US20170357750A1

US20170357750A1 - Method for evaluating drug sensitivity and disease vulnerability by analyzing cyclic amp responsive element binding protein gene

Info

Publication number: US20170357750A1
Application number: US15/645,739
Authority: US
Inventors: Kazutaka Ikeda; Daisuke Nishizawa; Kenichi Fukuda
Original assignee: Tokyo Metropolitan Institute of Medical Science
Current assignee: Tokyo Metropolitan Institute of Medical Science
Priority date: 2011-09-30
Filing date: 2017-07-10
Publication date: 2017-12-14
Also published as: WO2013047912A1; EP2787076B1; US20140243237A1; JPWO2013047912A1; JP6325254B2; EP2787076A4; EP2787076A1; US9727691B2

Abstract

The present invention provides a method for evaluating (predicting, etc.) an individual difference (the tendency of every individual) in terms of drug sensitivity and disease vulnerability, comprising using a gene polymorphism of a cyclic AMP responsive element binding protein gene or the like. The method for evaluating drug sensitivity and the method for evaluating disease vulnerability according to the present invention comprise associating a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism with the drug sensitivity and disease vulnerability of an individual.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of copending application Ser. No. 14/347,500, filed on Mar. 26, 2014, which is the National Phase under 35 U.S.C. §371 of PCT International Application No. PCT/JP2012/076054, filed on Oct. 1, 2012, which claims priority under 35 U.S.C. §119(a) to Patent Application No. 2011-217104, filed in JAPAN on Sep. 30, 2011, all of which are hereby expressly incorporated by reference into the present application.

TECHNICAL FIELD

The present invention relates to a method for evaluating drug sensitivity and disease vulnerability, comprising analyzing a cyclic AMP responsive element binding protein (cAMP responsive element binding protein; CREB) gene. Specifically, the present invention relates to a method for evaluating drug sensitivity and disease vulnerability, comprising associating a gene polymorphism of a CREB gene or a haplotype constituted by the gene polymorphism with the drug sensitivity and disease vulnerability of an individual. More specifically, the present invention relates to a method for evaluating a tendency in the presence or absence of the drug sensitivity and disease vulnerability of an individual, based on the results from the analysis of the above-described gene polymorphism or haplotype.

BACKGROUND ART

Pain is a pathology which is most frequently observed in the medical field, and it is often the case that the pain accompanying a disease is serious for the patient rather than the disease itself. The pain sensation plays an important role in terms of a biological warning system, however, excessive pain would significantly decrease QOL (quality of life) unless it is properly controlled. Recently, the importance of pain control has been recognized, and palliative care including pain therapy has remarkably progressed, and there is a tendency of increasing the frequency and amount of use of various analgesics.
It has been previously known that narcotic analgesics including morphine as a representative example act on a protein known as an “opioid receptor,” so as to cause analgesic action. The opioid receptor includes three types of receptors, a μ-type opioid receptor, a δ-type opioid receptor, and a κ-type opioid receptor, and all of these receptors are related to analgesic action. Since these receptors are Gi/o protein-coupled receptors, they activate a GIRK channel and suppression of a calcium channel through the mediation of a Gi-o protein. In addition, the receptors suppress adenylate cyclase (Non Patent Literature 1: Pierce K. et al., Seven-transmembrane receptors, Nat Rev Mol Cell Biol, (2002) 3: 639-650; Non Patent Literature 2: Bokoch G M. et al., Purification and properties of the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase, J Biol Chem, (1984) 259: 3560-3567). Activation of adenylate cyclase activates cyclic AMP-dependent protein kinase, and it causes activation of a cyclic AMP responsive element binding protein (CREB) through phosphorylation of the serine residue at position 133 of the protein (Non Patent Literature 3: Gonzalez G A. et al., Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133, Cell, (1989) 59: 675-680). The activated CREB binds to a CREB-binding protein acting as an activation cofactor (Non Patent Literature 4: Chrivia J C, et al., Phosphorylated CREB binds specifically to the nuclear protein CBP., Nature, (1993) 365: 855-859), and it binds to the cyclic AMP responsive element of genomic DNA, thereby promoting gene expression (Non Patent Literature 5: Montminy M R. et al., Identification of a cyclic-AMP-responsive element within the rat somatostatin gene., Proc Natl Acad Sci USA, (1986) 83: 6682-6686).

SUMMARY OF INVENTION

The objective of the present invention is to provide a method for evaluating (predicting, etc.) an individual difference (the tendency of every individual) in terms of drug sensitivity and disease vulnerability, comprising using a gene polymorphism of a cyclic AMP responsive element binding protein (CREB) gene or the like.
The present inventors focused on the cyclic AMP responsive element binding protein (CREB) gene and conducted extensive examinations based on conventional findings and clinical data. As a result, the inventors identified several useful gene polymorphisms by analyzing the association of each CREB gene polymorphism with sensitivity to drugs such as analgesics, and with disease vulnerability including pain sensitivity. Thereafter, the inventors found linkage disequilibrium among the thus identified gene polymorphisms, and we also revealed a significant correlation between drug sensitivity and disease vulnerability (more specifically, a change in the required administration amount of an analgesic and a change in the threshold value of pain sensitivity due to a difference in specific CREB gene polymorphisms), thereby accomplishing the present invention.
Thus, the present invention relates to the following:
1. A method for evaluating drug sensitivity, comprising associating a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism with an individual drug sensitivity.
The above-described evaluation method includes a method for evaluating a tendency in the presence or absence of an individual drug sensitivity based on the results from the analysis of the gene polymorphism or the haplotype.
2. The method according to 1 above, comprising the following steps: (1) a step of performing linkage disequilibrium analysis and haplotype analysis on a healthy subject and selecting gene polymorphisms in a linkage disequilibrium block; (2) a step of analyzing the association between the genotypes of the gene polymorphisms and drug sensitivity in a test subject; and (3) a step of using the gene polymorphism that has been significantly associated with drug sensitivity in the test subject for evaluation of the drug sensitivity.
3. A method for evaluating disease vulnerability, comprising associating a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism with an individual disease vulnerability.
The above-described evaluation method includes a method for evaluating a tendency in the presence or absence of an individual disease vulnerability is evaluated based on the results from the analysis of the gene polymorphism or the haplotype.
4. The method according to 3 above, comprising the following steps: (1) a step of performing linkage disequilibrium analysis and haplotype analysis on a healthy subject and selecting gene polymorphisms in a linkage disequilibrium block; (2) a step of analyzing the association between the genotypes of the gene polymorphisms and pain sensitivity; and (3) a step of using the gene polymorphism that has been significantly associated with pain sensitivity in the test subject for evaluation of the disease vulnerability.
5. The method according to 3 or 4 above, wherein the disease vulnerability is pain sensitivity or vulnerability to substance dependence (in particular, vulnerability to drug dependence).
6. The method according to any one of 1 to 5 above, wherein the gene polymorphism is at least one selected from the group consisting of a single nucleotide polymorphism, an insertion polymorphism, a deletion polymorphism, and a nucleotide repeat polymorphism.
7. The method according to any one of 1 to 6 above, wherein the gene polymorphism is at least one selected from among: rs16839837, rs2360969, rs10932200, rs2253206, rs2551640, rs11904814, rs16839883, rs6740584, rs3770704, rs2254137, rs2551645, rs2551946, rs4234080, rs2952768, rs2709386, rs7591784, and rs7594560 of a CREB1 subtype gene (which is a CREB1 gene as a subtype of the CREB gene (the same shall apply hereafter); rs1243872, rs2145925, rs2025126, rs1885373, rs1885374, GA007473, rs2295794, rs4879926, GA007477, rs867194, rs11541908, rs741917, rs7862485, rs2756894, rs2249250, rs2295795, rs877365, rs2737273, rs2295797, rs2295798, rs1534847, rs7873822, rs2737274, rs10972567, rs3763630, rs10814274, rs3750434, rs1570246, GA025684, rs1570248, rs1570249, rs34478611, rs1322045, rs1951432, GA025687, rs10814275, rs10758320, rs4878628, rs10758321, and rs10758322 of a CREB3 subtype gene (which is a CREB3 gene as a subtype of the CREB gene (the same shall apply hereafter); rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs2175738, rs17156579, rs17156603, rs17642145, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs7794304, rs6952227, rs42695, rs1029897, rs4722793, rs10233653, rs6955105, rs17156685, rs17156694, rs17156699, rs177572, rs177573, rs177574, rs177576, rs13437706, rs177580, rs177581, rs12666636, rs177584, rs177585, rs216715, rs10951197, rs160335, rs1008262, rs310353, rs310359, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs41348, rs886816, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs2066979, rs10486591, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955, and rs10228137 of a CREB5 subtype gene (which is a CREB5 gene as a subtype of the CREB gene (the same shall apply hereafter); and rs1153711, rs1153702, rs7583431, rs1153699, rs2302663, rs3845744, rs212349, rs212347, rs12693057, rs1153685, rs212360, rs212361, rs2072538, rs1205399, rs1153676, rs7566401, rs7578569, rs3755490, rs13388308, rs11888507, rs10497434, rs268214, rs166531, rs268228, rs268229, rs268230, rs268231, rs10497435, rs1982235, rs268237, rs13030474, and rs268174 of an ATF2 subtype gene (which is an ATF2 gene as an alias of a CREB2 gene that is a subtype of the CREB gene (the same shall apply hereafter)).
8. The method according to any one of 1 to 7 above, wherein the haplotype is at least one selected from the following table.
It is to be noted that haplotypes constituted by a combination of any given number and type of various gene polymorphisms according to 7 above can also be selected as haplotypes used in the evaluation method and the like of the present invention.

TABLE 1

Gene name CREB1

	Linkage disequilibrium block No.
	1
	Gene polymorphism name ()

1

2

3

12

13

17

Haplotype No.

Tag

4

5

6

7

8

9

10

11

Tag

14

15

16

Tag

H1

C

A

G

T

C

A

T

H2

C

A

G

A

T

A

C

T

A

T

C

T

G

T

H3

C

T

C

A

G

A

T

C

A

T

H4

T

C

A

G

A

T

A

C

T

A

T

C

A

T

G

C

H5

C

A

G

A

T

A

C

T

A

T

A

C

A

T

H6

T

C

A

G

A

T

A

C

T

A

T

C

T

G

T

H7

T

C

A

G

A

T

A

C

T

A

T

C

A

T

G

T

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

() 1~17: (in this order)rs16839837, rs2360969, rs10932200, rs2253206, rs2551640, rs11904814, rs16839883, rs6740584, rs3770704, rs2254137, rs2551645, rs2551946, rs4234080, rs2952768, rs2709386, rs7591784 and rs7594560

TABLE 2

Gene name CREB3

	Linkage disequilibrium block No.
	1
	Gene polymorphism name ()

1

2

3

9

11

Haplotype No.

Tag

4

5

6

7

8

Tag

10

Tag

12

13

14

15

16

17

18

19

20

21

22

23

24

H1

G

C

G

T

A

T

C

T

C

G

C

T

G

T

G

T

C

A

G

A

H2

G

C

G

T

A

T

C

G

C

T

G

T

G

T

C

A

G

A

H3

T

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

H4

T

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

H5

T

C

G

C

T

C

A

G

C

A

T

C

A

T

A

C

H6

T

G

T

A

T

C

T

C

T

G

A

T

G

C

G

T

C

A

G

C

H7

T

C

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

	Linkage disequilibrium block No.
	2
	Gene polymorphism name ()

25

26

29

36

38

39

Haplotype No.

Tag

27

28

Tag

30

31

32

33

34

35

Tag

37

Tag

40

H8

C

G

C

G

A

T

A

C

G

C

H9

C

T

A

T

G

T

A

G

A

G

A

T

C

A

T

H10

T

C

G

C

T

G

A

G

A

T

C

G

C

H11

C

T

A

G

T

A

G

A

G

A

T

C

G

T

H12

C

G

C

T

G

A

G

A

T

C

G

C

H13

C

T

A

T

G

T

A

G

A

G

A

G

T

C

A

T

H14

C

G

C

G

A

T

A

C

T

G

C

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

() 1~40: (in this order)rs1243872, rs2145925, rs2025126, rs1885373, rs1885374, GA007473, rs2295794, rs4879926, GA007477, rs867194, rs11541908, rs741917, rs7862485, rs2756894, rs2249250, rs2295795, rs877365, rs2737273, rs2295797, rs2295798, rs1534847, rs7873822, rs2737274, rs10972567, rs3763630, rs10814274, rs3750434, rs1570246, GA025684, rs1570248, rs1570249, rs34478611, rs1322045, rs1951432, GA025687, rs10814275, rs10758320, rs4878628, rs10758321 and rs10758322

TABLE 3

Gene name CREB5

Linkage disequilibrium block No.

1

5

6

9

10

Gene polymorphism name ()

1

2

5

6

7

8

9

11

13

14

15

16

17

18

Haplotype (H) No.

Tag

3

4

Tag

H No.

Tag

10

Tag

12

H No.

Tag

H No.

Tag

H No.

Tag

H1

C

T

A

G

C

H6

C

G

C

A

G

H11

T

G

H14

T

A

H17

G

T

H2

G

C

G

A

G

T

H7

C

A

C

G

H12

C

H15

C

A

H18

G

H3

G

C

G

A

T

H8

C

G

C

G

H13

C

G

H16

C

G

H19

A

G

H4

C

G

A

G

T

H9

T

A

C

G

A

. . .

H5

G

C

G

A

G

C

H10

C

A

C

G

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

Linkage disequilibrium block No.

11

12

13

15

16

Gene polymorphism name ()

19

20

21

22

23

24

25

26

27

28

29

30

31

Haplotype (H) No.

Tag

H No.

Tag

H No.

Tag

H No.

Tag

H No.

Tag

32

33

H20

T

A

H24

G

C

H28

A

H31

G

T

C

H35

C

G

C

H21

T

G

H25

G

C

T

H29

G

A

H32

A

C

T

H36

T

C

A

G

C

H22

C

G

H26

A

C

T

H30

G

H33

A

T

H37

C

A

G

C

H23

C

A

H27

G

T

. . .

H34

A

T

C

H38

T

C

A

T

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

Linkage disequilibrium block No.

25

26

27

32

Gene polymorphism name ()

34

35

36

37

38

39

40

42

43

44

45

47

50

Haplotype (H) No.

Tag

H No.

Tag

H No.

Tag

41

H No.

Tag

46

Tag

48

49

Tag

H39

T

A

H42

G

A

G

H47

A

H49

A

T

C

G

T

G

T

H40

C

G

H43

G

A

H48

G

H50

A

T

C

G

T

G

T

C

H41

T

G

H44

A

G

A

. . .

H51

A

T

A

C

G

T

C

. . .

H45

G

A

G

A

H52

C

T

A

C

T

C

T

C

H46

G

A

H53

C

T

A

C

T

G

C

T

C

. . .

H54

A

C

G

T

G

T

H55

A

T

A

G

T

G

T

C

	Haplotypes which are estimated to occur at a frequency of less than 1%

Linkage disequilibrium block No.

33

35

40

Gene polymorphism name ()

51

52

53

54

57

58

59

Haplotype (H) No.

Tag

55

56

H No.

Tag

H No.

Tag

60

61

62

63

64

H56

T

G

C

A

H61

C

T

H64

G

C

T

C

H57

T

A

T

G

C

H62

T

C

H65

A

T

C

A

H58

G

A

C

G

C

H63

T

. . .

H59

T

A

G

C

A

. . .

H60

T

A

C

G

C

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

() 1~64: (in this order) rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs6952227, rs42695, rs1029897, rs10233653, rs6955105, rs17156699, rs177572, rs177573, rs177580, rs177581, rs12666636, rs177584, rs177585, rs1008262, rs310353, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955 and rs10228137

TABLE 4

Gene name ATF2

Linkage disequilibrium block No.

1

2

Gene polymorphism name ()

1

2

3

4

5

Haplotype No.

Tag

6

7

8

9

10

11

12

13

14

15

16

H1

T

C

A

G

C

A

G

C

G

T

C

A

H2

G

C

A

G

T

G

A

T

A

C

A

C

G

H3

T

C

G

A

T

G

A

T

A

C

A

C

G

H4

T

A

G

A

T

G

A

T

A

C

A

C

G

T

G

A

T

A

C

A

C

G

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

	Linkage disequilibrium block No.
	2
	Gene polymorphism name ()

21

28

Haplotype No.

17

18

19

20

Tag

22

23

24

25

26

27

Tag

29

30

31

H5

C

T

C

G

A

C

A

C

T

G

T

H6

T

C

T

A

G

A

C

T

G

C

G

C

H7

T

C

T

A

G

A

C

T

C

G

C

H8

T

C

T

A

G

A

C

T

G

C

G

C

H9

T

C

T

C

G

A

C

A

C

T

G

T

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

() 1~31: (in this order)rs1153711, rs1153702, rs7583431, rs2302663, rs3845744, s212349, rs212347, rs12693057, rs1153685, rs212360, rs212361, rs2072538, rs1205399, rs1153676, rs7566401, rs7578569, rs3755490, rs13388308, rs11888507, rs10497434, rs268214, rs166531, rs268228, rs268229, rs268230, rs268231, rs10497435, rs1982235, rs268237, rs13030474 and rs268174

9. A method for determining the type, amount, and/or frequency of administration of a drug to be administered to an individual, comprising using the result from the evaluation by the method according to any one of 1 to 8 above as an index.
10. A method for predicting a side effect of a drug to be administered to an individual, comprising using the result from the evaluation by the method according to any one of 1 to 8 above as an index.
11. The method according to any one of 1, 2, 5, 9, and 10 above, wherein the drug is an opioid receptor function modulator and/or a cyclic AMP responsive element binding protein function modulator.
12. The method according to 11 above, wherein the opioid receptor function modulator is at least one selected from the group consisting of methamphetamine, methylenedioxymethamphetamine, amphetamine, dextroamphetamine, dopamine, morphine, DAMGO, codeine, methadone, carfentanil, fentanyl, heroin, cocaine, naloxone, naltrexone, nalorphine, levallorphan, pentazocine, pethidine, buprenorphine, oxycodone, hydrocodone, levorphanol, etorphine, dihydroetorphine, hydromorphone, oxymorphone, tramadol, diclofenac, indomethacin, ethanol, methanol, diethyl ether, propanol, butanol, flupirtine, laughing gas, F3 (1-chloro-1,2,2-trifluorocyclobutane), halothane, estradiol, dithiothreitol, thioridazine, pimozide, fluoxetine, paroxetine, desipramine, imipramine, clomipramine, tetramide, isoflurane, ginsenoside, ifenprodil, bupivacaine, tertiapin, clozapine, haloperidol, SCH23390, and cocaine; and the cyclic AMP responsive element binding protein function modulator is at least one selected from the group consisting of phosphodiesterase 4 (PDE4), calcineurin, protein kinase A, protein kinase C, p90 ribosome S6 kinase 1 (RSK1), calmodulin kinase, glycogen synthase kinase 3β, and CREB-regulated transcription coactivator 1 (CRTC1).
13. The method according to any one of 1 to 12 above, comprising using an oligonucleotide consisting of a nucleotide sequence of at least 10 nucleotides comprising the 51^stnucleotide of the nucleotide sequence represented by any one of SEQ ID NOS: 1 to 172, or a complementary nucleotide sequence thereto, which can specifically hybridize to a DNA fragment comprising a gene polymorphism of a cyclic AMP responsive element binding protein gene.
14. The method according to 13 above, wherein the oligonucleotide spans a length of 10 to 150 nucleotides.
15. The method according to 13 or 14 above, wherein the oligonucleotide is selected from the group consisting of the nucleotide sequence represented by any one of SEQ ID NOS: 1 to 38 and a complementary nucleotide sequence thereto.
16. A gene polymorphism marker for evaluating a tendency in the presence or absence of an individual drug sensitivity, comprising a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism. Examples of the drug applied herein are the same as those described in 11 and 12 above.
17. A gene polymorphism marker for evaluating a tendency in the presence or absence of an individual disease vulnerability, comprising a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism.
According to the present invention, there can be provided: a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism, which is capable of evaluating an individual difference in terms of drug sensitivity and disease vulnerability; a method for evaluating drug sensitivity and disease vulnerability, comprising using the gene polymorphism or the haplotype; etc. According to this evaluation method, it becomes possible to readily know or predict a proper prescribed amount, a proper prescribed schedule, and the like, associated with a narcotic drug such as morphine, and hence the method is extremely useful for personalized pain therapy, drug dependence therapy and the like.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a schematic view showing gene polymorphisms identified regarding a CREB1 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of D′ that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of D′ between rs16839837 and rs2551640 is 0.91.

FIG. 2 is a schematic view showing gene polymorphisms identified regarding a CREB1 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of r²that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of r²between rs16839837 and rs2551640 is 0.12.

FIG. 3 is a schematic view showing gene polymorphisms identified regarding a CREB3 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of D′ that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of D′ between rs1243872 and rs2025126 is 0.97.

FIG. 4 is a schematic view showing gene polymorphisms identified regarding a CREB3 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of r²that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of r²between rs1243872 and rs2025126 is 0.67.

FIG. 5 is a schematic view showing gene polymorphisms identified regarding a CREB5 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of D′ that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of D′ between rs4722778 and rs177479 is 0.93.

FIG. 6 is a schematic view showing gene polymorphisms identified regarding a CREB5 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of r²that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of r²between rs4722778 and rs177479 is 0.85.

FIG. 7 is a schematic view showing gene polymorphisms identified regarding a ATF2 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of D′ that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of D′ between rs1153711 and rs1153700 is 0.86.

FIG. 8 is a schematic view showing gene polymorphisms identified regarding a ATF2 subtype gene and a linkage disequilibrium found among them. In the figure, high-color squares indicate SNPs showing strong linkage with each other. In addition, the square found at the intersection of squares continued from each SNP to the lower left direction or the lower right direction indicates the calculation value (percentage) of r²that is an index of the linkage disequilibrium of a SNP and another SNP. For example, the calculation value of r²between rs1153711 and rs1153700 is 0.14.

FIG. 9 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs10932200) of a CREB1 subtype gene on the required administration amount (μg/kg) of analgesic in 24 hours after surgery, in all patients who were administered with analgesic in the surgery (orthognathic surgery).

FIG. 10 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs10932200) of a CREB1 subtype gene on the measurement results (sec) of pain sensitivity before surgery, in all patients who were administered with analgesic in the surgery (orthognathic surgery).

FIG. 11 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs10932200) of a CREB1 subtype gene on the measurement results (mm) of VAS (the intensity of pain on a visual analogue scale) 24 hours after surgery, in male patients who were administered with analgesic in the surgery (orthognathic surgery).

FIG. 12 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs7583431) of an ATF2 subtype gene on the measurement results (sec) of a difference in threshold of pain perception latency before surgery, in all patients who were administered with analgesic in the surgery (orthognathic surgery).

FIG. 13 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs2952768) of a CREB1 subtype gene on the required administration amount (μg/kg) of analgesic in 24 hours after surgery, in all patients who were administered with analgesic in the surgery (orthognathic surgery).

FIG. 14 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs2952768) of a CREB1 subtype gene on the required administration amount (μg/kg) of analgesic in 24 hours after surgery, in all patients who were administered with analgesic in the surgery (abdominal surgery).

FIG. 15 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs2952768) of a CREB1 subtype gene on the reward dependence (RD) score (average) of Temperament and Character Inventory (TCI) in all healthy subjects.

FIG. 16 is a graph showing the effect (overall average±standard deviation) of a polymorphism (rs2952768) of a CREB1 subtype gene on the expression level of a CREB1 subtype gene in postmortem brain tissue donors at the Stanley Foundation Brain Bank.

DESCRIPTION OF EMBODIMENT

Hereinafter, the present invention will be described in detail. However, the scope of the invention is not limited to the description, and changes and modifications can be made therein without departing from the spirit of the invention other than the following examples.
It is to be noted that the present specification includes all of the contents as disclosed in the specification and/or drawings of Japanese Patent Application No. 2011-217104 (filed on Sep. 30, 2011), which is a priority document of the present application. Moreover, all publications cited in the present specification, including prior art documents and patent literatures such as patent laid-open applications or patent publications, are incorporated herein by reference in their entirety.

1. Outline of the Present Invention

(1) Cyclic AMP Responsive Element Binding Protein

Cyclic AMP responsive element binding protein (cAMP responsive element binding protein; CREB) is a protein, which includes a signaling system downstream of G protein-coupled receptor such as a μ-type opioid receptor, is activated depending on an intracellular cyclic AMP concentration, and binds to a cyclic AMP responsive element (cAMP responsive element) of nuclear genomic DNA, so that it is associated with regulation of gene expression. The cyclic AMP responsive element binding protein is present in various tissues and/or organs such as cardiac muscle, various types of smooth muscle, fat cells, skeletal muscle and brain, and it is also associated with neurogenesis, memory, dependence, etc., through the expression of various genes.
Narcotic analgesics including morphine as a representative example act on a protein known as an “opioid receptor,” so as to cause analgesic action. The opioid receptor includes three types of receptors, a μ-type opioid receptor, a δ-type opioid receptor, and a κ-type opioid receptor, and all of these receptors are related to analgesic action. Since these receptors are Gi/o protein-coupled receptors, they activate a GIRK channel and suppression of a calcium channel through the mediation of a Gi-o protein. In addition, the receptors suppress adenylate cyclase. Activation of adenylate cyclase activates cyclic AMP-dependent protein kinase, and it causes activation of a cyclic AMP responsive element binding protein (CREB) through phosphorylation of the serine residue at position 133 of the protein. The activated CREB binds to a CREB-binding protein acting as an activation cofactor, and it binds to the cyclic AMP responsive element of genomic DNA, thereby promoting gene expression.
Herein, the cyclic AMP responsive element binding protein will be described. The cyclic AMP responsive element binding protein is distributed in various tissues and/or organs including brain and heart, and plays an important role for neurogenesis, memory, the expression of dependence, etc., through the expression of various types of genes. The cyclic AMP responsive element binding protein has a domain structure comprising a Q-rich domain, a kinase-inducible domain (KID), a basic region/leucine zipper (bZIP), etc. The protein binds to genomic DNA.
The cyclic AMP responsive element binding protein functions as a dimer formed from two subunits. The type of a receptor subtype is broadly classified into CREB1, CREB3, CREB5, ATF2 and the like, and their homologs have also been known. These subtypes are expressed in various tissues and/or organs including brain and heart.

(2) Gene Polymorphism

The present inventors identified gene polymorphisms (such as SNP) of the subtypes CREB1, CREB3, CREB5 and ATF2 (CREB2) capable of constituting a cyclic AMP responsive element binding protein in healthy subjects (FIGS. 1 to 8). Further, a linkage disequilibrium analysis was carried out as needed, and a block exhibiting a strong linkage disequilibrium (a haplotype block) was identified.
Here, the linkage equilibrium means a case where the relationship between two gene polymorphisms on the chromosome is independent, and the linkage disequilibrium means a case where a gene polymorphism is linked to the other gene polymorphism thereby deviating from the equilibrium situation according to Mendel's law of independence. Further, the haplotype means a genetic structure of such as genes or gene polymorphisms located in the vicinity of each other in one allele of a set of alleles (a gene derived from one of the parents).
Gene polymorphisms or the like located in the vicinity on a genome are inherited in a haplotype block. In other words, a haplotype also refers to a combination of the arrangement of the same gene in this haplotype block.
In the case where several gene polymorphisms appear in association with a certain phenotype in the cyclic AMP responsive element binding protein genes, even if not all the respective gene polymorphisms are typed, by analyzing several gene polymorphisms constituting a haplotype, a relationship between the genotype and the phenotype of a patient can be elucidated.
The present inventors analyzed the cyclic AMP responsive element binding protein CREB1 subtype gene, and as a result, they found 4 and 6 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, and also found 7 gene polymorphisms in the intron region (see Table 5).
In addition, with regard to the CREB3 subtype gene, the inventors found 25 and 14 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, and also found 1 gene polymorphism in the intron region (see Table 5).
Moreover, with regard to the CREB5 subtype gene, the inventors found 9 and 5 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, and also found 241 gene polymorphisms in the intron region and 2 gene polymorphisms (rs2190305 and rs3735566) in the noncoding region of exon (see Table 6).
Furthermore, with regard to the ATF2 subtype gene, the inventors found 11 and 6 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, and also found 16 gene polymorphisms in the intron region and 1 gene polymorphism (rs10497434) in the noncoding region of exon (see Table 5).
According to the present invention, by analyzing gene polymorphisms of the cyclic AMP responsive element binding protein gene or haplotypes constituted by the gene polymorphisms, an individual difference in phenotypes regarding sensitivity to drugs (i.e. drug sensitivity), such as the effectiveness of a drug, the side effects of a drug, and an effective duration of a drug (e.g. the required number of administration of analgesic, the total amount of analgesic, prolongation of stimulant-induced psychosis, etc.), and in phenotypes regarding the development of a disease including pain sensitivity, vulnerability to substance dependence (in particular, vulnerability to drug dependence), etc., can be easily evaluated. The results of evaluating drug sensitivity and disease vulnerability can be important information for determining the administration number, amount, type or the like of drugs to be administered to an individual, and predicting side effects. Therefore, the present invention provides a method for evaluating drug sensitivity and disease vulnerability based on the results obtained by analyzing gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms, and specifically, a method for evaluating (specifically, knowing in advance or predicting) a tendency in the presence or absence of drug sensitivity and disease vulnerability (more specifically, the presence or absence of the genetic factor thereof) in an individual (an individual person). In addition, the present invention also provides a gene polymorphism marker used for evaluating (specifically, knowing in advance or predicting) a tendency in the presence or absence of drug sensitivity and disease vulnerability (more specifically, the presence or absence of the genetic factor thereof) in an individual (an individual person), wherein the gene polymorphism marker comprises gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms.
In particular, because morphine, a stimulant or the like may cause a big social problem depending on the usage, it is important to know in advance an appropriate amount of drugs to be administered to an individual before administering the drugs. Therefore, the present invention is extremely useful for personalized pain therapy or drug dependence therapy.
Moreover, according to the present invention, by analyzing gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms, an individual difference in terms of dependence-prone personality can be easily evaluated. The results of evaluating dependence-prone personality can be information for determining whether reward dependence is high or low about the personality of an individual. Therefore, the present invention provides a method for evaluating dependence-prone personality based on the results obtained by analyzing gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms, and specifically, a method for evaluating (specifically, knowing in advance or predicting) a tendency in the presence or absence of dependence-prone personality (high reward dependence or low reward dependence) (more specifically, the presence or absence of the genetic factor thereof) in an individual (an individual person). In addition, the present invention also provides a gene polymorphism marker used for evaluating (specifically, knowing in advance or predicting) a tendency in the presence or absence of dependence-prone personality (high reward dependence or low reward dependence) (more specifically, the presence or absence of the genetic factor thereof) in an individual (an individual person), wherein the gene polymorphism marker comprises gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms. Herein, with regard to the types of a gene polymorphism and a haplotype constituted by the gene polymorphism that can be applied to the above-described evaluation of dependence-prone personality, the same gene polymorphisms and haplotypes as those that can be applied to the above-described method for evaluating drug sensitivity and disease vulnerability can be used. Moreover, with regard to oligonucleotides used for the above described evaluation of dependence-prone personality as well, the same oligonucleotides as those used for the above-described method for evaluating drug sensitivity and disease vulnerability can be used.
Furthermore, according to the present invention, by analyzing gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms, an individual difference in terms of the expression level of a CREB1 gene can be easily evaluated. The results of evaluating the expression level of a CREB1 gene can be information for predicting whether the expression level of a CREB1 gene in an individual is high or low. Therefore, the present invention provides a method for evaluating the high or low expression level of a CREB1 gene based on the results obtained by analyzing gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms, and specifically, a method for evaluating (specifically, knowing in advance or predicting) a tendency in the presence or absence of the high or low expression level of a CREB1 gene (more specifically, the presence or absence of the genetic factor thereof) in an individual (an individual person). In addition, the present invention also provides a gene polymorphism marker used for evaluating (specifically, knowing in advance or predicting) a tendency in the presence or absence of the high or low expression level of a CREB1 gene (more specifically, the presence or absence of the genetic factor thereof) in an individual (an individual person), wherein the gene polymorphism marker comprises gene polymorphisms of the cyclic AMP responsive element binding protein or haplotypes constituted by the gene polymorphisms. Herein, with regard to the types of a gene polymorphism and a haplotype constituted by the gene polymorphism that can be applied to the above-described evaluation of the high or low expression level of a CREB1 gene, the same gene polymorphisms and haplotypes as those that can be applied to the above-described method for evaluating drug sensitivity and disease vulnerability can be used. Moreover, with regard to oligonucleotides used for the above described evaluation of the high or low expression level of a CREB1 gene as well, the same oligonucleotides as those used for the above-described method for evaluating drug sensitivity and disease vulnerability can be used.

2. Gene Polymorphism of Cyclic AMP Responsive Element Binding Protein Gene

The human cyclic AMP responsive element binding protein gene polymorphisms of the present invention mainly include single nucleotide polymorphisms (hereinafter also referred to as “SNP”), however it is not limited to this, and insertion polymorphisms, deletion polymorphisms, and nucleotide repeat polymorphisms can also be included.
The single nucleotide polymorphism [SNP (SNPs)] means a gene polymorphism caused by substitution of a specific one nucleotide of a gene with another nucleotide. The insertion/deletion polymorphism means a gene polymorphism caused by deletion/insertion of one or more nucleotides.
Further, the nucleotide repeat polymorphism means a gene polymorphism caused by a difference in the number of repeats of nucleotide sequence. The nucleotide repeat polymorphism is divided into a microsatellite polymorphism (the number of nucleotides: about 2 to 4 nucleotides) and a VNTR (variable number of tandem repeat) polymorphism (repeated nucleotides: several to several tens of nucleotides) according to the difference in the number of repeated nucleotides, and the number of repeats varies depending on individuals.
The information of human cyclic AMP responsive element binding protein gene polymorphisms (SNPs in the CREB1 subtype gene, the CREB3 subtype gene, the CREB5 subtype gene and the ATF2 subtype gene observed on the genome of Japanese healthy subjects) elucidated by the present invention is shown in Table 5 and Table 6. The gene polymorphisms shown in Table 5 and Table 6 include the cyclic AMP responsive element binding protein gene polymorphisms of the present invention.

TABLE 5

CREB1 gene polymorphism	CREB3 gene polymorphism	ATF2 gene polymorphism

	Gene	Major		Gene	Major		Gene	Major
	polymorphism	allele:minor		polymorphism	allele:minor		polymorphism	allele:minor
Position	name	allele	Position	name	allele	Position	name	allele

5′ Flanking	rs16839837	C:T	5′ Flanking	rs1243872	G:T	5′ Flanking	rs268174	C:T
region	rs2360969	C:T	region	rs2145925	C:T	region	rs13030474	G:T
	rs10932200	A:C		rs2025126	G:A		rs268237	C:T
	rs2253206	G:A		rs1885373	T:C		rs1982235	T:G
Intron	rs2551640	A:G		rs1885374	A:C		rs10497435	T:C
	rs11904814	T:G		GA007473	T:C		rs268231	C:A
	rs16839883	A:G		rs2295794	T:C		rs268230	A:C
	rs6740584	C:T		rs4879926	T:C		rs268229	G:A
	rs3770704	T:C		GA007477	C:T		rs268228	A:G
	rs2254137	A:C		rs867194	T:C		rs166531	T:C
	rs2551645	T:C		rs11541908	C:T		rs268214	T:C
3′ Flanking	rs2551946	C:A		rs741917	C:T	Exon	rs10497434	T:C
region	rs4234080	C:A		rs7862485	G:A	Intron	rs11888507	C:T
	rs2952768	T:C		rs2756894	C:A		rs13388308	C:T
	rs2709386	G:A		rs2249250	T:G		rs3755490	T:C
	rs7591784	G:A		rs2295795	G:A		rs7578569	G:A
	rs7594560	T:C		rs877365	T:C		rs7566401	C:A
				rs2737273	G:A		rs1153676	C:A
				rs2295797	T:C		rs1205399	A:C
				rs2295798	C:T		rs2072538	C:T
				rs1534847	A:G		rs212361	A:G
				rs7873822	G:T		rs212360	T:C
				rs2737274	G:A		rs1153685	A:G
				rs10972567	A:C		rs12693057	G:A
				rs3763630	C:T		rs212347	T:C
			Intron	rs10814274	C:T		rs212349	T:C
			3′ Flanking	rs3750434	G:A		rs3845744	A:G
			region	rs1570246	G:T		rs2302663	G:A
				GA025684	G:C	3′ Flanking	rs35507277	T:G
				rs1570248	T:C	region	rs1153699	G:T
				rs1570249	G:A		rs1153700	C:G
				rs34478611	G:A		rs7583431	A:C
				rs1322045	A:G		rs1153702	T:C
				rs1951432	G:A		rs1153711	T:G
				GA025687	A:T
				rs10814275	A:G
				rs10758320	T:C
				rs4878628	C:T
				rs10758321	G:A
				rs10758322	C:T

TABLE 6

CREB5 gene polymorphism

	Gene	Major
	polymorphism	allele:minor
Position	name	allele

5′ Flanking	rs4722778	C:G
region	rs177479	T:C
	rs177480	A:G
	rs11981754	A:G
	rs177486	G:A
	rs177498	C:T
	rs849322	A:G
	rs177505	T:G
	rs2175738	G:A
Intron	rs4719932	A:C
	rs10258745	C:T
	rs1013900	G:T
	rs6955393	G:A
	rs6953880	A:G
	rs17156573	T:C
	rs6960209	C:T
	rs17156577	T:C
	rs7811922	A:C
	rs6973453	T:C
	rs17156579	C:T
	rs1073298	T:C
	rs6961801	C:T
	rs6977728	C:A
	rs6978238	C:T
	rs13230543	C:A
	rs12673465	A:G
	rs10251129	T:C
	rs2391656	T:C
	rs6971345	A:G
	rs17156603	A:G
	rs7806362	C:A
	rs17642145	T:C
	rs10229500	C:T
	rs10243659	C:A
	rs4722785	G:A
	rs16874503	C:T
	rs11772815	G:A
	rs6958133	G:A
	rs16874525	C:T
	rs17715174	G:C
	rs10242868	T:G
	rs12700884	G:A
	rs17156635	G:A
	rs10239606	C:T
	rs16874528	G:A
	rs7799687	C:A
	rs714218	G:A
	rs1860759	A:G
	rs997908	G:A
	rs12112050	C:T
	rs2191827	A:G
	rs4498447	T:C
	rs10254657	G:A
	rs6953524	C:T
	rs10239810	A:G
	rs17156649	G:A
	rs1811248	T:G
	rs887623	T:C
	rs740988	A:G
	rs7794304	T:C
	rs42694	A:G
	rs6952227	G:A
	rs42695	C:T
	rs1029897	T:C
	rs42699	A:C
	rs4722793	C:A
	rs735101	T:C
	rs10233653	G:A
	rs6955105	G:A
	rs2286841	C:A
	rs979915	C:T
	rs7794347	C:T
	rs16874562	G:T
	rs17156685	A:G
	rs174024	C:T
	rs6949786	G:A
	rs7793437	A:G
	rs3757677	T:C
	rs6462085	T:G
	rs17717216	T:C
	rs17156694	G:A
	rs17156699	A:G
	rs177572	T:C
	rs177573	T:C
	rs6977204	A:G
	rs177574	A:G
	rs177576	T:C
	rs177578	G:A
	rs13437706	C:T
	rs177580	C:T
	rs177581	C:T
	rs12666636	C:A
	rs177584	G:A
	rs177585	C:T
	rs177588	G:A
	rs6462088	G:A
	rs7796539	C:T
	rs1859020	A:G
	rs1011384	A:G
	rs6462090	G:T
	rs12671247	T:C
	rs217508	T:C
	rs4719936	G:T
	rs217509	G:T
	rs217510	T:C
	rs17718257	G:A
	rs149591	C:A
	rs1910553	C:A
	rs217517	G:A
	rs217519	G:A
	rs2391668	T:G
	rs4722804	G:T
	rs618776	A:G
	rs217503	C:T
	rs217513	C:T
	rs65264	C:T
	rs441355	G:T
	rs2391670	C:T
	rs2391671	A:G
	rs216708	A:G
	rs11980665	C:T
	rs11980669	C:T
	rs11984308	T:C
	rs160346	G:A
	rs150607	A:G
	rs177594	G:A
	rs6969064	A:G
	rs150610	A:G
	rs216715	T:C
	rs10951197	T:C
	rs12539927	A:G
	rs216720	A:G
	rs17156823	G:A
	rs2078980	G:A
	rs216730	T:G
	rs13228899	G:T
	rs160335	G:A
	rs10951200	G:A
	rs10486588	G:A
	rs216735	G:A
	rs216737	C:T
	rs216743	G:A
	rs216744	A:G
	rs216747	C:T
	rs1976489	A:G
	rs150613	C:T
	rs17156878	G:A
	rs767834	C:G
	rs4722820	G:A
	rs160337	C:A
	rs160338	G:A
	rs1008262	T:C
	rs310353	G:A
	rs310359	T:C
	rs310361	C:T
	rs13233942	A:G
	rs310338	T:C
	rs41273	G:A
	rs1637457	A:G
	rs17156919	G:A
	rs41276	A:G
	rs160375	A:G
	rs917275	A:G
	rs160342	A:G
	rs160343	T:C
	rs41295	C:T
	rs160357	A:G
	rs41298	G:A
	rs41305	G:A
	rs41307	C:T
	rs10228740	A:G
	rs3888613	G:A
	rs41320	C:T
	rs41321	A:G
	rs41322	A:G
	rs7780656	G:T
	rs41327	A:G
	rs42322	T:C
	rs41333	A:G
	rs9655280	A:G
	rs9655281	G:A
	rs4719945	A:G
	rs6945988	A:G
	rs10258405	T:G
	rs10243376	G:A
	rs41334	T:C
	rs10245004	C:T
	rs41339	G:T
	rs982947	C:T
	rs982950	A:G
	rs16874653	A:G
	rs41346	G:T
	rs41348	A:G
	rs9969149	C:T
	rs6968464	G:A
	rs886816	G:A
	rs757980	A:G
	rs41351	G:A
	rs9691873	A:C
	rs17157048	A:C
	rs6462098	T:C
	rs10951201	C:A
	rs13311248	G:C
	rs12540480	T:C
	rs10265166	G:T
	rs7798774	T:C
	rs7799246	T:C
	rs6972081	T:C
	rs7777929	T:C
	rs12533079	T:G
	rs7806547	G:A
	rs6462100	G:A
	rs6979352	C:T
	rs6950574	A:G
	rs4722835	A:C
	rs9648352	A:G
	rs879593	A:C
	rs879591	G:T
	rs2299110	C:T
	rs2237349	C:T
	rs2066979	T:C
	rs10486589	A:G
	rs10486591	G:A
	rs6462103	C:T
	rs721993	C:T
	rs2237351	T:C
	rs740315	G:A
	rs2237353	A:C
	rs2073537	T:C
	rs4722844	G:T
	rs17730621	C:T
	rs2282907	G:A
	rs10238623	G:A
	rs2299116	C:A
	rs2299117	T:C
	rs2237355	A:G
	rs2237360	T:G
	rs2237361	T:C
	rs2237362	T:C
	rs7791555	G:T
	rs2237364	A:G
	rs2282909	T:G
	rs2282910	C:T
	rs2282911	T:C
	rs1544470	A:G
	rs1964240	A:C
	rs17669844	T:C
	rs886750	A:G
	rs12531253	G:A
	rs10951205	A:G
Exon	rs2190305	A:G
	rs3735566	G:A
3′ Flanking	rs11975539	G:A
region	rs6462107	C:T
	rs2190306	T:C
	rs4719955	T:C
	rs10228137	C:A

In Table 5 and Table 6, “CREB1” (italic form) indicates a CREB1 subtype gene (a CREB1 gene as a subtype of the cyclic AMP responsive element binding protein (CREB) gene), “CREB3” (italic form) indicates a CREB3 subtype gene (a CREB3 gene as a subtype of the CREB gene), “CREB5” (italic form) indicates a CREB5 subtype gene (a CREB5 gene as a subtype of the CREB gene), and “ATF2” (italic form) indicates an ATF2 subtype gene (an ATF2 gene that is an alias of a CREB2 gene as a subtype of the CREB gene).
“Position” means a position on the genome of a cyclic AMP responsive element binding protein gene, and indicates a 5′ flanking region, a 3′ flanking region, intron, and exon.
“Gene polymorphism name” is the name of SNP at a position on the genome, and it has been registered in the dbSNP database (which is accessible from the NCBI website dbSNP Short Genetic Variations) (the same shall apply in the present specification). Basically, the ID “rs” is given before four or more digit numbers, so that the type of SNP can be identified.
“Major allele” indicates an allele occurring in the majority of the genomes of Japanese healthy subjects, and “minor allele” indicates an allele occurring in the minority of the genomes of Japanese healthy subjects.
In the present invention, a method of obtaining gene polymorphism information is as follows, for example.
(1) Genomic DNA is purified from a blood specimen collected from a human using the phenol method and the like. At this time, a commercially available genomic DNA extraction kit such as GFX Genomic Blood DNA Purification Kit (manufactured by GE Healthcare Bio-Sciences KK) or a device may be used.
(2) Then, the obtained genomic DNA is dissolved in TF buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) and the concentration of the obtained solution is adjusted to 100 ng/μl.
(3) Total genome genotyping is carried out by an infinium assay II method or the like, using iScan system (manufactured by Illumina, San Diego, Calif.), in accordance with the protocols provided by the manufacturer.
(4) Total genome genotyping data are analyzed using BeadStudio Genotyping module v3.3.7 (Illumina) or the like, and the quality of gene polymorphism data of each sample is evaluated (quality control).
(5) Based on these total genome genotyping data, gene polymorphism included in the gene regions and flanking regions of a gene of interest are selected, using the annotation information of the name of the gene of interest as a key, and all information regarding such gene polymorphisms is extracted using an output function of BeadStudio Genotyping module v3.3.7 (Illumina) or the like.
The present invention provides an oligonucleotide, which contains any one of CREB1 subtype gene polymorphisms (rs16839837, rs2360969, rs10932200, rs2253206, rs2551640, rs11904814, rs16839883, rs6740584, rs3770704, rs2254137, rs2551645, rs2551946, rs4234080, rs2952768, rs2709386, rs7591784 and rs7594560), CREB3 subtype gene polymorphisms (rs1243872, rs2145925, rs2025126, rs1885373, rs1885374, GA007473, rs2295794, rs4879926, GA007477, rs867194, rs11541908, rs741917, rs7862485, rs2756894, rs2249250, rs2295795, rs877365, rs2737273, rs2295797, rs2295798, rs1534847, rs7873822, rs2737274, rs10972567, rs3763630, rs10814274, rs3750434, rs1570246, GA025684, rs1570248, rs1570249, rs34478611, rs1322045, rs1951432, GA025687, rs10814275, rs10758320, rs4878628, rs10758321 and rs10758322), CREB5 subtype gene polymorphisms (rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs2175738, rs17156579, rs17156603, rs17642145, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs7794304, rs6952227, rs42695, rs1029897, rs4722793, rs10233653, rs6955105, rs17156685, rs17156694, rs17156699, rs177572, rs177573, rs177574, rs177576, rs13437706, rs177580, rs177581, rs12666636, rs177584, rs177585, rs216715, rs10951197, rs160335, rs1008262, rs310353, rs310359, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs41348, rs886816, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs2066979, rs10486591, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955 and rs10228137), and ATF2 subtype gene polymorphisms (rs1153711, rs1153702, rs7583431, rs1153699, rs2302663, rs3845744, rs212349, rs212347, rs12693057, rs1153685, rs212360, rs212361, rs2072538, rs1205399, rs1153676, rs7566401, rs7578569, rs3755490, rs13388308, rs11888507, rs10497434, rs268214, rs166531, rs268228, rs268229, rs268230, rs268231, rs10497435, rs1982235, rs268237, rs13030474 and rs268174), and which is capable of being specifically hybridized to a DNA fragment containing a gene polymorphism of the cyclic AMP responsive element binding protein gene. The gene polymorphic site is the 51^stnucleotide in the nucleotide sequence represented by any one of SEQ ID NOS: 1 to 172.
It is preferred that the oligonucleotide of the present invention has at least 10 nucleotides, preferably 10 to 150 nucleotides, more preferably 10 to 45 nucleotides, further more preferably 14 to 25 nucleotides.
Examples of the oligonucleotide of the present invention include oligonucleotides having a nucleotide sequence represented by any one of SEQ ID NOS: 1 to 172 containing the above-mentioned gene polymorphism of the cyclic AMP responsive element binding protein gene or a nucleotide sequence complementary to the nucleotide sequence (Tables 7 to 10).
The oligonucleotides of the present invention can be used as a probe or a primer specific to a cyclic AMP responsive element binding protein gene in the detection of cyclic AMP responsive element binding protein gene polymorphism described in the below-mentioned 6.

TABLE 7

		Gene
Gene		polymorphism		SEQ
name	Position	name	Sequence	ID NO:

CREB1	5′ Flanking region	rs16839837	ATTTAAAGAAGACCAGCAGAAAAATATTTATGAACTTATTTTCAACTTGT	1
			[T/C]CCCATTTTTGAACTTTTTTATCAGTGAAGAAATGGAAACATTTTT
			TCAAT
		rs2360969	ATGAAAAACTGGGGATGAGGGCCAGTCATCTGTATTTCAACAAGTCTTGC	2
			[T/C]GGTGATTCCGATGCACGCCATAGCGTGAGAACCAGTATAGCAATA
			AAACC
		rs10932200	AATAGGGAGAGCAAAAGAGCAAAGAGGTGGTTGTTCGGTGATCAATTTCC	3
			[A/C]CCAGAGTAGTAAGGAAAGGCCTCACAGAAACAGGAGCATTTGAGC
			AAAGA
		rs2253206	ACAAATAATGAGAAGTAGGAATTGGAAAAGAAAGTGATAAGTTACAGTTA	4
			[A/G]GTAGGAGGAATGGGTGACAGAAAAAAATTCCAGGGGAAGGGAAGG
			GCATG
	Intron	rs2551640	TTATAATACCTTATACAGTGCCTGCCCATCACTTGACTCTTATGGGTTCA	5
			[A/G]CATAGGAGTCAGCATGCAGCAAATTCAAGCTTTACTTCTGGGACT
			TGGGG
		rs11904814	AAGATAGTGTTGTGCATGTAAAGATCTAAGAACTTGATATTTCTATGAAA	6
			[T/G]CACAATGACTGAGCAATAGTCCTTTGCCTTAGTTTTTATTCCATT
			GAGTG
		rs16839883	CGAGGGATAGTACTTAAGTTTCCAAAGGACCATATATAGGTTTAGGAAAC	7
			[A/G]TCGAATATTACCATTGTTTTGATTGGTTCTAGTTACTTTATAGTT
			TATTT
		rs6740584	GTACTTAAGTTTCCAAAGGACCATATATAGGTTTAGGAAACATCGAATAT	8
			[T/C]ACCATTGTTTTGATTGGTTCTAGTTACTTTATAGTTTATTTTAAA
			ATTTC
		rs3770704	AAAATTTAATTTAAAAATTAGATGATTTATTTGGAAGAAGCATTTTTAGA	9
			[T/C]AGGTGGCAATATCCTCTCTAGACAATTCTCCCTGTAGGGGTCAAG
			CTTTT
		rs2254137	AAACCTTTAACTTAAAATTAGAAGCAAGTCTGATCAAGAAGTCTCAAGCA	10
			[A/C]AGGCTGAGTAGTAATATTTAAGACAACACTGCTTACTAAAGAAAA
			GAGTT
		rs2551645	AGGTCATATGTACTAAAACAGTTTATCCAAAAAGGCCTTTCTAAGACACA	11
			[T/C]TATTTTCAAACTCAAAAGTCAAAAACAAAGAAAAAATTCTTATGG
			AACCA

	3′ Flanking region	rs2551946	GTGTTTATGTAATACATATATAATCACTGAAAAATTACTGAATTGTATGA	12
			[A/C]AGTAATGTAAGTGAAAATACTTGTTCTTTAAGTGGTAAGTTAAAG
			TTGTT
		rs4234080	ATTTCTGCAACCCAAATTCCGTGGTCTCCTCATAGGGCACGAGGGCCATT	13
			[A/C]CGCCTGCACCCCGCCCTCTGCTCAGACCTGCCGTGCAAAAGAATC
			CTGGG
		rs2952768	CTCTGTCTCAAAAAAAAAAAAAATAGTGCTTTTTACTTTTATCTGAATGA	14
			[T/C]TGAAATGTCCTTTTCCCAATCCTATGATGCCTGACTGCAAAATAA
			TGGTA
		rs2709386	AACACTGACTTCCTATCACTGACTGTAAATATACAACTGATACATTATCA	15
			[A/G]TTTTCTTGTTATCTTTAACGTGAAAGCAGTATAGAGAGAGTGTGT
			TCAAA
		rs7591784	CTGGCTGTCCAGTCCCCACTCCACACCACAGAGCAACACCTAGCCAAAGA	16
			[A/G]GGTAGGTAAGAAAAGCTAAACACCCAGGGATATGAAACCAGCCTT
			CACAG
		rs7594560	TGGCAATTTTTGAATAAAAAGATTAACTACTAATTCTGAGGCAGTGGAGA	17
			[T/C]TGAGGAGAATAAGAAAGATGGCCAGCACTGCTTGCTTCTCTGGCT
			GTCCA

TABLE 8

		Gene
		polymorphism		SEQ
Gene name	Position	name	Sequence	ID NO:

Gene name	5′	rs1243872	AGTCCTCCTGTTCCCTGTGAGAGCACTTCAAGTGCTGGGGCCAG	18
	Flanking		GTCTGA[T/G]GCAGCTCTGATTCCTACACAGCAAAGCCTGGCC
	region		CAGGTAAGGGGATGGGA
Gene name		rs2145925	GGAGACGCGGTGTGTGTAGGGGCGCTACTAAGATTTGGAGGCTA	19
			CTGGGA[T/C]GGAAGCGGAATAACATAAAAGGGACAGTACAGT
			CAAGGGTACTGGTGGGA
Gene name		rs2025126	GTTGCCAACCTAAGACGACTGTCAGAGAGTCCATGCATTATGGA	20
			CAGAGT[A/G]GCTGGATGGGGACGTGGAAGCACACAGCATCCA
			CCTCTCTGACTGCCTCT
Gene name		rs1885373	CCCCCTCTAACAATAAATTCTCACACCTGGGTCAAGCTCTTCAC	21
			TTTTAG[T/C]GCTACCCTAGGCAGGAGTGTCCCAACCCAATCA
			AGAGCGGTGCCTACCCG
Gene name		rs1885374	AATGTCCTGGTAGCCACAAGGAACTGTGGTAGCTGCTGTCCCAG	22
			CAGCAG[A/C]AGAACGACACTTTGTGAGGCTTAGGTGCTGGCC
			AGGGAGGTGAGGATAAC
Gene name		GA007473	GATAGTATCAAAAAACGGTGAAGAGAGCTGATGAGGCTGTGGGG	23
			ACTGGC[T/C]GGAAGCTGCTGGCAGGGTGGAGTGGGCTGGGGC
			CCCGGCAGATTCAGATC
Gene name		rs2295794	AGCTTTAGGCCCCGCAGATCCCTACAGTTTCTCTCCCACTATGT	24
			TCTGGC[T/C]CAAAGCTGCCTCACGGAAATGCCTCAAGGATTT
			CTACCTTGCAAGCCCGA
Gene name		rs4879926	GAACTTGGACTTTTACATAATATGTGAAAGTCATAAAATATTTT	25
			GATGTA[T/C]GGGTTCAATCTGCAAACATTTATTAAAGATCAG
			CTAGGTGGCAGATTCTA
Gene name		GA007477	CTCCCAGCTGCGTTTTGCACCAGGACCTTGGTGTCCTCCACCAG	26
			CACCTT[T/C]GCAGTCTTCAGGATGCCCTCCCTGAGGGAGGGC
			CCAGCTTAGTCAGATCT
Gene name		rs867194	CATAACCCCATACAGGCCACATGGTAATCCACGGCCCTCTGATT	27
			CCCACA[T/C]TCAAGCATAAAGTGCTCCTCCCCTTCTCCCCAC
			TGTGCTTAACACAATCC
Gene name		rs11541908	GTGGATTACCAAACAACTATGGTGCGGACAGCCAAGGCCATTGC	28
			AGTGAC[T/C]GTTCAGGAGATGGTGAGTTTGGGCGAGTCCCAG
			AGGACTGCCCTCGGAGA
Gene name		rs741917	TGCCCATCCTCCATTCTGCCACAATGTATGCCCCCCAGCCACAC	29
			TGGTTC[T/C]CCATCCCTCAATACCTCATGCTTGTAATTAGCT
			TCTTGATGGAGTCTGAG
Gene name		rs7862485	CAGAGATACCCAAAGATGAACTGGGCATGGGAGAGGAAAGACAT	30
			ACTAAT[A/G]GAGAAACCATAAGAGGGCATGTGGGAGAGTAAG
			CTCGAACATCTACAGAG
Gene name		rs2756894	ATGTTGGGTCTTAGAGTGAAAAGTATGGCTTACTGTAAGTAGCA	31
			GTAAAA[A/C]GTTTGAGAGCCATATATAAATACACACCTTTGT
			GCACACAAGCAAAGCCT
Gene name		rs2249250	GAGCTTCTTGAAATGTCCCAGTGCTAGGAGGAAGCTGCAAGGTG	32
			AGAGGG[T/G]AAGTCAGACAGAAGAGTGGGGAATGATGCAGGG
			AGAAGTCTGGTAAAGGA
Gene name		rs2295795	CTGGCATTCTTTGACTCCTACGTTCCCCCACCCCCTACCGTCCT	33
			CCTACC[A/G]AGTCACTCAGGAGTCGCTTGCTGGCATCTCCAA
			CTGCCCTCAGGGCATTA
Gene name		rs877365	CCAGCTCCCATTTTCCTACCTCCCTCACAATATGCCCCATGCCT	34
			GGCTCT[T/C]TGCCCACATACCTGCATAATTCTCATTGCCCTG
			GGCAACCTCTCCCAGTA
Gene name		rs2737273	CCTCCCGAAGTGTTAGGATTGCAGGCATGAGCCACCACACCTGG	35
			CCTAAA[A/G]TTATTTTTTAATTGACATAATTTTACATATTCA
			TGAGGTACATAGTGACA
Gene name		rs2295797	ATGGGGAAGAATTTAGCAAAGAGTTTCATATCACAGCTAAGGAA	36
			TTAAGG[T/C]TGGATGCTAACTCTAACGAGAGAGAATTATGGG
			GACACTGGAAAGGTTGA
Gene name		rs2295798	TCAGATTATGCTACCTCATTTGATCCTATCAGTCCTACACGGGG	37
			CAAGTA[T/C]TGTTATCCCAAATCAGAGGTAAATAAAAGATTA
			CAAGAATATTGAACTGA
Gene name		rs1534847	ATGTAGTTCAGTATCTCTGCCCCTACCCCCATCTCTGAAGCAAG	38
			CATGTC[A/G]CTCTTTTTTGAGATTATCTGAAGAATTTTGCTG
			CAGTAGCCAGAGGGAAA
Gene name		rs7873822	TTATCTGGTCAACTCTTGTGTTTTGGAGAAGGGGAAATAGACTC	39
			TGGGAG[T/G]CCAGGAAACATTTTCAAGACAGGCCAGAGAAAG
			GACCCAGTCCCTGTAAC
Gene name		rs2737274	TCTCACCAGAGGCTTCCGTGCTTGAGGAGGAAGGGGGTGTCTAA	40
			GTGTCC[A/G]GAGGAAATGGGGGGAGACATGCAGTTTCAGCTT
			AGTGTGAAGGGTCCTTT
Gene name		rs10972567	ACAAGTATGAGTAGAAGCTAGCTCATTCCTCCTTTGGCCTGAGA	41
			ACTTTG[A/C]TCCCTTTTCCATTGTGTTTGATGGAACAGCAAC
			TCCCCACTGCCGTGTCC
Gene name		rs3763630	GGTCCCTATTTCCCACCTATGTTGTCTGTAAACAACACAGTCCA	42
			GAATCT[T/C]TGTCCCCTAACTGTGGTGGCCACAGCAAGGGCC
			TTGGGCTTAGAGAATGG

Gene name	Intron	rs10814274	TTTTTTTAATAGTTTAAAATGGTCTGGCTTGTTAGGGTTAACAC	43
			CTGGTC[T/C]GTGGAGGCATTCAGAAAGAATCTGAATGCCTGT
			TGGTCAGGGAAGCTGTA

Gene name	3′	rs3750434	AAGGAACATGTACCTCCCAAGATGGAAAGGATTTGGGGGTTCAG	44
	Flanking		CAGAGT[A/G]GGATCATCAAATGAATCCCAGTGCAAGTCTACT
	region		GACTTTGGTGGGTGGAG
Gene name		rs1570246	ATTCCAGATGCGGGCGCCGGTCGTTGTTAGGTATCGTCCCGGAG	45
			GGCCGG[T/G]CGTTGGGGAAAGCTTAAATGAGCTGGTGTTTCA
			GTGGAGCCGGGGAGCTC
Gene name		GA025684	TTGGCTGGGGAGGCGCTGGAGTGTGTAGTGACCGTCACCAACCC	46
			CCTTCC[C/G]CCCACGGCCACTTCTGCATCCAGGTGGGGATGC
			TGGCACTGAAGGTGGTG
Gene name		rs1570248	TTTAGCCATCTCATGTTAGAATCTAAAACCCTAACCTCTACTCT	47
			CATCTC[T/C]GTTCCCTCTCAGCATTACCTCTCCACTCATTCT
			TTCTCTAGGCCTTCAGG
Gene name		rs1570249	GAGGGTGTAATGGATCCTGATTCCTTATACACACTCCCAGACAT	48
			ACCCAC[A/G]TCTAGCCTCTGACCCGGAACAGTTTCTCAGACC
			TTCAACCTCTTCCTGTT
Gene name		rs34478611	AGTACGCACTATCCCCGTATTTAGTTTGTCTTTCCTGTTTCACA	49
			GCTGGA[A/G]GAAGCCTGGGTATTTTGACACGGGATCATCTGT
			AAGGCCCCATCCTCCCT
Gene name		rs1322045	TGGAGGGGGCACTGGACTGGGCACTTCCCCAGCAAGGAGGCAGG	50
			AGGGGC[A/G]AGGGCCCCCAGGTGGTCCCCAGATCTCTTCCCT
			GACCTGGAGAGAAGGAA
Gene name		rs1951432	CTGGCGCGCAGGTCCCGGAGGGGGCGGCTGGCGCGCACTACACG	51
			CTTGGG[A/G]ACAAGGAAAACATCCGCCGGAGGCCCGGCCGGG
			CGGCGCTCCAGCCTCGG
Gene name		GA025687	GTCGTGGTGTCGCTACGGGCGCGAAACGGACACTGAACACAGTC	52
			TGACTG[A/T]ATGGAGGCAGGTGGGGAGGGATCCCCTGGGAGA
			ACTTGGCGGGCCGAGAG
Gene name		rs10814275	TGCATTGCCTTTGGTCCCAAACAAGCAAATCTGGGTCAATTAAT	53
			GAAAAA[A/G]AAAAGAAAAGAAAAGAAAATGTCTTACTTTGGG
			CCCTGTTGCACTCTCCC
Gene name		rs10758320	TTGGATAACAAATTAACCTCCATTTCCACTGGACAGAGAACTCA	54
			TTCTTC[T/C]GGTATGTTTCAGAAGGCTAATGGAGCAAGGATA
			ACCTTATATTACTAATG
Gene name		rs4878628	TGGCTAGTGTTTTTTGTATCCTGCATAAGAAATCTTCCCTTACA	55
			CCAGGT[T/C]ACAAAGATTTTTTTCCTACATTTTCTCCTATAT
			CTAAAAGTTTTATGATT
Gene name		rs10758321	AGACATCATTAAATTCATCATGGCATTCTTTCTTGCTGAGCCTG	56
			GACATA[A/G]CCTGGTAAGACTAGAACTAGATAATAGGAAAAG
			AAATGTAGACATTAAGT
Gene name		rs10758322	TGGGAGAGGCTGGCATCAAATTACTCCTCTGTTTTTCTCTCTTG	57
			GTGACC[T/C]AGCAGGTGTTTAGGACAATGACGACTACTCATG
			TGGAACCTTTGCAGTCA

TABLE 9

		Gene		SEQ		Gene		SEQ
Gene		polymorphism		ID		polymorphism		ID
name	Position	name	Sequence	NO:	Position	name	Sequence	NO:

CRE	5′	rs4722778	AATCACCATTTTATGTGAAC	58	Intron	rs177584	GGGTGACGTAAGGGGGTGCA	100
B5	Flanking		AAATTGAAGTCTTTATAGCA				GAGATTCCCACTTGGGTTTA
	region		TTCTTAATTT[C/G]GTTTC				TGCTGGCCTC[A/G]TCTTT
			TGAAAGACATTTAGATAATT				GACTGGCTCTGTCATGTTGC
			GGGCAATTTAACAAAAGAGT				CCTTGTGGGGTCCTGTTTTC
			ATGTTC				ATTAA
		rs177479	CAAGTTTCATCCACGTTGTA	59		rs177585	TTTTGCACGGTAAATGCTTC	101
			GCGTGTATCAAAACTTCAAT				GTAAACGTCAGCTATTCATT
			TAATATTCTT[T/C]TATAT				AGTGAGGTGT[T/C]GGGGA
			GGGTATGTTACATTTTGTTT				GTTGTCGGGGGAAGAGAGAG
			ATCCATTCATCAGTTGGTAG				GAGAAAGAAGGAAGTGAGAG
			ACATG				GGGAG
		rs177480	GCTGCTGTGGACATTTGCAT	60		rs216715	AAAAAGAAAGAGCCAGCCTT	102
			ACAAGTTTTTTTGTGTGGAA				TAAGGAAACGGGAAGTCAAA
			ATATGTTTTC[A/G]ATTCT				GCTTGTGTAA[T/C]GAAGC
			CTTGGGAATATACCTAGGAC				AAGACCAGACTTTTTAAATC
			TGGAATGGGTCATTTGGAAA				TACCTCCCTTAACCTTTATA
			CTACG				AACAT
		rs11981754	ATTCAAAAATAACAGGATTG	61		rs10951197	ATGGTGCTACATAGGCTGGC	103
			TGAAATATCCAACTAAAATC				TTAACATCTTTTTTTGAAAT
			ATATTTGAAA[A/G]TGGTC				AAAAACCAAG[T/C]GTAAA
			CAGGAATCCCCAAATAACTT				CATGAGTCAGAATGACAGGG
			TTATGCATGTTATATGAAGA				CATATGCAGGACTCCAACAT
			TAAAT				TTACT
		rs177486	CCTTCCTTTCAGCATGCAGA	62		rs160335	TGAATTTGATGCTGTTCTCT	104
			ATTGAACTTGGCTCTGAAGT				TGGTCTTTTTCACAACTGAA
			AAAACAATAC[A/G]GGTTT				ACATTGGGCC[A/G]TTGGT
			TTGAGTGATCCAGCAGCTGT				GGGACGTTCTGTGCCTTGAA
			TCTACTTTGGTGAGAGTTTT				ACTTTTAATACGTGCAGCTC
			CTTCT				CATCT
		rs177498	GTCCTCAATTACATCTTTGT	63		rs1008262	GTCTTCTCCCCAAGAGGCCA	105
			GAGAATCAAATGTGATAAGG				CCTTTTTGACCAGGTGACTC
			CATAACACTC[T/C]TGGCA				TCCTCAGTGA[T/C]GATAT
			TGGTGGCTTTAGATATTAAC				GGTGCAATTTTTATGAGATT
			AACTCTTGCTATGTTGGTTG				TTGGGATGTGAAGCAGCTCT
			TGCTT				GTAGA
		rs2175738	ATTAGTTTCTGGCTATTGCA	64		rs310353	TGCTAACAGTGCCCTTGGGG	106
			GCTAATTCTCGGGTAAAGAA				AATGTTTGGAGGGACTTGAT
			TTTGAATGGC[A/G]TTCTA				TCCAGATCAG[A/G]AAAGA
			GTATTGCATTTTACCTAGAC				TAAACAGTGATCTGGAGGGT
			TACACTGTTACAGAATTGTG				CTGGTTTAGATGCAAGTCAT
			TGTAG				ATTTC
	Intron	rs17156579	TATCACAGGGTTCTTTGTTG	65		rs310359	CACCCTTTACATACCTGTGT	107
			GCTATTTATTGACCCATCTT				CCCTGGATCTTCCTTTCTCC
			CTCTCAGGCA[T/C]GTATA				ATGGTCCTCA[T/C]AGCCT
			TTCTCTGGGCAAGTATAGAC				CTCTTCTTTTACACTTACCT
			TCACAAGTGCCTGGAGTCCC				CTCCTTGAGCTCCCTGATGT
			TCCTC				GCCTT
		rs17156603	ATCAGTACATCAAACAACTC	66		rs41273	TGGTTCTGACAAGAAAAAGA	108
			AATTAACAAATGCTTGCATC				AAGTATTCATATTTGGTGGA
			TGCAATGTTC[A/G]TTATA				CGTGGTGGTA[A/G]GTAAA
			ATACAGCATCATAGTTGCAG				CTACTAATTTGTAAACATTG
			AATTAAAATGGCAAGATTAT				GAAATTTTTACTTTAAGTGA
			AAAAC				GAGCA
		rs17642145	GGAAGTAGGACCACCATCGG	67		rs1637457	AGCTGTTGAGCACACTCGCC	109
			CCCATACAACTTAAGTCCAA				TGTGGTTGACAGGACTCTGG
			TATATAGACT[T/C]TTAAC				CACAAGTGCC[A/G]TGGAG
			CTATGTCAGTGTGAATAGTT				GATGATGTTAGAGAGGTGGA
			GCCTGCTTGACCAGGGACTT				CACATGGGGTCAGAAGAGGA
			TAATT				AGGAG
		rs10029500	TCCAGTCAGATGACTATTTG	68		rs17156919	ATTCACCGCATACATTCACG	110
			TTCAAATATTTATTCTACTA				CAAAGGGGAAAATTTACTGC
			CATGACACAC[T/C]GTGCT				CTAAACAGAG[A/G]GACCT
			GGACACTTCAGAGATAGCTG				AAATCCCCCAGGCTAAATAA
			TGAGTTTTGCTTCCTGTGTG				ACCCAATGGAAACACAAGAA
			GTAGC				CTGCA
		rs10243659	GTGGGATGCAGGACAAAGTG	69		rs41276	TAAAAAGGTTTCTTCAAATG	111
			TTTACTTTTGTCTTTCAGAG				AAAAATGGATGGCTGAGCTG
			TCAAAATGGG[A/C]AAGGT				CTAATGGCCC[A/G]GTAAC
			TAACACAAGGAGTAAACTAA				CTAAAAATTTAACTCTTCCC
			GAAAATATATCCATATCCAT				TAATGCTCAGGGACCTCAGG
			ATTCA				TAAGG
		rs4722785	AACTGGCTTCAGCCAATTAC	70		rs160375	TTTTTGTCCTTTATTATTTT	112
			TATACCTGTTTCCTCTGGCT				TTGAATTACTTTGCTTTATT
			ATAGTGATTG[A/G]TTCAG				TTTCATGTGT[A/G]AAAAC
			GGAGAGGCCCTTAATCTAGT				ACCATATGGTGGCCACAGTG
			AGCTGTTGAGATGGAAAAAA				GGAAGCCAGGTCCTCTGCAC
			AAACA				TAAGA
		rs16874503	GAGATAGTTCCTATTCAGGA	71		rs917275	ATAGGAGGTTAGGTATGGTT	113
			CACACACCCAGTGCTTGCAG				CTGACATTGCAATATTCTCT
			ATCCATACTG[T/C]TAGAC				TCAAGTTAAC[A/G]GCAGG
			TACGTGAAGGAGGAAGAAAG				CATTTGTTACATGCTCAGAG
			ATGTTTGCAAAGGAGCCAAG				AATTTTATGATTTATAAAGA
			GGGGA				ACTTT
		rs11772815	ATCCATACTGCTAGACTACG	72		rs41348	TTCATTTACGTTATCAACTT	114
			TGAAGGAGGAAGAAAGATGT				AATTAATTTATTTATAAAAT
			TTGCAAAGGA[A/G]CCAAG				TTCCATGACC[A/G]TAGGA
			GGGGAAAGCAGGTTGCCTGC				TGACCACGTAGAAGTGTGGA
			ACCAAGATCAGACTGTCTCT				CTATGGATCACTAGCATCAA
			TGTGT				AATCT
		rs6958133	GATGTTTGCAAAGGAGCCAA	73		rs886816	CAGGAAAGCCATCTTTACAT	115
			GGGGGAAAGCAGGTTGCCTG				CACCTCTATTTAAAGCACAG
			CACCAAGATC[A/G]GACTG				GGTCCCTTTT[A/G]CCTAT
			TCTCTTGTGTTCTTTGATAA				GTCACTGAAAAACAGCAGAA
			CTCTGAGATTTTCCTTTCCT				GCCTGGTATCTAGTGGATTC
			ATTCC				ACCCC
		rs16874525	CTTTCTTCCCATCTATTAAT	74		rs17157048	ATGCAATCTGATTTATTTCC	116
			GAGCATGAACTACATCCTGG				ATGGATTCTGAGCTAGGAAT
			CCTTTAATCA[T/C]TGATA				CGCAATTGGG[A/C]ATCTC
			TCATTTCATATATACTTTTT				CAGAACCAATGGGGATTTTT
			CATTATCCTCATCTCTCCTT				GCTGTAGGACCATCGTTCTT
			TGCTT				TTCTG
		rs17715174	CATTTTTCATAGAGTCTTTG	75		rs6462098	TTTTCCAAAGCAGCTTAATG	117
			GCATTGGGTTGGACAATGAT				TAGAACAATAGGGCCAAGAA
			GGAAATTAGT[C/G]AGTTT				GGGGTTTTTT[T/C]GCTCT
			TACTCAGACAAGGTCCCTTC				GAAAAATACCGAGTCCCCTG
			CTTTGAGGAATTTATCCTCC				CCCAAGAGCTCCAGTGCCTC
			ATAAT				CCTCC
		rs6953524	TGGTATTTCCAGGGAAGAAT	76		rs10951201	CCCCAAATACCACATTTGTT	118
			ACATTAGTAATGCAGGCTTG				TGCAAGTAGGAATAGGACTG
			GGTAACCACT[T/C]GCAGC				TCTGAGGAAT[A/C]ATTTG
			TCACCTCACTACTGAGCAAT				AGAAACTGAGCCAGTCACTC
			GACGTGGAATTGGAGCTGGT				TCTTTGGCAACATGCAGGGC
			ATCAC				CACCA
		rs10239810	ATCAGAGCCTGAGCCAATAA	77		rs13311248	TTGTGTGCCAAGCTCAGTGC	119
			TATGAGCTGTCTTCTTGGAT				TCTCAAATATTCTCCCTTCA
			AGCTTGGGCT[A/G]GGCTC				GCCTAGAAGA[C/G]AGACT
			CACAACAGAAGAAGCTGGGG				GGTACCTGCTGTAAGGGGTC
			CAAATTGGCTCTGTTGCTGA				TGGCATGGAGAGAAAGCCGG
			GACCT				CTTCC
		rs17156649	TTCTGGCAGTGTGAACTTCA	78		rs12540480	TATTTTCTACAGCAGATCAC	120
			ATGGCCCACATAATTTTTTT				TCATCTCTTAAATAGATTAT
			GACCTAATGT[A/G]TAAAC				GCATTGATCG[T/C]CTTCA
			ATTTTACCTCATGTGTAGAA				AAGGGCTAAGCACACTCAAA
			ATAGGGACAATGGTACTACC				ATATTCTCTAAAGTCATTCT
			TCGTG				CATGC
		rs1811248	ATCCTCATTTTAAAGGGAAG	79		rs10265166	TCCCTAGAAAGCAAGTCAGA	121
			GAAACCAATGAGAGTGAAAT				CAGGGACAAGTCTATTTTTT
			TTAAGAAACA[T/G]ATCAG				AAGAGCCCAA[T/G]AAGAG
			ATTATTGGGAAATGGAGTAT				GAAATTTCAAAATCTCTATT
			TCTTCCCAGAGCTCCTCAAA				AGCCATTTAATTGTTTTACA
			ATATC				CTATT
		rs887623	GAGTACCCCTTAACTCAGTG	80		rs7798774	CATCATTATCTGCCAGCCTT	122
			AGGTAGACACCCAAAAGCAA				CTCTAATGTCTCCCCCATGG
			CCATCCTGCA[T/C]TTTTT				GCTAAAGAAG[T/C]CTTAT
			TCCGTGAGCATTAATAAAGT				TTCCTTTACTTTTCCCATTA
			CTATTGTTCATTGTAGAATG				AGTCTTCCCTTCCGGCTTTT
			TTCTG				TAGTA
		rs740988	TCCCATCTTTCTCACCATTA	81		rs7799246	TCAGCCTTGTTAGGCAATGC	123
			ACATGTACACATTATGCCTA				CCCTTTCCTTGTTCATGTTT
			ACACGAATCC[A/G]CCAAT				CCTTGGAGAA[T/C]AAGTG
			CCCTTGCAGCCACTGGCATG				ATCCTCTCAGCACGCTATCA
			CTCATTGGTCTCTGCCTCCA				CTTTATCATTAAGAATAGAA
			GACCC				CTTGA
		rs7794304	TCCTTGCTTACTTCTTTCTC	82		rs6972081	TTGGTTAATAAATGAATCAA	124
			AATCACGCATAATGCCTCAA				GCTGACTGCATGACTAATTC
			CTCTTAGAGC[T/C]GGCAT				AGATTAATGG[T/C]GCAGA
			TTGTTGTATCAGTCCTAATA				AATCAGTCACTAAAGAAGCC
			ACTCTTGAGGTATCTCTGAA				AAAAAAAGTTTGCTTTAATA
			ATCAG				GTCTT
		rs6952227	TAAAGACTTGGAAAGTGTCA	83		rs12533079	GCTTGACAGTAAGATTTGGT	125
			CATTGTAGTACAGTGGGGTT				TCGGAACATGAGCTCATTCA
			TTCTCCTGAT[A/G]GCTAC				CAAAAAGATA[T/G]GGGTA
			AATTTACATGCCAGGAGCCC				ATAAGACGTCTTTTAAAAAT
			TGTAAGCCCTCTAGCATTTT				ATGGGTCAGGCAGCTTTCTC
			CTTGA				GTGTT
		rs42695	CATAATTTTATCAAAATTTT	84		rs7806547	CTTCAAGAGTCTTTGAGATG	126
			TTTCACATACGTTGGCATGG				CCTATAGGCTCATCTGTTCA
			TCTTCAGACC[T/C]GTGGT				TTACAAGATG[A/G]TGAAA
			AATAATCACACCTCTCTTAA				TGGAGAGCCTGAAAGTTAAG
			CGGGTGGCGTGCTGATCAAA				AGTCTTTTCCCCCAGTCAAT
			TAAGT				AACTT
		rs1029897	CTGTTGAGTGTTCAAGTCTG	85		rs6462100	CAGATCTTCTGAAGACCTGA	127
			ATTTGGCTTCACCAAGAATA				GAAAGGACAGCAGGGTGGAG
			GAACAATGTT[T/C]CTAAA				AGACCCCTTC[A/G]CACCT
			AAGTTTGTCATGAAGAGAAG				TCCAGACGAAAGCACTGGCC
			CCCATTTAGAAATTCATCCT				TGAGGATAGGCTTGCCCAAG
			CTAAC				GGCAA
		rs4722793	GGGAGGTGTATTAACTTTTG	86		rs6979352	ACCCCTAGACAGGAAAACAT	128
			CCTATGGAGCTAGTAACAGG				CCTTCGGGGGGAAAAATGAG
			TAGAACCGGG[A/C]TTCTT				GACATGAAAT[T/C]GCTTG
			TTTTTTCATCATTTTTTATT				CTGCGGTGCCTATCATTCTG
			ATGTAAAATATATATAACAA				TTAAGGACAGTGAAAACACA
			AATTT				GTCTG
		rs10233653	AAGTTCAGAGTAACTTCCCA	87		rs6950574	TATTCAGAATCCAAACATAT	129
			GATTTTAAATATTCTGTGTC				AGGGATCTCAAATAATCCTT
			ATGTAAGAAC[A/G]AGGAG				TCCCTTCTAT[A/G]CACTA
			GAATCGCTGATCAATTAGGT				CTAATTAGCTTGATCGATAT
			TTAAAAGCTACTGAAATTCT				CATTAGGAAATTATTATTAT
			CAAGA				AATCC
		rs6955105	CGGCTGTCAAATCTCTTGCT	88		rs4722835	ACATTAAGACCGGAGGATAT	130
			GTCTGCTGCCTTTCCTCTCA				CAACAAATTTGGTTGACTGA
			GCATGTGAGC[A/G]TGGAG				GCCACATCCT[A/C]TACCT
			CTGGGGGTCTGGTGGATCCT				ATCTGACTCAGTCTATCCAC
			GTCAATCATATGTCTGTGGG				CTGTGAAGGAGACTTTAAGA
			CAGCA				CCTAG
		rs17156685	TTTATCATATGTATCTCCAG	89		rs2066979	TGGTGAATGAAAGGCAGTGC	131
			CTTGCACCTCTCTCCTTGGC				AGAGACTGCCTCTCTTTTTG
			AATGGCCTTC[A/G]CTGCA				AGGATGTTTG[T/C]TACAG
			CCTTTGACATTTTCCAACTG				AGCCTTGGTGTCAGATAATC
			CGCCTTTGACATTTTCCTCT				ATGTAACAAGCACTGGATTG
			AGATG				GCAAG
		rs17156694	CTCAGACTTTCTTTGATGGA	90		rs10486591	GGATTCATTTTCTGAAGAAT	132
			GCCAGCCTCCTTGAAAGCAG				TAAGTCAACAGACATGGCTT
			TTATTTTTAG[A/G]TGTTC				CACAATGCAC[A/G]TATTG
			CAACAGCCATCTATCTTACA				GATTCCTTTTGGGGGTCAGA
			AAAGGATTTTCTCTTCAGAT				GCAGACTCAGAGCTCTGAGA
			AGGCT				GGCTT
		rs17156699	AGAGGAGCTCAGTCAATGGT	91		rs721993	GAGAGCCTCAGCTTCCCAGT	133
			GAGATCGAATCTTTGGACCT				TGCTTGCTGGACCCTAAAGC
			CCTTTGGACC[A/G]CCGGA				TGTAAGAACT[T/C]TGTGA
			ATGAAATCACACGTTCCCTA				AACTTGAATGTTTCTTTTTT
			CAATAACAAGAGAAGCTGTT				TAACCAAGGTAAGGAATTTA
			ATTTT				ATGCC
		rs177572	CTCACCTTATTTCCATGATG	92		rs2237351	GGAGGTACCTTCATCCTTGA	134
			CTTGGTTGTCAGTGAACGCA				GAAGAGAGACTTCAGTATCT
			GATATTGGAG[T/C]TAAGG				GTGGAACAAG[T/C]GAAGC
			CCAGTGTTTGTCCCAGGGCC				TAGAACTTGGCATCGGAGCA
			CCAGATCCAACTGGAGTGAA				TAGTGCTGAGCAAAGAAGCC
			TATTA				TCTAC
		rs177573	TTTCTATGGGTGCTATTAAG	93		rs3735566	TCTCTTAAACTCCCTCCACT	135
			CATATAAAATTTTTTTTCAA				CAACACAACTGATACCTTTC
			AAGGACTGAG[T/C]TGAGT				ATTATCTCCT[A/G]TAGTG
			TGGTATGCCACTGTGAACAG				TCTGTGGCATTGGTATTCTA
			TAACTTCATCACTTGGAAGA				AAGGAGAAAACTAGAATCTA
			TCCGA				ATGAG
		rs177574	ATTTCCTCATGGTAGCATTT	94		rs11975539	TTGTTGTTTTTGTTGCCACC	136
			GAACCAAGCCTTTAAGTAGA				ACAAGAGCAAAGGTATTTCC
			ACAAGATTTT[A/G]CTAAA				TATTTTGTTT[A/G]AATTT
			CATAGAAGGCAGAAAGGGCA				GTCACTAAGATCTAAAACAG
			CTGCTGACTAGCTATTTGAA				TGGACACACAATGGGCACAC
			GAAAA				AACAA
		rs177576	TCATTGGTGGATCTGGAGAG	95		rs6462107	ATGCCTCCAGGCTTATGTTC	137
			TAGCTGACCTGAAAACAGTC				TTAGTCTAATACTCAGCCCT
			TTCATCTTTC[T/C]GCCAA				TAGCTCACAA[T/C]GGAAT
			AATAATTTTAACACTTAAAA				CATCAATCCCAGCAACTAGA
			AAAATTTTTTTGAGAAGGTA				TATTGGGACAGGGAACCTAG
			CTAGA				AGAGT
		rs13437706	CTGAGACACAGTGGGCCTTG	96		rs2190306	TTGATAACCTAGTTTAGTAT	138
			GAAATGGCAGTTCCCATAGG				CCTATGAGTGCCTTAAATAC
			GAGTCCTGCA[T/C]GAGCC				AGAGGATGCT[T/C]AATGA
			ATGAAGGCGAGAAGCCAAGG				AAATTTATAGACTGCCCGCT
			CTTTGCATGCTATGCTTTGG				CAGCAGCTCACTGGGATTG
			GTGTG				AATAT
		rs177580	TGCAGATCTGATGACAGTAC	97		rs4719955	ATCTGCACATGCCAGTGGTC	139
			ATCCACACCCTGTCGCTTTC				TGAATAACAGAAGGAGTCCT
			CCTGCCAAGA[T/C]GAACT				TCCAAGGCCA[T/C]CCTGA
			GTAGCCGTCAGAGCCTCCAT				CCTGCAGCCATGTTGGTGTA
			TCTGCTCCCCACACCCATCC				GGAACTGTCTCCAGGGAGCC
			AGTGA				AAAGT
		rs177581	TGACAGTACATCCACACCCT	98		rs10228137	CTAGAAAATATCCGTCTCTT	140
			GTCGCTTTCCCTGCCAAGAC				GTTCTAGCAGCCATAGGTAA
			GAACTGTAGC[T/C]GTCAG				ATGACAATGG[A/C]GACGC
			AGCCTCCATTCTGCTCCCCA				TACTGAAAAATCACAACTCG
			CACCCATCCAGTGACCATCC				TGTGTTCTAAAATGACCACA
			ACTAA				AAGGG
		rs1266	TCTTCTGCACTCCAGCCTCT
			CCCTCTACTTCCCTCCTTTT
			TGCTTCAGCC[A/C]GAGGC
			AGATGGCAGACATGGATACA
			CATTTATGGATTGGCTGATG
			TGTCT

TABLE 10

		Gene
		polymorphism		SEQ
Gene name	Position	name	Sequence	ID NO:

ATF2	5′	rs268174	ACGCGTGTATGTTTTACAATATACATCTCTCATAATATCAACTGAAGCAA[T/C]	141
	Flanking		ATTTAATGTTTCAGTCTACCACAGATCATTTATTTTCTAGCAAATGTCTT
	region	rs13030474	ACATGTGAGTTGAAGTTACTCCATGAAGCCCCTAAGAATGTGCAGAAAAG[T/G]	142
			GATTGATTCAAATGGATCATTCTTTCTTTTCCATTACCTTTTTTTTTTCC
		rs268237	TGGACTGACTTATATAAAAAATTAGAGAAAAATACAAATTAGTACACATT[T/C]	143
			CAGGACAAAGTTGTGTGATGCACTAAGGGAAATCGCATTAGAAAAGAGAT
		rs1982235	TGAGGAGGAAGCAAGAAAGAAGCCAAGATCCACAGTGGCTGCTTCCAAGT[T/G]	144
			GCATATGGACTAGTTGCTTGTGGCAGGGAGAGACATGGGTTCCGAATCCC
		rs10497435	AAATAGAGATAATTCACGTGTACTGTTCAACAAGCAATTATTCATATAGT[T/C]	145
			TCTCAAGTACTCAATTCTAACCAAGAACATGGTGTCCTGTGGTGTCTACA
		rs268231	GTAATTCTAACAAATGGCTAATGGAAGTGATATCAACACGTCAACATAAA[A/C]	146
			GATTAAACATCTAGAATGCCCTGCTAAGAAGATGGCTGGGGACTGAACTC
		rs268230	GAAAGAAGGGTCATTCACTACTTAACAGGAAACTAGGGTCCCCAGCAAAG[A/C]	147
			GAAGATATTTATTTCAAGGAACCTGGAAAATGGTTCCAGAAGTATGGCTA
		rs268229	ACAAAAAGAAAAACTGTAGATTCACCCCGGCAGAGAGGACTAAACAGATT[A/G]	148
			ACTTTTGATATGAGTTGGCTGCAGGATAGTGGGCCTTTTTCTTCACGTTG
		rs268228	CCCTGTGCCCTCCACTTACCTTCCCAGGAGGCGGCGGCGGCACGGGCTGC[A/G]	149
			GCAGAGGTCGAAGGAGTGGGACTCAATGCGCAAGCGCGGTCCGGCTCTTA
		rs166531	GGTGAGCTCCGGAAAGGCTGCTAGAGGGAAAGCAGGATGGGTCCTCCGAG[T/C]	150
			CCAGCCCCAGGAGCCGGGTGTCTCCGTTTCCGTCACTTCCCAGCACTAGG
		rs268214	TTCTGGAGGGGGCGGGACCAGAGGGCCCAAGGAGCGTTACTTCTGTAAAC[T/C]	151
			CGGAGCTGTGGAAGACTGTGATTGGCTGTCGGCTGGAGGAGGGCGCGGGT
	Exon	rs10497434	TTCTAGCTGGTGGGCCATGAGCTTTATTTACTCTGCTTCCAGGAATACCT[T/C]	152
			AGCTGTTATCAATAAGCAGTCCTTTCTCAAGTTTCCATCTAGTACCCTTA
	Intron	rs11888507	CCCCTCTTAAAGAGCTTGATCTGCCAACATTGGAGAAAAGGGCAATCCTA[T/C]	153
			ATATCCATGATCCTGACATACCTGCCTCAGGTAAACTAGGGGAGATACTG
		rs13388308	TTTCCCTCCTCCTATCCCACCATGGGCTGGATTCTTCATTTCACATCCTA[T/C]	154
			AAAAACTCAGCATAATTTCCAGGTTTGAAATGGCAACTTTCTCTCTGTCT
		rs3755490	AAATTGCATTTTTCATAATTTTGTTCATAAATGAAGTTTCAAGAATGTCA[T/C]	155
			GCTCAGAAAAATTTGGTAATTCTTGTGGGGAAATGTGTAACTAGCCAAAG
		rs7578569	ATAAATTTGGGAAATGTTGAATGTGTAGGCTTCATTTCACAGGACTTTTC[A/G]	156
			TGACCTTAATGTTATGTCAATTAAGGATTCATAACTTTAAAAAATGCCCC
		rs7566401	ATACTTTCTAAAGCTCAGTTGCACTATTGAAGAAAAAGCAGAATTTCTTG[A/C]	157
			CAAAAGTTTCCTGGGTTTTTTTTCATCCTAACTCTAAAATTTTACAGAAT
		rs1153676	GAAGGTAACTGTTAATAATCCAAACAAAAGATGATGATGGTTTGGGCTTA[A/C]	158
			GTGGTGTCACTGAACACAGACATAGAGGATGAGATTCAGGTCTGATAAAA
		rs1205399	GCACTCCAGCCTGAGTGACAAAGAGAAAGACTGTCCAAAAACAACAACAA[A/C]	159
			AAAAAAAGAATTACAGTCAGGTGCAGTGACTCACGCCTGTAATCCCAACA
		rs2072538	TTAGGTTTTCCCTGTCCCCAGTAAGCAGATCTAGTTCTCTTTTGCTGTTG[T/C]	160
			AGGTTTGCCAGTTAATTATTGGATTGTACTGGACTCACATTCAGAGCATG
		rs212361	ACAAGATACAGTTATGTAAATACCTATGCTTAGGTGGCAATCTAAAACTT[A/G]	161
			TTTATATGTGTTTCTTTGATTGAAAACTTTTGCTTTTTAATGCCAATGCT
		rs212360	GAAGCTTAATTTCTGCTACTCAGAGTTACATTTGTATATTTTTATGCCTA[T/C]	162
			CAAGGATTGGAGGCTTCTTAGAAGTGTATACTGCTCCTTCTCTCCCCATG
		rs1153685	CTTATTTCAGTTGCTTTTCATAATAGTACTTATTCTATCAGTTTGACGGA[A/G]	163
			AAACAAAGGCTTAGGAAGATTCTTAGTAAAAGCTTCAAATGTAAGTATTA
		rs12693057	ATTAGAAGCACAGTCTCCATTTTTAAAGTAGCAGCTCAGTTCACTCTGAC[A/G]	164
			GTATTTCACTGACGTAGCCTAAGGCTATAGGTAATGGAACATTACTCACT
		rs212347	TCTTATCAAAAAAGAAGGACATTACAAAAAGGAAAAGGCACAATTAACCT[T/C]	165
			TAAAATGCTGAAAACAAAAGAATCTCATTCTTTGGGAAAACATTTAGCAG
		rs212349	ACGGAATCTTTTAAATTAAAAAATATTGCCCATTCTGATGAAACTGCTTA[T/C]	166
			AATGACTACAAGTAAAGATGGTGGCCATTAAGTTTTATCGTGAGCACCTG
		rs3845744	CCAGTTTTAGCACTGAAAGTCCTGCTTCCTAAGAAGACCCCTCAGTCGTG[A/G]	167
			GAAAACCATGACAGTTAGTCACCCCAACAGTTAAGTAATATAAAACCTGA
		rs2302663	TTATCAGCAGCTGGGTGGAAAAAAGAAAAATTATTCATTTTCCTAAAATC[A/G]	168
			GTAAGAATGCACCAGTATGCTGAGGCAATACACAGAGTAAAAAGTTAGAA
	3′	rs1153699	GTGGGTTTGATTCTTGTCCTAGTCTAGCCTCAGTTTTGGGCAGGCACTGC[T/G]	169
	Flanking		TTGGGGTGGGGCTTTCTCAAATATCCTGCCCCTTTTCCAGTAGCAGGAAA
	region	rs7583431	ATCCTTTCTGTGTGTCTCCTCTTGTGGCTACACTTGACGGGCCATATTAT[A/C]	170
			AAAGAATACAAAACAATAGTACAGACAGGTAAATGTTTATGCCTAGAAAT
		rs1153702	TGTTGTTCATATTTTAAAAAAATTCTTAGCCATTATCTCTTCAAATAACA[T/C]	171
			GTTTGCCAAGTTCTCAATATGATATTGTTCCATAGATCTTGGATGCTGTG
		rs1153711	TGTGTGTCTTGCAGCAGCTGGATGAAGGTTCTGTAAATGTATGCTACGTC[T/G]	172
			GTTGAGTCCATGGTGTAGTTTAAGTCTGATAATTTTTGTTGATTTTTTTT

In Table 7 to Table 10 (SEQ ID NOS: 1 to 172), 101 nucleotides are shown, and a gene polymorphic site is shown at the 51st nucleotide. For example, one represented by “A/G” means a gene polymorphism associated with transitions between “A” and “G”, and “C/T” means a gene polymorphism associated with transitions between “C” and “T”.

3. Haplotype Analysis

In the present invention, by using SNP among the above-mentioned gene polymorphisms, a haplotype can be constructed. The SNP to become a target of a haplotype analysis may be any as long as its gene polymorphism frequency is 0.5% or higher, preferably, those with a gene polymorphism frequency of 1%, more preferably those with a gene polymorphism frequency of 5% or higher can be selected. Further, SNP to become a target of a haplotype analysis may be a full or partial sequence thereof.
The haplotype analysis can be carried out using various computer programs, and for example, Haploview (available from the following website: Broad Institute) (the same shall apply hereafter); Barrett J C, Fry B, Mailer J, Daly M J. Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics. 2005 Jan. 15 [PubMed ID: 15297300] Whitehead Institute for Biomedical Research Cambridge, Mass. 02142, USA.) can be used.
As an example of the haplotype analysis, among cyclic AMP responsive element binding protein gene polymorphisms in Japanese healthy subjects found as in the above-mentioned 2, with regard to the 17 sites of SNPs which are CREB1 subtype gene polymorphisms, the 40 sites of SNPs which are CREB3 subtype gene polymorphisms, the 64 sites of SNPs which are CREB5 subtype gene polymorphisms, and the 31 sites of SNPs which are ATF2 subtype gene polymorphisms, a haplotype was estimated for each linkage disequilibrium block (haplotype block), using Haploview. The estimated haplotypes are shown in Tables 11 to 14. It is to be noted that the “Tag” shown in the tables indicates a Tag SNP that is a typical gene polymorphism in the linkage disequilibrium block.

TABLE 11

Gene name CREB1

	Linkage disequilibrium block No.
	1
	Gene polymorphism name ()

1

2

3

12

13

17

Haplotype No.

Tag

4

5

6

8

9

10

11

Tag

14

15

16

Tag

H1

C

A

G

T

C

A

T

H2

C

A

G

A

T

A

C

T

A

T

C

T

G

T

H3

C

T

C

A

G

A

T

C

A

T

H4

T

C

A

G

A

T

A

C

T

A

T

C

A

T

G

C

H5

C

A

G

A

T

A

C

T

A

T

A

C

A

T

H6

T

C

A

G

A

T

A

C

T

A

T

C

T

G

T

H7

T

C

A

G

A

T

A

C

T

A

T

C

A

T

G

T

TABLE 12

Gene name CREB3

	Linkage disequilibrium block No.
	1
	Gene polymorphism name ()

1

2

3

9

11

Haplotype No.

Tag

4

5

6

7

8

Tag

10

Tag

12

13

14

15

16

17

18

19

20

21

22

23

24

H1

G

C

G

T

A

T

C

T

C

G

C

T

G

T

G

T

C

A

G

A

H2

G

C

G

T

A

T

C

G

C

T

G

T

G

T

C

A

G

A

H3

T

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

H4

T

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

H5

T

C

G

C

T

C

A

G

C

A

T

C

A

T

A

C

H6

T

G

T

A

T

C

T

C

T

G

A

T

G

C

G

T

C

A

G

C

H7

T

C

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

	Linkage disequilibrium block No.
	2
	Gene polymorphism name ()

25

26

29

36

38

39

Haplotype No.

Tag

27

28

Tag

30

31

32

33

34

35

Tag

37

Tag

40

H8

C

G

C

G

A

T

A

C

G

C

H9

C

T

A

T

G

T

A

G

A

G

A

T

C

A

T

H10

T

C

G

C

T

G

A

G

A

T

C

G

C

H11

C

T

A

G

T

A

G

A

G

A

T

C

G

T

H12

C

G

C

T

G

A

G

A

T

C

G

C

H13

C

T

A

T

G

T

A

G

A

G

A

G

T

C

A

T

H14

C

G

C

G

A

T

A

C

T

G

C

TABLE 13

Gene name CREB5

Linkage disequilibrium block No.

1

5

6

9

10

Gene polymorphism name ()

1

2

5

6

7

8

9

11

13

14

15

16

17

18

Haplotype (H) No.

Tag

3

4

Tag

H No.

Tag

10

Tag

12

H No.

Tag

H No.

Tag

H No.

Tag

H1

C

T

A

G

C

H6

C

G

C

A

G

H11

T

G

H14

T

A

H17

G

T

H2

G

C

G

A

G

T

H7

C

A

C

G

H12

C

H15

C

A

H18

G

H3

G

C

G

A

T

H8

C

G

C

G

H13

C

G

H16

C

G

H19

A

G

H4

C

G

A

G

T

H9

T

A

C

G

A

. . .

H5

G

C

G

A

G

C

H10

C

A

C

G

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

Linkage disequilibrium block No.

11

12

13

15

16

Gene polymorphism name ()

19

20

21

22

23

24

25

26

27

28

29

30

31

Haplotype (H) No.

Tag

H No.

Tag

H No.

Tag

H No.

Tag

H No.

Tag

32

33

H20

T

A

H24

G

C

H28

A

H31

G

T

C

H35

C

G

C

H21

T

G

H25

G

C

T

H29

G

A

H32

A

C

T

H36

T

C

A

G

C

H22

C

G

H26

A

C

T

H30

G

H33

A

T

H37

C

A

G

C

H23

C

A

H27

G

T

. . .

H34

A

T

C

H38

T

C

A

T

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

Linkage disequilibrium block No.

25

26

27

32

Gene polymorphism name ()

34

35

36

37

38

39

40

42

43

44

45

47

50

Haplotype (H) No.

Tag

H No.

Tag

H No.

Tag

41

H No.

Tag

46

Tag

48

49

Tag

H39

T

A

H42

G

A

G

H47

A

H49

A

T

C

G

T

G

T

H40

C

G

H43

G

A

H48

G

H50

A

T

C

G

T

G

T

C

H41

T

G

H44

A

G

A

. . .

H51

A

T

A

C

G

T

C

. . .

H45

G

A

G

A

H52

C

T

A

C

T

C

T

C

H46

G

A

H53

C

T

A

C

T

G

C

T

C

. . .

H54

A

C

G

T

G

T

H55

A

T

A

G

T

G

T

C

	Haplotypes which are estimated to occur at a frequency of less than 1%

Linkage disequilibrium block No.

33

35

40

Gene polymorphism name ()

51

52

53

54

57

58

59

Haplotype (H) No.

Tag

55

56

H No

Tag

H No.

Tag

60

61

62

63

64

H56

T

G

C

A

H61

C

T

H64

G

C

T

C

H57

T

A

T

G

C

H62

T

C

H65

A

T

C

A

H58

G

A

C

G

C

H63

T

. . .

H59

T

A

G

C

A

. . .

H60

T

A

C

G

C

. . .	Haplotypes which are estimated to occur at a frequency of less than 1%

() 1~64: (in this order)rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs6952227, rs42695, rs1029897, rs10233653, rs6955105, rs17156699, rs177572, rs177573, rs177580, rs177581, rs12666636, rs177584, rs177585, rs1008262, rs310353, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955 and rs10228137

TABLE 14

Gene name ATF2

Linkage disequilibrium block No.

1

2

Gene polymorphism name ()

1

2

3

4

5

Haplotype No.

Tag

6

7

8

9

10

11

12

13

14

15

16

H1

T

C

A

G

C

A

G

C

G

T

C

A

H2

G

C

A

G

T

G

A

T

A

C

A

C

G

H3

T

C

G

A

T

G

A

T

A

C

A

C

G

H4

T

A

G

A

T

G

A

T

A

C

A

C

G

T

G

A

T

A

C

A

C

G

21

28

Haplotype No.

17

18

19

20

Tag

22

23

24

25

26

27

Tag

29

30

31

H5

C

T

C

G

A

C

A

C

T

G

T

H6

T

C

T

A

G

A

C

T

G

C

G

C

H7

T

C

T

A

G

A

C

T

C

G

C

H8

T

C

T

A

G

A

C

T

G

C

G

C

H9

T

C

T

C

G

A

C

A

C

T

G

T

Further, from the genotype information of cyclic AMP responsive element binding protein (CREB) genes of the respective individuals in a population, a haplotype frequency in the population is calculated, and a linkage disequilibrium analysis can be carried out based on the thus obtained haplotype frequency. The D′ value and r2 value, which indicate measures of linkage disequilibrium, can be calculated based on the following definition.

Definition

It is assumed that there are SNP A and SNP B, and the respective alleles are represented by A and a, and B and b. The four haplotypes formed by SNP A and SNP B are represented by AB, Ab, aB and ab, and the respective haplotype frequencies are represented by PAB, PAb, PaB and Pab.
D=P _AB ×P _ab −P _Ab ×P _aB(In the case of D>0)
D′=(P _AB ×P _ab −P _Ab ×P _aB)/Minimum(((P _AB +P _aB)×(P _aB +P _ab)),((P _AB +P _Ab)×(P _Ab +P _ab))) (In the case of D<0)
D′=(P _AB ×P _ab −P _Ab ×P _aB)/Minimum(((P _AB +P _aB)×(P _AB +P _Ab)),((P _aB +P _ab)×(P _Ab +P _ab)))
r ²=(P _AB ×P _ab −P _Ab ×P _aB)²/[(P _AB +P _Ab)(P _AB +P _aB)(P _aB +P _ab)(P _Ab +P _ab)]
[However, Minimum (((P_AB+P_aB)×(P_aB+P_ab)), ((P_AB+P_Ab)×(P_Ab+P_ab))) means that a smaller value among (P_AB+P_aB)×(P_aB+P_ab) and (P_AB+P_Ab)×(P_Ab+P_ab) is adopted.]
Further, a haplotype block can be estimated from the results of the linkage disequilibrium analysis. As for the haplotype block, a linkage block can be estimated from the results of the haplotype analysis by using, for example, Haploview.
When a specific SNP in the estimated haplotype blocks is examined, the information of SNPs indirectly linked to each other in the same block can be obtained. That is, when a gene polymorphism of the cyclic AMP responsive element binding protein gene (specifically, a CREB1 subtype gene, a CREB3 subtype gene, a CREB5 subtype gene, or an ATF2 subtype gene) is examined, it is not necessary to analyze all the SNPs, and it is only necessary to perform typing for several specific SNPs, for example, representative SNPs such as a Tag SNP.
4. Correlation of Cyclic AMP Responsive Element Binding Protein Gene Polymorphism with Drug Sensitivity and Disease Vulnerability
It is considered that when a gene polymorphism occurs in the cyclic AMP responsive element binding protein gene, the function or expression level of the cyclic AMP responsive element binding protein might change. Therefore, there is a correlation between a cyclic AMP responsive element binding protein gene polymorphism and various phenotypes associated with the cyclic AMP responsive element binding protein in some cases.
Here, as the phenotype, a phenotype associated with sensitivity to drugs (drug sensitivity) and a phenotype associated with occurrence of a disease (disease vulnerability) can be exemplified. As the drug sensitivity, an efficacy of drugs, a side effect of drugs, duration of efficacy of drugs and the like can be exemplified. As the disease vulnerability, pain sensitivity, vulnerability to substance dependence (in particular, vulnerability to drug dependence) and the like can be exemplified.
In the present invention, the type of the aforementioned drug is not particularly limited, and preferred examples of the drug include opioid receptor function modulators and cyclic AMP responsive element binding protein function modulators. Examples of such modulators include various drugs acting directly or indirectly on the opioid receptor or the cyclic AMP responsive element binding protein. Specific examples of various drugs acting directly or indirectly on the opioid receptor include a stimulant such as methamphetamine, a dopamine receptor agonist, a dopamine receptor antagonist, a m-, κ-, or δ-opioid receptor agonist, a m-, κ-, or δ-opioid receptor antagonist, and the like. Specific examples of various drugs acting directly or indirectly on the cyclic AMP responsive element binding protein include phosphorylated enzyme, a coactivator, a PDE4 inhibitor, dephosphorylated enzyme, an agonist for each subtypes of the cyclic AMP responsive element binding protein, an antagonist for each subtypes of the cyclic AMP responsive element binding protein, and the like.
Examples of the opioid receptor function modulator include morphine, DAMGO, codeine, methadone, carfentanil, fentanyl, heroin, cocaine, naloxone, naltrexone, nalorphine, levallorphan, pentazocine, pethidine, buprenorphine, oxycodone, hydrocodone, levorphanol, etorphine, dihydroetorphine, hydromorphone, oxymorphone, tramadol, diclofenac, indomethacin, flurbiprofen axetil, marcain, ethanol, methanol, diethyl ether, propanol, butanol, flupirtine, laughing gas, F3 (1-chloro-1,2,2-trifluorocyclobutane), halothane, estradiol, dithiothreitol, thioridazine, pimozide, fluoxetine, paroxetine, desipramine, imipramine, clomipramine, tetramide, isoflurane, ginsenoside, ifenprodil, bupivacaine, tertiapine, clozapine, haloperidol, SCH23390, cocaine, and the like. In particular, morphine, pentazocine, pethidine, buprenorphine, diclofenac, indomethacin, flurbiprofen axetil and marcain are preferred, and morphine, fentanyl and pentazocine are more preferred.
Preferred examples of the cyclic AMP responsive element binding protein function modulator include phosphodiesterase 4 (PDE4), calcineurin, protein kinase A, protein kinase C, p90 ribosome S6 kinase 1 (RSK1), calmodulin kinase, glycogen synthase kinase 3β, and CREB-regulated transcription coactivator 1 (CRTC1).
The correlation between a cyclic AMP responsive element binding protein gene polymorphism and a phenotype can be examined as described in the following (1) to (4), for example.
(1) A gene polymorphism in a linkage disequilibrium block estimated as a result of a linkage disequilibrium analysis and a haplotype analysis in healthy subjects is selected. For example, a Tag SNP which is a typical gene polymorphism is selected as a cyclic AMP responsive element binding protein gene polymorphism for analyzing a correlation with a phenotype.
(2) Then, a gene polymorphism frequency of the gene polymorphism in test subjects (patients) is analyzed. In the case where a correlation between a gene polymorphism and disease vulnerability is examined, a comparison is made in terms of gene polymorphisms between the test subjects and the healthy subjects. It is effective to use a statistical technique such as a chi-square test in the comparison.
Here, the test subjects are classified into groups depending on the difference in phenotypes, and a comparison may be made in terms of gene polymorphism frequencies or genotypes between healthy subjects and test subjects in each group. In the case where the phenotype associated with the occurrence of a disease is a stimulant-induced psychotic-like symptom, it can be classified, for example, according to a period of time from the start of the use of a stimulant to the occurrence of delusion or hallucination, a period of duration of delusion or hallucination after termination of the use thereof, the presence or absence of the relapse, and the presence or absence of multiple drug abuse.
(3) If there is a gene polymorphism significantly linked to drug sensitivity in the test subjects, the gene polymorphism can be used for evaluating the genetic predisposition to drug sensitivity. Further, if there is a gene polymorphism with a significant difference in the gene polymorphism frequency between the healthy subjects and the test subjects, the gene polymorphism can be used for evaluating the genetic predisposition to disease vulnerability.
However, it is suggested that a tendency to gene polymorphism would be affected by the race, birthplace or the like, therefore, it is preferred that in a group showing a similar gene polymorphism to that of a population used for finding an associated gene polymorphism (such as SNP), the above-mentioned evaluation using the gene polymorphism is carried out.
Specific examples of the correlation between a cyclic AMP responsive element binding protein gene polymorphism and a phenotype will be shown in the following (1) to (4).
(1) In the correlation with the measurement results of the required total administration amount of analgesic in 24 hours after surgery, in the case of patients who had a minor allele (C) of the CREB1 subtype gene polymorphism (rs10932200) and underwent the surgery, the required administration amount (logarithmic transformation) of analgesic after the surgery was statistically significantly high in correlation with the number of alleles which they had. Thus, by analyzing the CREB1 subtype gene polymorphism (rs10932200), the sensitivity to analgesic can be more easily predicted.
(2) In the correlation with the measurement results of pain perception latency due to finger immersion in ice water before surgery, the presence or absence of a minor allele (C) of the CREB1 subtype gene polymorphism (rs10932200) and pain perception latency (logarithmic transformation) showed a significant correlation. Thus, by analyzing the CREB1 subtype gene polymorphism (rs10932200), the sensitivity to pain can be more easily predicted.
(3) In the correlation with the measurement results of the scale of the intensity of pain (VAS: on visual analogue scale) 24 hours after surgery, in the case of patients who had a minor allele (C) of the CREB1 subtype gene polymorphism (rs10932200) and underwent the surgery, the value of VAS (logarithmic transformation) was statistically significantly high in correlation with the number of alleles which they had. Thus, by analyzing the CREB1 subtype gene polymorphism (rs10932200), the sensitivity to pain or analgesic after the surgery can be more easily predicted.
(4) In the correlation with the measurement results of a difference in threshold of pain perception latency due to finger immersion in ice water before surgery, in the case of patients who had a minor allele (C) of the ATF2 subtype gene polymorphism (rs7583431) and underwent the surgery, a difference in threshold of pain perception latency (logarithmic transformation) due to finger immersion in ice water was statistically significantly short in correlation with the number of alleles which they had. Thus, by analyzing the ATF2 subtype gene polymorphism (rs7583431), the sensitivity to analgesic before the surgery can be more easily predicted.

5. Use of Analysis Results

As in the above-mentioned 4, the correlation between a cyclic AMP responsive element binding protein gene polymorphism and a phenotype analyzed can be used as an index in a method of predicting sensitivity to various drugs associated with the opioid receptor and the cyclic AMP responsive element binding protein and also to pain, a method of selecting a method of treating or preventing a disease associated with the opioid receptor and the cyclic AMP responsive element binding protein, a method of determining an appropriate administration amount of therapeutic drugs, a method of predicting side effects, or the like.
Further, by using the gene polymorphism or the method of the present invention, it is possible to evaluate drug sensitivity and disease vulnerability in different races. The subjects are not particularly limited, and examples thereof include Japanese, Europeans, Americans and the like. In the present invention, however, they are preferably Japanese or those having a similar gene polymorphism tendency to that of Japanese.

6. Detection of Gene Polymorphism

A genome sample of a test subject can be extracted from the blood, saliva, skin or the like, however, the origin is not limited to these as long as a genome sample can be collected therefrom. The extraction and purification methods of genomic DNA are publicly well known. For example, genomic DNA is purified from a specimen such as the blood, saliva, skin or the like collected from a human using the phenol method or the like. At this time, a commercially available genomic DNA extraction kit such as GFX Genomic Blood DNA Purification Kit (manufactured by GE Healthcare Bio-Sciences KK) or a device may be used. In the case where SNP to be detected is present in an exon, mRNA or total RNA may be extracted instead of genomic DNA.
In the detection of a cyclic AMP responsive element binding protein gene polymorphism in a genome sample, the above-mentioned oligonucleotide of the present invention can be used as a probe or a primer. Hereinafter, an example of the gene polymorphism detection method will be described.

(1) Detection of Gene Polymorphism by PCR Method

In order to amplify a test sample by PCR, it is preferred that a high fidelity DNA polymerase, for example, KOD Dash polymerase (manufactured by TOYOBO) is used. A primer to be used is designed such that a target SNP in the test sample can be amplified and synthesis is carried out. It is preferred that a gene polymorphism or a strand complementary thereto is contained at a given position between the forward and reverse primers. After completion of the amplification reaction, detection of the amplified products is carried out, and the presence or absence of a gene polymorphism is determined by a sequence method or the like.

(2) Detection of Gene Polymorphism by Sequencing Method

The gene polymorphism of the present invention can also be detected by a sequencing method based on the dideoxy method. As a sequencer to be used for the sequencing, a commercially available ABI series (Applied Biosystems (Life Technologies)) can be used.

(3) Detection of Gene Polymorphism Using DNA Microarray

A DNA microarray is a microarray in which oligonucleotide probes have been immobilized on a support, and includes a DNA chip, a Gene chip, a microchip, a bead array and the like. First, a polynucleotide of a test sample is isolated and amplified by PCR, and then labeled with a fluorescent reporter group. Then, a labeled DNA/mRNA, or total RNA is incubated along with an array.
Then, this array is inserted in a scanner, and a hybridization pattern is detected. The data of the hybridization is collected as emitted light from the fluorescent reporter group bound to the probe array (i.e., incorporated in a target sequence). A probe which is completely identical with the target sequence generates a stronger signal than those having a region which is not identical with the target sequence. Because the sequence and the position of each probe on the array are known, the sequence of the target polynucleotide reacted with the probe array can be determined based on the complementarity.

(4) Detection of Gene Polymorphism by TaqMan PCR Method

The TaqMan PCR method is a method utilizing an allele specific oligonucleotide (also referred to as TaqMan probe) labeled with fluorescence and PCR with Taq DNA polymerase. The allele specific oligonucleotide is an oligonucleotide containing a gene polymorphic site. The allele specific oligonucleotide to be used in the TaqMan PCR method can be designed based on the above-mentioned gene polymorphism information.

(5) Detection of Gene Polymorphism by Invader Method

The invader method is a method of detecting a gene polymorphism by subjecting an allele specific oligonucleotide and a template to hybridization. A kit for carrying out the invader method is commercially available (for example, NanoInvader® Array (manufactured by BML, Inc.)), and it is possible to easily detect a gene polymorphism by this method.

7. Kit

The present invention provides a kit for evaluating drug sensitivity and disease vulnerability. The kit for detecting a gene polymorphism of the present invention includes one or more components necessary for carrying out the present invention.
For example, the kit of the present invention preferably includes a component for storing or supplying an enzyme and/or a reaction component necessary for detecting a gene polymorphism. Such a component is not limited, and examples thereof include the oligonucleotide of the present invention, an enzyme buffer solution, dNTP, a reagent for control (such as a tissue sample or a target oligonucleotide for a positive or negative control), a reagent for labeling and/or detection, a solid phase support, a written instruction manual and the like. Further, the kit of the present invention may be a partial kit including only a part of the necessary components. In this case, a user can prepare the other components.
The kit of the present invention can be provided as a microarray in which the above-mentioned oligonucleotide has been immobilized on a support. The microarray is one in which the oligonucleotide of the present invention has been immobilized on a support, and includes a DNA chip, a Gene chip, a microchip, a bead array and the like.
The kit of the present invention preferably includes an oligonucleotide which contains a cyclic AMP responsive element binding protein gene polymorphism found in the present invention and is capable of being specifically hybridized to a DNA fragment containing the gene polymorphism.
In the case where a gene polymorphism is determined using the kit of the present invention, for example, the blood is collected before drugs are applied to patients or the like (for example, before surgery, at the time of occurrence of cancer pain or the like), and DNA containing a cyclic AMP responsive element binding protein 1 gene is isolated. Then, this gene is reacted with an oligonucleotide in the kit, and thereby a genotype is determined.
From the determined genotype and gene polymorphism, a dosage regimen such as the type or administration amount of the drugs can be designed. As a result, an effect of the drugs suitable for an individual can be obtained, which is useful in the personalized medicine. For example, in the case of using morphine, it becomes possible to obtain an analgesic effect suitable for an individual, and also to suppress the side effects to the minimum.
Hereinafter, the present invention will be described in more detail with reference to Examples. However, the present invention is not limited to these.

Example 1

(SNP Analysis)

Genomic DNA was extracted from the blood of humans (127 Japanese healthy subjects) by a standard method, and gene polymorphisms were identified in four subtypes (CREB1, CREB3, CREB5, and ATF2) of a human cyclic AMP responsive element binding protein.
With regard to the CREB1 subtype gene, an entire exon region, 5′ and 3′ flanking regions, and an intron region were analyzed. In the case of the CREB1 subtype gene, 7 gene polymorphisms in an intron region were identified in the Japanese samples. Further, 4 and 6 gene polymorphisms were found in the 5′ and 3′ flanking regions, respectively (see Table 15). As a result of linkage disequilibrium analysis, 1 linkage disequilibrium block was found in a region ranging from the 5′ flanking region to the 3′ flanking region (see FIG. 1 and FIG. 2). It was found that rs16839837, rs2360969, rs10932200, rs2551946, rs4234080 and rs7594560 were suitable as Tag SNPs representing this linkage disequilibrium block.
Further, in the same manner as above, with regard to the CREB3 subtype gene, an entire exon region, 5′ and 3′ flanking regions, and an intron region were analyzed. In the case of the CREB3 subtype gene, 1 gene polymorphism in an intron region, and 25 and 14 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, were identified in the Japanese samples (see Table 15). As a result of linkage disequilibrium analysis, 1 linkage disequilibrium block was found in the 5′ flanking region, and 1 linkage disequilibrium block was found in a region ranging from the 5′ flanking region to the 3′ flanking region (see FIG. 3 and FIG. 4). It was found that rs1243872, rs2145925, rs2025126, GA007477, rs11541908, rs3763630, rs10814274, rsGA025684, rs10814275, rs4878628 and rs10758321 were suitable as Tag SNPs representing this linkage disequilibrium block.
Further, in the same manner as above, with regard to the CREB5 subtype gene, an entire exon region, 5′ and 3′ flanking regions, and an intron region were analyzed. In the case of the CREB5 subtype gene, 2 gene polymorphisms in a noncoding region of exon, 241 gene polymorphisms in an intron region, and 9 and 5 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, were identified in the Japanese samples (see Table 16). As a result of linkage disequilibrium analysis performed on some of the aforementioned gene polymorphisms, 1 linkage disequilibrium block was found in the 5′ flanking region, 15 linkage disequilibrium blocks were found in the intron region, and 1 linkage disequilibrium block was found in a region ranging from the noncoding region of exon to the 3′ flanking region (see FIG. 5 and FIG. 6). It was found that rs4722778, rs177479, rs177486, rs177498, rs10229500, rs10243659, rs4722785, rs11772815, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs6952227, rs42695, rs1029897, rs10233653, rs6955105, rs17156699, rs177572, rs177573, rs177580, rs177581, rs12666636, rs1008262, rs310353, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs17157048, rs6462098, rs10951201, rs13311248, rs10265166, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs721993, rs2237351 and rs3735566 were suitable as Tag SNPs representing this linkage disequilibrium block.
Further, in the same manner as above, with regard to the ATF2 subtype gene, an entire exon region, 5′ and 3′ flanking regions, and an intron region were analyzed. In the case of the ATF2 subtype gene, 1 gene polymorphism in a noncoding region of exon 1, 16 gene polymorphisms in an intron region, and 11 and 6 gene polymorphisms in the 5′ and 3′ flanking regions, respectively, were identified in the Japanese samples (see Table 15). As a result of linkage disequilibrium analysis, 1 linkage disequilibrium block was found in a region ranging from the 5′ flanking region to the intron region, and 1 linkage disequilibrium block was found in the 3′ flanking region (see FIG. 7 and FIG. 8). It was found that rs1153711, rs1153702, rs7583431, rs2302663, rs3845744, rs268214 and rs1982235 were suitable as Tag SNPs representing this linkage disequilibrium block.

TABLE 15

CREB1 gene polymorphism	CREB3 gene polymorphism	ATF2 gene polymorphism

	Gene	Major	Minor		Gene	Major	Minor		Gene	Major	Minor
	polymorphism	allele:minor	allele		polymorphism	allele:minor	allele		polymorphism	allele:minor	allele
Position	name	allele	frequency	Position	name	allele	frequency	Position	name	allele	frequency

5′	rs16839837	C:T	0.232	5′	rs1243872	G:T	0.449	5′	rs268174	C:T	0.201
Flanking	rs2360969	C:T	0.126	Flanking	rs2145925	C:T	0.39	Flanking	rs13030474	G:T	0.008
region	rs10932200	A:C	0.315	region	rs2025126	G:A	0.366	region	rs268237	C:T	0.189
	rs2253206	G:A	0.315		rs1885373	T:C	0.055		rs1982235	T:G	0.398
Intron	rs2551640	A:G	0.323		rs1885374	A:C	0.051		rs10497435	T:C	0.189
	rs11904814	T:G	0.319		GA007473	T:C	0.051		rs268231	C:A	0.197
	rs16839883	A:G	0.181		rs2295794	T:C	0.453		rs268230	A:C	0.197
	rs6740584	C:T	0.307		rs4879926	T:C	0.051		rs268229	G:A	0.197
	rs3770704	T:C	0.181		GA007477	C:T	0.358		rs268228	A:G	0.205
	rs2254137	A:C	0.323		rs867194	T:C	0.028		rs166531	T:C	0.189
	rs2551645	T:C	0.319		rs11541908	C:T	0.283		rs268214	T:C	0.189
3′	rs2551946	C:A	0.02		rs741917	C:T	0.028	Exon	rs10497434	T:C	0.154
Flanking	rs4234080	C:A	0.193		rs7862485	G:A	0.051	Intron	rs11888507	C:T	0.154
region	rs2952768	T:C	0.331		rs2756894	C:A	0.445		rs13388308	C:T	0.154
	rs2709386	G:A	0.331		rs2249250	T:G	0.425		rs3755490	T:C	0.146
	rs7591784	G:A	0.331		rs2295795	G:A	0.374		rs7578569	G:A	0.154
	rs7594560	T:C	0.181		rs877365	T:C	0.445		rs7566401	C:A	0.154
					rs2737273	G:A	0.413		rs1153676	C:A	0.154
					rs2295797	T:C	0.374		rs1205399	A:C	0.169
					rs2295798	C:T	0.008		rs2072538	C:T	0.146
					rs1534847	A:G	0.37		rs212361	A:G	0.154
					rs7873822	G:T	0.417		rs212360	T:C	0.154
					rs2737274	G:A	0.051		rs1153685	A:G	0.154
					rs10972567	A:C	0.445		rs12693057	G:A	0.154
					rs3763630	C:T	0.276		rs212347	T:C	0.154
				Intron	rs10814274	C:T	0.409		rs212349	T:C	0.154
				3′	rs3750434	G:A	0.402		rs3845744	A:G	0.291
				Flanking	rs1570246	G:T	0.385		rs2302663	G:A	0.146
				region	GA025684	G:C	0.323	3′	rs35507277	T:G	0.008
					rs1570248	T:C	0.268	Flanking	rs1153699	G:T	0.472
					rs1570249	G:A	0.409	region	rs1153700	C:G	0.425
					rs34478611	G:A	0.323		rs7583431	A:C	0.366
					rs1322045	A:G	0.268		rs1153702	T:C	0.476
					rs1951432	G:A	0.26		rs1153711	T:G	0.209
					GA025687	A:T	0.264
					rs10814275	A:G	0.067
					rs10758320	T:C	0.264
					rs4878628	C:T	0.134
					rs10758321	G:A	0.394
					rs10758322	C:T	0.402

TABLE 16

CREB5 gene polymorphism

	Gene	Major
	polymorphism	allele:minor	Minor allele
Position	name	allele	frequency

5′ Flanking	rs4722778	C:G	0.256
region	rs177479	T:C	0.252
	rs177480	A:G	0.242
	rs11981754	A:G	0.008
	rs177486	G:A	0.161
	rs177498	C:T	0.248
	rs849322	A:G	0.437
	rs177505	T:G	0.201
	rs2175738	G:A	0.161
Intron	rs4719932	A:C	0.016
	rs10258745	C:T	0.031
	rs1013900	G:T	0.021
	rs6955393	G:A	0.035
	rs6953880	A:G	0.02
	rs17156573	T:C	0.035
	rs6960209	C:T	0.02
	rs17156577	T:C	0.039
	rs7811922	A:C	0.035
	rs6973453	T:C	0.201
	rs17156579	C:T	0.055
	rs1073298	T:C	0.201
	rs6961801	C:T	0.319
	rs6977728	C:A	0.106
	rs6978238	C:T	0.102
	rs13230543	C:A	0.248
	rs12673465	A:G	0.343
	rs10251129	T:C	0.327
	rs2391656	T:C	0.333
	rs6971345	A:G	0.327
	rs17156603	A:G	0.398
	rs7806362	C:A	0.173
	rs17642145	T:C	0.008
	rs10229500	C:T	0.138
	rs10243659	C:A	0.476
	rs4722785	G:A	0.488
	rs16874503	C:T	0.004
	rs11772815	G:A	0.331
	rs6958133	G:A	0.13
	rs16874525	C:T	0.461
	rs17715174	G:C	0.354
	rs10242868	T:G	0.205
	rs12700884	G:A	0.476
	rs17156635	G:A	0.189
	rs10239606	C:T	0.374
	rs16874528	G:A	0.008
	rs7799687	C:A	0.189
	rs714218	G:A	0.327
	rs1860759	A:G	0.327
	rs997908	G:A	0.476
	rs12112050	C:T	0.484
	rs2191827	A:G	0.484
	rs4498447	T:C	0.327
	rs10254657	G:A	0.449
	rs6953524	C:T	0.492
	rs10239810	A:G	0.307
	rs17156649	G:A	0.079
	rs1811248	T:G	0.13
	rs887623	T:C	0.252
	rs740988	A:G	0.276
	rs7794304	T:C	0.457
	rs42694	A:G	0.035
	rs6952227	G:A	0.291
	rs42695	C:T	0.244
	rs1029897	T:C	0.409
	rs42699	A:C	0.024
	rs4722793	C:A	0.405
	rs735101	T:C	0.409
	rs10233653	G:A	0.421
	rs6955105	G:A	0.465
	rs2286841	C:A	0.228
	rs979915	C:T	0.012
	rs7794347	C:T	0.248
	rs16874562	G:T	0.283
	rs17156685	A:G	0.087
	rs174024	C:T	0.268
	rs6949786	G:A	0.26
	rs7793437	A:G	0.016
	rs3757677	T:C	0.016
	rs6462085	T:G	0.016
	rs17717216	T:C	0.016
	rs17156694	G:A	0.441
	rs17156699	A:G	0.268
	rs177572	T:C	0.331
	rs177573	T:C	0.417
	rs6977204	A:G	0.48
	rs177574	A:G	0.047
	rs177576	T:C	0.327
	rs177578	G:A	0.012
	rs13437706	C:T	0.374
	rs177580	C:T	0.382
	rs177581	C:T	0.169
	rs12666636	C:A	0.224
	rs177584	G:A	0.169
	rs177585	C:T	0.169
	rs177588	G:A	0.098
	rs6462088	G:A	0.437
	rs7796539	C:T	0.004
	rs1859020	A:G	0.378
	rs1011384	A:G	0.236
	rs6462090	G:T	0.004
	rs12671247	T:C	0.154
	rs217508	T:C	0.398
	rs4719936	G:T	0.004
	rs217509	G:T	0.197
	rs217510	T:C	0.193
	rs17718257	G:A	0.004
	rs149591	C:A	0.051
	rs1910553	C:A	0.22
	rs217517	G:A	0.193
	rs217519	G:A	0.173
	rs2391668	T:G	0.382
	rs4722804	G:T	0.189
	rs618776	A:G	0.378
	rs217503	C:T	0.382
	rs217513	C:T	0.236
	rs65264	C:T	0.394
	rs441355	G:T	0.189
	rs2391670	C:T	0.362
	rs2391671	A:G	0.362
	rs216708	A:G	0.425
	rs11980665	C:T	0.173
	rs11980669	C:T	0.173
	rs11984308	T:C	0.173
	rs160346	G:A	0.37
	rs150607	A:G	0.276
	rs177594	G:A	0.106
	rs6969064	A:G	0.169
	rs150610	A:G	0.094
	rs216715	T:C	0.287
	rs10951197	T:C	0.39
	rs12539927	A:G	0.185
	rs216720	A:G	0.051
	rs17156823	G:A	0.252
	rs2078980	G:A	0.492
	rs216730	T:G	0.236
	rs13228899	G:T	0.201
	rs160335	G:A	0.496
	rs10951200	G:A	0.047
	rs10486588	G:A	0.461
	rs216735	G:A	0.287
	rs216737	C:T	0.031
	rs216743	G:A	0.055
	rs216744	A:G	0.055
	rs216747	C:T	0.031
	rs1976489	A:G	0.496
	rs150613	C:T	0.169
	rs17156878	G:A	0.232
	rs767834	C:G	0.437
	rs4722820	G:A	0.177
	rs160337	C:A	0.047
	rs160338	G:A	0.075
	rs1008262	T:C	0.323
	rs310353	G:A	0.299
	rs310359	T:C	0.217
	rs310361	C:T	0.169
	rs13233942	A:G	0.339
	rs310338	T:C	0.323
	rs41273	G:A	0.169
	rs1637457	A:G	0.22
	rs17156919	G:A	0.299
	rs41276	A:G	0.417
	rs160375	A:G	0.067
	rs917275	A:G	0.067
	rs160342	A:G	0.264
	rs160343	T:C	0.083
	rs41295	C:T	0.094
	rs160357	A:G	0.492
	rs41298	G:A	0.236
	rs41305	G:A	0.402
	rs41307	C:T	0.425
	rs10228740	A:G	0.126
	rs3888613	G:A	0.205
	rs41320	C:T	0.323
	rs41321	A:G	0.402
	rs41322	A:G	0.335
	rs7780656	G:T	0.063
	rs41327	A:G	0.343
	rs42322	T:C	0.331
	rs41333	A:G	0.331
	rs9655280	A:G	0.063
	rs9655281	G:A	0.063
	rs4719945	A:G	0.307
	rs6945988	A:G	0.413
	rs10258405	T:G	0.185
	rs10243376	G:A	0.185
	rs41334	T:C	0.429
	rs10245004	C:T	0.189
	rs41339	G:T	0.317
	rs982947	C:T	0.181
	rs982950	A:G	0.181
	rs16874653	A:G	0.28
	rs41346	G:T	0.02
	rs41348	A:G	0.386
	rs9969149	C:T	0.339
	rs6968464	G:A	0.02
	rs886816	G:A	0.181
	rs757980	A:G	0.035
	rs41351	G:A	0.083
	rs9691873	A:C	0.098
	rs17157048	A:C	0.087
	rs6462098	T:C	0.122
	rs10951201	C:A	0.154
	rs13311248	G:C	0.146
	rs12540480	T:C	0.063
	rs10265166	G:T	0.043
	rs7798774	T:C	0.087
	rs7799246	T:C	0.059
	rs6972081	T:C	0.287
	rs7777929	T:C	0.037
	rs12533079	T:G	0.079
	rs7806547	G:A	0.157
	rs6462100	G:A	0.134
	rs6979352	C:T	0.012
	rs6950574	A:G	0.13
	rs4722835	A:C	0.13
	rs9648352	A:G	0.028
	rs879593	A:C	0.138
	rs879591	G:T	0.15
	rs2299110	C:T	0.228
	rs2237349	C:T	0.272
	rs2066979	T:C	0.276
	rs10486589	A:G	0.039
	rs10486591	G:A	0.272
	rs6462103	C:T	0.118
	rs721993	C:T	0.154
	rs2237351	T:C	0.091
	rs740315	G:A	0.004
	rs2237353	A:C	0.238
	rs2073537	T:C	0.217
	rs4722844	G:T	0.181
	rs17730621	C:T	0.327
	rs2282907	G:A	0.24
	rs10238623	G:A	0.193
	rs2299116	C:A	0.091
	rs2299117	T:C	0.154
	rs2237355	A:G	0.366
	rs2237360	T:G	0.268
	rs2237361	T:C	0.217
	rs2237362	T:C	0.106
	rs7791555	G:T	0.272
	rs2237364	A:G	0.26
	rs2282909	T:G	0.366
	rs2282910	C:T	0.366
	rs2282911	T:C	0.366
	rs1544470	A:G	0.362
	rs1964240	A:C	0.293
	rs17669844	T:C	0.008
	rs886750	A:G	0.354
	rs12531253	G:A	0.272
	rs10951205	A:G	0.374
Exon	rs2190305	A:G	0.374
	rs3735566	G:A	0.039
3′ Flanking	rs11975539	G:A	0.039
region	rs6462107	C:T	0.039
	rs2190306	T:C	0.043
	rs4719955	T:C	0.047
	rs10228137	C:A	0.051

In Table 15 and Table 16, there is not found any polymorphism causing amino acid substitution, namely, a polymorphism in which the type of amino acid after translation is changed depending on the gene polymorphism allele.
Moreover, in Table 15 and Table 16, “minor allele frequency” means the ratio of a minor allele. It is to be noted that the number of healthy subjects used as test subjects was 127.

(Haplotype Construction)

As some examples of haplotype analysis, with regard to the 17 sites of SNPs which are CREB1 subtype gene polymorphisms, the 40 sites of SNPs which are CREB3 subtype gene polymorphisms, the 83 sites of SNPs which are CREB5 subtype gene polymorphisms, and the 23 sites of SNPs which are ATF2 subtype gene polymorphisms, as shown in Table 15 and Table 16, among the cyclic AMP responsive element binding protein gene polymorphisms in Japanese healthy subjects, a haplotype was estimated for each linkage disequilibrium block (haplotype block), using Haploview. The estimated haplotypes are shown in Tables 17, 18, 19 and 20. It is to be noted that the “Tag” used in each table means a Tag SNP that is a representative gene polymorphism in the linkage disequilibrium block.

TABLE 17

Gene name CREB1

	Linkage disequilibrium block No.
	1
	Gene polymorphism name ()

		1	2	3								12	13			17
Haplotype No.	Frequency (%)	Tag	Tag	Tag	4	5	6	7	8	9	10	11	Tag	Tag	14	15	16	Tag

H1

16.9

C

A

G

T

C

A

T

H2

42.5

C

A

G

A

T

A

C

T

A

T

C

T

G

T

H3

12.2

C

T

C

A

G

A

T

C

A

T

H4

17.7

T

C

A

G

A

T

A

C

T

A

T

C

A

T

G

C

H5

1.6

C

A

G

A

T

A

C

T

A

T

A

C

A

T

H6

3.5

T

C

A

G

A

T

A

C

T

A

T

C

T

G

T

H7

1.2

T

C

A

G

A

T

A

C

T

A

T

C

A

T

G

T

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

100.0%

() 1~17: (in this order)rs16839837, rs2360969, rs10932200, rs2253206, rs2551640, rs11904814, rs16839883, rs6740584, rs3770704, rs2254137, rs2551645, rs2551946, rs4234080, rs2952768, rs2709386, rs7591784 and rs7594560

TABLE 18

Gene name CREB3

		Linkage disequilibrium block No.
		1
Hap-		Gene polymorphism name ()

lotype

Frequen-

1

2

3

9

11

No.

cy (%)

Tag

4

5

6

7

8

Tag

10

Tag

12

13

14

15

16

17

18

19

20

21

22

23

24

H1

18.9

G

C

G

T

A

T

C

T

C

G

C

T

G

T

G

T

C

A

G

A

H2

35.4

G

C

G

T

A

T

C

G

C

T

G

T

G

T

C

A

G

A

H3

25.6

T

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

H4

8.3

T

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

H5

4.3

T

C

G

C

T

C

A

G

C

A

T

C

A

T

A

C

H6

2.8

T

G

T

A

T

C

T

C

T

G

A

T

G

C

G

T

C

A

G

C

H7

1.2

T

C

A

T

A

T

C

T

C

T

C

G

A

G

A

C

A

C

G

T

G

C

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

100.0%

		Linkage disequilibrium block No.
		2
Hap-		Gene polymorphism name ()

lotype

Frequen-

25

26

29

36

38

39

No.

cy (%)

Tag

27

28

Tag

30

31

32

33

34

35

Tag

37

Tag

40

H8

12.6

C

G

C

G

A

T

A

C

G

C

H9

29.8

C

T

A

T

G

T

A

G

A

G

A

T

C

A

T

H10

25.9

T

C

G

C

T

G

A

G

A

T

C

G

C

H11

2.0

C

T

A

G

T

A

G

A

G

A

T

C

G

T

H12

6.4

C

G

C

T

G

A

G

A

T

C

G

C

H13

6.7

C

T

A

T

G

T

A

G

A

G

A

G

T

C

A

T

H14

12.2

C

G

C

G

A

T

A

C

T

G

C

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

100.0%

() 1~40: (in this order)rs1243872, rs2145925, rs2025126, rs1885373, rs1885374, GA007473, rs2295794, rs4879926, GA007477, rs867194, rs11541908, rs741917, rs7862485, rs2756894, rs2249250, rs2295795, rs877365, rs2737273, rs2295797, rs2295798, rs1534847, rs7873822, rs2737274, rs10972567, rs3763630, rs10814274, rs3750434, rs1570246, GA025684, rs1570248, rs1570249, rs34478611, rs1322045, rs1951432, GA025687, rs10814275, rs10758320, rs4878628, rs10758321 and rs10758322

TABLE 19

Gene name CREB5

Linkage disequilibrium block No.

1

5

6

9

10

Hap-	Gene polymorphism name ()

lotype

Frequen-

1

2

5

6

H

Frequen-

7

8

9

11

H

Frequen-

13

14

H

Frequen-

15

16

H

Frequen-

17

18

(H) No.

cy (%)

Tag

3

4

Tag

No.

cy (%)

Tag

10

Tag

12

No.

cy (%)

Tag

No.

cy (%)

Tag

No.

cy (%)

Tag

H1

73.2

C

T

A

G

C

H6

33.1

C

G

C

A

G

H11

46.1

T

G

H14

49.2

T

A

H17

87.0

G

T

H2

6.7

G

C

G

A

G

T

H7

33.8

C

A

C

G

H12

35.4

C

H15

20.1

C

A

H18

5.1

G

H3

16.1

G

C

G

A

T

H8

16.5

C

G

C

G

H13

18.5

C

G

H16

30.7

C

G

H19

7.9

A

G

H4

1.2

C

G

A

G

T

H9

12.6

T

A

C

G

A

. . .

H5

1.2

G

C

G

A

G

C

H10

2.0

C

A

C

G

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

	100.0%								100.0%								100.0%				100.0%				100.0%

Linkage disequilibrium block No.

11

12

13

15

16

Hap-	Gene polymorphism name ()

lotype

Frequen-

19

20

H

Frequen-

21

22

23

H

Frequen-

24

25

H

Frequen-

26

27

28

H

Frequen-

29

30

31

(H) No.

cy (%)

Tag

No.

cy (%)

Tag

No.

cy (%)

Tag

No.

cy (%)

Tag

No.

cy (%)

Tag

32

33

H20

71.2

T

A

H24

40.1

G

C

H28

42.1

A

H31

26.8

G

T

C

H35

60.6

C

G

C

H21

3.6

T

G

H25

6.4

G

C

T

H29

4.3

G

A

H32

33.1

A

C

T

H36

21.3

T

C

A

G

C

H22

24.0

C

G

H26

28.7

A

C

T

H30

53.5

G

H33

25.2

A

T

H37

1.2

C

A

G

C

H23

1.2

C

A

H27

24.0

G

T

. . .

H34

15.0

A

T

C

H38

16.9

T

C

A

T

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

	100.0%				100.0%					100.0%				100.0%					100.0%

Linkage disequilibrium block No.

25

26

27

32

Hap-	Gene polymorphism name ()

lotype

Frequen-

34

35

H

Frequen-

36

37

38

39

H

Frequen-

40

H

Frequen-

42

43

44

45

47

50

(H) No.

cy (%)

Tag

No.

cy (%)

Tag

No.

cy (%)

Tag

41

No.

cy (%)

Tag

46

Tag

48

49

Tag

H39

29.9

T

A

H42

41.7

G

A

G

H47

93.3

A

H49

58.7

A

T

C

G

T

G

T

H40

32.3

C

G

H43

29.9

G

A

H48

6.7

G

H50

13.0

A

T

C

G

T

G

T

C

H41

37.8

T

G

H44

16.9

A

G

A

. . .

H51

5.4

A

T

A

C

G

T

C

. . .

H45

6.3

G

A

G

A

H52

4.3

C

T

A

C

T

C

T

C

H46

5.1

G

A

H53

4.3

C

T

A

C

T

G

C

T

C

. . .

H54

12.2

A

C

G

T

G

T

H55

1.2

A

T

A

G

T

G

T

C

Haplotypes which are estimated to occur at a frequency of less than 1%

	100.0%				100.0%						100.0%				100.0%

Linkage disequilibrium block No.

33

35

40

Hap-	Gene polymorphism name ()

lotype

Frequen-

51

52

53

54

H

Frequen-

57

58

H

Frequen-

59

(H) No.

cy (%)

Tag

55

56

No.

cy (%)

Tag

No.

cy (%)

Tag

60

61

62

63

64

H56

83.4

T

G

C

A

H61

84.6

C

T

H64

94.5

G

C

T

C

H57

1.2

T

A

T

G

C

H62

9.1

T

C

H65

3.9

A

T

C

A

H58

7.5

G

A

C

G

C

H63

6.3

T

. . .

H59

2.8

T

A

G

C

A

. . .

H60

4.3

T

A

C

G

C

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

	100.0%	100.0%	100.0%

() 1~64: (in this order)rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs6952227, rs42695, rs1029897, rs10233653, rs6955105, rs17156699, rs177572, rs177573, rs177580, rs177581, rs12666636, rs177584, rs177585, rs1008262, rs310353, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955 and rs10228137

TABLE 20

Gene name ATF2

Linkage disequilibrium block No.

1

2

Gene polymorphism name ()

1

2

3

4

5

Haplotype No.

Frequency (%)

Tag

6

7

8

9

10

11

12

13

14

15

16

H1

26.3

T

C

A

G

C

A

G

C

G

T

C

A

H2

20.9

G

C

A

G

T

G

A

T

A

C

A

C

G

H3

36.2

T

C

G

A

T

G

A

T

A

C

A

C

G

H4

16.2

T

A

G

A

T

G

A

T

A

C

A

C

G

T

G

A

T

A

C

A

C

G

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

100.0%

		Linkage disequilibrium block No.
		2
		Gene polymorphism name ()

21

28

Haplotype No.

Frequency (%)

17

18

19

20

Tag

22

23

24

25

26

27

Tag

29

30

31

H5

14.6

C

T

C

G

A

C

A

C

T

G

T

H6

8.8

T

C

T

A

G

A

C

T

G

C

G

C

H7

40.3

T

C

T

A

G

A

C

T

C

G

C

H8

28.3

T

C

T

A

G

A

C

T

G

C

G

C

H9

4.3

T

C

T

C

G

A

C

A

C

T

G

T

. . .

Haplotypes which are estimated to occur at a frequency of less than 1%

100.0%

() 1~31: (in this order)rs1153711, rs1153702, rs7583431, rs2302663, rs3845744, s212349, rs212347, rs12693057, rs1153685, rs212360, rs212361, rs2072538, rs1205399, rs1153676, rs7566401, rs7578569, rs3755490, rs13388308, rs11888507, rs10497434, rs268214, rs166531, rs268228, rs268229, rs268230, rs268231, rs10497435, rs1982235, rs268237, rs13030474 and rs268174

As shown in Table 17, at least 7 haplotypes were estimated as the haplotype of CREB1 subtype gene polymorphism in the Japanese healthy subjects, and among these, there were 6 haplotypes observed at a high frequency of 3% or higher (haplotype Nos. H1 to H6). Incidentally, specific description regarding haplotypes which are estimated to occur at a frequency of less than 1% was omitted from Table 17.
In addition, as shown in Table 18, at least 14 haplotypes were estimated as the haplotype of CREB3 subtype gene polymorphism in the Japanese healthy subjects, and among these, there were 11 haplotypes observed at a high frequency of 3% or higher (haplotype Nos. H1 to H5, H8 to H10, and H12 to H14). Incidentally, specific description regarding haplotypes which are estimated to occur at a frequency of less than 1% was omitted from Table 18.
Moreover, as shown in Table 19, at least 65 haplotypes were estimated as the haplotype of CREB5 subtype gene polymorphism in the Japanese healthy subjects, and among these, there were 57 haplotypes observed at a high frequency of 3% or higher (haplotype Nos. H1 to H3, H6 to H9, H11 to H22, H24 to H36, H38 to H54, H56, H58, and H60 to H65). Incidentally, specific description regarding haplotypes which are estimated to occur at a frequency of less than 1% was omitted from Table 19.
Furthermore, as shown in Table 20, at least 9 haplotypes were estimated as the haplotype of ATF2 subtype gene polymorphism in the Japanese healthy subjects, and among these, there were 9 haplotypes observed at a high frequency of 3% or higher (haplotype Nos. H1 to H9). Incidentally, specific description regarding haplotypes which are estimated to occur at a frequency of less than 1% was omitted from Table 20.
As well as an analysis of the haplotype frequencies in the haplotype analysis shown in Tables 17, 18, 19 and 20, a linkage disequilibrium analysis was carried out. The results are shown in FIG. 1 to FIG. 8. The linkage disequilibrium among the CREB1 subtype gene polymorphisms in the Japanese healthy subjects is shown in FIG. 1 and FIG. 2. In addition, the linkage disequilibrium among the CREB3 subtype gene polymorphisms in the Japanese healthy subjects is shown in FIG. 3 and FIG. 4. Moreover, the linkage disequilibrium among the CREB5 subtype gene polymorphisms in the Japanese healthy subjects is shown in FIG. 5 and FIG. 6. Furthermore, the linkage disequilibrium among the ATF2 subtype gene polymorphisms in the Japanese healthy subjects is shown in FIG. 7 and FIG. 8.
A linkage disequilibrium block was estimated from the results of the linkage disequilibrium analysis (FIG. 1 to FIG. 8) using Haploview.
In FIG. 1, a D′ value, which is an index of a linkage disequilibrium between SNP and SNP, is calculated, and the resulting value (two places of decimals) is written in the square at the intersection of squares continued from each SNP to the lower left direction or the lower right direction. Further, an r²value, which is a more stringent index of the linkage disequilibrium, is calculated in the same manner, and the resulting value is written in the same square as defined above in FIG. 2. It is to be noted that the square in which no numbers are written indicates that the D′ or r²value is 1. Also, the same shall apply to FIGS. 3 and 4, FIGS. 5 and 6, and FIGS. 7 and 8.
When focusing attention on the D′ values in FIG. 1, a complete linkage disequilibrium (D′=1) was observed in many combinations of gene polymorphisms. Further, when focusing attention on the r²values in FIG. 2, it was found that several gene polymorphisms showed a strong linkage disequilibrium (r²=1). It was found that suitable Tag SNPs representing these linkage disequilibrium blocks are rs16839837, rs2360969, rs10932200, rs2551946, rs4234080 and rs7594560.
In addition, when focusing attention on the D′ values in FIG. 3, a complete linkage disequilibrium (D′=1) was observed in many combinations of gene polymorphisms. Further, when focusing attention on the r²values in FIG. 4, it was found that several gene polymorphisms showed a strong linkage disequilibrium (r²=1). It was found that suitable Tag SNPs representing these linkage disequilibrium blocks are rs1243872, rs2145925, rs2025126, GA007477, rs11541908, rs3763630, rs10814274, rsGA025684, rs10814275, rs4878628 and rs10758321.
When focusing attention on the D′ values in FIG. 5, a complete linkage disequilibrium (D′=1) was observed in many combinations of gene polymorphisms. Further, when focusing attention on the r²values in FIG. 6, it was found that several gene polymorphisms showed a strong linkage disequilibrium (r²=1). It was found that suitable Tag SNPs representing these linkage disequilibrium blocks are rs4722778, rs177479, rs177486, rs177498, rs10229500, rs10243659, rs4722785, rs11772815, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs6952227, rs42695, rs1029897, rs10233653, rs6955105, rs17156699, rs177572, rs177573, rs177580, rs177581, rs12666636, rs1008262, rs310353, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs17157048, rs6462098, rs10951201, rs13311248, rs10265166, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs721993, rs2237351 and rs3735566.
In addition, when focusing attention on the D′ values in FIG. 7, a complete linkage disequilibrium (D′=1) was observed in many combinations of gene polymorphisms. Further, when focusing attention on the r²values in FIG. 8, it was found that several gene polymorphisms showed a strong linkage disequilibrium (r²=1). It was found that suitable Tag SNPs representing these linkage disequilibrium blocks are rs1153711, rs1153702, rs7583431, rs2302663, rs3845744, rs268214 and rs1982235.
Further, a linkage disequilibrium block was estimated from the results of the linkage disequilibrium analysis (FIG. 1 to FIG. 8) using Haploview. As a result, with regard to SNP in the CREB1 subtype gene shown in FIG. 1 and FIG. 2, one linkage disequilibrium block was confirmed in a region ranging from the 5′ flanking region to the 3′ flanking region. In a similar manner, with regard to SNP in the CREB3 subtype gene shown in FIG. 3 and FIG. 4, one linkage disequilibrium block was confirmed in the 5′ flanking region, and one linkage disequilibrium block was confirmed in a region ranging from the 5′ flanking region to the 3′ flanking region. In a similar manner, with regard to SNP in the CREB5 subtype gene shown in FIG. 5 and FIG. 6, one linkage disequilibrium block was confirmed in the 5′ flanking region, 15 linkage disequilibrium blocks were confirmed in the intron region, and one linkage disequilibrium block was confirmed in a region ranging from the noncoding region of exon to the 3′ flanking region. In a similar manner, with regard to SNP in the ATF2 subtype gene shown in FIG. 7 and FIG. 8, one linkage disequilibrium block was confirmed in a region ranging from the 5′ flanking region to the intron region, and one linkage disequilibrium block was confirmed in the 3′ flanking region.

Example 2

<Correlation Between CREB1 Subtype Gene Polymorphism (rs10932200) and Required Administration Amount of Analgesic>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and the required administration amount of analgesic was examined. Genomic DNA was extracted from the blood of 247 patients undergoing surgery (orthognathic surgery), and one gene polymorphism (rs10932200) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and the required administration amount of analgesic after the surgery was analyzed.
Incidentally, as the analgesic, fentanyl, which is mainly administered intravenously through a PCA (patient-controlled analgesia) pump, was used.
As a result, as shown in the following Table 21 and FIG. 9, in the case of patients who had a minor allele (C) of the CREB1 subtype gene polymorphism (rs10932200) and underwent the surgery, the required administration amount (logarithmic transformation) of analgesic after the surgery was statistically significantly high in correlation with the number of alleles which they had. Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs10932200), the sensitivity to analgesic can be predicted.
Using the median (2.222 (μg/kg)) of the required administration amounts of fentanyl in 24 hours after the surgery as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “high analgesic sensitivity group” and a “low analgesic sensitivity group,” respectively, and the groups were then stratified in terms of the rs10932200 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 54% and 46% of patients were determined to belong to the high analgesic sensitivity group and the low analgesic sensitivity group, respectively, in the A/A patient group. In contrast, in the C/C patient group, 23% and 77% of patients were determined to belong to the high analgesic sensitivity group and the low analgesic sensitivity group, respectively.

TABLE 21

Effect of CREB1 rs10932200 polymorphism on required amount
of analgesic administered in 24 hours after surgery in
patients treated with analgesic in surgery (orthognathic surgery)
(descriptive statistics by gender)

				Number of test
CREB1		Average		subjects
rs10932200	Gender		Standard deviation	(subjects)

A/A	F	2.47	2.06	77
	M	2.42	2.57	36
	Total	2.46	2.23	113
A/C	F	3.23	2.65	75
	M	2.05	1.97	33
	Total	2.87	2.51	108
C/C	F	4.37	3.32	13
	M	4.94	2.70	13
	Total	4.65	2.98	26
Total	F	2.96	2.50	165
	M	2.67	2.54	82
	Total	2.87	2.51	247

 Required administration amount (μg/kg) of analgesic in 24 hours after surgery

Example 3

<Correlation Between CREB1 Subtype Gene Polymorphism (rs10932200) and Pain Sensitivity>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 247 patients undergoing surgery (orthognathic surgery), and one gene polymorphism (rs10932200) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and the measurement of pain perception latency due to finger immersion in ice water before the surgery was analyzed.
As a result, as shown in the following Table 22 and FIG. 10, in the case of patients who had a minor allele (C) of the CREB1 subtype gene polymorphism (rs10932200) and underwent the surgery, the measurement result (logarithmic transformation) of pain perception latency was statistically significantly low in correlation with the number of alleles which they had. Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs10932200), the sensitivity to pain can be predicted.
Using the median (15 sec) of the measurement results of pain perception latency due to finger immersion in ice water before the surgery as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “high pain sensitivity group” and a “low pain sensitivity group,” respectively, and the groups were then stratified in terms of the rs10932200 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 45% and 55% of patients were determined to belong to the high pain sensitivity group and the low pain sensitivity group, respectively, in the A/A patient group. In contrast, in the A/C or C/C patient group, 55% and 45% of patients were determined to belong to the high pain sensitivity group and the low pain sensitivity group, respectively.

TABLE 22

Effect of CREB1 rs10932200 polymorphism on
measurement results of pain sensitivity before surgery in
patients treated with analgesic in surgery (orthognathic surgery)
(descriptive statistics by gender)

A/A	F	20.95	18.21	76
	M	27.96	32.07	36
	Total	23.20	23.65	112
A/C	F	17.83	15.13	75
	M	21.91	17.12	33
	Total	19.07	15.80	108
C/C	F	12.07	8.03	14
	M	17.73	9.86	13
	Total	14.80	9.24	27
Total	F	18.78	16.33	165
	M	23.90	24.28	82
	Total	20.48	19.43	247

 Measurement (sec) of pain perception latency due to finger immersion in ice water before surgery

Example 4

<Correlation Between CREB1 Subtype Gene Polymorphism (rs10932200) and Pain Sensitivity>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 247 patients undergoing surgery (orthognathic surgery), and one gene polymorphism (rs10932200) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and the measurement of the scale of the intensity of pain (VAS: on visual analogue scale) 24 hours after the surgery was analyzed.
As a result, as shown in the following Table 23 and FIG. 11, in correlation with the measurement result (logarithmic transformation) of the scale of the intensity of pain (VAS) 24 hours after the surgery, patients who had a minor allele (T) of the CREB1 subtype gene polymorphism (rs10932200) and underwent the surgery had a value of VAS (logarithmic transformation) that was statistically significantly high in correlation with the number of alleles which they had. Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs10932200), the sensitivity to pain or analgesic after the surgery can be more easily predicted.
Using the median (24 mm) of the VAS values 24 hours after the surgery as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “low pain sensitivity group” and a “high pain sensitivity group,” respectively, and the groups were then stratified in terms of the rs10932200 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 54% and 46% of patients were determined to belong to the low pain sensitivity group and the high pain sensitivity group, respectively, in the A/A patient group. In contrast, in the A/C or C/C patient group, 47% and 53% of patients were determined to belong to the low pain sensitivity group and the high pain sensitivity group, respectively.

TABLE 23

Effect of CREB1 rs10932200 polymorphism on measurement results
of VAS (intensity of pain on visual analogue scale) after surgery
in patients administered with analgesic in surgery (orthognathic surgery)
(descriptive statistics by gender)

				Number of test
			Standard	subjects
CREB1 rs10932200	Gender	Average 	deviation	(subjects)

A/A	F	24.17	19.35	77
	M	21.94	18.59	36
	Total	23.46	19.06	113
A/C	F	28.51	24.49	75
	M	26.33	20.06	33
	Total	27.84	23.15	108
C/C	F	37.31	17.78	13
	M	34.31	21.78	13
	Total	35.81	19.54	26
Total	F	27.18	21.92	165
	M	25.67	19.92	82
	Total	26.68	21.25	247

 Measurement (mm) of scale of intensity of pain by VAS after surgery

Example 5

<Correlation Between ATF2 Subtype Gene Polymorphism (rs7583431) and Analgesic Effect of Fentanyl>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 247 patients undergoing surgery (orthognathic surgery), and one gene polymorphism (rs7583431) in the ATF2 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and the analgesic effect of fentanyl that was evaluated based on the measurement of a difference in threshold of pain perception latency due to finger immersion in ice water after administration of an analgesic before the surgery was analyzed.
As a result, as shown in the following Table 24 and FIG. 12, in correlation with the measurement result (logarithmic transformation) of a difference in threshold of pain perception latency due to finger immersion in ice water before the surgery, patients who had a minor allele (C) of the ATF2 subtype gene polymorphism (rs7583431) and underwent the surgery had a difference in threshold of pain perception latency (logarithmic transformation) due to finger immersion in ice water before the surgery that was statistically significantly low in correlation with the number of alleles which they had. Accordingly, by analyzing the ATF2 subtype gene polymorphism (rs7583431), the sensitivity to analgesic before surgery can be more easily predicted.
Using the median (13 sec) of the measurement results of a difference in threshold of pain perception due to finger immersion in ice water before the surgery as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “low analgesic sensitivity group” and a “high analgesic sensitivity group,” respectively, and the groups were then stratified in terms of the rs7583431 polymorphism of the ATF2 gene. As a result, in terms of this polymorphism, 40% and 60% of patients were determined to belong to the low analgesic sensitivity group and the high analgesic sensitivity group, respectively, in the A/A patient group. In contrast, in the C/C patient group, 71% and 29% of patients were determined to belong to the low analgesic sensitivity group and the high analgesic sensitivity group, respectively.

TABLE 24

Effect of ATF2 rs7583431 polymorphism on measurement results of
difference in threshold of pain sensitivity before surgery in patients
treated with analgesic in surgery (orthognathic surgery)
(descriptive statistics by gender)

				Number of test
ATF2				subjects
rs7583431	Gender	Average 	Standard deviation	(subjects)

A/A	F	26.69	32.69	64
	M	34.78	36.28	32
	Total	29.39	33.96	96
A/C	F	26.97	36.11	76
	M	28.73	42.97	44
	Total	27.62	38.60	120
C/C	F	14.24	25.36	25
	M	12.00	19.07	6
	Total	13.81	24.00	31
Total	F	24.93	33.49	165
	M	29.87	39.24	82
	Total	26.57	35.50	247

 Measurement (sec) of difference in threshold of pain perception latency due to finger immersion in ice water before surgery

Example 6

In the same manner as Examples 2 to 5 above, a correlation between each gene polymorphism of the cyclic AMP responsive element binding protein gene, and each of the required administration amount of analgesic, pain sensitivity, and the analgesic effect of fentanyl was examined. Genomic DNA was extracted from the blood of 355 patients undergoing surgery (orthognathic surgery), and gene polymorphisms in the CREB1, CREB3, CREB5 and ATF2 subtype genes (Tag SNPs in linkage disequilibrium blocks and individual SNPs outside of the linkage disequilibrium blocks) were determined. Then, a correlation between these results of determination of the gene polymorphisms, and each of the required administration amount of analgesic in 24 hours after the surgery, pain sensitivity before the surgery, pain sensitivity (VAS) 24 hours after the surgery and before the surgery, and the analgesic effect of fentanyl, was analyzed.
Incidentally, as the analgesic, fentanyl, which is mainly administered intravenously through a PCA (patient-controlled analgesia) pump, was used.
As a result, the gene polymorphisms of the CREB1, CREB3, CREB5 and ATF2 subtype genes showed a statistically significant correlation with any phenotype of the required administration amount of analgesic in 24 hours after the surgery, pain sensitivity before the surgery, pain sensitivity (VAS) 24 hours after the surgery and before the surgery, and the analgesic effect of fentanyl. Accordingly, by analyzing these gene polymorphisms, the sensitivity to analgesic, pain sensitivity, and the analgesic effect of fentanyl can be predicted.
The results of the present example, as well as the results of Examples 2 to 5, are collectively shown in the following Table 25.

TABLE 25

Tag SNPs in linkage disequilibrium (LD) blocks and SNPs outside of LD blocks found to have significant association with phenotypes

Gene

Chromosome

Minor allele

Statistics

region	No.	LD block No.	Tag SNP	Position	frequency	Phenotype	N	BETA	SE	R²	P

CREB1	2	1	rs16839837	208079256	0.232	Required amount of fentanyl	247	−0.1347	0.06718	0.01613	0.04613
						administered in 24 hours after
						surgery
CREB1	2	1	rs2360969	208081241	0.126	Required amount of fentanyl	353	0.1453	0.07182	0.01153	0.04377
						administered in 24 hours after
						surgery
CREB1	2	1	rs10932200	208095930	0.315	Required amount of fentanyl	247	0.2211	0.0597	0.05301	0.000263
						administered in 24 hours after
						surgery
CREB1	2	1	rs10932200	208095930	0.315	VAS 24 hours after surgery	247	0.3084	0.1305	0.02228	0.01891
CREB1	2	1	rs10932200	208095930	0.315	pain perception latency before	247	−0.14	0.06043	0.02145	0.02129
						surgery
CREB1	2	1	rs4234080	208197346	0.193	Required amount of fentanyl	247	−0.2078	0.07133	0.03348	0.003906
						administered in 24 hours after
						surgery
CREB3	9	1	rs2145925	35679373	0.39	Required amount of fentanyl	246	−0.1258	0.05884	0.01838	0.03358
						administered in 24 hours after
						surgery
CREB3	9	1	rs2025126	35686625	0.366	Required amount of fentanyl	247	−0.1281	0.06011	0.01821	0.03402
						administered in 24 hours after
						surgery
CREB3	9	2	rs4878628	35756561	0.134	VAS 24 hours after surgery	253	−0.4888	0.1739	0.0305	0.005343
CREB5	7	1	rs4722778	28278588	0.256	Required amount of fentanyl	126	−0.2062	0.09913	0.03371	0.03961
						administered in 24 hours after
						surgery
CREB5	7	—	rs2175738	28304605	0.161	Required amount of fentanyl	126	−0.3309	0.1141	0.06356	0.0044
						administered in 24 hours after
						surgery
CREB5	7	—	rs2175738	28304605	0.161	VAS 24 hours after surgery	126	−0.5459	0.2322	0.04267	0.0203
CREB5	7	—	rs17156579	28327642	0.055	Required amount of fentanyl	253	−0.306	0.1293	0.02184	0.01868
						administered in 24 hours after
						surgery
CREB5	7	—	rs17156603	28348671	0.398	pain perception latency before	253	−0.1105	0.05533	0.01564	0.04692
						surgery
CREB5	7	—	rs17642145	28355789	0.008	pain perception latency before	247	−0.7505	0.319	0.02209	0.01943
						surgery
CREB5	7	5	rs4722785	28356666	0.488	Required amount of fentanyl	253	−0.1142	0.05564	0.01651	0.04114
						administered in 24 hours after
						surgery
CREB5	7	6	rs16874525	28357707	0.461	VAS 24 hours after surgery	252	0.2455	0.121	0.01621	0.04345
CREB5	7	10	rs17156649	28386945	0.079	Analgesic effect of fentanyl	247	−0.6983	0.2917	0.02285	0.01743
						before surgery
CREB5	7	—	rs7794304	28394916	0.457	pain perception latency before	354	−0.1154	0.05005	0.01487	0.02175
						surgery
CREB5	7	12	rs1029897	28400928	0.409	pain perception latency before	353	−0.09985	0.04979	0.01133	0.04568
						surgery
CREB5	7	—	rs4722793	28404054	0.405	pain perception latency before	252	−0.1225	0.05698	0.01817	0.03246
						surgery
CREB5	7	13	rs10233653	28406845	0.421	pain perception latency before	354	−0.119	0.05004	0.0158	0.01796
						surgery
CREB5	7	13	rs6955105	28409814	0.465	pain perception latency before	354	−0.1222	0.0481	0.01802	0.01147
						surgery
CREB5	7	—	rs17156685	28431959	0.087	pain perception latency before	247	−0.2359	0.1053	0.02007	0.02598
						surgery
CREB5	7	—	rs17156694	28451480	0.441	pain perception latency before	253	−0.1379	0.05492	0.02449	0.0127
						surgery
CREB5	7	15	rs177572	28460820	0.331	pain perception latency before	253	0.1454	0.06446	0.01988	0.02492
						surgery
CREB5	7	—	rs177574	28461566	0.047	Analgesic effect of fentanyl	247	0.7803	0.3685	0.01798	0.03521
						before surgery
CREB5	7	—	rs177576	28463632	0.327	Analgesic effect of fentanyl	247	0.4179	0.1582	0.02771	0.008764
						before surgery
CREB5	7	—	rs13437706	28465016	0.374	Analgesic effect of fentanyl	253	0.3854	0.1501	0.02559	0.01083
						before surgery
CREB5	7	16	rs177580	28465195	0.382	Analgesic effect of fentanyl	253	−0.3177	0.1503	0.01749	0.03554
						before surgery
CREB5	7	16	rs12666636	28465748	0.224	VAS 24 hours after surgery	354	−0.2569	0.1182	0.01325	0.03036
CREB5	7	—	rs216715	28539407	0.287	VAS 24 hours after surgery	354	−0.2213	0.1069	0.01202	0.03923
CREB5	7	—	rs10951197	28540048	0.39	VAS 24 hours after surgery	253	−0.248	0.1181	0.01726	0.03676
CREB5	7	—	rs160335	28554342	0.496	VAS 24 hours after surgery	247	0.2437	0.122	0.01602	0.04693
CREB5	7	25	rs310353	28603649	0.299	VAS 24 hours after surgery	247	−0.2846	0.1258	0.02047	0.02451
CREB5	7	—	rs310359	28606311	0.217	Required amount of fentanyl	126	0.2233	0.1071	0.0339	0.03904
						administered in 24 hours after
						surgery
CREB5	7	26	rs1637457	28615963	0.22	pain perception latency before	253	−0.1327	0.06564	0.01603	0.04422
						surgery
CREB5	7	—	rs41348	28683373	0.386	pain perception latency before	354	−0.1254	0.04985	0.01765	0.01235
						surgery
CREB5	7	—	rs886816	28689932	0.181	Required amount of fentanyl	354	0.1596	0.06074	0.01924	0.008961
						administered in 24 hours after
						surgery
CREB5	7	32	rs17157048	28703823	0.087	Required amount of fentanyl	354	−0.2175	0.09867	0.01361	0.02816
						administered in 24 hours after
						surgery
CREB5	7	32	rs10951201	28711867	0.154	Required amount of fentanyl	354	−0.2078	0.07776	0.01988	0.007896
						administered in 24 hours after
						surgery
CREB5	7	32	rs10265166	28713821	0.043	Analgesic effect of fentanyl	253	0.9373	0.4605	0.01624	0.04285
						before surgery
CREB5	7	32	rs6972081	28715571	0.287	pain perception latency before	354	−0.1289	0.05762	0.01402	0.02588
						surgery
CREB5	7	33	rs6462100	28720620	0.134	Required amount of fentanyl	354	−0.2022	0.08	0.01782	0.01194
						administered in 24 hours after
						surgery
CREB5	7	—	rs2066979	28730634	0.276	pain perception latency before	250	−0.1395	0.06557	0.01792	0.0344
						surgery
CREB5	7	—	rs10486591	28733403	0.272	pain perception latency before	354	−0.1231	0.05973	0.01192	0.04007
						surgery
CREB5	7	35	rs721993	28745985	0.154	Required amount of fentanyl	354	−0.1698	0.07741	0.01348	0.02898
						administered in 24 hours after
						surgery
CREB5	7	40	rs3735566	28831314	0.039	Required amount of fentanyl	247	−0.2901	0.1445	0.01618	0.04582
						administered in 24 hours after
						surgery
ATF2	2	1	rs1153702	175629003	0.476	Analgesic effect of fentanyl	354	0.2496	0.1214	0.01188	0.04044
						before surgery
ATF2	2	1	rs7583431	175630628	0.366	Analgesic effect of fentanyl	247	−0.5442	0.1582	0.04606	0.000685
						before surgery
ATF2	2	—	rs1153699	175631032	0.472	Analgesic effect of fentanyl	247	0.3865	0.1524	0.02558	0.01184
						before surgery
ATF2	2	—	rs1153699	175631032	0.472	pain perception latency before	247	0.1173	0.0577	0.01659	0.04312
						surgery
ATF2	2	2	rs268214	175741461	0.189	Analgesic effect of fentanyl	247	0.4065	0.1863	0.01907	0.03004
						before surgery

Example 7

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Required Administration Amount of Analgesic>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and the required administration amount of analgesic was examined. Genomic DNA was extracted from the blood of 354 patients undergoing surgery (orthognathic surgery), and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and the required administration amount of analgesic after the surgery was analyzed.
Incidentally, as the analgesic, fentanyl, which is mainly administered intravenously through a PCA (patient-controlled analgesia) pump, was used.
As a result, as shown in the following Table 26 and FIG. 13, in the case of patients who had a minor allele (C) of the CREB1 subtype gene polymorphism (rs2952768) and underwent the surgery, the required administration amount (logarithmic transformation) of analgesic after the surgery was statistically significantly high in correlation with the number of alleles which they had. Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs2952768), the sensitivity to analgesic can be predicted.
Using the median (2.268 (μg/kg)) of the required administration amount of fentanyl in 24 hours after the surgery as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “high analgesic sensitivity group” and a “low analgesic sensitivity group,” respectively, and the groups were then stratified in terms of the rs2952768 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 53% and 47% of patients were determined to belong to the high analgesic sensitivity group and the low analgesic sensitivity group, respectively, in the T/T or T/C patient group. In contrast, in the C/C patient group, 22% and 78% of patients were determined to belong to the high analgesic sensitivity group and the low analgesic sensitivity group, respectively.

TABLE 26

Effect of CREB1 rs2952768 polymorphism on required amount
of analgesic administered in 24 hours after surgery in patients
treated with analgesic in surgery (orthognathic surgery)
(descriptive statistics by gender)

				Number of test
CREB1				subjects
rs2952768	Gender	Average 	Standard deviation	(subjects)

T/T	F	2.78	2.43	101
	M	2.38	2.46	58
	Total	2.64	2.44	159
T/C	F	2.99	2.42	108
	M	2.38	2.50	52
	Total	2.79	2.46	160
C/C	F	4.80	3.05	20
	M	5.08	2.55	15
	Total	4.92	2.81	35
Total	F	3.05	2.53	229
	M	2.71	2.62	125
	Total	2.93	2.57	354

 Required amount (μg/kg) of analgesic administered in 24 hours after surgery

Example 8

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Required Administration Amount of Analgesic>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and the required administration amount of analgesic was examined. Genomic DNA was extracted from the blood or the oral mucosa of 112 patients undergoing surgery (abdominal surgery), and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and the required administration amount of analgesic after the surgery was analyzed.
Incidentally, as the analgesic, analgesics such as pentazocine and pethidine, which are mainly administered intravenously, buprenorphine, diclofenac and indomethacin, which are mainly administered as a suppository, flurbiprofen axetil, which is injected by intravenous infusion, as well as epidural morphine and marcain were used.
Incidentally, the total amount of each analgesic in terms of fentanyl means the total amount of analgesic (mg) in the case where the amount of each administered analgesic is converted to a value corresponding to the potency equivalent to fentanyl. The conversion of the amount of each analgesic to a value corresponding to the potency of fentanyl was carried out by setting a potency equivalent to 0.3 mg of fentanyl at 90 mg of pentazocine, 360 mg in the case of pethidine (Opystan), 1 mg in the case of buprenorphine (Lepetan), 300 mg in the case of diclofenac (Voltaren), 300 mg in the case of flurbiprofen axetil (Ropion), and 6 mg in the case of epidural morphine.
As a result, as shown in the following Table 27 and FIG. 14, in the case of patients who did not have a major allele (T) of the CREB1 subtype gene polymorphism (rs2952768) and underwent the surgery, the required administration amount (logarithmic transformation) of analgesic after the surgery was statistically significantly higher compared with patients having the aforementioned allele (T). Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs2952768), the sensitivity to each analgesic can be predicted.
Using the median (2.453 (μg/kg)) of the required total administration amount of each analgesic in terms of fentanyl in 24 hours after the surgery as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “high analgesic sensitivity group” and a “low analgesic sensitivity group,” respectively, and the groups were then stratified in terms of the rs2952768 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 52% and 48% of patients were determined to belong to the high analgesic sensitivity group and the low analgesic sensitivity group, respectively, in the T/T or T/C patient group. In contrast, in the C/C patient group, 33% and 67% of patients were determined to belong to the high analgesic sensitivity group and the low analgesic sensitivity group, respectively.

TABLE 27

Effect of CREB1 rs2952768 polymorphism on required amount
of analgesic administered in 24 hours after surgery in patients
treated with analgesic in surgery (abdominal surgery)
(descriptive statistics by gender)

T/T	F	0.93	1.35	18
	M	0.43	0.63	26
	Total	0.63	1.01	44
T/C	F	0.67	1.32	28
	M	0.78	1.01	28
	Total	0.72	1.17	56
C/C	F	1.12	1.34	6
	M	1.78	1.61	6
	Total	1.45	1.45	12
Total	F	0.81	1.32	52
	M	0.73	1.00	60
	Total	0.77	1.15	112

 Required total amount (μg/kg) of analgesic administered in terms of fentanyl in 24 hours after surgery

Example 9

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Severity of Drug Dependence in Methamphetamine-Dependent Patients>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and vulnerability to drug dependence associated with the severity of drug dependence was examined. Genomic DNA was extracted from the blood of 194 methamphetamine-dependent patients, and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. The patients were classified based on the presence or absence of the abuse of many drugs, and a comparison was made among them. The results are shown in the following Table 28. Here, the methamphetamine-dependent patients were classified into two groups, namely, a patient group involving the abuse of drugs other than stimulants (two or more types) and a patient group involving the abuse of a single drug (only one type).
As a result, as is clear from the following Table 28, a significant difference in genotype frequency was observed in terms of the gene polymorphism (rs2952768), and in methamphetamine-dependent patients having a major allele (T), the number of patients who abused many drugs was statistically significantly higher compared with patient who did not have the aforementioned allele T.
From the above results, it was demonstrated that drug sensitivity associated with the severity of stimulant dependence can be easily predicted by determining genotype frequency in the cyclic AMP responsive element binding protein gene polymorphism.

TABLE 28

Comparison of genotype and allele frequencies in rs2952768 polymorphism in methamphetamine-dependent patients
classified based on presence or absence of abuse of many drugs
Gene polymorphism name: CREB1 rs2952768

	(Number of samples)	Genotype frequency (%)	Allele frequency
Sample name	Abuse of many drugs	T/T T/C C/C	T C

Methamphetamine- dependent patients	No (53) Yes (141)	$\begin{matrix} \begin{matrix} 19 & 22 & 12 \\ (35.8 %) & (41.5 %) & (22.6 %) \\ 58 & 67 & 16 \end{matrix}) \begin{matrix} (P = 0.137; genotype) \\ (P = 0.503; dominant model) \\ * (P = 0.046; recessive model) \end{matrix} \\ (41.1 %) (47.5 %) (11.3 %) \end{matrix}$	$\begin{matrix} 0.556 & 0.444 \\ 0.644 & 0.356 \end{matrix}) (P = 0.133; allele)$

Each model indicates hereditary mode regarding minor allele (C);
*. P < 0.05.

Example 10

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Severity of Drug Dependence in Alcohol-Dependent Patients>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and vulnerability to drug dependence associated with the severity of drug dependence was examined. Genomic DNA was extracted from the blood of 436 alcohol-dependent patients, and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. The patients were classified based on the presence or absence of drug abuse, and a comparison was made among them. The results are shown in the following Table 29. Here, the alcohol-dependent patients were classified into two groups, namely, a patient group with drug abuse (one or more types) and a patient group without drug abuse (only alcohol ingestion).
As a result, as is clear from the following Table 29, a significant difference in genotype and allele frequencies was observed in terms of the gene polymorphism (rs2952768), and it was demonstrated that the frequency of major allele (T) was statistically significantly higher in the patient group with drug abuse than in the patient group without drug abuse.
From the above results, it was demonstrated that vulnerability to drug dependence associated with the severity of drug dependence can be easily predicted by determining genotype and allele frequencies in the cyclic AMP responsive element binding protein gene polymorphism.

TABLE 29

Comparison of genotype and allele frequencies in rs2952768 polymorphism in alcohol-dependent patients
classified based on presence or absence of drug abuse
Gene polymorphism name: CREB1 rs2952768

	(Number of samples)	Genotype frequency (%)	Allele frequency
Sample name	Drug abuse	T/T T/C C/C	T C

Alcohol- dependent patients	No (391) Yes (45)	$\begin{matrix} \begin{matrix} 172 & 166 & 53 \\ (44.0 %) & (42.5 %) & (13.6 %) \\ 25 & 18 & 2 \end{matrix}) \begin{matrix} (P = 0.142; genotype) \\ (P = 0.140; dominant model) \\ † (P = 0.097; recessive model) \end{matrix} \\ (55.6 %) (40.0 %) (4.4 %) \end{matrix}$	$\begin{matrix} 0.652 & 0.348 \\ 0.756 & 0.244 \end{matrix}) * (P = 0.049; allele)$

Each model indicates hereditary mode regarding minor allele (C);
*. P < 0.05.
†. 0.05 ≦ P < 0.1.

Example 11

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Severity of Drug Dependence in Eating Disorder Patients>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and vulnerability to drug dependence associated with the severity of drug dependence was examined. Genomic DNA was extracted from the blood of 221 patients with eating disorder, and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. The patients were classified based on the presence or absence of drug dependence, and a comparison was made among them. The results are shown in the following Table 30. Here, the eating disorder patients were classified into two groups, namely, a patient group with a complication of drug dependence and a patient group without a complication of drug dependence.
As a result, as is clear from the following Table 30, a significant difference in genotype and allele frequencies was observed in terms of the gene polymorphism (rs2952768), and it was demonstrated that the frequency of major allele (T) was statistically significantly higher in the patient group with a complication of drug dependence than in the patient group without a complication of drug dependence.
From the above results, it was demonstrated that vulnerability to drug dependence associated with the severity of drug dependence can be easily predicted by determining genotype and allele frequencies in the cyclic AMP responsive element binding protein gene polymorphism.

TABLE 30

Comparison of genotype and allele frequencies in rs2952768 polymorphism in patients with eating disorder
classified based on presence or absence of complication of drug dependence
Gene polymorphism name: CREB1 rs2952768

	(Number of samples)	Genotype frequency (%)	Allele frequency
Sample name	Complication of drug	T/T T/C C/C	T C

Eating disorder patients	No (200) Yes (21)	$\begin{matrix} \begin{matrix} 85 & 93 & 22 \\ (42.5 %) & (46.5 %) & (11.0 %) \\ 14 & 6 & 1 \end{matrix}) \begin{matrix} (P = 0.103; genotype) \\ * (P = 0.034; dominant model) \\ (P = 0.705; recessive model) \end{matrix} \\ (66.7 %) (28.6 %) (4.8 %) \end{matrix}$	$\begin{matrix} 0.658 & 0.343 \\ 0.810 & 0.190 \end{matrix}) * (P = 0.046; allele)$

Each model indicates hereditary mode regarding minor allele (C);
*. P < 0.05.

Example 12

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Severity of Substance Dependence in Eating Disorder Patients>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and vulnerability to substance (alcohol) dependence associated with the severity of substance (alcohol) dependence was examined. Genomic DNA was extracted from the blood of 221 patients with eating disorder, and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. The patients were classified based on the presence or absence of alcohol dependence, and a comparison was made among them. The results are shown in the following Table 31. Here, the eating disorder patients were classified into two groups, namely, a patient group with a complication of alcohol dependence and a patient group without a complication of alcohol dependence.
As a result, as is clear from the following Table 31, a significant difference in genotype and allele frequencies was observed in terms of the gene polymorphism (rs2952768), and it was demonstrated that the frequency of major allele (T) was statistically significantly higher in the patient group with a complication of alcohol dependence than in the patient group without a complication of alcohol dependence.
From the above results, it was demonstrated that vulnerability to substance (alcohol) dependence associated with the severity of substance (alcohol) dependence can be easily predicted by determining genotype and allele frequencies in the cyclic AMP responsive element binding protein gene polymorphism.

TABLE 31

Comparison of genotype and allele frequencies in rs2952768 polymorphism in patients with eating disorder
classified based on presence or absence of complication of alcohol dependence
Gene polymorphism name: CREB1 rs2952768

	(Number of samples)	Genotype frequency (%)	Allele frequency
Sample name	Complication of alcohol	T/T T/C C/C	T C

Eating disorder patients	No (151) Yes (70)	$\begin{matrix} \begin{matrix} 61 & 72 & 18 \\ (40.4 %) & (47.7 %) & (11.9 %) \\ 38 & 27 & 5 \end{matrix}) \begin{matrix} (P = 0.136; genotype) \\ † (P = 0.053; dominant model) \\ (P = 0.279; recessive model) \end{matrix} \\ (54.3 %) (38.6 %) (7.1 %) \end{matrix}$	$\begin{matrix} 0.642 & 0.358 \\ 0.736 & 0.264 \end{matrix}) † (P = 0.052; allele)$

Each model indicates hereditary mode regarding minor allele (C);
†. 0.05 ≦ P < 0.1.

Example 13

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Reward Dependence>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and dependence-prone personality associated with reward dependence was examined. Genomic DNA was extracted from the oral mucosa of 495 healthy subjects, and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and dependence-prone personality was analyzed.
It is to be noted that, as a test of dependence-prone personality, a reward dependence (RD) score (average) in the Temperament and Character Inventory (TCI) was used.
As a result, as is shown in the following Table 32 and FIG. 15, healthy subjects having a minor allele (C) in the CREB1 subtype gene polymorphism (rs2952768) had an RD score (logarithmic transformation) that was statistically significantly low in correlation with the number of alleles which they had. Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs2952768), dependence-prone personality can be predicted.
Using the median (0.667) of the RD score as a reference, a patient group with a value smaller than the reference and a patient group with a value larger than the reference were defined as a “low reward dependence group” and a “high reward dependence group,” respectively, and the groups were then stratified in terms of the rs2952768 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 43% and 57% of healthy subjects were determined to belong to the low reward dependence group and the high reward dependence group, respectively, in the T/T patient group. In contrast, in the C/C patient group, 58% and 42% of healthy subjects were determined to belong to the low reward dependence group and the high reward dependence group, respectively.
From the above results, it was demonstrated that dependence-prone personality associated with reward dependence can be easily predicted by determining genotype and allele frequencies in the cyclic AMP responsive element binding protein gene polymorphism.

TABLE 32

Effect of CREB1 rs2952768 polymorphism on
reward dependence (RD) score (average)
(descriptive statistics by gender)

				Number of test
CREB1		Average		subjects
rs2952768	Gender		Standard deviation	(subjects)

T/T	F	0.72	0.18	98
	M	0.66	0.18	122
	Unknown	0.60	0.09	2
	Total	0.68	0.18	222
T/C	F	0.70	0.18	103
	M	0.62	0.20	98
	Unknown	0.78	0.08	3
	Total	0.66	0.19	204
C/C	F	0.68	0.22	37
	M	0.58	0.18	32
	Total	0.63	0.21	69
	F	0.70	0.19	238
	M	0.63	0.19	252
	Unknown	0.71	0.12	5
	Total	0.67	0.19	495

 Average in dimension of average score in each subscale

Example 14

<Correlation Between CREB1 Subtype Gene Polymorphism (rs2952768) and Expression Level of CREB1 Gene>
A correlation between a cyclic AMP responsive element binding protein gene polymorphism and a gene expression level was examined. Genomic DNA was extracted from the blood of 100 postmortem brain tissue donors at the Stanley Foundation Brain Bank, and one gene polymorphism (rs2952768) in the CREB1 subtype gene was determined. Then, a correlation between these results of determination of the gene polymorphism and a gene expression level was analyzed.
Incidentally, as the gene expression level, the value of the relative mRNA expression level of CREB1, which was standardized with the value of the mRNA expression level of a β-actin gene (ACTB), was used.
As a result, as shown in the following Table 33 and FIG. 16, in the case of subjects who did not have a major allele (T) of the CREB1 subtype gene polymorphism (rs2952768), the mRNA expression level (logarithmic transformation) of CREB1 was statistically significantly higher compared with subjects having the aforementioned allele (T). Accordingly, by analyzing the CREB1 subtype gene polymorphism (rs2952768), the gene expression level can be predicted.
Using the median (0.0145) of the relative mRNA expression level as a reference, a subject group with a value smaller than the reference and a subject group with a value larger than the reference were defined as a “low gene expression group” and a “high gene expression group,” respectively, and the groups were then stratified in terms of the rs2952768 polymorphism of the CREB1 gene. As a result, in terms of this polymorphism, 52% and 48% of subjects were determined to belong to the low gene expression group and the high gene expression group, respectively, in the T/T or T/C subject group. In contrast, in the C/C subject group, 30% and 70% of subjects were determined to belong to the low gene expression group and the high gene expression group, respectively.
From the above results, it was demonstrated that the tendency of the expression level of a CREB1 gene (whether the gene tends to be expressed at a high level or at a low level) can be easily predicted by determining genotype and allele frequencies in the cyclic AMP responsive element binding protein gene polymorphism.

TABLE 33

Effect of CREB1 rs2952768 polymorphism on gene expression level
in postmortem brain tissue donors at Stanley Foundation Brain Bank
(descriptive statistics)

			Number of test
			subjects
CREB1 rs2952768	Average 	Standard deviation	(subjects)

T/T	0.015345	0.006618	48
T/C	0.015667	0.005267	42
C/C	0.023360	0.023537	10
Total	0.016282	0.009396	100

 Relative mRNA expression level value of CREB1 standardized with mRNA expression level value of β-actin gene (ACTB)

INDUSTRIAL APPLICABILITY

According to the present invention, it becomes possible to provide: a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism, which can evaluate an individual difference in terms of drug sensitivity and disease vulnerability; a method for evaluating drug sensitivity and disease vulnerability using the gene polymorphism or the haplotype; and the like. According to this evaluation method, it becomes possible to readily know or predict a proper prescribed amount, a proper prescribed schedule associated with a narcotic drug such as morphine, and the like, and hence the method is extremely useful for personalized pain therapy, drug dependence therapy and the like.

Claims

1. A method for evaluating drug sensitivity, comprising associating a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism with an individual drug sensitivity.

2. The method according to claim 1, wherein a tendency in the presence or absence of an individual drug sensitivity is evaluated based on the results from the analysis of the gene polymorphism or the haplotype.

3. The method according to claim 1, comprising the following steps:

(1) a step of performing linkage disequilibrium analysis and haplotype analysis on a healthy subject and selecting gene polymorphisms in a linkage disequilibrium block;

(2) a step of analyzing the association between the genotypes of the gene polymorphisms and drug sensitivity in a test subject; and

(3) a step of using the gene polymorphism that has been significantly associated with drug sensitivity in the test subject for evaluation of the drug sensitivity.

4. A method for evaluating disease vulnerability, comprising associating a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism with an individual disease vulnerability.

5. The method according to claim 4, wherein a tendency in the presence or absence of an individual disease vulnerability is evaluated based on the results from the analysis of the gene polymorphism or the haplotype.

6. The method according to claim 4, comprising the following steps:

(2) a step of comparing the frequency of gene polymorphisms in a test subject with the frequency of gene polymorphisms in the healthy subject; and

(3) a step of using the gene polymorphism that has a significant difference in the gene polymorphism frequency between the test subject and the healthy subject for evaluation of the disease vulnerability.

7. The method according to claim 4, wherein the disease vulnerability is pain sensitivity or vulnerability to substance dependence.

8. The method according to claim 1 or 4, wherein the gene polymorphism is at least one selected from the group consisting of a single nucleotide polymorphism, an insertion polymorphism, a deletion polymorphism, and a nucleotide repeat polymorphism.

9. The method according to claim 1 or 4, wherein the gene polymorphism is at least one selected from among: rs16839837, rs2360969, rs10932200, rs2253206, rs2551640, rs11904814, rs16839883, rs6740584, rs3770704, rs2254137, rs2551645, rs2551946, rs4234080, rs2952768, rs2709386, rs7591784, and rs7594560 of a CREB1 subtype gene; rs1243872, rs2145925, rs2025126, rs1885373, rs1885374, GA007473, rs2295794, rs4879926, GA007477, rs867194, rs11541908, rs741917, rs7862485, rs2756894, rs2249250, rs2295795, rs877365, rs2737273, rs2295797, rs2295798, rs1534847, rs7873822, rs2737274, rs10972567, rs3763630, rs10814274, rs3750434, rs1570246, GA025684, rs1570248, rs1570249, rs34478611, rs1322045, rs1951432, GA025687, rs10814275, rs10758320, rs4878628, rs10758321, and rs10758322 of a CREB3 subtype gene; rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs2175738, rs17156579, rs17156603, rs17642145, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs7794304, rs6952227, rs42695, rs1029897, rs4722793, rs10233653, rs6955105, rs17156685, rs17156694, rs17156699, rs177572, rs177573, rs177574, rs177576, rs13437706, rs177580, rs177581, rs12666636, rs177584, rs177585, rs216715, rs10951197, rs160335, rs1008262, rs310353, rs310359, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs41348, rs886816, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs2066979, rs10486591, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955, and rs10228137 of a CREB5 subtype gene; and rs1153711, rs1153702, rs7583431, rs1153699, rs2302663, rs3845744, rs212349, rs212347, rs12693057, rs1153685, rs212360, rs212361, rs2072538, rs1205399, rs1153676, rs7566401, rs7578569, rs3755490, rs13388308, rs11888507, rs10497434, rs268214, rs166531, rs268228, rs268229, rs268230, rs268231, rs10497435, rs1982235, rs268237, rs13030474, and rs268174 of an ATF2 subtype gene.

10. The method according to claim 1 or 4, wherein the haplotype is at least one selected from the following table.

TABLE 1 Gene name CREB1 Linkage disequilibrium block No. 1 Gene polymorphism name () 1 2 3 12 13 17 Haplotype No. Tag Tag Tag 4 5 6 7 8 9 10 11 Tag Tag 14 15 16 Tag H1 C C C A G G G T C C C C C C A A T H2 C C A G A T A C T A T C C T G G T H3 C T C A G G A T T C C C C C A A T H4 T C A G A T A C T A T C A T G G C H5 C C A G A T A C T A T A C C A A T H6 T C A G A T A C T A T C C T G G T H7 T C A G A T A C T A T C A T G G T . . . Haplotypes which are estimated to occur at a frequency of less than 1% () 1~17: (in this order)rs16839837, rs2360969, rs10932200, rs2253206, rs2551640, rs11904814, rs16839883, rs6740584, rs3770704, rs2254137, rs2551645, rs2551946, rs4234080, rs2952768, rs2709386, rs7591784 and rs7594560

TABLE 2 Gene name CREB3 Linkage disequilibrium block No. 1 Gene polymorphism name () 1 2 3 9 11 Haplotype No. Tag Tag Tag 4 5 6 7 8 Tag 10 Tag 12 13 14 15 16 17 18 19 20 21 22 23 24 H1 G C G T A T T T C T C C G C T G T G T C A G G A H2 G C G T A T T T T T C C G C T G T G T C A G G A H3 T T A T A T C T C T T C G A G A C A C C G T G C H4 T T A T A T C T C T C C G A G A C A C C G T G C H5 T C G C C C C C C T C C A A G G C A T C A T A C H6 T T G T A T C T C C C T G A T G C G T C A G G C H7 T C A T A T C T C T T C G A G A C A G C G T G C . . . Haplotypes which are estimated to occur at a frequency of less than 1% Linkage disequilibrium block No. 2 Gene polymorphism name () 25 26 29 36 38 39 Haplotype No. Tag Tag 27 28 Tag 30 31 32 33 34 35 Tag 37 Tag Tag 40 H8 C C G G G C G G G A T A C C G C H9 C T A T G T A G A G A A T C A T H10 T C G G C T G A A G A A T C G C H11 C T A G G T A G A G A A T C G T H12 C C G G C T G A A G A A T C G C H13 C T A T G T A G A G A G T C A T H14 C C G G G C G G G A T A C T G C . . . Haplotypes which are estimated to occur at a frequency of less than 1% () 1~40: (in this order)rs1243872, rs2145925, rs2025126, rs1885373, rs1885374, GA007473, rs2295794, rs4879926, GA007477, rs867194, rs11541908, rs741917, rs7862485, rs2756894, rs2249250, rs2295795, rs877365, rs2737273, rs2295797, rs2295798, rs1534847, rs7873822, rs2737274, rs10972567, rs3763630, rs10814274, rs3750434, rs1570246, GA025684, rs1570248, rs1570249, rs34478611, rs1322045, rs1951432, GA025687, rs10814275, rs10758320, rs4878628, rs10758321 and rs10758322

TABLE 3 Gene name crEB5 Linkage disequilibrium block No. 1 5 6 9 10 Gene polymorphism name () 1 2 5 6 7 8 9 11 13 14 15 16 17 18 Haplotype (H) No. Tag Tag 3 4 Tag Tag H No. Tag Tag Tag 10 Tag 12 H No. Tag Tag H No. Tag Tag H No. Tag Tag H1 C T A A G C H6 C C G C A G H11 T G H14 T A H17 G T H2 G C G A G T H7 C A A C G G H12 C C H15 C A H18 G G H3 G C G A A T H8 C C G C G G H13 C G H16 C G H19 A G H4 C C G A G T H9 T A A C G A . . . . . . . . . H5 G C G A G C H10 C C A C G G . . . Haplotypes which are estimated to occur at a frequency of less than 1% Linkage disequilibrium block No. 11 12 13 15 16 Gene polymorphism name () 19 20 21 22 23 24 25 26 27 28 29 30 31 Haplotype (H) No. Tag Tag H No. Tag Tag Tag H No. Tag Tag H No. Tag Tag Tag H No. Tag Tag Tag 32 33 H20 T A H24 G C C H28 A A H31 G T C H35 C C C G C H21 T G H25 G C T H29 G A H32 A C T H36 T C A G C H22 C G H26 A C T H30 G G H33 A T T H37 C C A G C H23 C A H27 G T T . . . H34 A T C H38 T T C A T . . . Haplotypes which are estimated to occur at a frequency of less than 1% Linkage disequilibrium block No. 25 26 27 32 Gene polymorphism name () 34 35 36 37 38 39 40 42 43 44 45 47 50 Haplotype (H) No. Tag Tag H No. Tag Tag Tag Tag H No. Tag 41 H No. Tag Tag Tag Tag 46 Tag 48 49 Tag H39 T A H42 G A G G H47 A A H49 A T C G T G T T T H40 C G H43 G A A A H48 G G H50 A T C G T G T T C H41 T G H44 A G G A . . . H51 A T A C C G T C C . . . H45 G A G A H52 C T A C T T C T C H46 G G G A H53 C T A C T G C T C . . . H54 A C C G T G T T T H55 A T A G T G T T C Haplotypes which are estimated to occur at a frequency of less than 1% Linkage disequilibrium block No. 33 35 40 Gene polymorphism name () 51 52 53 54 57 58 59 Haplotype (H) No. Tag Tag Tag Tag 55 56 H No. Tag Tag H No. Tag 60 61 62 63 64 H56 T G G C A A H61 C T H64 G G C T T C H57 T A A T G C H62 T C H65 A A T C C A H58 G A A C G C H63 T T . . . H59 T A G C A A . . . H60 T A A C G C . . . Haplotypes which are estimated to occur at a frequency of less than 1% () 1~64: (in this order)rs4722778, rs177479, rs177480, rs11981754, rs177486, rs177498, rs10229500, rs10243659, rs4722785, rs16874503, rs11772815, rs6958133, rs16874525, rs17715174, rs6953524, rs10239810, rs17156649, rs1811248, rs887623, rs740988, rs6952227, rs42695, rs1029897, rs10233653, rs6955105, rs17156699, rs177572, rs177573, rs177580, rs177581, rs12666636, rs177584, rs177585, rs1008262, rs310353, rs41273, rs1637457, rs17156919, rs41276, rs160375, rs917275, rs17157048, rs6462098, rs10951201, rs13311248, rs12540480, rs10265166, rs7798774, rs7799246, rs6972081, rs12533079, rs7806547, rs6462100, rs6979352, rs6950574, rs4722835, rs721993, rs2237351, rs3735566, rs11975539, rs6462107, rs2190306, rs4719955 and rs10228137

TABLE 4 Gene name ATF2 Linkage disequilibrium block No. 1 2 Gene polymorphism name () 1 2 3 4 5 Haplotype No. Tag Tag Tag Tag Tag 6 7 8 9 10 11 12 13 14 15 16 H1 T C A A G C C A G C G T C A A A H2 G C A G G T T G A T A C A C C G H3 T T C G A T T G A T A C A C C G H4 T T A G A T T G A T A C A C C G G G T T G A T A C A C C G . . . Haplotypes which are estimated to occur at a frequency of less than 1% Linkage disequilibrium block No. 2 Gene polymorphism name () 21 28 Haplotype No. 17 18 19 20 Tag 22 23 24 25 26 27 Tag 29 30 31 H5 C T T C C C G A C A C T T G T H6 T C C T T T A G A C T G C G C H7 T C C T T T A G A C T T C G C H8 T C C T T T A G A C T G C G C H9 T C C T C C G A C A C T T G T . . . Haplotypes which are estimated to occur at a frequency of less than 1% () 1~31: (in this order)rs1153711, rs1153702, rs7583431, rs2302663, rs3845744, s212349, rs212347, rs12693057, rs1153685, rs212360, rs212361, rs2072538, rs1205399, rs1153676, rs7566401, rs7578569, rs3755490, rs13388308, rs11888507, rs10497434, rs268214, rs166531, rs268228, rs268229, rs268230, rs268231, rs10497435, rs1982235, rs268237, rs13030474 and rs268174

11. A method for determining the type, amount, and/or frequency of administration of a drug to be administered to an individual, comprising using the result from the evaluation by the method according to claim 1 or 4 as an index.

12. A method for predicting a side effect of a drug to be administered to an individual, comprising using the result from the evaluation by the method according to claim 1 or 4 as an index.

13. The method according to claim 1 or 7, wherein the drug is an opioid receptor function modulator and/or a cyclic AMP responsive element binding protein function modulator.

14. The method according to claim 13, wherein the opioid receptor function modulator is at least one selected from the group consisting of methamphetamine, methylenedioxymethamphetamine, amphetamine, dextroamphetamine, dopamine, morphine, DAMGO, codeine, methadone, carfentanil, fentanyl, heroin, cocaine, naloxone, naltrexone, nalorphine, levallorphan, pentazocine, pethidine, buprenorphine, oxycodone, hydrocodone, levorphanol, etorphine, dihydroetorphine, hydromorphone, oxymorphone, tramadol, diclofenac, indomethacin, flurbiprofen axetil, marcaine, ethanol, methanol, diethyl ether, propanol, butanol, flupirtine, laughing gas, F3 (1-chloro-1,2,2-trifluorocyclobutane), halothane, estradiol, dithiothreitol, thioridazine, pimozide, fluoxetine, paroxetine, desipramine, imipramine, clomipramine, tetramide, isoflurane, ginsenoside, ifenprodil, bupivacaine, tertiapin, clozapine, haloperidol, SCH23390, and cocaine; and the cyclic AMP responsive element binding protein function modulator is at least one selected from the group consisting of phosphodiesterase 4 (PDE4), calcineurin, protein kinase A, protein kinase C, p90 ribosome S6 kinase 1 (RSK1), calmodulin kinase, glycogen synthase kinase 3β, and CREB-regulated transcription coactivator 1 (CRTC1).

15. The method according to claim 1 or 4, comprising using an oligonucleotide consisting of a nucleotide sequence of at least 10 nucleotides comprising the 51^stnucleotide of the nucleotide sequence represented by any one of SEQ ID NOS: 1 to 172, or a complementary nucleotide sequence thereto, which can specifically hybridize to a DNA fragment comprising a gene polymorphism of a cyclic AMP responsive element binding protein gene.

16. The method according to claim 15, wherein the oligonucleotide spans a length of 10 to 150 nucleotides.

17. The method according to claim 15, wherein the oligonucleotide is selected from the group consisting of the nucleotide sequence represented by any one of SEQ ID NOS: 1 to 172 and a complementary nucleotide sequence thereto.

18. A gene polymorphism marker for evaluating a tendency in the presence or absence of an individual drug sensitivity, comprising a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism.

19. A gene polymorphism marker for evaluating a tendency in the presence or absence of an individual disease vulnerability, comprising a gene polymorphism of a cyclic AMP responsive element binding protein gene or a haplotype constituted by the gene polymorphism.