアレイ（マイ クロチップ)を用いた方法等により測定することが可能であり、ここに挙げた方法はい ずれも公知の方法で行うことができる。遺伝子の発現は、上記遺伝子の一部から転 写されたメッセンジャー RNA(mRNA)の量を測定すればよぐ該 mRNAにハイブリダィ ズするヌクレオチドプローブまたはプライマーの使用により測定することができる。  [0041] In order to measure the increase or decrease in the expression of these genes, the degree of gene expression may be measured using nucleotides containing all or part of the base sequences of the above genes as probes or primers. The level of gene expression can be measured by the Northern plot method, RT-PCR method, real-time PCR method, in situ hybridization method, microarray (microchip) method, etc. Any of the methods mentioned can be carried out by known methods. The expression of a gene can be measured by using a nucleotide probe or primer that hybridizes to the mRNA as long as the amount of messenger RNA (mRNA) transcribed from a part of the gene is measured.

[0042] プローブまたはプライマーの塩基長は、 10〜50bp、好ましくは 15〜25bpである。 [0042] The base length of the probe or primer is 10 to 50 bp, preferably 15 to 25 bp.

[0043] 被験体が 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症に罹患して ヽる 力どうかの検出 (診断)、被験体が 2型糖尿病、肥満症、糖尿病性合併症または動脈 硬化症に罹患するリスクを有して 、るかどうかの評価のためには試料サンプルとして 、肝臓生検サンプル、血清、血漿等を用いればよい。 [0043] Detecting whether the subject is suffering from type 2 diabetes, obesity, diabetic complications or arteriosclerosis (diagnosis), subject is type 2 diabetes, obesity, diabetic complications or artery A liver biopsy sample, serum, plasma or the like may be used as a sample sample for evaluating whether or not there is a risk of suffering from sclerosis.

[0044] HbAlc病態関連遺伝子群、 HOMA-R病態関連遺伝子群、 MCR病態関連遺伝子 群および BMI病態関連遺伝子群に属する遺伝子の 1種以上の発現が正常人に比較 して亢進または減弱している場合に、被験体が 2型糖尿病、肥満症、糖尿病性合併 症または動脈硬化症に罹患していると診断することができ、被験体が 2型糖尿病、肥 満症、糖尿病性合併症または動脈硬化症に罹患するリスクを有して ヽると評価するこ とができる。この際、 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症の病態 と関連して発現が亢進する遺伝子の場合は、その亢進の程度を定量することにより、 また、 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症の病態と関連して発 現が減弱する遺伝子の場合は、その減弱の程度を定量することにより、より正確に、 被験体が 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症に罹患していると 診断することができ、被験体が 2型糖尿病、肥満症、糖尿病性合併症または動脈硬 化症に罹患するリスクを有していると評価することができる。例えば、遺伝子の発現が より亢進している力、またはより減弱していれば、 2型糖尿病、肥満症、糖尿病性合併 症または動脈硬化症の重篤度が高!、と判断し得るし、また遺伝子の発現がより亢進 しているか、またはより減弱していれば、 2型糖尿病、肥満症、糖尿病性合併症また は動脈硬化症に罹るリスクが高いと評価することができる。  [0044] The expression of one or more genes belonging to the HbAlc disease-related gene group, the HOMA-R disease-related gene group, the MCR disease-related gene group, and the BMI disease-related gene group is increased or attenuated compared to normal individuals. In some cases, the subject can be diagnosed as having type 2 diabetes, obesity, diabetic complications or arteriosclerosis, and the subject has type 2 diabetes, hypertrophy, diabetic complications or arteries. Can be assessed as having a risk of suffering from sclerosis. In this case, in the case of a gene whose expression is increased in association with type 2 diabetes, obesity, diabetic complications or arteriosclerosis pathology, by quantifying the degree of the enhancement, type 2 diabetes, obesity A gene whose expression is attenuated in relation to the pathology of diabetes mellitus, diabetic complications or arteriosclerosis, the quantification of the extent of the attenuation makes the subject more accurate, such as type 2 diabetes, obesity, Can be diagnosed as suffering from diabetic complications or arteriosclerosis and assessed that the subject is at risk of suffering from type 2 diabetes, obesity, diabetic complications or arteriosclerosis can do. For example, if the expression of a gene is more enhanced or attenuated, it can be determined that type 2 diabetes, obesity, diabetic complications, or arteriosclerosis is more serious! In addition, if the gene expression is more enhanced or attenuated, it can be evaluated that the risk of developing type 2 diabetes, obesity, diabetic complications or arteriosclerosis is high.

[0045] さらに、遺伝子の発現を直接測定するばかりでなぐ遺伝子発現産物を測定しても よい。例えば、上記遺伝子がコードするタンパク質に対する抗体を用いることにより遺 伝子発現産物を測定することができる。本発明は、 2型糖尿病患者または肥満症患 者の肝臓と正常人の肝臓との間で、病態ごとに発現が変動する遺伝子がコードする タンパク質、該タンパク質に対する抗体、これらを含む試薬等も包含する。  [0045] Further, gene expression products may be measured as well as directly measuring gene expression. For example, the gene expression product can be measured by using an antibody against the protein encoded by the gene. The present invention also includes a protein encoded by a gene whose expression varies depending on the pathological condition between the liver of a type 2 diabetic or obese patient and the liver of a normal person, an antibody against the protein, a reagent containing these, and the like. To do.

[0046] 本発明は、 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症の病因解明の ための、 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症の検出（診断)のた めの、および 2型糖尿病、肥満症、糖尿病性合併症または動脈硬化症に罹患するリ スクを評価するための、ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病、肥満症、 糖尿病性合併症または動脈硬化症患者の肝臓組織で発現が亢進または減弱してい る遺伝子の塩基配列力 なるヌクレオチドまたはその一部配列を含むヌクレオチドを 含む 2型糖尿病関連遺伝子および Zまたは肥満症関連遺伝子の発現を測定するた めの試薬を包含する。該試薬は、前記遺伝子の塩基配列からなるヌクレオチドまたは その一部配列を含むヌクレオチドをプローブまたはプライマーとして含む試薬であり、 また前記遺伝子の塩基配列からなるヌクレオチドまたはその一部配列を含むヌクレオ チドを固相化したマイクロアレイ等の基板である。 [0046] The present invention provides detection (diagnosis) of type 2 diabetes, obesity, diabetic complications or arteriosclerosis for elucidation of the etiology of type 2 diabetes, obesity, diabetic complications or arteriosclerosis. For human type 2 diabetes, obesity, diabetic complications or atherosclerosis, compared to liver tissue of normal humans, human type 2 diabetes, obesity, Increased or attenuated expression in liver tissue of patients with diabetic complications or arteriosclerosis And a reagent for measuring the expression of type 2 diabetes-related genes and Z or obesity-related genes, which include nucleotides comprising a nucleotide sequence or a partial sequence thereof. The reagent is a reagent comprising a nucleotide comprising the nucleotide sequence of the gene or a nucleotide comprising a partial sequence thereof as a probe or primer, and a nucleotide comprising the nucleotide sequence of the gene or a nucleotide comprising a partial sequence thereof is immobilized. A phased substrate such as a microarray.

[0047] マイクロアレイは、前記遺伝子の塩基配列力 なるヌクレオチドまたはその一部配列 を含むヌクレオチドを適当な基板上に固定ィ匕することにより作製することができる。  [0047] The microarray can be prepared by immobilizing a nucleotide having the nucleotide sequence of the gene or a nucleotide containing a partial sequence thereof on an appropriate substrate.

[0048] 固定基板としては、ガラス板、石英板、シリコンウェハーなどが挙げられる。基板の 大きさとしては、例えば 3.5mm X 5.5mm、 18mm X 18mm, 22mm X 75mmなどが挙げら れる力 これは基板上のプローブのスポット数やそのスポットの大きさなどに応じて様 々に設定することができる。ポリヌクレオチドまたはその断片の固定ィ匕方法としては、 ヌクレオチドの荷電を利用して、ポリリジン、ポリエチレンィミン、ポリアルキルァミンな どのポリ陽イオンで表面処理した固相担体に静電結合させたり、アミノ基、アルデヒド 基、エポキシ基などの官能基を導入した固相表面に、アミノ基、アルデヒド基、 SH基、 ピオチンなどの官能基を導入したヌクレオチドを共有結合により結合させることもでき る。固定ィ匕は、アレイ機を用いて行えばよい。  [0048] Examples of the fixed substrate include a glass plate, a quartz plate, and a silicon wafer. For example, 3.5mm x 5.5mm, 18mm x 18mm, 22mm x 75mm, etc. as the size of the board This is set according to the number of probe spots on the board and the size of the spots. can do. As a method for immobilizing a polynucleotide or a fragment thereof, using a nucleotide charge, it can be electrostatically bound to a solid support surface-treated with a polycation such as polylysine, polyethyleneimine or polyalkylamine, Nucleotides introduced with functional groups such as amino group, aldehyde group, SH group, and pyotin can be covalently bonded to the solid phase surface introduced with functional groups such as amino group, aldehyde group, and epoxy group. Fixing may be performed using an array machine.

[0049] 前記試薬は、 HbAlc病態関連遺伝子群、 HOMA-R病態関連遺伝子群、 MCR病態 関連遺伝子群および BMI病態関連遺伝子群に属する 62遺伝子の中の少なくとも 1 種、好ましくは少なくとも 2種、さらに好ましくは少なくとも 5種、さらに好ましくは少なく とも 10種、さらに好ましくは少なくとも 20種、さらに好ましくは少なくとも 30種、さらに好 ましくは全部を含んでいる。また、 HbAlc病態関連遺伝子群、 HOMA-R病態関連遺 伝子群、 MCR病態関連遺伝子群および BMI病態関連遺伝子群のそれぞれの群に属 する遺伝子の中から少なくとも 1種、好ましくは少なくとも 2種、さらに好ましくは 5種、 さらに好ましくは少なくとも 10種、さらに好ましくは全部を含んで ヽる。  [0049] The reagent is at least one of 62 genes belonging to the HbAlc disease state-related gene group, HOMA-R disease state-related gene group, MCR disease state-related gene group and BMI disease state-related gene group, preferably at least two types, Preferably, it contains at least 5, more preferably at least 10, more preferably at least 20, more preferably at least 30, more preferably all. In addition, at least one, preferably at least two of the genes belonging to the HbAlc disease-related gene group, HOMA-R disease-related gene group, MCR disease-related gene group, and BMI disease-related gene group, More preferably, it includes 5 types, more preferably at least 10 types, and even more preferably all.

[0050] 上記のように、これらの遺伝子は、糖'脂質代謝、糖尿病性合併症、動脈硬化症な どの病態を作るキー分子となり得る。従って、本発明の遺伝子および遺伝子を含む 試薬は、糖'脂質代謝、糖尿病性合併症、動脈硬化症などの病態を作るキー分子の 解析のためのツールとして用 、得る。 [0050] As described above, these genes can serve as key molecules for pathological conditions such as glycolipid metabolism, diabetic complications, and arteriosclerosis. Therefore, the gene of the present invention and the reagent containing the gene are key molecules that produce pathologies such as sugar lipid metabolism, diabetic complications, and arteriosclerosis. Use as a tool for analysis, get.

[0051] また、上記試薬を含む 2型糖尿病、肥満症、糖尿病性合併症もしくは動脈硬化症の 検出 (診断薬)、または 2型糖尿病、肥満症、糖尿病性合併症もしくは動脈硬化症に 罹るリスクを評価するための検出薬も本発明に包含される。  [0051] In addition, detection of type 2 diabetes, obesity, diabetic complications or arteriosclerosis (diagnostic agent) containing the above reagents, or risk of suffering from type 2 diabetes, obesity, diabetic complications or arteriosclerosis Detection agents for assessing are also encompassed by the present invention.

[0052] さらに、本発明は、ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病、肥満症、糖尿 病性合併症または動脈硬化症患者の肝臓組織で発現が亢進または減弱している遺 伝子を用いて 2型糖尿病、肥満症、糖尿病性合併症もしくは動脈硬化症の治療薬、 または 2型糖尿病、肥満症、糖尿病性合併症もしくは動脈硬化症の予防薬をスクリー ユングする方法を包含する。例えば、候補薬剤を被験体に投与し、あるいは候補薬 剤を前記遺伝子と接触させ、前記遺伝子の発現が亢進させるかまたは減弱させる薬 剤を、治療薬または予防薬として選択することができる。  [0052] Furthermore, in the present invention, expression is increased or decreased in the liver tissue of patients with human type 2 diabetes, obesity, diabetic complications or arteriosclerosis, as compared to liver tissue of normal humans. How to use the gene to screen for type 2 diabetes, obesity, diabetic complications or arteriosclerosis treatment, or type 2 diabetes, obesity, diabetic complications or arteriosclerosis prophylaxis. Include. For example, a drug that increases or decreases the expression of the gene can be selected as a therapeutic or prophylactic agent by administering the candidate drug to a subject or bringing the candidate drug into contact with the gene.

実施例  Example

[0053] 本発明を以下の実施例によって具体的に説明するが、本発明はこれらの実施例に よって限定されるものではない。  [0053] The present invention will be specifically described with reference to the following examples, but the present invention is not limited to these examples.

[0054] サンプノレ [0054] Sampu Nore

SAGE法を用いた解析をおこなうために、胃癌、胆嚢癌、大腸癌等の悪性腫瘍に対 して外科的治療を必要とした、 2型糖尿病患者 5名、耐糖能正常患者 5名から肝組織 を得た (表 1)。肝組織を肝の癌非浸潤部から外科的に切除し、液体窒素にて速やか に凍結した。  Liver tissue from 5 patients with type 2 diabetes and 5 patients with normal glucose tolerance who needed surgical treatment for malignant tumors such as gastric cancer, gallbladder cancer, and colon cancer in order to perform analysis using the SAGE method (Table 1). The liver tissue was surgically excised from the non-cancerous part of the liver and immediately frozen in liquid nitrogen.

[表 1] 解析に用いた 2 糖尿病及び耐糖能正常患者の眩床生化学的特徴 GT DM  [Table 1] 2 Diabetes biochemical characteristics of patients with diabetes and normal glucose tolerance used in the analysis GT DM

n  n

Age (years) 62. fi±H. 8 BO. 2± 1 1. Q  Age (years) 62.fi ± H. 8 BO. 2 ± 1 1. Q

B i (kg/m2) 24. 2±i 7 2 &, 2±a. B i (kg / m 2 ) 24. 2 ± i 7 2 &, 2 ± a.

casual p t aiJina gl ucose (ing/dL) 1 ! 3±5. 0 173+G4*  casual p t aiJina gl ucose (ing / dL) 1! 3 ± 5. 0 173 + G4 *

HbAlc iK] 6. 2±1. 1 8. l±2. (T  HbAlc iK] 6. 2 ± 1. 1 8. l ± 2. (T

AST (IU/L) 3 28+24  AST (IU / L) 3 28 + 24

ALT (1U/L) 2S±21 29+ 18  ALT (1U / L) 2S ± 21 29+ 18

(画 i 44+24  (Picture i 44 + 24

Data are mean±SD, AST, aspartate t raiisHJiiiB se; ALT, al anine auiinot rans feiase ; Data are mean ± SD, AST, aspartate t raiisHJiiiB se; ALT, al anine auiinot rans feiase;

, gamiiia-gJ ut aiyl trans ferase. *p vs. 定量的リアルタイム PCR法をおこなうために、 2型糖尿病患者 21名、耐糖能正常患 者 11名から肝組織を得た (表 2)。経皮的超音波ガイド下肝生検にて肝組織を獲得し た後、液体窒素にて速やかに凍結した。 , gamiiia-gJ ut aiyl trans ferase. * p vs. To perform quantitative real-time PCR, liver tissue was obtained from 21 patients with type 2 diabetes and 11 patients with normal glucose tolerance (Table 2). Liver tissue was obtained by percutaneous ultrasound-guided liver biopsy, and then immediately frozen in liquid nitrogen.

[表 2]  [Table 2]

NGT DM NGT DM

11 21  11 21

Age (years) 43. S+12. 7 53. 0±9. 7  Age (years) 43. S + 1 2. 7 53. 0 ± 9.7

26. 5÷4. 6 24. 4±4. 1  26. 5 ÷ 4. 6 24. 4 ± 4.1

casual plasm glucose dng/dL) 90+8 143±50l casual plasm glucose dng / dL) 90 + 8 143 ± 50 l

HbAlc [%) 5, 1±0. 5 7. 4±1. 6'  HbAlc (%) 5, 1 ± 0. 5 7. 4 ± 1.6.

AST 36+21  AST 36 + 21

ALT (IUA) 79±81 35±21  ALT (IUA) 79 ± 81 35 ± 21

aspartate transaminase ; ALT, alanine  aspartate transaminase; ALT, alanine

[0056] 2型糖尿病群と耐糖能正常群で年齢、 BM 肝機能に有意差はなカゝつた。全症例 において B、 C型肝炎関連検査に異常はな力つた。糖尿病群症例は、アメリカ糖尿病 学会の糖尿病診断及び分類基準に基づ ヽて 2型糖尿病と診断した。全症例から書 面にてインフォームドコンセントを獲得した。本研究は実験プロトコールについて金沢 大学倫理委員会の承認を得たのち、ヘルシンキ宣言に沿って施行した。 [0056] There was no significant difference in age and BM liver function between the type 2 diabetes group and the normal glucose tolerance group. In all cases, abnormalities were observed in tests related to hepatitis B and C. Diabetes group cases were diagnosed as type 2 diabetes based on the Diabetes Diagnosis and Classification Criteria of the American Diabetes Association. Informed consent was obtained from all cases in writing. This study was conducted in accordance with the Declaration of Helsinki after obtaining approval from the Kanazawa University Ethics Committee for the experimental protocol.

[0057] SAGE法  [0057] SAGE method

肝組織から抽出した全 RNAを FastTrac mRNA抽出キット（Invitrogen, San Diego, C A)を用いてアデ-ル化した。各サンプルにっき 2.5 gの RNAを SAGE法に用いた。 S AGE法のプロトコールは過去の報告に従った（Yamashita T. et al.， Biochem Biophys Res Commun 269: 110—116, 2000; Yamashita T. et al., Biochem Biophys Res Comm un 282:647-654, 2001)。 SAGEライブラリーを ABI PRISM 377 DNAシーケンサーと Bi gDyeターミネータ一サイクルシーケンスキット（PE Biosystems, Foster city, CA)を用 いてシーケンスした。シーケンスしたファイルを SAGE 2000ソフトウェア（www. sagenet. org)を用いて解析した。  Total RNA extracted from liver tissue was adenylated using FastTrac mRNA extraction kit (Invitrogen, San Diego, CA). For each sample, 2.5 g of RNA was used in the SAGE method. The protocol for the S AGE method followed previous reports (Yamashita T. et al., Biochem Biophys Res Commun 269: 110-116, 2000; Yamashita T. et al., Biochem Biophys Res Comm un 282: 647-654, 2001). The SAGE library was sequenced using the ABI PRISM 377 DNA sequencer and the BigDye terminator single cycle sequencing kit (PE Biosystems, Foster city, CA). Sequenced files were analyzed using SAGE 2000 software (www.sagenet.org).

[0058] 定量的リアルタイム PCR法 [0058] Quantitative real-time PCR

二重鎖 cDNAを定量的リアルタイム PCR法のための铸型として用いた。この目的の ために、 ABI Prism 7700シーケンスディテクシヨンシステム（Applied Biosystems, Fost er City, Calif., USA)を使用した。プライマーと TaqManプローブのセットは Applied Bi osystems専: Cのものである (Assays— on— Demand gene expression product)。異なつ 7こ サンプルにおける PCRに利用可能な DNA量の変動を補正する目的で、標的配列の 遺伝子発現を内因性コントロールであるベータァクチン RNA (TaqMan Control Reage nt Kit, Appleid Biosystems)の発現量で標準化した。 PCR条件は、 50°C2分間および 95°C10分間を 1サイクル、その後 95°C15秒間と 60°C1分間を 40サイクルとした。 Double-stranded cDNA was used as a cage for quantitative real-time PCR. For this purpose ABI Prism 7700 Sequence Detection System (Applied Biosystems, Foster City, Calif., USA) was used. Primer and TaqMan probe set is from Applied Biosystems: C (Assays—on—Demand gene expression product). In order to compensate for variations in the amount of DNA available for PCR in 7 different samples, the gene expression of the target sequence was standardized with the expression level of the endogenous control beta-actin RNA (TaqMan Control Reagent Kit, Appleid Biosystems). . PCR conditions were 50 ° C for 2 minutes and 95 ° C for 10 minutes for 1 cycle, and then 95 ° C for 15 seconds and 60 ° C for 1 minute for 40 cycles.

[0059] Gene Ontologyによる細胞内局在の推定  [0059] Estimation of intracellular localization by Gene Ontology

Gene Ontologyし onsortium (http://www.geneontology.org/)の 己載に ¾づさ、 |P】 定した遺伝子を転写産物の細胞内局在によって分類した。そのなかで、「細胞外」の クラスに分類された遺伝子群を分泌蛋白コード遺伝子としてより詳細に検討した。  Based on the self-publishing of Gene Ontology onsortium (http://www.geneontology.org/), | P] the identified genes were classified by the intracellular localization of the transcripts. Among them, we examined in more detail the genes classified into the “extracellular” class as secretory protein-coding genes.

[0060] HOMA-R  [0060] HOMA-R

各症例におけるインスリン抵抗性、特に肝臓におけるインスリン抵抗性を評価する ために、各症例の HOMA-Rを計算した。 HOMA-Rは過去の報告（Matthews DR et al ., Diabetologia 28:412-419, 1985)に従い、各症例の空腹時血糖値（mg/dL) X空腹 時の血中インスリン濃度; z U/ml) /405から計算した。  To evaluate insulin resistance in each case, especially in the liver, HOMA-R for each case was calculated. HOMA-R is based on previous reports (Matthews DR et al., Diabetologia 28: 412-419, 1985). Fasting blood glucose level (mg / dL) X fasting blood insulin concentration in each case; z U / ml ) Calculated from / 405.

[0061] MCR [0061] MCR

骨格筋におけるインスリン抵抗性を正確に評価するために、人工脾臓を用いた高ィ ンスリン正常血糖下クランプ法（DeFronzo RA et al., Am J Physiol 237:E214-223, 19 79)を用いて、 Metabolic clearance rate (MCR)を計算した。  In order to accurately evaluate insulin resistance in skeletal muscle, a high-insulin normoglycemic clamp method using an artificial spleen (DeFronzo RA et al., Am J Physiol 237: E214-223, 19 79) Metabolic clearance rate (MCR) was calculated.

[0062] 統計学的検定方法 [0062] Statistical test method

2型糖尿病患者群と耐糖能正常患者群の臨床的特徴の比較は unpaired-t testを 用いた。 BMI、 HOMA-R, MCRと各遺伝子の発現レベルとの関係の解析は線形相関 解析でおこなった。 V、ずれも P〈0.05を有意とした。  The unpaired-t test was used to compare the clinical characteristics of patients with type 2 diabetes and normal glucose tolerance. The relationship between BMI, HOMA-R, MCR and the expression level of each gene was analyzed by linear correlation analysis. For V and deviation, P <0.05 was considered significant.

[0063] 〔結果〕 [0063] [Result]

SAGE解析  SAGE analysis

SAGE法を用いて、 144,901タグ（2型糖尿病患者群 44280タグ、耐糖能正常患者群 1 00,621タグ）の遺伝子を同定した。 Basic local alignment search tool program (BLAST http://www.ncbi.nlm.nih.gov/BLAST/)を用いてタグ配列から相当遺伝子を解析し た結果、 2型糖尿病患者群力 5221遺伝子、耐糖能正常患者群カゝら 6511遺伝子の 発見 同定しァこ。 Gene Ontology Consortium (http:// www. geneontology.org/)の己 載を用いて遺伝子を転写産物の細胞内局在別に分類したところ、 232遺伝子が"細 胞外分泌蛋白"をコードする遺伝子であった。このうちで、 2型糖尿病患者群で耐糖 能正常患者群に比較して 1.5倍以上に発現亢進している 63遺伝子、 1.5倍以下に発 現低下している 114遺伝子を同定した。上記の遺伝子について、定量的リアルタイム PCR法を用いて各症例における遺伝子発現量を測定し、 BMI、 H0MA-R、 MCRといつ た臨床的パラメーターと比較検討した。 Using the SAGE method, the genes of 144,901 tags (type 2 diabetes patient group 44280 tag, normal glucose tolerance patient group 100,621 tag) were identified. Basic local alignment search tool program (BLAST (http://www.ncbi.nlm.nih.gov/BLAST/), and the corresponding gene was analyzed from the tag sequence. Discover and identify. Using the gene ontology consortium (http://www.geneontology.org/), the genes were classified according to the subcellular localization of transcripts. As a result, 232 genes were genes encoding "extracellular secretory proteins". It was. Of these, 63 genes that were upregulated 1.5 times or more in the type 2 diabetes patient group and 114 genes that were expressed 1.5 times or less in comparison with the group with normal glucose tolerance were identified. For the above genes, the amount of gene expression in each case was measured using quantitative real-time PCR, and compared with clinical parameters such as BMI, H0MA-R, and MCR.

[0064] 発現レベルと HbAlc値との間に有意な相関を認める遺伝子群の同定 [0064] Identification of genes that show a significant correlation between expression level and HbAlc level

発現レベルと HbAlc値との間に有意な相関を認める 14遺伝子を同定した (表 3)。こ れらの遺伝子は、糖代謝を正あるいは負に制御する作用、あるいは高血糖や高イン スリン血症、酸ィヒストレス等を感知して発現が制御されるものと考えられる。これらの 中に糖尿病自体、あるいは合併症の病態を作り出して Lヽる遺伝子が含まれて ヽること が期待される。  We identified 14 genes that showed a significant correlation between expression levels and HbAlc values (Table 3). The expression of these genes is thought to be controlled by positively or negatively controlling glucose metabolism, or by detecting hyperglycemia, hyperinsulinemia, acid stress, etc. These genes are expected to contain diabetes mellitus itself or genes that can cause complications.

[¾3] 発現レベルと HbAlcの間に有 な相闊を認める逸伝子群  [¾3] An anecdote group with a significant correlation between expression level and HbAlc

[0065] 発現レベルと BMIとの聞に有意な相関を認める遺伝子群の同定 [0065] Identification of genes that have a significant correlation between expression level and BMI

発現レベルと BMIとの聞に有意な相関を認める 29遺伝子を同定した (表 4)。これら の遺伝子は、全身の脂肪蓄積量と関連してその発現が変動していると考えられる。こ れらの中に、細胞の脂質代謝やエネルギー代謝を調節する遺伝子、あるいはインス リン低抗性に関連した病態を形成する遺伝子が含まれていることが期待される。 We identified 29 genes that showed a significant correlation between expression level and BMI (Table 4). These genes are thought to vary in their expression in relation to systemic fat accumulation. This Among these, genes that regulate lipid metabolism and energy metabolism of cells or genes that form pathologies related to insulin resistance are expected.

[表 4] レベルと BMIとの に》意な ffi 13 める 群  [Table 4] Level and BMI

Figure imgf000028_0001
Figure imgf000028_0001

Figure imgf000028_0002
Figure imgf000028_0002

発現レベルと HOMA-Rとの間に有意な相関を認める遺伝子群の同定 Identification of genes that have a significant correlation between expression level and HOMA-R

発現レベルと HOMA-Rとの間に有意な相関を認める 6遺伝子を同定した (表 5)。こ れらの遺伝子は、肝臓におけるインスリン抵抗性と関連してその発現が変動している 、これらの中に、細胞の脂質代謝やエネルギー代謝を調節する遺伝子、あるいはィ ンスリン抵抗性に関連した病態を形成する遺伝子が含まれていることが期待される。  Six genes were identified that showed a significant correlation between expression levels and HOMA-R (Table 5). These genes vary in their expression in relation to insulin resistance in the liver. Among these genes are genes that regulate cell lipid metabolism and energy metabolism, or pathologies related to insulin resistance. It is expected to contain genes that form

[表 5] 発現レベルと HOMA Rの間に有意な相閣 認める遺伝子群 [Table 5] Significant gene group between expression level and HOMA R

DM/NGT, p value  DM / NGT, p value

Symbol Name  Symbol Name

by SAGE vs.HO A coefficient  by SAGE vs. HO A coefficient

Hs.l 64226 THBSl thrombospondin 1 0.D5 0.002 0.67  Hs.l 64226 THBSl thrombospondin 1 0.D5 0.002 0.67

Hs.181301 CTSS cat e sin S 0.OD7 0  Hs.181301 CTSS cat e sin S 0.OD7 0

Hs.28220S PO 1 paraaxonase 1 2.46 0.C22 0.523  Hs.28220S PO 1 paraaxonase 1 2.46 0.C22 0.523

def nsin beta  def nsin beta

Hs.465627 AP2K2 MAP2K2 mitogen activated protein kinase kinase 0,01 0.C37 0.481  Hs.465627 AP2K2 MAP2K2 mitogen activated protein kinase kinase 0,01 0.C37 0.481

Hs.93304 PLA2G7 phospho lipase A2, group VH (platelet-activating 2.Z7 0.04  Hs.93304 PLA2G7 phospho lipase A2, group VH (platelet-activating 2.Z7 0.04

factor aeetvlhvdrolase, o!esma} 発現レベルと MCRとの間に有意な相関を認める遺伝子群の同定  factor aeetvlhvdrolase, o! esma} Identification of genes that show a significant correlation between expression level and MCR

発現レベルと MCRとの間に有意な相関を認める 21遺伝子を同定した（表 6)。これら の遺伝子は、骨格筋を代表とする末梢組織のインスリン抵抗性と関連してその発現 が変動している。これらの中に、臓器間のインスリンシグナルや糖'脂質の流れを制 御する遺伝子、あるいはインスリン低抗性に関連した病態を形成するが含まれて Lヽる ことが期待される。  We identified 21 genes that showed a significant correlation between expression levels and MCR (Table 6). The expression of these genes varies in association with insulin resistance in peripheral tissues such as skeletal muscle. Among these, genes that control insulin signals between sugars and the flow of sugars and lipids, and pathologies related to insulin resistance are expected to be used.

[¾6] 発現レベルと MCRとの間に有意な相閱を認める 子群 [¾6] Offspring group showing significant correlation between expression level and MCR

DM/NGT.hy p value correlation  DM / NGT.hy p value correlation

Unigene Symbol Name  Unigene Symbol Name

SAGE vs-MCR coefficient  SAGE vs-MCR coefficient

THBS1 UirDm aspDndin 1 0.05 0.OO3 -0.772  THBS1 UirDm aspDndin 1 0.05 0.OO3 -0.772

Hs.401316 IGFSP1 IGFBP1 insulin like emwtfi factor binding protein 1 0.01 0.009 0.711  Hs.401316 IGFSP1 IGFBP1 insulin like emwtfi factor binding protein 1 0.01 0.009 0.711

Hs.43857 SU F2 suEfa asB 2 0.57 0.01 -0.7C7  Hs.43857 SU F2 suEfa asB 2 0.57 0.01 -0.7C7

Hs.306051 WNT5B wingless type WTV integration site family member 0.05 0.01 0706  Hs.306051 WNT5B wingless type WTV integration site family member 0.05 0.01 0706

5B 5B

.8230 ADAMTS1 ADA TS1 a disintegrin like and metal lop rate ase 0.05 0.O1 -0.687  .8230 ADAMTS1 ADA TS1 a disintegrin like and metal lop rate ase 0.05 0.O1 -0.687

(reprolysin type) with thrombospondin ty e 1 motif 1  (reprolysin type) with thrombospondin ty e 1 motif 1

Hs.275775 SEPP1 selenoprotein P, plasma, 1r B.33 0-021 -0.B5B  Hs.275775 SEPP1 selenoprotein P, plasma, 1r B.33 0-021 -0.B5B

CHAD !herirk 45,17 0.022 0.G49  CHAD! Herirk 45,17 0.022 0.G49

Hs-l 5831S DCN dccorin 0.38 0.024 -0  Hs-l 5831S DCN dccorin 0.38 0.024 -0

81301 CTSS cathepsin Ξ 2.27 0.026 -0.638  81301 CTSS cathepsin Ξ 2.27 0.026 -0.638

Hs.154078 LBP lip opolysac ch ng binding protein 0.44 0.02S -0.629 Hs.154078 LBP lipopolysac ch ng binding protein 0.44 0.02S -0.629

.2125S1 MMP24 MMP24 matrix nrK alloproteinase 24 (membrane 0,05 0.03 0.623  .2125S1 MMP24 MMP24 matrix nrK alloproteinase 24 (membrane 0,05 0.03 0.623

inserted)  inserted)

Hs.62886 SPARCL1 SPARC-like \ (mas . he in) 6.S2 Q.034 -0.S13  Hs.62886 SPARCL1 SPARC-like \ (mas .he in) 6.S2 Q.034 -0.S13

Hs.10844 L G1 leucine~rich alpha - 2 - glycoprotein 1 0.1 S □ 036 0.BQ9  Hs.10844 L G1 leucine ~ rich alpha-2-glycoprotein 1 0.1 S □ 036 0.BQ9

CUorflOn chromosome 14 open reading frame 100 45.00 0.048 -0.579  CUorflOn chromosome 14 open reading frame 100 45.00 0.048 -0.579

Hs.279761 C14 rf123 chromosome 14 open reading frame 123 6,80 0.04 -0.5S8  Hs.279761 C14 rf123 chromosome 14 open reading frame 123 6,80 0.04 -0.5S8

Hs.109052 C14orf2 cbromosamc 14 open reading frame 2 6.77 0.026 0.63 &  Hs.109052 C14orf2 cbromosamc 14 open reading frame 2 6.77 0.026 0.63 &

H3.Z29B1 GIBorfID CIBorflO chromosome Ifl open reading frame 1C 0.03 -0.635  H3.Z29B1 GIBorfID CIBorflO chromosome Ifl open reading frame 1C 0.03 -0.635

Hs.50S741 C5orf13 chramosome 5 open reading frame 13 225 0021 -0.682  Hs.50S741 C5orf13 chramosome 5 open reading frame 13 225 0021 -0.682

Hs.214043 C6orn9 open reading "frame 75 2300 0.044 -0.615  Hs.214043 C6orn9 open reading "frame 75 2300 0.044 -0.615

Hs.44468 C6arf80 CBorfBQ chromosome 6 open reading frame SO 1.00 0,007 0,756  Hs.44468 C6arf80 CBorfBQ chromosome 6 open reading frame SO 1.00 0,007 0,756

Hs.44BS34 C9orf10 ehromosomc 9 open reading frame 10 0.55 0033 0.716 以上の実施例が示すように、 2型糖尿病患者の肝臓で発現が減弱もしくは亢進して おり、その発現レベルが血糖コントロール、肥満、肝臓'骨格筋のインスリン低抗性と 相関する分泌蛋白コード遺伝子群を同定した。これらの遺伝子にコードされる肝由来 分泌蛋白は、細胞内のインスリンシグナル、糖'脂質代謝等に影響を及ぼすと思われ 、全身の脂肪蓄積、インスリン低抗性ゃ動脈硬化発症に関連している可能性がある、 これらの分泌蛋白の機能を明らかにすることは、 2型糖尿病や肥満症の病因解明な らびに新規治療法の開発につながることが期待される。 Hs.44BS34 C9orf10 ehromosomc 9 open reading frame 10 0.55 0033 0.716 As shown in the above examples, expression is attenuated or enhanced in the liver of type 2 diabetic patients, and its expression level is glycemic control, obesity, liver 'skeleton Muscle insulin resistance Correlated secretory protein coding genes were identified. The liver-derived secretory protein encoded by these genes is thought to affect intracellular insulin signals, sugar and lipid metabolism, etc., and is related to systemic fat accumulation and the development of insulin hypotonia and arteriosclerosis. It is expected that elucidating the functions of these secreted proteins may lead to elucidation of the etiology of type 2 diabetes and obesity and the development of new therapies.

本明細書で引用した全ての刊行物、特許および特許出願をそのまま参考として本 明細書にとり入れるものとする。  All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

Claims

請求の範囲 The scope of the claims ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者または肥満症患者の肝臓組 織で発現が亢進または減弱している遺伝子であって、分泌タンパク質をコードしてい る以下の遺伝子からなる群力 選択される少なくとも 1つの遺伝子の塩基配列からな るヌクレオチドまたはその一部配列を含むヌクレオチドを含む 2型糖尿病関連遺伝子 および Zまたは肥満症関連遺伝子の発現を測定するための試薬。  A gene that is upregulated or attenuated in the liver tissue of human type 2 diabetic or obese patients compared to normal human liver tissue and consists of the following genes encoding secreted proteins Group force A reagent for measuring the expression of a type 2 diabetes-related gene and a Z or obesity-related gene comprising a nucleotide consisting of a base sequence of at least one selected gene or a nucleotide containing a partial sequence thereof. (1) poliovirus receptor  (1) poliovirus receptor (2) lectin, galactoside- Dinding, soluble, 8 (galectin 8)  (2) lectin, galactoside- Dinding, soluble, 8 (galectin 8) (3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen) (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2) (6) nucleoDindin 2  (6) nucleoDindin 2 (7) matrix metalloproteinase 11 (stromelysin 3)  (7) matrix metalloproteinase 11 (stromelysin 3) (8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma) (9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2 (10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4 (11) chromosome 10 open reading frame 10  (11) chromosome 10 open reading frame 10 (12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74 (13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA (14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1 (15) bone morphogenetic protein 2  (15) bone morphogenetic protein 2 (16) leukocyte cell-derived chemotaxin 2  (16) leukocyte cell-derived chemotaxin 2 (17) adipose differentiation- related protein  (17) adipose differentiation- related protein (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA (19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 (20) GNAS complex locus (21) AHSG alpha 2 HS glycoprotein (20) GNAS complex locus (21) AHSG alpha 2 HS glycoprotein (22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16 (23) afamin  (23) afamin (24) LTBPl latent transforming growth factor beta binding protein 1  (24) LTBPl latent transforming growth factor beta binding protein 1 (25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha (26) lysozyme (renal amyloidosis)  (26) lysozyme (renal amyloidosis) (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)  (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (28) histidine- rich glycoprotein (28) histidine-rich glycoprotein (29) FST follistatin  (29) FST follistatin (30) glypican 3  (30) glypican 3 (31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15 (33) SPINT2 serine protease inhibitor Kunitz type 2  (33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A  (34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A (35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123 (36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32 (37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13 (38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10 (39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107 (40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38 (41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58 (42) thrombospondin 1  (42) thrombospondin 1 (43) cathepsin S  (43) cathepsin S (44) paraoxonase 1  (44) paraoxonase 1 (45) DEFBl defensin beta 1  (45) DEFBl defensin beta 1 (46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2 (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) (48) IGFBP1 insulin like growth factor binding protein 1 (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) (48) IGFBP1 insulin like growth factor binding protein 1 (49) sulfatase 2  (49) sulfatase 2 (50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B (51) ADAMTS1 a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTS1 a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1 (52) selenoprotein P， plasma, In  (52) selenoprotein P, plasma, In (53) chondroadherin  (53) chondroadherin (54) decorin  (54) decorin (56) lipopolysaccharide binding protein  (56) lipopolysaccharide binding protein (57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin) (58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1 (59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100 (60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10 (61) chromosome 6 open reading frame 79  (61) chromosome 6 open reading frame 79 (62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80 ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱がヒト血中 HbAlc値と相関している以下の遺伝子からなる群から選 択される少なくとも 1つの遺伝子である請求項 1記載の 2型糖尿病関連遺伝子および A gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetes patient is selected from the group consisting of the following genes whose up- or down-expression is correlated with human blood HbAlc levels: The type 2 diabetes-related gene according to claim 1, which is at least one gene and Zまたは肥満症関連遺伝子の発現を測定するための試薬。 Reagent for measuring the expression of Z or obesity related gene. (1) poliovirus receptor  (1) poliovirus receptor (2) lectin, galactoside- binding, soluble, 8 (galectin 8)  (2) lectin, galactoside-binding, soluble, 8 (galectin 8) (3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen) (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2) (6) nucleobindin 2  (6) nucleobindin 2 (7) matrix metalloproteinase 11 (stromelysin 3)  (7) matrix metalloproteinase 11 (stromelysin 3) (8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma) (9) chromosome 14 open reading frame 2 (10) chromosome 14 open reading frame 4 (9) chromosome 14 open reading frame 2 (10) chromosome 14 open reading frame 4 (11) chromosome 10 open reading frame 10  (11) chromosome 10 open reading frame 10 (12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74 (13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA (14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1 ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱が HOMA-Rと相関している以下の遺伝子からなる群から選択され る少なくとも 1つの遺伝子である請求項 1記載の 2型糖尿病関連遺伝子および Zまた は肥満症関連遺伝子の発現を測定するための試薬。  At least one selected from the group consisting of the following genes whose expression is up- or down-regulated in the liver tissue of human type 2 diabetic patients and whose up- or down-expression is correlated with HOMA-R: The reagent for measuring the expression of type 2 diabetes-related gene and Z or obesity-related gene according to claim 1, which is a gene. (42) thrombospondin 丄  (42) thrombospondin 丄 (43) cathepsin S  (43) cathepsin S (44) paraoxonase 1  (44) paraoxonase 1 (45) DEFB1 defensin beta 1  (45) DEFB1 defensin beta 1 (46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2 (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱が MCRと相関している以下の遺伝子からなる群から選択される少 なくとも 1つの遺伝子である請求項 1記載の 2型糖尿病関連遺伝子および Zまたは肥 満症関連遺伝子の発現を測定するための試薬。  (47) Phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) The reagent for measuring the expression of type 2 diabetes-related gene and Z or hypertrophy-related gene according to claim 1, which is at least one gene selected from the group consisting of the following genes: (42) thrombospondin 丄  (42) thrombospondin 丄 (43) cathepsin S  (43) cathepsin S (48) IGFBP1 insulin like growth factor binding protein 1  (48) IGFBP1 insulin like growth factor binding protein 1 (49) sulfatase 2  (49) sulfatase 2 (50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1 (52) selenoprotein P， plasma, In (53) chondroadherin (52) selenoprotein P, plasma, In (53) chondroadherin (54) decorin  (54) decorin (56) lipopolysaccharide binding protein  (56) lipopolysaccharide binding protein (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin) (58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1 (59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100 (35) chromosome 14 open reading frame 123 (35) chromosome 14 open reading frame 123 (9) chromosome 14 open reading frame 2 (9) chromosome 14 open reading frame 2 (60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10 (37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13 (61) chromosome 6 open reading frame 79  (61) chromosome 6 open reading frame 79 (62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80 (38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10 ヒト肥満症患者の肝臓糸且織で発現が亢進または減弱して 、る遺伝子が、発現の亢 進または減弱が BMIと相関している以下の遺伝子からなる群から選択される少なくと も 1つの遺伝子である請求項 1記載の 2型糖尿病関連遺伝子および Zまたは肥満症 関連遺伝子の発現を測定するための試薬。  A gene whose expression is up- or down-regulated in liver thread and tissue of a human obese patient is selected from the group consisting of the following genes whose up- or down-expression is correlated with BMI: The reagent for measuring the expression of type 2 diabetes-related gene and Z or obesity-related gene according to claim 1, which is a gene. (15) bone morphogenetic protein 2  (15) bone morphogenetic protein 2 (16) leukocyte cell-derived chemotaxin 2  (16) leukocyte cell-derived chemotaxin 2 (17) adipose differentiation- related protein  (17) adipose differentiation- related protein (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA (19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 (20) GNAS complex locus  (20) GNAS complex locus (21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein (22) chemo ine (C- C motif) ligand 16  (22) chemo ine (C- C motif) ligand 16 (23) afamin (24) LTBP1 latent transforming growth factor beta binding protein 1 (23) afamin (24) LTBP1 latent transforming growth factor beta binding protein 1 (25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha (26) lysozyme (renal amyloidosis)  (26) lysozyme (renal amyloidosis) (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy)  (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (28) histidine- rich glycoprotein (28) histidine-rich glycoprotein (29) FST follistatin  (29) FST follistatin (30) glypican 3  (30) glypican 3 (31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15 (33) SPINT2 serine protease inhibitor Kunitz type 2  (33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A  (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A (35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123 (14) chromosome 8 open reading frame 1 (14) chromosome 8 open reading frame 1 (36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32 (37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13 (38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10 (39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107 (40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38 (41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58 少なくとも 5つの遺伝子を含む請求項 1〜5のいずれ力 1項に記載の 2型糖尿病関 連遺伝子および Zまたは肥満症関連遺伝子の発現を測定するための試薬。  The reagent for measuring expression of type 2 diabetes-related gene and Z or obesity-related gene according to any one of claims 1 to 5, comprising at least 5 genes. ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進ま たは減弱している遺伝子の塩基配列からなるヌクレオチドまたはその一部配列を含 むヌクレオチドを固相化したマイクロアレイを含む請求項 1〜6のいずれ力 1項に記載 の 2型糖尿病関連遺伝子および Zまたは肥満症関連遺伝子の発現を測定するため の試薬。 ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進ま たは減弱している遺伝子の塩基配列からなるヌクレオチドまたはその一部配列を含 むヌクレオチドをターゲットとしたプローブまたはプライマーを含む請求項 1〜6のい ずれか 1項に記載の 2型糖尿病関連遺伝子および Zまたは肥満症関連遺伝子の発 現を測定するための試薬。 Compared with normal human liver tissue, nucleotides comprising the nucleotide sequence of a gene whose expression is up-regulated or attenuated in the liver tissue of human type 2 diabetics or a nucleotide containing a partial sequence is immobilized. A reagent for measuring the expression of type 2 diabetes-related gene and Z or obesity-related gene according to any one of claims 1 to 6, comprising a microarray. Compared with liver tissue of normal human subjects, nucleotides consisting of the nucleotide sequence of a gene whose expression is increased or attenuated in liver tissue of human type 2 diabetic patients or nucleotides containing a partial sequence thereof were targeted. A reagent for measuring the expression of a type 2 diabetes-related gene and Z or obesity-related gene according to any one of claims 1 to 6, comprising a probe or a primer. ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進ま たは減弱している遺伝子であって、分泌タンパク質をコードしている以下の遺伝子か らなる群から選択される少なくとも 1つの遺伝子について、被検体における発現を測 定し、該遺伝子の発現が正常人に比較して亢進または減弱している場合に前記被 検体が 2型糖尿病、肥満症、糖尿病性合併症もしくは動脈硬化症に罹患しているか 、または罹患するリスクがあると判断する方法。  A gene whose expression is increased or attenuated in the liver tissue of human type 2 diabetic patients compared to the liver tissue of normal humans, from the group consisting of the following genes encoding secreted proteins: For at least one selected gene, the expression in the subject is measured, and when the expression of the gene is increased or decreased compared to a normal person, the subject is type 2 diabetes, obesity, diabetic A method of determining whether you are suffering from or at risk of suffering from a complication or arteriosclerosis. (1) poliovirus receptor  (1) poliovirus receptor (2) lectin, galactoside- Dinding, soluble, 8 (galectin 8)  (2) lectin, galactoside- Dinding, soluble, 8 (galectin 8) (3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen) (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2) (6) nucleoDindin 2  (6) nucleoDindin 2 (7) matrix metalloproteinase 11 (stromelysin 3)  (7) matrix metalloproteinase 11 (stromelysin 3) (8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma) (9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2 (10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4 (11) chromosome 10 open reading frame 10  (11) chromosome 10 open reading frame 10 (12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74 (13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA (14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1 (15) bone morphogenetic protein 2  (15) bone morphogenetic protein 2 (16) leukocyte cell-derived chemotaxin 2 (17) adipose differentiation- related protein (16) leukocyte cell-derived chemotaxin 2 (17) adipose differentiation- related protein (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA (19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 (20) GNAS complex locus  (20) GNAS complex locus (21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein (22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16 (23) afamin  (23) afamin (24) LTBPl latent transforming growth factor beta binding protein 1  (24) LTBPl latent transforming growth factor beta binding protein 1 (25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha (26) lysozyme (renal amyloidosis)  (26) lysozyme (renal amyloidosis) (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (28) histidine- rich glycoprotein (28) histidine-rich glycoprotein (29) FST follistatin  (29) FST follistatin (30) glypican 3  (30) glypican 3 (31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15 (33) SPINT2 serine protease inhibitor Kunitz type 2  (33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A  (34) TNFRSFIA tumor necrosis factor receptor superfamily member 1A (35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123 (36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32 (37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13 (38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10 (39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107 (40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38 (41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58 (42) thrombospondin 1  (42) thrombospondin 1 (43) cathepsin S (44) paraoxonase 1 (43) cathepsin S (44) paraoxonase 1 (45) DEFB1 defensin beta 1  (45) DEFB1 defensin beta 1 (46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2 (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) (48) IGFBP1 insulin like growth factor binding protein 1 (48) IGFBP1 insulin like growth factor binding protein 1 (49) sulfatase 2  (49) sulfatase 2 (50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B (51) ADAMTS1 a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1  (51) ADAMTS1 a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1 (52) selenoprotein P， plasma, In  (52) selenoprotein P, plasma, In (53) chondroadherin  (53) chondroadherin (54) decorin  (54) decorin (56) lipopolysaccharide binding protein  (56) lipopolysaccharide binding protein (57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin) (58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1 (59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100 (60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10 (61) chromosome 6 open reading frame 79  (61) chromosome 6 open reading frame 79 (62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80 ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱がヒト血中 HbAlc値と相関している以下の遺伝子からなる群から選 択される少なくとも 1つの遺伝子である請求項 9記載の方法。  A gene whose expression is up- or down-regulated in the liver tissue of a human type 2 diabetic patient is selected from the group consisting of 10. The method of claim 9, wherein the method is at least one gene. (1) poliovirus receptor  (1) poliovirus receptor (2) lectin, galactoside- binding, soluble, 8 (galectin 8)  (2) lectin, galactoside-binding, soluble, 8 (galectin 8) (3) ficolin (collagen/ fibrinogen domain containing) 3 (Hakata antigen)  (3) ficolin (collagen / fibrinogen domain containing) 3 (Hakata antigen) (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (5) midkine (neurite growth-promoting factor 2)  (5) midkine (neurite growth-promoting factor 2) (6) nucleobindin 2 (7) matrix metalloproteinase 11 (stromelysin 3) (6) nucleobindin 2 (7) matrix metalloproteinase 11 (stromelysin 3) (8) glutathione peroxidase 3 (plasma)  (8) glutathione peroxidase 3 (plasma) (9) chromosome 14 open reading frame 2  (9) chromosome 14 open reading frame 2 (10) chromosome 14 open reading frame 4  (10) chromosome 14 open reading frame 4 (11) chromosome 10 open reading frame 10  (11) chromosome 10 open reading frame 10 (12) chromosome 6 open reading frame 74  (12) chromosome 6 open reading frame 74 (13) chromosome 9 open reading frame 83,SNF7 domain containing 2 (SNF7DC2), m RNA  (13) chromosome 9 open reading frame 83, SNF7 domain containing 2 (SNF7DC2), mRNA (14) chromosome 8 open reading frame 1  (14) chromosome 8 open reading frame 1 ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱が HOMA-Rと相関している以下の遺伝子からなる群から選択され る少なくとも 1つの遺伝子である請求項 9記載の方法。  At least one selected from the group consisting of the following genes whose expression is up- or down-regulated in the liver tissue of human type 2 diabetic patients and whose up- or down-expression is correlated with HOMA-R: The method according to claim 9, which is a gene. (42) thrombospondin 丄  (42) thrombospondin 丄 (43) cathepsin S  (43) cathepsin S (44) paraoxonase 1  (44) paraoxonase 1 (45) DEFB1 defensin beta 1  (45) DEFB1 defensin beta 1 (46) MAP2K2 mitogen activated protein kinase kinase 2  (46) MAP2K2 mitogen activated protein kinase kinase 2 (47) phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱が MCRと相関している以下の遺伝子からなる群から選択される少 なくとも 1つの遺伝子である請求項 9記載の方法。  (47) Phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) The method according to claim 9, wherein the gene is at least one gene selected from the group consisting of: (42) thrombospondin 丄  (42) thrombospondin 丄 (43) cathepsin S  (43) cathepsin S (48) IuFBPl insulin Ike growth factor binding protein 1  (48) IuFBPl insulin Ike growth factor binding protein 1 (49) sulfatase 2  (49) sulfatase 2 (50) wingless type MMTV integration site family member 5B  (50) wingless type MMTV integration site family member 5B (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1 (52) selenoprotein P， plasma, In (51) ADAMTSl a disintegrin like and metalloprotease (reprolysin type) with thrombo spondin type 1 motif 1 (52) selenoprotein P, plasma, In (53) chondroadherin  (53) chondroadherin (54) decorin  (54) decorin (56) lipopolysaccharide binding protein  (56) lipopolysaccharide binding protein (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (57) SPARC— like 1 (mast9, hevin)  (57) SPARC— like 1 (mast9, hevin) (58) leucine- rich alpha- 2- glycoprotein 1  (58) leucine- rich alpha- 2- glycoprotein 1 (59) chromosome 14 open reading frame 100  (59) chromosome 14 open reading frame 100 (35) chromosome 14 open reading frame 123 (35) chromosome 14 open reading frame 123 (9) chromosome 14 open reading frame 2 (9) chromosome 14 open reading frame 2 (60) C18orfl0 chromosome 18 open reading frame 10  (60) C18orfl0 chromosome 18 open reading frame 10 (37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13 (61) chromosome 6 open reading frame 79  (61) chromosome 6 open reading frame 79 (62) C6orf80 chromosome 6 open reading frame 80  (62) C6orf80 chromosome 6 open reading frame 80 (38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10 ヒト 2型糖尿病患者の肝臓組織で発現が亢進または減弱して!/、る遺伝子が、発現 の亢進または減弱が BMIと相関している以下の遺伝子からなる群から選択される少な くとも 1つの遺伝子である請求項 9記載の方法。  A gene whose expression is up- or down-regulated in the liver tissue of human type 2 diabetes patients is selected from the group consisting of the following genes whose up- or down-expression is correlated with BMI: The method according to claim 9, which is a gene. (15) bone morpnogenetic protein 2  (15) bone morpnogenetic protein 2 (16) leukocyte cell-derived chemotaxin 2  (16) leukocyte cell-derived chemotaxin 2 (17) adipose differentiation- related protein  (17) adipose differentiation- related protein (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA  (18) Homo sapiens dystonin (DST), transcript variant leA, mRNA (19) serine (or cysteine) proteinase inhibitor, clade F (alpha- 2 antiplasmin, pigment epithelium derived factor), member 1  (19) serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 (20) GNAS complex locus  (20) GNAS complex locus (21) AHSG alpha 2 HS glycoprotein  (21) AHSG alpha 2 HS glycoprotein (22) chemokine (C- C motif) ligand 16  (22) chemokine (C- C motif) ligand 16 (23) afamin (24) LTBP1 latent transforming growth factor beta binding protein 1 (23) afamin (24) LTBP1 latent transforming growth factor beta binding protein 1 (25) complement component 4 binding protein, alpha  (25) complement component 4 binding protein, alpha (26) lysozyme (renal amyloidosis)  (26) lysozyme (renal amyloidosis) (4) MMP24 matrix metalloproteinase 24 (membrane inserted)  (4) MMP24 matrix metalloproteinase 24 (membrane inserted) (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (27) prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (28) histidine- rich glycoprotein (28) histidine-rich glycoprotein (29) FST follistatin  (29) FST follistatin (30) glypican 3  (30) glypican 3 (31) glycosylphosphatidylinositol specific phospholipase Dl  (31) glycosylphosphatidylinositol specific phospholipase Dl (32) GDF15 growth differentiation factor 15  (32) GDF15 growth differentiation factor 15 (33) SPINT2 serine protease inhibitor Kunitz type 2  (33) SPINT2 serine protease inhibitor Kunitz type 2 (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A  (34) TNFRSF1A tumor necrosis factor receptor superfamily member 1A (35) chromosome 14 open reading frame 123  (35) chromosome 14 open reading frame 123 (14) chromosome 8 open reading frame 1 (14) chromosome 8 open reading frame 1 (36) chromosome 14 open reading frame 32  (36) chromosome 14 open reading frame 32 (37) chromosome 5 open reading frame 13  (37) chromosome 5 open reading frame 13 (38) chromosome 9 open reading frame 10  (38) chromosome 9 open reading frame 10 (39) C10orfl07 chromosome 10 open reading frame 107  (39) C10orfl07 chromosome 10 open reading frame 107 (40) Clorf38 chromosome 1 open reading frame 38  (40) Clorf38 chromosome 1 open reading frame 38 (41) C10orf58 chromosome 10 open reading frame 58  (41) C10orf58 chromosome 10 open reading frame 58 少なくとも 5つの遺伝子について被検体における発現を測定する請求項 9〜13の いずれか 1項に記載の方法。  The method according to any one of claims 9 to 13, wherein expression in a subject is measured for at least 5 genes. ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進ま たは減弱している遺伝子の塩基配列からなるヌクレオチドまたはその一部配列を含 むヌクレオチドを固相化したマイクロアレイを用いる請求項 9〜14のいずれ力 1項に 記載の方法。  Compared with normal human liver tissue, nucleotides comprising the nucleotide sequence of a gene whose expression is up-regulated or attenuated in the liver tissue of human type 2 diabetics or a nucleotide containing a partial sequence is immobilized. The method according to any one of claims 9 to 14, wherein the microarray is used. ヒト正常人の肝臓組織と比較して、ヒト 2型糖尿病患者の肝臓組織で発現が亢進ま たは減弱している遺伝子の塩基配列からなるヌクレオチドまたはその一部配列を含 むヌクレオチドをターゲットとしたリアルタイム PCRにより該遺伝子の発現を解析するこ とを含む請求項 9〜 14のいずれか 1項に記載の方法。 Compared with liver tissue of normal humans, it contains nucleotides consisting of the base sequence of a gene whose expression is increased or attenuated in liver tissue of human type 2 diabetic patients, or a partial sequence thereof. 15. The method according to any one of claims 9 to 14, which comprises analyzing the expression of the gene by real-time PCR targeting the nucleotides that are present. 請求項 1〜8のいずれか 1項に記載の 2型糖尿病関連遺伝子および Zまたは肥満 症関連遺伝子の発現を測定するための試薬を含む 2型糖尿病、肥満症、糖尿病性 合併症および動脈硬化症の検出またはリスク判定のためのキット。  A type 2 diabetes, obesity, diabetic complication and arteriosclerosis comprising a reagent for measuring the expression of the type 2 diabetes related gene and Z or obesity related gene according to any one of claims 1 to 8 Kit for detection or risk assessment.
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